Humoral immune reactions on the hepatocellular plasma membrane in the experimental chronic active hepatitis in rabbits

Zur Induktion einer experimentellen chronisch aktiven Hepatitis (CAH) wurden Kaninchen mit humanem leberspezifischem Protein plus komplettem Freundschen Adjuvans langzeitimmunisiert. Nach 23wochiger Versuchsdauer waren im Serum Antikorper gegen allogene hepatocellulare Membranantigene mit Hilfe isolierter normaler Kaninchenhepatocyten bzw. in der passiven Hamagglutination mit leberspezifischem Membranprotein als Antigen nachweisbar. Gleichzeitig zeigten isolierte Hepatocyten dieser Tiere in vivo-fixiertes IgG an den Plasmamembranen in Form eines gemischt linear-granularen Fluoreszenzmusters. Mit der Raji-Zell-Technik wurden im Serum der Tiere Immunkomplexe gefunden. Eine chronisch aktive He…

Infection of murine hepatocyte cultures by herpes simplex virus (HSV) 1 and 2

A study was undertaken of the interaction between liver cells and Herpes Simplex Virus (HSV) in vitro. Hepatocytes were obtained from HSV-resistant (C57/B16) and from HSV-susceptible (BALB/c, A/J, C3H) mouse strains and cultured according to standard methods. Each culture was infected with several strains of HSV-type 1 or of HSV-type 2, respectively. The multiplicity of infection was 5. The cytopathic effect was evaluated by light- and electron-microscopy. The number of infectious particles was determined using rabbit kidney or Vero cell cultures. All evaluations were made at different time intervals after infection. No difference concerning the replication rate of HSV-1 and 2 in isolated h…

Complement component C3: molecular basis of the C3*S025 variant and evidence for molecular heterogeneity of other variants.

Complement component 3 (C3) is the central molecule of the complement system. It displays a number of polymorphic variants with, as yet, unclear functional consequences. We have investigated a number of rare C3 variants by PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) analysis and could identify the molecular basis of a C3*S025 variant. The decreased electrophoretic mobility of this protein is caused by the exchange of a neutral serine residue to an arginine residue (positively charged). This exchange is unlikely to have functional consequences as it maps to the C-terminus of the alpha-chain. C3 variants appear to have originated from various independent mutat…

Experimentelle Untersuchungen zur Speciesspezifität von Anti-Gamma-Globulinfaktoren

Nach Charakterisierung des Rheumafaktors bei der chronischen Polyarthritis durch Waaler [16], Rose [14] sowie Waller u. Vaughan [17] konnten Rheumafaktor-ahnliche Substanzen auch bei Fallen von Tuberkulose, Syphilis, Endokarditis lenta, bei Kollagenosen, Sarkoidosen, bei Lebererkrankungen und im Serum von Normalpersonen (Ubersich in [8]) beobachtet und zum Teil an Hand bestimmter Reaktionen vom Rheumafaktor abgegrenzt werden [3, 8, 19]. Es handelt sich bei den Faktoren um Antikorper vom 19 S-Typ [5], die gegen Human-Gamma-Globulin gerichtet sind und z. T. mit Gamma-Globulin anderer Species kreuzreagieren. Zur Entstehung der Rheumafaktoren nahmen Najjar [13] und Milgrom [10] unabhangig von e…

Azathioprine, Mycophenolate Mofetil, and Methotrexate Specifically Modulate Cytokine Production by T Cells

IgG Subclass Distribution of Autoantibodies to Glomerular Basement Membrane in Goodpasture’s Syndrome Compared to Other Autoantibodies

The IgG subclass distribution of autoantibodies to glomerular basement membrane (anti-GBM antibodies) was investigated and compared to the distribution of liver-kidney microsomal (LKM) autoantibodies in chronic active hepatitis, to antimitochondrial autoantibodies (AMA) in primary biliary cirrhosis, and to the subclass distribution of total serum IgG within a healthy population. Solid phase assays for the demonstration of these autoantibodies were performed with four mouse monoclonal antibodies specific for each human subclass to provide quantitative data for the autoantibodies. In addition, the subclass distribution of total IgG in these sera was analyzed. IgG1 accounted for 75% of the tot…

Spontaneous and antibody-dependent cellular immune reactions to ethanol-altered hepatoma cells

— Spontaneous cell-mediated cytotoxicity (SCMC), antibody-dependent cellular cytotoxicity (ADCC) and proliferative lymphocyte stimulation in alcoholic liver disease (ALD) were investigated. Peripheral blood lymphocytes (PBL) from eight patients with advanced ALD and nine normal controls were tested against hepatoma cells (PLC/PRF/5) as targets. Target cells were grown in either normal culture medium or medium supplemented with 1 and 5% ethanol, respectively, for 24 to 48 h. Ethanol-exposed hepatoma cells exhibited profound and characteristic morphological alterations. Ethanol preincubation, however, proved to be without effect on immune reactions. Provided that hepatoma cells are an appropr…

Heat shock protein-peptide complexes for use in vaccines

Abstract The heat shock proteins gp96, HSP70, and HSP90 are complexed to a diverse array of cellular proteins and peptides as a consequence of their chaperone functions. There is good experimental evidence that vaccination with these heat shock protein-peptide complexes elicit immune responses against chaperoned peptide antigens. As shown with gp96, this requires internalization of the heat shock protein-peptide complexes by macrophages and processing of the chaperoned peptides for class I restricted presentation. Via this process, primarily CD8+ antigen-specific T cells are primed by gp96 vaccination. This might represent a general mechanism for priming of MHC-class I restricted T cells by…

Quantitative and functional analysis of core-specific T-helper cell and CTL activities in acute and chronic hepatitis B

Aims/background CD4+ T-helper cell (Th) responses to hepatitis B virus (HBV) core antigen (HBc) are increased during exacerbations in acute and chronic hepatitis B (AHB, CHB) and might influence the induction of CD8+ cytotoxic T lymphocytes (CTL) that are important for viral clearance. Methods HBc-specific proliferative responses and cytokine release of blood mononuclear cells (PBMC) were studied in patients with AHB or CHB, as well as responders and non-responders to interferon-alpha treatment (IFN-R, IFN-NR), by [3H]-thymidine-uptake, enzyme-linked immunosorbent assay (ELISA) and Elispot assay and were compared to peptide HBc18 27-specific CTL precursor frequencies among CD8+ T cells deri…

Regulation of the neutralizing anti-hepatitis B surface (HBs) antibody response in vitro in HBs vaccine recipients and patients with acute or chronic hepatitis B virus (HBV) infection

Abstract Antibodies directed to the HBs antigen indicate viral clearance and the development of life-long immunity in patients that recovered from HBV infection. In HBs antigen vaccine recipients anti-HBs antibodies provide protective immunity. However, little is known about the regulation of this HBs-specific antibody response. The existence of anti-HBs-secreting B cells was demonstrated using the highly sensitive ELISPOT technique compared with conventional ELISA in serum and cell culture supernatants. In the peripheral blood of patients with acute self-limited hepatitis B, HBs-specific B cells were demonstrated with a high frequency despite undetectable anti-HBs serum antibodies. HBV-imm…

Duplexsonographische Untersuchungen zur Pathogenese der lienalen Hämodynamik bei Leberzirrhose

AIM OF STUDY The haemodynamic role of the spleen in portal hypertension remains unclear. Duplex sonography was undertaken prospectively to discover the relationship of splenomegaly and splenic vein flow to type and severity of liver cirrhosis and portal vein flow, as well as to the degree of oesophageal varices. PATIENTS AND METHODS 89 patients (54 men, 35 women; mean age 52 [26-81] years), diagnosed in the second half of 1993 as having liver cirrhosis were consecutively included in the study. In 40 patients the cirrhosis was in stage A (according to Child-Pugh classification), in 31 in stage B and in 18 in stage C. RESULTS Portal vein flow fell significantly with increasing degree of cirrh…

Monoklonale Gammopathie bei HIV-Infektion

A lambda-light chain-IgA plasmocytoma, accompanied by a changing clinical picture of fever, nocturnal perspirations and weight loss, developed in a 46-year-old homosexual male with AIDS, stage IV (classification according to the Centers for Disease Control). He had been suffering from recurrent Salmonella septicaemia. Serum protein electrophoresis demonstrated marked elevation of the beta- and gamma-fractions (44% and 24%, respectively). There were 15% plasma cells in the differential blood count and in the bone marrow smear. Immunoelectrophoresis demonstrated free lambda-light chains. The IgA concentration in cerebrospinal fluid was raised to 202 mg/l, and there was an IgA paraproteinaemia…

Chirurgie bei Hepatitis

Virushepatitiden gehoren weltweit zu den wichtigsten Infektionskrankheiten. Allein uber 300 Millionen chronische HBsAg-Trager und etwa die gleiche Anzahl chronischer Hepatitis-C-Virustrager werden geschatzt. Nach der Tuberkulose steht die Virushepatitis in der Statistik der infektios bedingten Berufskrankheiten an zweiter Stelle. Insbesondere Teile Asiens, Afrikas, Lateinamerikas sowie der Mittelmeerraum und der Nahe Osten werden zu den Hochendemiegebieten der Virushepatitis gezahlt. Aufgrund der immunologischen und molekularen Differenzierung sind heute die Virushepatitiden von A bis E einschlieslich G weitgehend charakterisiert. Wahrend die enteral ubertragene Hepatitis A und E nur akute …

Funktionelle Beziehung zwischen Leber und Niere: Teil II. Wasserexkretion und hepatorenales Syndrom*

Chemotherapy in advanced pancreatic cancer

Patients with advanced adenocarcinomas of the pancreas have an exceptionally poor prognosis. Modest activity has been demonstrated with single agents (response rates of 25% at best with 5-fluorouracil [5-FU] and mitomycin). Better results have not been obtained by combination chemotherapy. Improvements in the palliation have been achieved by treatment with 5-FU, folinic acid (FA), and interferon-alpha-2A (IFN-alpha) weekly in the context of a phase II trial. Of 57 evaluable patients, eight (14%) had a partial response (PR), eight (14%) a minor response (MR), and 28 (49%) had no change of disease (NC). The median survival time was 10 mo for patients with progressive disease. Twenty-two out o…

HLA-B27-restricted cytotoxic T lymphocyte responses to arthritogenic enterobacteria or self-antigens are dominated by closely related TCRBV gene segments. A study in patients with reactive arthritis.

Identification of the T-cell receptors (TCR) used by synovial cytotoxic T lymphocytes (CTL) of patients with reactive arthritis (ReA) may be crucial to better understanding the pathogenetic mechanism underlying the HLA-B27 association of spondylarthropathies. The authors, therefore, sequenced 25 TCRB chains from HLA-B27-restricted CD8+ CTL clones and two clonal lines specific for self- or Yersinia enterocolitica antigen isolated from synovial fluids of 3 HLA-B27+ patients with ReA and PBL of one healthy HLA-B27+ individual. Fourteen non-HLA-B27-restricted CTL served as controls. Both autoreactive and Y. enterocolitica specific HLA-B27-restricted CTL used a highly limited set of VB genes wit…

Investigations on the pathogenesis of active chronic hepatitis

Es wird uber die Ergebnisse von Sensibilisierungen bei Kaninchen mit nativen und alterierten homologen sowie heterologen organgebundenen Leberproteinen, Serumproteinen und Antigen-Antikorper-Komplexen berichtet. — Die neonatal erworbene Immuntoleranz gegenuber loslichem organgebundenem Leberprotein ist mit alteriertem homologem oder heterologem Protein zerstorbar. Organspezifische Autoantikorper bzw. spezifisch-sensibilisierte Zellen konnten bei Verwendung von haptenisierten homologen Proteinen nach 2–3 Monaten, bei Verwendung von heterologen Proteinen nach 6 Monaten nachgewiesen werden. — Der Befund einer aktiven chronischen Hepatitis fand sich nach langdauernder Sensibilisierung mit klein…

Expression of the gene of the alpha-smooth muscle-actin isoform in rat liver and in rat fat-storing (ITO) cells.

Fat storing cells (FSCs) in the liver represent the main site of vitamin A deposition in the body. These cells are considered to play an important role during scar formation and fibrogenesis in the liver. The putative descent of FSCs from the fibroblastic or from the myofibroblastic system have not been determined yet by morphological or immunohistochemical studies. To further define the origin of these liver cells, we analysed the pattern of expression of three structural proteins: vimentin, desmin and the α-smooth muscle (SM)-actin isoform in FSCs of the rat liver, in smooth muscle cells (SMCs) from the aorta and in rat skin fibroblasts. FSCs were studied by immunohistochemical methods im…

Tolerance towards resident intestinal flora in mice is abrogated in experimental colitis and restored by treatment with interleukin-10 or antibodies to interleukin-12

There is now increasing evidence that hyperresponsiveness towards intestinal flora is a crucial event in the pathogenesis of inflammatory bowel disease (IBD). In support of this hypothesis, we recently described in humans that tolerance exists towards indigenous intestinal flora but is broken in active IBD lesions. In the present study, we have attempted to transfer this model into mice from different genetic backgrounds (BALB/c, SJL/J, C3H/HeJ). We found that mononuclear cells from spleen, small bowel and large bowel of mice do not proliferate, i.e. are tolerant when exposed to bacterial sonicates derived from autologous intestine (BsA) but do proliferate, i.e. are immune when exposed to b…

Sinusoidal endothelial liver cells in vitro release endothelin — Augmentation by transforming growth factorβ and Kupffer cell-conditioned media

Endothelin is the most potent vasoconstrictor peptide known today. Using a radioimmunoassay for endothelin, we measured immunoreactive endothelin in culture media of guinea pig sinusoidal endothelial liver cells and human umbilical vein endothelial cells. A time-dependent release of immunoreactive endothelin by confluent sinusoidal endothelial liver cells in culture was found. Sinusoidal endothelial liver cells produced similar amounts of immunoreactive endothelin as umbilical vein endothelial cells, about 900 pg/microgram DNA per 24 h. In the presence of transforming growth factor beta a dose-dependent increase of immunoreactive endothelin release was measured. The maximal increase of 50% …

Epstein Barr virus-transformed lymphocytes of patients with Wegener's granulomatosis produce antibodies against the cytoplasm of human neutrophil granulocytes (ANCA)

Phenotypical analysis and cytokine release of liver-infiltrating and peripheral blood T lymphocytes from patients with chronic hepatitis of different etiology.

Cytokines released by infiltrating T cells may contribute to the hepatic injury in chronic hepatitis. Therefore, we characterized peripheral blood- and liver-infiltrating T cells from patients with chronic hepatitis of different etiology and determined the T cell phenotypes and the cytokine release. Liver tissue and peripheral blood-derived T cells from patients with autoimmune hepatitis and primary biliary cirrhosis predominantly expressed CD4-molecules and the alpha- and beta-chains of the T cell receptor (TCR). In chronic viral hepatitis B and C, liver- and blood-derived T cells were preferentially CD8+ T cells expressing the alpha beta TCR. Mitogenic stimulation with irradiated Daudi ly…

Hepatitis C virus antibody secretion in vitro by peripheral blood lymphocytes.

A recombinant polypeptide corresponding to a virus-specific cDNA clone (c100-3) serves as the antigen for a hepatitis C virus (HCV) antibody assay. Previous investigations have shown an 80% prevalence of HCV antibodies in sera of patients suffering from post-transfusional chronic hepatitis non-A, non-B, but positive results were also obtained for 30 to 70% of sera from patients with chronic hepatitis B or autoimmune hepatitis. In this study we show that HCV antibodies are secreted by peripheral blood lymphocytes (PBL) in vitro. PBL from 12/35 patients with chronic non-A, non-B hepatitis and 1/6 patients with chronic active hepatitis B spontaneously secreted HCV antibodies in cell culture su…

Glomerulonephritis und Hepatitis-B-Virus-Infektion

The significance of the pre-S region of the hepatitis B virus

Histokompatibilitäts-(HLA) Antigene bei Patienten mit HBsAg-positiver und -negativer chronisch aktiver Hepatitis und gesunden Trägern von HBsAg und Anti-HBs

Eine Analyse der Literatur und eigene Befunde uber die Haufigkeit von 23 HLA-Antigenen bei Patienten mit Hepatitis-Oberflachen-Antigen (HBsAg)-positiver und -negativer chronisch aktiver Hepatitis (CAH) und gesunden Tragern von HBsAg und Antikorpern gegen HBsAg (Anti-HBs) erlauben die folgenden Schlusfolgerungen: 1. Bei Patienten mit CAH und HBsAg-Persistenz findet sich eine normale Haufigkeit von HLA-B8 (und -A1). 2. Auch bei Patienten mit HBsAg-negativen Formen einer CAH ohne Autoimmunphanomene (AutoAk) ist die HLA-B8-Haufigkeit normal. 3. Nur Patienten mit HBsAg-negativer CAH mit AutoAk haben eine statistisch signifikant grosere Haufigkeit von HLA-B8 (und -A1) (p < 0,01 nach Korrektion fu…

Acute hepatic failure: limitations of medical treatment and indications for liver transplantation.

The function of the soluble IL-6 receptor in vivo.

Interleukin-6 (IL-6) is considered an important mediator of acute inflammatory responses. Moreover, IL-6 functions as a differentiation and growth factor of hematopoietic precursor cells, B-cells, T-cells, keratinocytes, neuronal cells, osteoclasts and endothelial cells. IL-6 exhibits its action via a receptor complex consisting of a specific IL-6 receptor (IL-6R) and a signal-transducing subunit (gp130). Soluble forms of both receptor components are generated by shedding and are found in patients with various diseases such as AIDS, rheumatoid arthritis and others. The function of the soluble IL-6R in vivo is unknown. To discriminate between the biologic function of hIL-6 alone and that of …

Die Vielgestaltigkeit der Virushepatitis

Bei der gemeinsamen Planung war von Anfang an klar, das nur einige Fragen und Themen aufgegriffen werden konnten, die uns auch fur den praktisch tatigen Internisten von besonderer Wichtigkeit erschienen. Aber es sollten auch Probleme aufgegriffen werden, die von grundsatzlicher Bedeutung zum Verstandnis neuerer Entwicklungen sind.

Modification of the immune response against hepatitis B virus by the human immunodeficiency virus.

Hepatitis B virus and the human immunodeficiency virus are similarly transmitted. Individuals with preexisting HIV infection have a higher chance to become HBsAg carriers than do anti-HIV negative persons. Cytotoxic T cells with specificity for HBcAg, that are under the control of HBcAg-specific helper T cells, are responsible for liver injury. There is good evidence that HIV infection lowers inflammatory activity, is associated with milder liver histology, high levels of viral replication and low seroconversion rates. In addition interferon alpha therapy is less effective in anti-HIV positive subjects. The immune response against HBsAg is helper T-cell dependent and vaccination against hep…

Scleroedema adultorum und Plasmocytom

Bei einem 37jahrigen Patienten mit einem im Bereich beider Arme und Schultern lokalisiertenScleroedema adultorum (Buschke) traten fast gleichzeitig sensomotorische Ausfalle der Schulter-Arm-Region auf. Diese waren durch einen pramedullar, extradural in Hohe von C 7/Th 1 gelegenen raumfordernden Prozes hervorgerufen, dessen histologische Untersuchung ein Plasmocytom ergab. Diese Diagnose wurde durch eine Paraproteinamie und eine Paraproteinurie bestatigt, die sich durch papierelektrophoretische und immunelektrophoretische Untersuchungen als Gamma-G-Gammopathie charakterisieren lies. Die Frage bleibt offen, ob es sich hierbei um ein mehr als zufalliges Zusammentreffen beider Erkrankungen hand…

Tubulointerstitielle-Nephritis-Uveitis-Syndrom (TINU-Syndrom)

Abstract For 10 weeks a 25-year-old man had been suffering from tiredness, fatigue, nausea and a 16 kg weight loss. Erythrocyte sedimentation rate (83/133 mm), serum C-reactive protein (5.5 mg/dl) and creatinine (5.05 mg/dl) were all elevated. He also had proteinuria (1120 mg daily), sterile leukocytosis and a creatinine clearance of 10 ml/min. Renal biopsy showed interstitial nephritis and bone marrow biopsy revealed non-caseous epithelioid-cell granulomas. 14 days after admission he developed acute iritis in the right eye. Other causes having been excluded, the diagnosis of tubulo-intestinal nephritis with uveitis (TINU syndrome) was made. The clinical symptoms and laboratory findings imp…

Hepatitis C virus (HCV) and autoimmune liver diseases

Anti-HCV tests were positive in 18–45% of sera from patients with autoimmune chronic active hepatitis. High gammaglobulin levels may result in false positive results, however, some sera show true positivity. PCR testing of such sera is necessary in order to determine whether HCV is directly involved in specific forms of the disease.

Antigen-presenting function and B7 expression of murine sinusoidal endothelial cells and Kupffer cells.

Abstract BACKGROUND & AIMS: Inflammatory liver disease as well as rejection of liver allografts are thought to be mediated by resident antigen- presenting cells in the liver. At the same time, in vivo antigen presentation in the liver appears to be a more tolerogenic than systemic antigen challenge. The aim of this study was to show and characterize the antigen-presenting capability of sinusoidal endothelial cells and Kupffer cells. METHODS: Purified murine sinusoidal endothelial cells and Kupffer cells were studied for their ability to serve as accessory cells and antigen-presenting cells by proliferation assays. They were also studied for their expression of interleukin 1 and the B7 costi…

Funktionelle Beziehung zwischen Leber und Niere: Teil I. Natriumexkretion*

Clinical significance of the polymerase chain reaction (PCR) assay in chronic HBV carriers

PCR was evaluated as a clinical tool for use in accurate identification of the specific etiologic agent in chronic HBV carriers. The method was found to be valuable in diagnosis and for monitoring therapy, as well as for elucidation of genotypic variants of HBV in chronic HBV cases. By this means an HBV defective variant with alterations in the preSl/preS2 sequence was detected and is consequently described here.

Expression and regulation by interferon-γ of the membrane-bound complement regulators CD46 (MCP), CD55 (DAF) and CD59 in gastrointestinal tumours

The membrane-bound complement inhibitors CD46 (membrane cofactor protein), CD55 (decay-accelerating factor) and CD59 (protectin) protect tumour cells against lysis by activated complement. In this study, a total of 14 (3 gastric, 3 colonic and 8 pancreatic) gastrointestinal tumour cell lines were examined for the expression of CD46, CD55 and CD59 with respect to the regulatory efficacy of interferon-gamma (IFN-gamma). The effects of IFN-gamma on mRNA and protein expression levels of CD46, CD55 and CD59 were evaluated by Northern blot hybridisation, RT-PCR, flow cytometry and immunostaining. In unstimulated cell lines, CD46 and CD59 transcripts were expressed at comparable levels, whereas th…

Anti-Endothelzell-Antikörper

Cholinesterase variants: rapid characterisation by PCR/SSCP and evidence for molecular homogeneity.

We have applied the technique of PCR-SSCP (polymerase chain reaction-single stranded conformation polymorphism) to characterise the molecular basis of cholinesterase deficiency and variants in a Jordanian family. PCR-SSCP proved to be a quick and sensitive method of screening cholinesterase variants in a clinical setting. An AG insertion at position 351 was found to cause a silent allele, for which the parents were heterozygous and three children homozygous. In addition, the father and two sons were heterozygous for an A to G transition at position 209, known to cause the dibucaine resistant variant. No linkage to the K variant was found, which has been reported previously in white populati…

Post-infantile giant cell hepatitis in patients with primary sclerosing cholangitis and autoimmune hepatitis.

In post-infancy, multinucleated giant cell hepatitis is rare. Various conditions and diseases associated with post-infantile giant cell hepatitis have been described, but the pathogenesis remains unknown. In this paper we review the case reports of four patients (3 male, 1 female; aged 22 to 32 years) with primary sclerosing cholangitis and autoimmune hepatitis. The follow-up ranges from five to seven years. All patients showed cholestasis and repeated elevation of hepatic transaminases. Patients with viral infections, metabolic disorders and toxic influences were excluded. Histopathology of liver tissue in all four patients revealed giant cell formation with up to 20 nuclei in 20-70% of al…

Therapieentscheidung beim Gallensteinleiden aus internistischer Sicht

Fur die Therapie des Gallensteinleidens lassen sich folgende Leitlinien festlegen: 1. Gallengangssteine lassen sich in uber 90% der Falle endoskopisch, haufig kombiniert mit mechanischer oder extrakorporaler Stoswellenlithotripsie (ESWL) behandeln. Steine <10 mm im O konnen ohne endoskopische Papillotomie (EPT), grosere Steine nach EPT und Lithotripsie extrahiert werden. Die ESWL ist bei grosen, zu harten oder multiplen Steinen sowie bei atypischer Anatomie indiziert. Patienten mit hohem OP-Risiko werden zunachst nur endoprothetisch versorgt (ca. 2%). Selten (5%) ist bei Gallengangssteinen eine OP notwendig. 2. Symptomatische Blasensteine werden in 80–85% der Falle chirurgisch behandelt. Nu…

A human renal cancer line as a new antigen source for the detection of antibodies to cytoplasmic and nuclear antigens in sera of patients with Wegener's granulomatosis.

Autoantibodies directed against cytoplasmic antigens of neutrophils (ANCA), especially proteinase 3 (C-ANCA), have proved to be a useful clinical tool to support the diagnosis or to monitor disease activity in Wegener's granulomatosis (WG). Till now, human neutrophil granulocytes have represented the major antigen source used to detect antibodies in WG by the immunofluorescence technique (IFT). We have tested serum samples of 164 patients with different connective tissue diseases (50 suffering from clinically active WG) performing IFT on a human renal cancer line (SK-RC11) and have found antibodies against the nuclear and cytoplasmic antigens in 39 patients. C-ANCA+ sera displayed a charact…

The asialoglycoprotein receptor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers.

As a putative mechanism of hepatitis B virus (HBV) uptake into hepatocytes the interaction between HBV and the hepatic, human-derived asialoglycoprotein receptor (ASGPR) was investigated. Sera from patients with different variations of hepatitis B surface antigen-(HBsAg) positive chronic hepatitis, HBV particles isolated from HBV carriers with high-titre viraemia and commercial HBsAg served as sources of HBV. ASGPR was affinity-purified from human liver. HBV that had bound to isolated ASGPR was either detected by radio-immunoassay using solid-phase bound ASGPR or enzyme immunoassay with biotin-ASGPR bound to immobilized HBV. Furthermore, binding and uptake of purified, 125I-labelled HBV par…

Autoimmune findings resembling connective tissue disease in a patient with Castleman's disease.

Multicentric angiofollicular lymphnode hyperplasia (multicentric Castleman's disease) may be associated with acute phase reaction and several autoimmune features. Since lymphadenopathy is a common feature in connective tissue disease, a clear distinction between the different disease entities may be difficult. We describe a 26-year-old male patient with predominant cervical lymphadenopathy, hepatosplenomegaly and polyserositis, diagnosed as collagen disease. He showed several autoimmune features including autoimmune haemolytic anaemia, cryoglobulinaemia, positive antinuclear and anti smooth muscle antibodies, serum immune complexes and a sensorimotor polyneuropathy. Under immunosuppressive …

Contrary roles of IL-4 and IL-12 on IL-10 production and proliferation of human tumour reactive T cells.

The cytokine profile of tumour reactive T cells is likely to play a central role in their function. However, little is known about how cytokine patterns of tumour reactive T cells can be regulated. Here, the authors investigated the influence of exogenous regulatory cytokines in addition to interleukin-2 (IL-2) on cytokine patterns and the proliferation of T cells recognizing an autologous sarcoma cell line. In this system, IL-4 and IL-12 showed the most polarizing influences on tumour reactive T cells. Exogenous IL-4 induced a predominant production of IL-4 while decreasing the interferon-gamma (IFN-gamma) and IL-10 production by tumour reactive T cells. It also stimulated the growth of tu…

Hepatitis C virus associated primary hepatocellular carcinoma in a noncirrhotic liver

The case of a 71-year-old man with a primary hepatocellular carcinoma in a non-cirrhotic liver is reported. There were no risk factors of hepatocellular carcinoma (HCC)-like liver cirrhosis, alcohol drinking, tobacco smoking, exposure to vinyl chloride, thorotrast, aflatoxin or alpha 1-antitrypsin deficiency. Serologically, the patient was positive for antibodies to the hepatitis B virus (anti-HBc, anti-HBs) and for anti-hepatitis C virus (HCV) antibodies. Virologically, positive and negative strands of HCV RNA could be detected in the patient's serum and tumorous liver tissue by reverse transcription polymerase chain reaction as a sign of persistent HCV replication. Histologically, the HCC…

Autoantibodies against the human asialoglycoprotein receptor: Effects of therapy in autoimmune and virus-induced chronic active hepatitis

The hepatic asialoglycoprotein receptor (ASGPR) was recently identified as a target antigen for both humoral and cellular immune response in inflammatory liver diseases. Thereby anti-ASGPR autoantibodies directed against human substrate were closely associated with autoimmune chronic active hepatitis. The present study compares the occurrence, titer and immunoglobulin classification of anti-human(h-)-ASGPR antibodies in 23 patients with newly diagnosed autoimmune chronic hepatitis before and after initiation of immunosuppressive therapy to 22 patients with autoimmune hepatitis in remission. Additionally, 1-year follow-up examinations of 42 patients with HBsAg-positive chronic hepatitis and …

T cell specificity and cross reactivity towards enterobacteria,Bacteroides,Bifidobacterium, and antigens from resident intestinal flora in humans

BACKGROUNDT cell responses to normal intestinal bacteria or their products may be important in the immunopathogenesis of chronic enterocolitis.AIMSTo investigate the T cell specificity and cross reactivity towards intestinal bacteria.PATIENTS/METHODST cell clones were isolated with phytohaemagglutinin from peripheral blood and biopsy specimens of inflamed and non-inflamed colon from five patients with inflammatory bowel disease (IBD) and two controls. T cell clones were restimulated with anaerobicBacteroides andBifidobacteria species, enterobacteria, and direct isolates of aerobic intestinal flora. T cell phenotype was analysed by single-cell immunocyte assay.RESULTSAnalysis of 96 T cell cl…

The role of the Thomsen-Friedenreich antigen as a tumor-associated molecule.

The Thomsen-Friedenreich antigen (Gal-GalNAc) represents a tumor-associated molecule, which is assumed to be one of the few chemically well-defined antigens with a proven association with malignancy. In order to analyze the role of the carbohydrate structure Gal-GalNAc for gastrointestinal tumors, we immunized Balb/c mice with MCF-7 breast tumor cells together with synthetic Gal-GalNAc linked to a BSA carrier. One monoclonal antibody (82-A6) was established which recognizes the Thomsen-Friedenreich antigen according to the biochemical and serological analysis presented here. In contrast to the studies performed in the past, immunohistochemical results using this antibody 82-A6 did not exhib…

Methotrexate specifically modulates cytokine production by T cells and macrophages in murine collagen-induced arthritis (CIA): a mechanism for methotrexate-mediated immunosuppression

SUMMARYImmunosuppressive therapy with methotrexate (MTX) has been established as effective treatment for patients with rheumatoid arthritis. To analyse the therapeutic potential and mechanisms of action of MTX, we determined serum cytokine levels and cytokine production by splenic T cells and macrophages in untreated and MTX-treated mice. Furthermore, we assessed the role of MTX in a murine model of experimental arthritis induced by collagen type II (CIA). MTX reduced spontaneous and IL-15-induced tumour necrosis factor (TNF) production by splenic T cells but not by macrophages from healthy mice in vitro in a dose-dependent manner. In contrast, interferon-gamma (IFN-γ) production was less s…

The endoscopic measurement of intravascular pressure and flow in oesophageal varices.

Schleimhaut-Polyp-Karzinom: Ein molekulargenetisches Modell der Entstehung kolorektaler Tumoren

Fat storing cells (FSC) of rat liver synthesize and secrete fibronectin

Summary Fat storing cells (FSC) of rat liver were isolated and kept in culture for up to 2 weeks. Freshly isolated cells and cells in culture were characterized functionally (vitamin A content) and morphologically. Their synthetic capacity for fibronectin was studied quantitatively by ELISA-measurement and qualitatively by biosynthetic labeling, immunoprecipitation and SDS-PAGE analysis. The synthesis product and the kinetics of the synthesis and secretion were compared with those obtained from culture of hepatocytes. FSC were shown to synthesize and secrete fibronectin under the culture conditions used. No fibronectin synthesis was detected during the first 3 days of culture. Thereafter th…

Natural killer cell activity as a prognostic parameter in the progression to AIDS.

Clonal analysis of liver-infiltrating T cells in patients with LKM-1 antibody-positive autoimmune chronic active hepatitis

SUMMARY Autoantibodies against microsomal antigen of liver and kidney (LKM-1) are diagnostic markers for a subgroup of autoimmune chronic active hepatitis (AI-CAH). Cytochrome P4S0dbl, now classified as cytochrome P450 IID6, is the major antigen of LKM-1 antibodies. Immunohistological studies suggest that hepatic injury in AI-CAH is mediated by liver-infiltrating T cells. In the present study the specificity and function of liver-infiltrating T cells was analysed at the clonal level. Phenotypical characterization of 189 T cell clones isolated from four liver biopsies of LKM-1 antibody-positive patients showed an enrichment of CD4+CD8- T cells. Five CD4+CD8- T cell clones proliferated specif…

Thymoma and pure red cell aplasia in a patient with systemic lupus erythematosus.

We present the case of a female patient with a diagnosis of systemic lupus erythematosus (SLE) at the age of 54 years. At the age of 63 years, she suffered from malignant thymoma and 3 years after removal of the thymoma a diagnosis of pure red cell aplasia (PRCA) was established. This is, to our knowledge, the first report of the occurrence of SLE, thymoma and PRCA in the same patient. The case is discussed with regard to the already known associations between these diseases.

Autoreactive liver-infiltrating T cells in primary biliary cirrhosis recognize inner mitochondrial epitopes and the pyruvate dehydrogenase complex.

Primary biliary cirrhosis (PBC) is characterized by lymphoid infiltrates in the portal tracts of the liver and the occurrence of antimitochondrial autoantibodies in serum directed against components of the pyruvate dehydrogenase complex and the other alpha-keto acid dehydrogenase complexes. These enzymes are located on the inner mitochondrial membrane. The destruction of the biliary tract in PBC is thought to be mediated by autoreactive liver-infiltrating T cells exerting cytotoxic activity or releasing certain lymphokines. In this study the reactivity of liver infiltrating T cells was shown to a bovine pyruvate dehydrogenase complex (PDH), a purified E2 subunit (PDH-E2) and a crude prepara…

T cell receptor gene rearrangements of T lymphocytes infiltrating the liver in chronic active hepatitis B and primary biliary cirrhosis (PBC): Oligoclonality of PBC-derived T cell clones

Immunological events are involved in the pathophysiology of chronic active hepatitis as indicated from the accumulation of T lymphocytes at the site of tissue damage. We generated T cell clones from liver biopsies of 3 patients with chronic active hepatitis B and 2 patients with primary biliary cirrhosis. These T cell clones (n = 84) were analyzed by means of T cell receptor (TcR) beta gene rearrangements to determine whether the infiltrate consists of a polyclonal or oligoclonal T cell population. The vast majority (62 of 64) of T cell clones from three different patients with chronic active hepatitis B showed no identical rearrangements of the TcR beta chain genes. In marked contrast, in …

Bile duct epithelia as target cells in primary biliary cirrhosis and primary sclerosing cholangitis.

Primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are chronic autoimmune-mediated diseases of the biliary tree, resulting in a loss of bile ducts. There are morphological features that clearly distinguish them from each other: in PBC, there is overt destruction of the bile ducts with disruption of the basement membrane; in PSC there is abundant periductular fibrosis with shrinkage and subsequent loss of the bile ducts. In order to see if the disparate histopathology is paralleled by different immunohistology we looked at a panel of epitopes on bile duct epithelia especially to see if biliary epithelial cells may present as targets for cell mediated immune response. In…

Antibodies to cytoskeletal proteins in patients with Crohn's disease

. The immunologic basis of inflammatory bowel disease has been the focus of interest of a series of studies on Crohn's disease and the process of immune sensitization at the gastrointestinal mucosal level is functionally poorly understood. To date only few contradictory reports concerning the incidence of autoantibodies in patients with this disease exist. The aim of this study was to investigate the sera drawn from 60 patients suffering from biopsy-proven Crohn's disease to evaluate the prevalence of autoantibodies against nuclear antigens and cytoskeletal proteins. Using standard methods, no anti-nuclear antibodies or antibodies to extractable nuclear antigens could be detected. All sera …

Sekundärer Immundefekt bei Niereninsuffizienz am Beispiel der Hepatitis B-Impfung

In dialysis patients the immune response to hepatitis B-vaccination is greatly impaired. In vitro the non-responders show a failure of the monocytes to support the process of primary T-cell activation. This defect results in a lack of interleukin 2-production and an enhanced sensitivity of the interleukin-2 receptor system. Addition of low doses of interleukin-2 fully reconstitutes the deficient immune response in vitro. Furthermore, the local application of low dose interleukin-2 during a standard vaccination with 40 µg hepatitis B-vaccine normalizes the non-responder state in vivo.

Indomethacin treatment in amphotericin B induced nephrogenic diabetes insipidus.

Nephrogenic diabetes insipidus (NDI) is a serious side effect of various drugs. Elevated renal prostaglandin E2 levels have been found in patients with lithium-induced NDI and have been implicated in the pathogenesis. We report the case of a patient who developed NDI following treatment with amphotericin B. Prostaglandin levels were elevated. Indomethacin had an antidiuretic effect and normalized prostaglandin levels.

Remission of severe rheumatoid arthritis following liver transplantation.

We present the case of a 32-year-old male who suffered from severe RA from the age of 21 years. After 9 years of active disease and poor response to therapy the patient developed severe hepatitis induced by the NSAID pirprofen. He went into fulminant hepatic failure necessitating emergency liver transplantation. Liver transplantation was followed by clinical and laboratory remission of his RA and he has remained virtually asymptomatic for more than 3.5 years. The possibility that this favourable clinical course was due to the immunosuppressive effect of the liver transplant rather than the ensuing immunosuppressive therapy is discussed.

Analysis of the in vitro cytokine production by liver-infiltrating T cells of patients with autoimmune hepatitis.

SUMMARY The pathogenic mechanisms underlying the development of autoimmune hepatitis (AIH) are still unclear. Since AIH is associated with the presence of various autoantibodics and certain HLA subtypes, it is likely that T and B cells play a major role in this disease. In this study we have determined the functional capacities of in vivo preactivated liver-infiltrating T cells (LTC) from patients with AIH. As controls we used LTC from patients with non-autoimmune hepatitis (non-AIH). Our results show that preactivated LTC from patients with AIH predominantly (190/255 clones) reside in the CD4+ population, whereas LTC in non-AIH are dominated by the CD8+ phenotype (148/254 clones). In view …

Zur Frage immunbiologischer Beziehungen zwischen Lupus erythematodes disseminatus und progressiver Sklerodermie

Nach einer kurzen Literaturbesprechung werden eigene Untersuchungen uber das Vorkommen antinuclearer Antikorper bei 15 Patienten mit Lupus erythematodes disseminatus, 5 Patienten mit Lupus erythematodes disseminatus kombiniert mit anderen immunologisch bedingten Erkrankungen sowie 12 Patienten mit progressiver Sklerodermie gegenubergestellt. Zur Bewertung der Befunde dienen Untersuchungen uber die Haufigkeit antinuclearer Faktoren bei primar chronischer Polyarthritis, Morbus Bechterew, akuten und chronischen Lebererkrankungen, einer inhomogenen Gruppe mit verschiedenen sogenannten Kollagenkrankheiten sowie klinisch gesunden Blutspendern. Die in samtlichen Gruppen nachweisbaren antinuclearen…

Adjuvante und palliative Therapie kolorektaler Karzinome

Membrane Expression of Autoantigens on Mechanically and Enzymatically Isolated Hepatocytes

Die Plasmamembranexpression von Leberzellmembran-Antigenen wurde mit indirekter Immunfluoreszenz (IF) und Immunelektronenmikroskopie (IELMI) an mechanisch und enzymatisch isolierten Kaninchenhepatozyten untersucht. 90 – 95 % der enzymatisch isolierten Hepatozyten, aber nur 40 – 60 % der mechanisch isolierten Hepatozyten waren intakt und vital. Eine Untergruppe der autoimmunen chronisch aktiven Hepatitis (CAH) weist zirkulierende Antikorper gegen ein mikrosomales Antigen aus Leber und Niere (LKM) auf. Die vermutete Membranexpression fur LKM-Antigene konnte durch unsere Untersuchungen mit IF und IELMI an vitalen Kaninchenhepatozyten nicht nachgewiesen werden. Der als Referenzantikorper verwen…

Autoimmune hepatitis

Autoimmune hepatitis (AIH) is a distinct form of acute and chronic inflammatory liver disease in which immune reactions against host antigens are found to be the major pathological mechanism. If left untreated it carries an unfavourable prognosis, and the diagnosis should be made as soon as possible. The diagnostic approach has been greatly facilitated by the establishment of a panel of marker autoantibodies, which do not define distinct therapeutic groups of AIH, but do allow a subgrouping based on differences in patient populations, some clinical features and prognosis. The characterization of organ-specific components of the liver cell surface as targets of cellular and humoral autoimmun…

Untersuchungen zur Frage organspezifischer Antigene der Leber

Es wird uber den Nachweis speciesunspezifischer bzw. organspezifischer Faktoren gegen mindestens ein Protein der loslichen Proteinfraktionen von Lebergeweben bei vier Patienten mit chronischer Hepatitis berichtet. Die Befunde konnten tierexperimentell bestatigt werden. Mit Rattenleber-, -nieren- bzw. -schilddrusenhomogenat immunisierte Kaninchen bildeten in den ersten 8–10 Tagen nach der letzten Immunisierung vorwiegend Makroglobulinantikorper, nach dem 20. Tag nach der Immunisierung vorwiegend bei 65° hitzestabile 7-S-Globulin-Antikorper. In den Leberantiseren liesen sich 7-S-Antikorper gegen mindestens ein Protein der loslichen Proteinfraktion von Rattenlebern abgrenzen. Eine ausfuhrliche…

The gene of hepatocyte growth factor is expressed in fat-storing cells of rat liver and is downregulated during cell growth and by transforming growth factor-β

Hepatocyte growth factor (HGF) has been detected in non-parenchymal cells but not in hepatocytes. We performed Northern blot analysis of total RNA extracted from rat hepatocytes, Kupffer cells, endothelial cells and fat-storing (Ito-) cells. Total RNA was extracted from fat-storing cells at different times after isolation and from cells treated with different amounts of transforming growth factor beta. The RNA was hybridized with HGF, fibronectin-, and alpha-actin-specific cDNA probes, consecutively. We found an abundant amount of HGF mRNA in freshly isolated fat-storing cells, but not in other liver cells. The amount of the HGF transcripts decreases significantly in FSC during the time of …

Hepatic transferrin receptors in hereditary hemochromatosis.

Secretory effect of azodisalicylate (azodisal sodium) on the short circuited mucosa of the rat ileum in vitro.

Azodisalicylate (ADS) is one of the newly developed substitutes of sulphasalazine consisting of two molecules of 5-amino-salicylic acid. Azodisalicylate caused diarrhoea in some patients, apparently caused by an antiabsorptive secretagogue action of this compound. The mechanism of this was studied in the short circuited isolated mucosa of the rat ileum. Mucosal addition of ADS increased the potential difference (PD) and short circuit current (Isc) at a concentration of 1.3.10(-4) mol/l (4 mg/dl) with maximal effects at 1.3.10(-3) mol/l (40 mg/dl). Epithelial resistance was only slightly decreased at the higher concentrations of 40 and 100 mg/dl. Serosal ADS had no effect on electrical param…

Tolerance exists towards resident intestinal flora but is broken in active inflammatory bowel disease (IBD)

SUMMARY Hyporesponsiveness to a universe of bacterial and dietary antigens from the gut lumen is a hallmark of the intestinal immune system. Since hyperresponsiveness against these antigens might be associated with inflammation, we studied the immune response to the indigenous intestinal microflora in peripheral blood, inflamed and non-inflamed human intestine. Lamina propria monocuclear cells (LPMC) isolated from inflamed intestine but not peripheral blood mononuclear cells (PBMC) of IBD patients with active inflammatory disease strongly proliferated after co-culture with sonicates of bacteria from autologous intestine (BsA), Proliferation was inhibitable by anti-MHC class II MoAb, suggest…

Cytotoxic effects of antibodies to proteinase 3 (C-ANCA) on human endothelial cells.

SUMMARY Autoantibodies directed against cytoplasmic antigens of neutrophils (ANCA), especially those with specificity for proteinase 3 (PR-3) and myeloperoxidase, are valuable markers for differential diagnosis and monitoring of disease activity in Wegener's granulomatosis (WG) and other vasculitides. Till now, several concepts concerning a direct role of antibodies against PR-3 in the pathogenesis of WG have been discussed. Recently we were able to show that these antibodies recognize PR-3 translocated into the membrane of human endothelial cells. The aim of this study was to investigate putative cytotoxic effects of antibodies to PR-3 on human endothelial cells. Antibodies were obtained b…

Human parvovirus B19 infection associated with severe acute perimyocarditis in a 34-year-old man

Rare association of herpes simplex virus IgM-specific antibodies and Guillain-Barré syndrome successfully treated with plasma exchange and immunosuppression

Herpes simplex virus (HSV) has been associated with various neurological disorders. In contrast, HSV infection is very rarely found in acute polyneuroradiculitis. In this report, a patient is described with a severe course of Guillain-Barr¿ syndrome (GBS). HSV IgM-specific antibodies and a rise of complement-fixation antibodies were detected. During the acute phase of neurologic syndrome, a nerve biopsy showed myelin damage and IgM deposits on the inner layer of the perineurium. Plasma exchange, in combination with immunosuppression, was successfully applied as a treatment in the relapsing course of GBS. Finally, after recovery, HSV-specific IgM antibodies disappeared.

Constitutive and inducible protein/DNA interactions of the interferon-γ promoter inin vivo CD45RA and CD45RO T helper subsets

Interferon-gamma (IFN-gamma) is a key cytokine of T lymphocytes with major regulatory functions in the immune system. To determine and compare protein/DNA interactions at the native IFN-gamma locus in T cells, we analyzed the human IFN-gamma promoter by ligation-mediated polymerase chain reaction (LM-PCR) techniques. Accordingly, Jurkat T cells and primary CD45RA and CD45R0 CD4+ T cell subsets isolated from peripheral blood using immunomagnetic beads were cultured and analyzed by LM-PCR. Constitutive and inducible protein/DNA interactions of the IFN-gamma promoter in vivo were detected in all T cells tested. Interestingly, an inducible footprint between -183 and -196 was consistently observ…

Anti-neutrophil cytoplasmic antibodies in a rat model of trinitrobenzenesulphonic acid-induced liver injury

Background In sera from patients with autoimmune liver diseases, e.g. primary sclerosing cholangitis (PSC) and autoimmune hepatitis, anti-neutrophil cytoplasmic antibodies (ANCAs) can be found. Until now, no animal model of ANCA induction in liver disease has been described. In this study, we describe a novel rat model of acute liver injury and subsequent ANCA production. Materials and methods The hapten reagent 2,4,6-trinitrobenzenesulphonic acid (TNBS) was injected into the portal vein of female Lewis rats. Two experimental groups were studied: group A (TNBS/ethanol) received different TNBS concentrations; control animals of group B (ethanol) were injected with 10% (v/v) ethanol/0.9% (w/v…

A p16INK4a-insensitive CDK4 mutant targeted by cytolytic T lymphocytes in a human melanoma.

A mutated cyclin-dependent kinase 4 (CDK4) was identified as a tumor-specific antigen recognized by HLA-A2. 1-restricted autologous cytolytic T lymphocytes (CTLs) in a human melanoma. The mutated CDK4 allele was present in autologous cultured melanoma cells and metastasis tissue, but not in the patient's lymphocytes. The mutation, an arginine-to-cysteine exchange at residue 24, was part of the CDK4 peptide recognized by CTLs and prevented binding of the CDK4 inhibitor p16INK4a, but not of p21 or of p27KIP1. The same mutation was found in one additional melanoma among 28 melanomas analyzed. These results suggest that mutation of CDK4 can create a tumor-specific antigen and can disrupt the ce…

The function of the soluble interleukin 6 (IL-6) receptor in vivo: sensitization of human soluble IL-6 receptor transgenic mice towards IL-6 and prolongation of the plasma half-life of IL-6.

Interleukin 6 (IL-6) is considered an important mediator of acute inflammatory responses. Moreover, IL-6 functions as a differentiation and growth factor of hematopoietic precursor cells, B cells, T cells, keratinocytes, neuronal cells, osteoclasts, and endothelial cells. IL-6 exhibits its action via a receptor complex consisting of a specific IL-6 receptor (IL-6R) and a signal transducing subunit (gp130). Soluble forms of both receptor components are generated by shedding and are found in patients with various diseases such as acquired immune deficiency syndrome, rheumatoid arthritis, and others. The function of the soluble (s)IL-6R in vivo is unknown. Since human (h)IL-6 acts on human and…

Circadian variations in antigen-specific proliferation of human T lymphocytes and correlation to cortisol production.

Cortisol is a well-known immunosuppressant when used therapeutically. The present investigation was set out to study if diurnal variations in endogenous cortisol production are reflected by changes in proliferative responses of human lymphocytes to either a mitogen (phytohemagglutinin-A, PHA) or an antigen (tetanus toxoid, TT) stimulus. The study included eight healthy volunteers. Blood was withdrawn at 0200, 0600, 1000, and 1800h for preparation of lymphocytes and determination of cortisol in plasma. Isolated cells were incubated without (baseline activity) or with inclusion of either 1 micrograms PHA or 10 micrograms TT. Proliferation was measured by labelling with 3H-thymidine for 16 h o…

Rat hepatocytes in primary culture synthesize and secrete cellular fibronectin

Fibronectins, involved in cell-matrix interactions and cell attachment, are glycoproteins which show a remarkable heterogeneity, due to alternative splicing. The type III-related domains, ED-A and ED-B, are present in cellular fibronectin in a variety of ratios whereas they are absent in circulating plasma fibronectin. Fibronectin synthesis by hepatocytes which are accepted as suppliers of plasma fibronectin was studied in primary cultures during a 6-day culture period. Using site-specific antibodies we demonstrate that rat hepatocytes are also able to synthesize and secrete fibronectin bearing the ED-A domain from Day 3 on after inoculation. By immunocytological characterization of the hep…

Antisense Phosphorothioate Oligonucleotides to the p65 Subunit of NF-κB Abrogate Fulminant Septic Shock Induced byS. typhimuriumin Mice

The aim of this study was to characterize the functional relevance of the transcription factor NF-κB in the pathogenesis of septic shock. BALB/c mice were infected with two wild-type (WT 1, WT 2) strains of S. typhimurium that induce NF-κB or an escape variant that lacks this ability (P21) at a dose of 1 × 109/animal, respectively. Furthermore, wild-type infected mice were treated with antisense oligonucleotides directed against NF-κB 24 h before and 3 or 6 h after infection, while mismatched oligonucleotides were used as controls. Subsequently, the clinical course, histological and immunological alterations were monitored. Infection with WT 1 and WT 2 strains led to lethal septic shock wit…

Expression of epithelial antigens EPM-1 and EXO-1 in normal, transitional, inflammatory and neoplastic colorectal mucosa

EPM-1 (a high molecular weight glycoprotein) and EXO-1 (a carbohydrate epitope expressed on polar neutral glycolipids and mucins) are two developmental antigens of normal and neoplastic human epithelia and were characterised by monoclonal antibodies. Their distribution was investigated in normal and pathological human colorectal mucosa. In normal mucosa, EPM-1 and EXO-1 showed characteristic expression patterns. EPM-1 was differentially expressed along the crypt villus axis with maximum at the crypt basis. EXO-1 was present throughout the whole mucosa. The characteristic gradient of EPM-1 expression along the crypt axis in normal mucosa was no longer detectable in benign polyps. Intact grad…

Hepatitis B virus and delta infection in male homosexuals

— Six hundred and sixty-six homosexuals were analysed in respect of hepatitis B virus and delta infections. Evidence of ongoing or recent hepatitis B virus infection was found in 450/666 (67.6%) homosexuals; 44 were HBsAg positive. Anti-delta was found in two HBsAg-positive homosexuals. Both individuals had a non-replicative form of HBV infection and biochemical evidence of liver disease. The study confirms that HBV infection is frequent in homosexuals and indicates that delta-infection is rare in male homosexuals.

Angiomyolipome der Nieren bei Morbus Bourneville-Pringle

Three cases with renal angiomyolipoma in Bourneville's disease are reported. In two patients advanced renal insufficiency developed, one being treated by chronic intermittent hemodialysis. In the third patient rupture of angiomyolipoma with severe retroperitoneal bleeding occurred. In one patient, tumorous enlargement initially was diagnosed only in the right kidney and preceded angiomyolipoma of the left kidney for several years. In this patient nephrectomy was performed because malignancy was assumed before diagnosis of Bourneville's disease was established. Occasionally, histology may suggest sarcomatous changes. However, the absence of distant metastasis and the overall long survival of…

Reactivity of infiltrating T lymphocytes with microbial antigens in Crohn's disease.

Intestinal T lymphocytes are normally unresponsive to microbial and recall antigens in vitro, whereas the same antigens induce strong immune responses in peripheral-blood-derived T cells. We obtained T lymphocytes from peripheral blood and from the non-inflamed and inflamed intestinal mucosa of 6 patients (3 male, 3 female; mean age 33 years) with Crohn's disease. The T cells were stimulated in vitro with a range of microbial antigens. Whereas T cells from normal mucosa were unresponsive, those from inflamed mucosa had a proliferative response comparable to that of the peripheral-blood-derived T cells. These findings suggest that physiologic unresponsiveness to luminal antigens is abrogated…

Die Duplexsonographie der Leber und des Pfortadersystems

□ Die Duplexsonographie ermoglicht die Beurteilung der Hamodynamik des gesamten portalvenosen und Lebervenengefassystems. Die Methode ist etabliert als „noninvasive Splenoportographie” und hat einen hohen diagnostischen Rang durch fruhzeitigen und schnellen Nachweis von Pfortader- und Milzvenenthrombosen, dem Budd-Chiari-Syndrom, der Darstellung portosystemischer Kollateralen und Veranderungen der Stromungsrichtung in der Pfortader.

Estimation of the frequency of self-reactive T cells in health and inflammatory diseases by limiting dilution analysis and single cell cloning.

Autoreactive T cells have recently been detected not only in autoimmune diseases but also in healthy individuals, but their frequency is thought to be low. The aim of our study was to estimate the frequency of self-reactive T cells by using limiting dilution analyses of peripheral blood lymphocytes. Assessment of self-reactivity in this study was defined as T-cell proliferation to autologous non-T cells in the absence of foreign antigens. When culture conditions were optimized by adding interleukin 2, healthy individuals showed a frequency of self-reactive T cells ranging from 1/60 to 1/600. These results were confirmed by using unseparated peripheral blood leukocytes or Epstein-Barr virus …

Translocation of the nuclear autoantigen La to cell surface: assembly and disassembly with the extracellular matrix.

La (SS-B) protein is known as one major antigenic target for autoantibodies from patients with certain autoimmune diseases such as Sjogren's syndrome or Lupus Erythematosus. La protein belongs to the so called "extractable nuclear antigens". Here we report that La antigen is not restricted to the nucleus as one might deduce from the exclusive nuclear staining pattern of patient anti-La antibodies but after stimulation of serum-starved cells with 10% fetal calf serum (FCS) appears and stays for at least 45 min at the outer surface of CV-1 cells being available for binding of anti-La antibodies. In addition we found that a minor part of La antigen associates with the extracellular fibronectin…

Lack of hepatitis B virus DNA sequences in sera from patients with acute and chronic liver diseases diagnosed as non-A, non-B-hepatitis

— The sera of 15 individuals with transfusion-associated acute or chronic non A, non B hepatitis, which lacked hepatitis B virus markers, were tested for hepatitis B virus DNA by dot blot hybridization test. Three sera of two patients positive in this test, however, also gave positive results when the labeled plasmid was used as probe instead of labeled HBV-DNA, indicating false positive results in the initial test. In conclusion, the data indicated that sera of patients with confirmed non A, non B hepatitis do not contain DNA-sequences in the serum hybridizing with HBV-DNA.

Differential effects of IL-10 on proliferation and cytokine production of human gamma/delta and alpha/beta T cells.

Gamma/delta TCR bearing T lymphocytes represent a T-cell subset whose functional relevance remains unclear. Nevertheless these T cells may play a role in the early immune response against bacteria. Until now the regulatory mechanisms on this response have not been investigated. The study described here evaluated the immunoregulatory effects of Interleukin-10 on gamma/delta and alpha/beta TCR-positive T-cell clones and freshly isolated peripheral-blood mononuclear cells (PBMC). IL-10 has been shown previously to inhibit lectin and antigen-induced proliferation and cytokine production by alpha/beta T cells. The results outlined below show that rhIL-10 strongly inhibits lectin-induced producti…

ELISA techniques for the detection of antimitochondrial antibodies

Die Charakterisierung des klinisch gesunden Hepatitis-B-Antigen (HBsAg)-Trägers

Wir haben 129 Blutspender, bei denen bei Routineuntersuchungen HBsAg im Serum festgestellt wurde, zur diagnostischen Klarung einer moglichen Lebererkrankung untersucht. Zwolf hatten, als sie zur klinischen und leberbioptischen Untersuchung aufgenommen wurden, schon kein HBsAg mehr im Serum. Keiner von diesen hatte klinisch oder histologisch Anhalt fur eine entzundliche Lebererkrankung. Zwei der 129 Patienten hatten eine milde, ikterische Hepatitis, verloren das Antigen wahrend des Verlaufes und wurden anti-HBs-positiv. Die ubrigen 115 Patienten, die klinisch gesund erschienen und keine fruhere ikterische Leberkrankheit hatten, blieben HBsAg-positiv wahrend des bisherigen Verlaufes von im Mi…

Detection of mitochondrial antibodies directed against the primary biliary cirrhosis (M2) antigen by an enzyme-linked immunosorbent assay (ELISA)

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of 1 subtype of mitochondrial antibodies (AMA) specific for chronic cholestatic inflammatory liver diseases. AMA were detected by ELISA in 16 of 16 patients with primary biliary cirrhosis (PBC) and in 2 of 31 patients with chronic active hepatitis. These 18 positive sera were positive for AMA by indirect immunofluorescence (IF) and by radioimmunoassay (RIA). No AMA were detected by ELISA in 2 patients with the pseudolupus erythematosus syndrome, who were positive for AMA by IF, 2 patients with secondary syphilis, positive for cardiolipin antibodies, 1 patient with systemic lupus erythematosus, positive for AMA by I…

GD3 ganglioside: A marker for the diagnosis and treatment of neuroectodermal tumors?

Liver membrane autoantibodies in chronic active hepatitis

Summary Target antigens relevant for immune reactions in inflammatory liver diseases should be expressed on the hepatocellular membrane. Using mechanically or enzymatically isolated rabbit hepatocytes, we evaluated the influence of cell integrity on the detection of membrane-expressed antigens by sera from patients with chronic hepatitis and by murine monoclonal antibodies. Our results provide evidence that target antigens of liver membrane autoantibodies (LMA) as well as liver kidney microsomal antibodies (LKM) are not expressed on the hepatocellular membrane of viable and intact isolated rabbit hepatocytes. However, LMA were detected in the sera of 56% of patients with autoimmune chronic …

Pneumatoceles and pneumothoraces complicating staphylococcal pneumonia: treatment by synchronous independent lung ventilation.

A 54 year old man with a staphylococcal sepsis developed staphylococcal pneumonia complicated by multiple pneumatoceles and bilateral tension pneumothoraces caused by bronchopleural fistulae. Excessive enlargement of the right sided pneumatoceles and a tension pneumothorax not improved by drainage led to mediastinal shift and compression of the right lung. Reversal of the mediastinal shift and closure of the bronchopleural fistulae was achieved by assisted independent lung ventilation.

Chronifizierte paranoid-halluzinatorische Psychose als Erstmanifestation einer HIV-Infektion?

In a 36-year-old patient an acute onset of psychosis occurred, probably due to HIV infection. For one year HIV-infection with reduced T4/T8 ratio had been known without clinical manifestation (stage IV B of the CDC-classification). He developed chronic delusional hallucinations, which persisted for more than one year in spite of adequate psychoactive drug therapy. So far AIDS-related dementia has not become evident. Focal lesions caused by opportunistic infections or tumour were excluded by computed tomography and magnetic resonance imaging. The latter revealed several small lesions and the brain scan showed a nonhomogeneous pattern of cerebral blood flow. CSF-examination disclosed a mild l…

Analysis of liver-specific protein LSP using murine monoclonal antibodies.

. We describe twenty murine monoclonal antibodies directed against different antigenic determinants of human and rabbit liver-specific protein LSP. Among them, nine were directed against liver-specific epitopes as judged from immunohistological studies. Immunoelectronmicroscopy revealed that seven of these monoclonals recognized membrane determinants differing in staining of distinct areas of the hepatocellular surface. Eleven antibodies were directed against intracellular structures. Western blot analysis showed that the epitopes detected were displayed on either single or multiple protein bands with apparent molecular weights between 24 000 and 60 000. Further differences were observed wi…

Immunochemical characterization of anti-acetylcholine receptor antibodies in primary biliary cirrhosis

Although the presence of anti-mitochondrial antibodies is the main characteristic of primary biliary cirrhosis (PBC), other autoantibodies have been described in this disease. This study employs immunoblot methods to test whether the sera of PBC patients also contain antibodies directed against nicotinic acetylcholine receptors (AChR). We show that the majority of patients' sera indeed react with AChR just as sera of myasthenic patients do. In contrast, however, these anti-AChR antibodies do not lead to significant clinical symptoms of myasthenia. In all cases studied, PBC sera recognized a protein with the molecular weight of the alpha-chain of acetylcholine receptor (40 kDa). In addition,…

The diagnostic significance of intrahepatocellular hepatitis-B-surface-antigen (HB s Ag), hepatitis-B-core-antigen (HB c Ag) and IgG for the classification of inflammatory liver diseases

Liver biopsies of patients with inflammatory liver diseases and clinically healthy HBsAg-carriers were examined for presence of intracellular HBsAg, HBcAg and IgG by direct immunofluorescence. The studies revealed the following results: 1. In most cases healthy HBsAg-carriers had HBsAg in the cytoplasm, but they did never show HBcAg in the nuclei of hepatocytes. 2. In the early phase some patients with HBsAg-positive acute hepatitis had HBcAg and/or HBsAg in their hepatocytes. In a normal course with complete recovery the immunoelimination may clear either phenomenon at variable stages of the disease. 3. Cases one year after complete recovery of acute virus B-hepatitis had no HB-components …

Minimalheparinisierung bei Dialysepatienten mit erhöhter Blutungsgefährdung

Abstract In 78 patients (47 men, 31 women; mean age 53 [22-78] years) 174 dialyses were undertaken within one week of a bleeding episode or a diagnostic or therapeutic procedure which may cause bleeding. Minimal anticoagulation with low molecular weight heparin (LMWH) was the aim, using a biocompatible dialyser. During the dialysis coagulation was controlled by global tests (Quick value/international normalized ratio [INR], partial thromboplastin time, thrombin time, antifactor Xa activity), by molecular markers of clotting activity (thrombin-antithrombin III complex [TAT], D-dimers), as well as measurement of elastase (elastase-alpha 1-protein inhibitor complex). The LMWH dosage averaged 9…

Effects of Th1 and Th2 cytokines on cytokine production and ICAM-1 expression on synovial fibroblasts

OBJECTIVES--To investigate the influence of the Th1 and Th2 lymphokines interleukins (IL)-4 and IL-13, interferon gamma (IFN gamma), and several monokines on the adhesion of mononuclear cells to synovial fibroblasts and intercellular adhesion molecule-1 (ICAM-1) expression and cytokine production of synovial fibroblasts in patients with osteoarthritis. METHODS--Synovial fibroblasts were isolated from patients with osteoarthritis and stimulated with IL-1 beta, IL-4, IL-6, IL-10, IL-12, IL-13, tumour necrosis factor alpha (TNF alpha), and IFN gamma. Subsequently, we determined the production of IL-1 alpha, IL-1 beta, IL-6, IL-10, IL-12, IFN alpha and TNF alpha, and the expression of ICAM-1 ly…

Human asialoglycoprotein receptor as an autoantigen in chronic hepatitis.

ASGPR may play a role in the pathogenesis of autoimmune chronic liver diseases. Several lines of evidence support this hypothesis. Antibodies to rabbit ASGPR could be found in various inflammatory liver diseases. In contrast, ASGPR preparations derived from human liver (h-ASGPR) were recognized predominantly by sera from patients with ai-CAH. Moreover, anti-h-ASGPR showed a close correlation to the inflammatory activity of ai-CAH both in terms of prevalence (88% in histologically proven active diseases), immunoglobulin class (IgM antibodies restricted to active inflammation) and behavior during treatment. Anti-h-ASGPR secretion could also be found in vitro, when PBL of patients were stimula…

NITROGLYCERINE DILATATION OF SPHINCTER OF ODDI FOR ENDOSCOPIC REMOVAL OF BILEDUCT STONES

Liver cell damage caused by monoclonal antibody against an organ-specific membrane antigen in vivo and in vitro

Summary Monoclonal antibodies have been raised against different antigenic determinants of normal rabbit hepatocytes. One antibody (2D3) recognized a liver-specific 43 kDa protein displayed exclusively on the basolateral portion of the hepatocellular membrane. Purified monoclonal antibodies were injected intravenously into rabits. Following the injection of antibody 2D3, a dose-dependent increase of liver enzyme activities in sera was observed. Within 8 h, marked morphological alterations of the hepatocytes, including multiple cell necroses, could be demonstrated by light and electron microscopy. When isolated vital rabbit hepatocytes in culture were used as targets, cytotoxic effects of th…

Cellular cytotoxicity against the human hepatoma cell line PLC/PRF/5 in patients with hepatitis B virus-induced chronic active hepatitis (CAH) and non-A, non-B (NANB) CAH

Untersuchungen zur Hepatitis B-Antigen(HBAg)-Fixation an peripheren Lymphocyten und isolierten Leberzellen bei Patienten mit entz�ndlichen Lebererkrankungen

Bei 127 Patienten mit verschiedenen entzundlichen Lebererkrankungen wurde die Fixation von HBAg an peripheren Lymphocyten untersucht. Bei 60 von diesen Patienten wurden im Parallelansatz die Fixation von HBAg an peripheren Lymphocyten und isolierten Leberzellen sowie die Fixation von IgG an isolierten Leberzellen gepruft.

Elevated serum E-selectin in patients with liver metastases of colorectal cancer

E-selectin, an endothelial cell adhesion molecule, mediates the initial step of leucocyte adhesion to activated vascular endothelium. The soluble isoform of E-selectin promotes angiogenesis in rat cornea. In the present study, we investigated whether leucocyte adhesion and angiogenesis are also involved in tumour progression and metastasis of colorectal cancer. Therefore, we determined the level of circulating soluble E-selectin in serum samples of 38 patients with colorectal cancer; 20 patients with non-metastatic and 18 patients with metastatic disease. Median levels of soluble E-selectin were found to be significantly higher in metastatic tumour disease (88.7 ng/ml, range 25-203 ng/ml) t…

Liver-infiltrating T helper cells in autoimmune chronic active hepatitis stimulate the production of autoantibodies against the human asialoglycoprotein receptor in vitro.

SUMMARYAutoantibodies against the human asialoglycoprotein receptor (ASGPR) occur in the sera orpaticnts with autoimmune liver disorders. Live-nfiltrating T cell clones that specifically recognize the ASGPR have been described in patients with autoimmune chronic active hepatitis (A-AH) and primary biliary cirrhosis (PBC). Recently, we have shown that peripheral blood mononuclcar cells (PBMC) from patients with A-AH or PBC but not chronic viral hepatitis secreted ant-SGPR antibodies in vitro. In this study we characterized the influence of live-nfiltrating T cells on the secretion of ASGP-pecific autoantibodies by autologous B cells in cell culture supernatants. T cell clones from liver biop…

?(1,3)Fucosyltransferase expression in E-selectin-mediated binding of gastrointestinal tumor cells

E-selectin recognizes the oncofetal antigen sialyl-Lewis X, which is highly expressed in adenocarcinoma. Five α(1,3)fucosyltransferases (FT) have been cloned that confer cell-surface expression of sialyl-Lewis X on transfected cells. We show here that 12/18 gastrointestinal-tumor cell lines bind specifically to immobilized E-selectin and that in sialyl-Lewis-X-positive cells binding is inhibited with a monoclonal antibody against sialyl-Lewis X. Using RT-PCR, we determined the expression of the α(1,3)fucosyltransferases III, IV, V, VI and VII in gastrointestinal tumor cells. Transcripts of FT IV and FT VII are abundantly expressed in all tested cells. Therefore no single fucosyltransferase …

Characterization of liver cytokeratin as a major target antigen of anti-SLA antibodies.

Abstract Anti-SLA antibodies characterize a newly defined subgroup of patients with autoimmune chronic active hepatitis. The aim of the present study was the immunochemical characterization of the target antigen(s) of anti-SLA antibodies. Anti-SLA-positive sera were found to contain high titres of anti-cytokeratin antibodies. In immunoblotting analyses with 100 000 × g supernatants of human liver homogenates (S-100) these sera recognized various proteins with a molecular mass of 40–60 kDa. These proteins were also recognized by monoclonal anti-cytokeratin antibodies. Two-dimensional co-electrophoresis and immunoblotting analysis of S-100 and liver cytokeratins showed that anti-SLA antibodie…

[Pseudoxanthoma elasticum (Grönblad-Strandberg syndrome) and rheumatoid arthritis].

A 72-year-old woman, not previously known to have coronary heart disease, was admitted to hospital with an acute anterior wall myocardial infarction. The history revealed that, when about 40 years of age, a coarse skin-fold and yellowish-white xanthoma-like efflorescences had been noted around her umbilicus, the inguinal regions and axillae. These changes subsequently developed into a pathognomonic picture of pseudoxanthoma elasticum (PE), which was a significant factor in the myocardial infarction. At the age of 69 years, rheumatoid arthritis (RA), stage II after Steinbrocker, had been diagnosed on the basis of morning stiffness, symmetrical arthritis in more than three joint regions and t…

Antigen-independent in vitro expansion of T cells does not affect the T cell receptor V beta repertoire.

Analysis of the variable chains (V alpha/V beta) of the specific T cell receptor (TCR) of organ-infiltrating T cells may provide further insights into the pathogenesis of many infectious diseases, malignancies, and autoimmune disorders. To determine the TCR V beta repertoire of these small T cell populations antigen-independent in vitro expansion is necessary but may select for certain T cell subpopulations. In this study various antigen independent T cell activation protocols were used to stimulate peripheral blood mononuclear cells (PBMC) of six healthy blood donors, and TCR V beta molecules were analyzed by flow cytometry and semiquantitative reverse-transcriptase polymerase chain reacti…

Organ-specificity and diagnostic value of cell-mediated immunity against a liver-specific membrane protein: Studies in hepatic and non-hepatic diseases

In chronic active hepatitis (CAH, n=58) 70% of the HBsAg negative and 48% of the HBsAg positive cases showed a CMI against human liver specific proteins (HLPI). Using HBsAg as antigen only 12% of the HBsAg negative and 24% of the HBsAg positive cases gave a CMI response. On the basis of HBsAg and autoantibodies in the serum CAH patients could be divided into 4 subgroups. A close correlation between CMI against HLPI, sex, ANA and HL-A-8 could be detected. In a follow-up study of patients with acute virus B hepatitis (n=62) CMI against HBsAg was detected in 60% of the cases in the acute phase of the disease but in 15% only 3-6 months after the onset of the illness (n=40). In patients who deve…

Bacteria-specific T-cell clones are selective in their reactivity towards different enterobacteria or H. pylori and increased in inflammatory bowel disease.

In the present study the authors investigated the T-cell response to different enterobacteria or Helicobacter pylori and tested the hypothesis that the frequency of bacteria-specific T cells is increased in the intestine of patients with active inflammatory bowel disease (IBD), i.e. Crohn's disease (CD) and ulcerative colitis (UC). The analysis of a large panel of T-cell clones (Tc) (n = 888) from peripheral blood, non-inflamed and inflamed intestine from IBD patients and control individuals shows that both peripheral blood and intestinal T-cell clones were selectively stimulated by either Salmonella typhimurium, Yersinia enterocolitica 03, Escherichia coli or Helicobacter pylori sonicates,…

Immunotherapy of Malignant Melanomas

A large number of antigens on human melanoma cells have been identified by means of monoclonal antibody technology [18, 21]. The availability of monoclonal antibodies has rapidly led to clinical investigations. Experience with monoclonal antibodies in the treatment of patients with melanoma is limited [4, 9, 10, 12, 13, 17, 19] because only small groups of patients have been treated so far. According to these studies, treatment with ganglioside antibodies appears most promising.

Electromagnetically generated extracorporeal shock waves for gallstone lithotripsy: in vitro experiments and clinical relevance.

. First generation shock wave sources have been proved to disintegrate gallstones effectively, but they require the immersion of the patient's body in a tank of water. A recently developed second generation shock wave source (Siemens-Lithostar, Erlangen, FRG) generates shock waves electromagnetically. It presents several novel features. In particular the waterbath can be omitted and due to lower shock wave pressure general anaesthesia is not required. In vitro studies showed that 36 out of 38 gallstones (11–30 mm in diameter) could be disintegrated. Two concrements resisting lithotripsy were pure white cholesterol stones. Independent of shape, size, and composition (cholesterol or pigment) …

Sustained elimination of hepatitis B virus from serum induced in a patient with chronic hepatitis B and advanced human immunodeficiency virus infection

A 48-year-old male patient was admitted with acquired immunodeficiency syndrome (stage III, Centers for Disease Control 1993) and viremic hepatitis B. Blood CD4 count was 15/microliters. Discontinuation of prednisolone, previously prescribed by the patient's family practitioner because of elevated liver enzymes, resulted in severe hepatitis (alanine aminotransferase > 300U/l). Administration of interferon-alpha (9 x 10(6) U s.c. 3 x weekly) was initiated. Serum markers of viral replication disappeared, and aminotransferase levels returned to normal within a few weeks. The patient's serum was found negative for HBsAg after 3 months. Immunohistochemical analysis of liver biopsies before and d…

Two tyrosinase nonapeptides recognized on HLA-A2 melanomas by autologous cytolytic T lymphocytes

A number of cytolytic T lymphocyte (CTL) clones derived from several melanoma patients have been found to recognize a majority of melanomas from HLA-A2 patients. We have reported previously that two such CTL clones recognize a product of the tyrosinase gene that is presented by HLA-A2. Here we show that one of these CTL clones recognizes a peptide encoded by the first nine amino acids of the putative signal sequence of tyrosinase. The other CTL clone recognizes a different tyrosinase peptide corresponding to amino acids 368-376. Both peptides contain consensus motifs of HLA-A2 binding peptides.

Kardiale Komplikationen der chronischen Polyarthritis mit Angiitis („malignant rheumatoid arthritis“)

Im Verlauf der chronischen Polyarthritis kann es zum Auftreten einer Angiitis kommen, deren Manifestationen entsprechend der Organbeteiligung sehr unterschiedlich ausfallen. Klinisch wird dabei am haufigsten eine Neuritis beobachtet, die auf eine Beteiligung der vasa nervorum hinweisen kann. Ferner kann eine lokalisierte Angiitis als Myositis oder im Bereich der Haut unter anderem als Purpura, Gangran oder Ulceration in Erscheinung treten. Bei einer generalisierten Vasculitis konnen aber auch verschiedene innere Organe und dabei das Herz mit seinen Coronargefasen beteiligt sein (BOCK, 1957; SOKOLOFF, 1966; KULKA, 1959; FASSBENDER, 1967).

Human endothelial cells express proteinase 3, the target antigen of anticytoplasmic antibodies in Wegener's granulomatosis

Abstract Autoantibodies directed against cytoplasmic antigens of neutrophils (ANCA), especially proteinase 3 (PR-3), have proved to be a useful clinical tool confirming the diagnosis or monitoring disease activity of Wegener's granulomatosis (WG). Although several concepts concerning the pathophysiologic potentials of ANCA have been discussed, only sparse data about ANCA-endothelium interactions have been available. In this study, we have investigated the expression of PR-3 in cytokine- treated human endothelial cells using purified anti-PR-3 antibodies of patients with WG, murine and human monoclonal anti-PR-3 antibodies as probes. We were able to show that tumor necrosis factor-alpha, int…

Untersuchungen �ber den Hemmfaktor der Phosphoglucose-Isomerase in Erythrocyten von Kranken mit florider Psoriasis vulgaris

Die Untersuchungen uber Herkunft und Beschaffenheit des Hemmkorpers der PGI von Kranken mit florider Psoriasis vulgaris haben ergeben, das es sich um ein nur in der Zelle, in diesem Falle im Erythrocyten, nicht aber im Serum solcher Kranker vorliegendes Protein handelt.

Alternative pathway activation of T cells by binding of CD2 to its cell-surface ligand.

Activation of resting T lymphocytes is initiated by the interaction of cell-surface receptors with their corresponding ligands. In addition to activation through the CD3 (T3)-Ti antigen-receptor complex1, recent experiments have demonstrated induction of T-cell proliferation through the CD2 (T11) molecule2–4, traditionally known as the erythrocyte(E)-receptor, through which T cells can bind red blood cells (RBC)5–7. This 'alternative pathway' of T-cell activation2 was observed in vitro in response to combinations of anti-CD2 monoclonal antibodies (mAbs) that bind to distinct epitopes of CD2, such as mAbs against T112 plus T113 (ref. 2). The physiological importance of this activation pathwa…

Metastasierendes Lungencarcinom aus juveniler Lungenadenomatose

Es wird uber einen Fall einer Lungenadenomatose bei einem erst 19 Jahre alten Mann berichtet, die zu seinen Lebzeiten bereits aufgrund klinischer Befunde und einer Lungenpunktion diagnostiziert werden konnte. Hervorhebenswert ist das ungewohnlich jugendliche Alter mit einer spateren Metastasierung in die thorakalen Lymphknoten. Es werden insgesamt 177 Falle einer Adenomatose der Weltliteratur zusammengestellt und hinsichtlich Altersverteilung, Tumorausbreitung und Verlaufsdauer analysiert.

Analysis of the precore DNA sequence and detection of precore antigen in liver specimens from patients with anti-hepatitis b e—positive chronic hepatitis

A number of naturally occurring hepatitis B virus (HBV) mutants unable to synthesize the hepatitis B e antigen (HBeAg) have been identified in patients characterized by HBV DNA and anti-HBe in their serum. Because the analysis of the HBV-associated DNA and antigens in the liver tissue is still not complete, we investigated the precore sequence of HBV DNA and its encoded proteins in the liver tissue of 32 patients positive for HBV DNA and anti-HBe in their serum. Three different groups of patients were identified. Group I (n = 14) was characterized by viral DNA sequences with a G-A transition in the distal precore gene region, thus creating a termination codon (TAG). Liver tissue from this g…

PENTAZOCINE HAMPERS BILE FLOW

Studies of the immunbiologic importance of soluble hepatic proteins

Antikorper gegen Mitochondrien- und Mikrosomenfraktionen aus Leber, Niere und Schilddruse von Mensch und Ratte sind komplett species- und organunspezifisch. Antikorper gegen entsprechende losliche Proteine der Leber sind unter Anwendung verschiedener serologischer Techniken zum Teil als organ- und im untersuchten Bereich auch als speciesspezifisch anzusprechen. Eine nahere Charakterisierung der loslichen Proteinfraktion gelang mittels Ultrazentrifugation, Chromatographie und Ammoniumsulfatfallung. Die organgebundenen Proteine waren im 150000 G Uberstand einer Ultrazentrifugation kernfreier Leberhomogenate enthalten. Chromatographisch fanden sich die Proteine im I. und II. Gipfel einer Gelfi…

Dot-immunobinding assay with the globular domain of collagen type IV for antiglomerular basement membrane antibodies

A dot-immunobinding assay for the detection of antiglomerular basement membrane antibodies has been developed. The globular domain NC1 of basement membrane collagen type IV was used as antigen. The assay proved to be specific, sensitive, and reproducible. Circulating antibodies in each of 12 sera from patients with florid Goodpasture's syndrome could be demonstrated, whereas sera from patients with Goodpasture's syndrome in clinical remission and various control sera showed no reactivity. The advantages of the dot-blot assay are: the usage of the purified Goodpasature target antigen NCI reduces unspecific binding of IgG; only minimal amounts of antigen are required to give a positive signal…

Cytolytic T-cell clones against an autologous human melanoma: specificity study and definition of three antigens by immunoselection.

Cytolytic T-lymphocyte (CTL) clones against an autologous melanoma (SK-MEL-29) were generated by mixed lymphocyte tumor culture and subsequent cloning of responder lymphocytes at limiting dilutions. These CTL clones lysed autologous melanoma but not autologous Epstein-Barr virus-transformed B cells and none of the allogeneic tumor targets included in the specificity analysis. The lysis of autologous melanoma targets could be inhibited by monoclonal antibodies against monomorphic HLA class I determinants. For proliferation of CTLs, the stimulation with the relevant target antigen on autologous tumor cells was essential. Immunoselection experiments carried out with two CTL clones revealed the…

Fatal cerebellar haemorrhage due to phenprocoumon poisoning.

A 32-year-old patient died of a cerebellar haemorrhage and the blood coagulation analysis before death suggested defective synthesis of vitamin K-dependent clotting factors due to vitamin K deficiency. The post-mortem toxicological examination of different tissues revealed phenprocoumon poisoning as the cause of death. The differential diagnosis of vitamin K deficiency and the toxicology of hydroxycoumarins are discussed.

Modification of Hepatitis B Virus Infection by Recombinant Leukocyte Alpha A Interferon

A defect in alpha interferon production in patients with chronic type B hepatitis offers a rationale principle for treating this disease with interferon. Two trials with interferon in chronic type B hepatitis indicate that this therapy achieves an elimination of HBsAg and HBeAg significantly higher than spontaneous. Our study and that of others indicate, however, that the response to interferon therapy is dependent on many variables including: the type of interferon, the interferon dose, duration of interferon treatment, sex, sexual preference in men and coinfection with other viruses. As of the multiple modes of action of interferon, a better understanding of viral replication, of the anti…

Zytokine bei chronisch-entzündlichen Darmerkrankungen

No Association between Mannose-Binding Lectin Alleles and Susceptibility to Chronic Hepatitis B Virus Infection in German Patients

Variants of the mannose-binding lectin (MBL) have been shown to be associated with low serum concentrations of the protein and to predispose to bacterial, fungal and viral infections. A recent small study on 33 Caucasian patients had suggested that a mutation at codon 52 of the MBL gene is associated with chronic hepatitis B virus (HBV) infection. Exon 1 of the MBL gene was amplified by PCR in 61 patients with chronic HBV infection, 28 patients with acute infection and in 60 controls. MBL variants were detected by subsequent restriction enzyme digestion and agarose gel electrophoresis. The occurrence of the codon 52 mutation in patients with chronic HBV infection did not differ significantl…

Immune Reactions in Liver Diseases

Chronic hepatitis is a heterogeneous syndrome with different underlying etiologies. Viruses (hepatitis B virus, non-A, non-B viruses), drugs and even alcohol are regarded as etiological agents. A further subgroup is autoimmune-type chronic hepatitis in which an immunogenetic predisposition is of major importance. Primary biliary cirrhosis has to be separated from these subgroups of chronic hepatitis.

Binding properties of mechanically and enzymatically isolated hepatocytes for IgG and C3.

— The presence of Fc and C3 receptors was studied on mechanically and enzymatically isolated rabbit, mouse and rat hepatocytes as well as on hepatocytes derived from primary cultures. The same cell preparations were used for studying the uptake of an antibody against nuclear antigens. Mechanically isolated hepatocytes were able to bind AIgG and activate complement. However, the same cells were not able to form rosettes with EA or with EAC. Enzymatically isolated hepatocytes did not bind AIgG or activate complement nor did they form rosettes with EA or with EAC. The mechanically isolated cells and the nonviable fraction of the enzymatically isolated cells showed a nuclear fluorescence when i…

Local administration of antisense phosphorothioate oligonucleotides to the p65 subunit of NF-kappa B abrogates established experimental colitis in mice.

Chronic intestinal inflammation induced by 2,4,6,-trinitrobenzene sulfonic acid (TNBS) is characterized by a transmural granulomatous colitis that mimics some characteristics of human Crohn's disease. Here, we show that the transcription factor NF-kappa B p65 was strongly activated in TNBS-induced colitis and in colitis of interleukin-10-deficient mice. Local administration of p65 antisense phosphorothioate oligonucleotides abrogated clinical and histological signs of colitis and was more effective in treating TNBS-induced colitis than single or daily administration of glucocorticoids. The data provide direct evidence for the central importance of p65 in chronic intestinal inflammation and …

Antibodies to proteinase 3 mediate expression of vascular cell adhesion molecule-1 (VCAM-1).

SUMMARY VCAM-1 was first identified as an adhesion molecule induced on human endothelial cells (HEC) by inflammatory cytokines such as IL-1, tumour necrosis factor (TNF), and lipopolysaccharide (LPS). The molecule binds to a variety of leucocytes, including B cells, T cells, basophils, eosinophils and monocytes. Vascular expression of VCAM-1 has been associated with a number of disease states, including rheumatoid arthritis and vasculitis. The detection of antineutrophil cytoplasmic antibodies (ANCA), especially to proteinase 3 (PR3), has become important in the diagnosis of Wegener’s granulomatosis (WG) and related vasculitides. Recently we were able to demonstrate a direct effect of anti-…

New cell lines of gastric and pancreatic cancer: distinct morphology, growth characteristics, expression of epithelial and immunoregulatory antigens.

Two new cell lines from stomach cancers and one from a pancreatic carcinoma are presented. MZ-GC-1 was established from a hepatic metastasis of a well differentiated gastric adenocarcinoma. MZ-GC-2 was derived from ascites induced by a poorly differentiated gastric adenocarcinoma. MZ-PC-1 originated from the pleural effusion of a moderately well differentiated pancreatic ductal adenocarcinoma. MZ-GC-1 cells were adherent and partially polarized, connected tightly via desmosomes. In contrast MZ-GC-2 cells consisted of slightly adherent or floating subpopulations and displayed no desmosomes. MZ-PC-1 cells were adherent and showed polarized growth, connected by apical junctional complexes. Cel…

Tenascin gene expression in rat liver and in rat liver cells. In vivo and in vitro studies.

Tenascin is a major glycoprotein constituent of the extracellular matrix with a strong affinity to fibronectin; its distribution is believed to be temporarily and spatially limited. Tenascin gene expression is increased during wound healing processes. As repair mechanisms in chronic liver diseases resemble wound healing we studied tenascin gene expression in rat liver and in isolated rat liver cells. In normal rat liver a tenascin specific antiserum stains sinusoidal cells with fiber-like prolongations, which at the same time are desmin-positive (ITO-cells). In the CCl4-acutely-damaged liver a strong tenascin staining is detected in cells located among the mononuclear cells of the inflammat…

Hepatozelluläre Karzinommarker bei einem primären Gallenblasenkarzinom

Eine Adenokarzinomzellinie, Mz-ChA-2, konnte kurzlich aus einem primaren Gallenblasenkarzinom, das lokal in die Leber eingewachsen war, in Gewebekultur etabliert werden (1). Zyto- und Histomorphologie dieser Tumorzellinie in vitro und nach Transplantation auf Nacktmause entsprechen dem Primartumor der 65jahrigen Patientin. Im Serum wurden leicht erhohte AFP-Werte bestimmt, wahrend die CEA-Werte normal waren. Im Gewebekulturuberstand dieser neuen Tumorzellinie MZ-ChA-2 konnte AFP nachgewiesen werden. Auf klonaler Ebene zeigten sich erhebliche quantitative Schwankungen in der AFP-Synthese. CEA konnte im Gewebekulturuberstand nicht nachgewiesen werden, dem Serumbefund bei der Patientin entspre…

Tumornekrosefaktor und Interferon als prognostische Marker der HIV-Infektion

Peripheral blood cells were obtained from patients at different stages of their human immunodeficiency virus (HIV) infection. It was found that the capacity to generate interferon alpha was reduced already at Walter Reed stage 2 (WR) while the interferon gamma capacity remained largely unaffected until WR stage 4. Endogenous tumor necrosis factor (TNF) alpha production increased as the HIV disease progressed. The data obtained add to our knowledge on destruction of the immune system by the HIV. Moreover TNF and acid labile interferon alpha might contribute to HIV replication and disease progression. Nevertheless the tests performed are too time-consuming to be introduced into routine analys…

Akutes Lungenversagen bei Malaria tropica in der Schwangerschaft: Erfolgreiche Behandlung durch extrakorporale CO2-Elimination

Eine 32jahrige Schwangere (26. Schwangerschaftswoche) erkrankte am 6. Tag nach der Ruckkehr von einem Indonesienaufenthalt mit Fieber bis 42°C, hamolytischer Anamie (Hamoglobin 7,6 g/dl) und Thrombozytopenie (7000/µl). Es wurde eine Malaria diagnostiziert; eine Prophylaxe hatte sie nicht durchgefuhrt. Unter antiparasitarer Chemotherapie mit Chloroquin, Chinin und Pyrimethamin kam es zur Elimination der Parasiten (Makrogametozyten von Plasmodium falciparum) aus dem peripheren Blut. Im weiteren Verlauf entwickelte sich jedoch ein progredientes Lungenversagen und eine therapierefraktare Pneumonie, die zu einem »adult respiratory distress syndrome« (ARDS; Morel-Stadium 4) fuhrten. Bei drohendem…

Induction of the anti-ergotypic response.

The injection of syngeneic activated T cells into rodents can induce a T cell response against activation markers of the T cells, ergotopes. The responding anti-ergotypic T cells have been shown to suppress experimental autoimmune encephalomyelitis (EAE). This paper reports the characteristics of the anti-ergotypic response. It was found that irradiated activated T cells were as good as untreated living activated T cells in inducing anti-ergotypic cells in vivo. Glutardialdehyde-fixed (0.3%) cells were poor stimulators in vivo and non-stimulatory in vitro. Dilution of glutardialdehyde to 0.003% before fixation preserved the stimulatory capacity in vitro. Fixation or irradiation of T cells a…

Methodology and significance of the detection of liver-kidney-microsomal (lkm) autoantibodies in autoimmune-type chronic active hepatitis

Liver-kidney-microsomal (LKM) autoantibodies are diagnostic markers for a subgroup of HBsAg-negative chronic active hepatitis, presumably owing to autoimmunity. They were originally detected by indirect immunofluorescence and can now be evaluated by radioimmunoassay, enzyme-linked immunosorbent assay, and immunoblotting. In immunoblotting LKM-positive sera react strongly with a 50-kilodalton (KD) polypeptide band of microsomes. In immunoelectron microscopy, LKM-positive sera show a binding with membranes of the endoplasmic reticulum. The LKM antigen was further identified on various isoenzymes of cytochrome P-450. Immunofluorescence is still the method of choice for screening sera routinely…

Endothelin-1 modulates the expression of adhesion molecules on fibroblast-like synovial cells (FLS)

Endothelin-1 is known to possess various biological properties. In the present study we have investigated the effects of Endothelin-1 (Et-1) on the expression of adhesion molecules ICAM-1, VCAM-1 and CD-44 by fibroblast-like synoviocytes. Cultured fibroblast-like synoviocytes were treated with Et-1 in the absence or presence of C1306, a specific endothelin-A-receptor antagonist. Cell surface expression of ICAM-1, VCAM-1 and CD-44 was determined by immunofluorescence studies, Cyto-ELISA and FACS-analysis. ICAM-1, VCAM-1 and CD-44 were constitutively expressed on cultured FLS. After incubation with Et-1 the expression of ICAM-1, VCAM-1 and CD-44 increased. The level of expression of adhesion …

Identification of the Yersinia enterocolitica urease beta subunit as a target antigen for human synovial T lymphocytes in reactive arthritis.

The local T-cell response to bacterial antigens is involved in the pathogenesis of reactive arthritis (ReA). Here, we have identified a 19-kDa antigen of Yersinia enterocolitica O:9 recognized by Yersinia-specific synovial fluid CD4+ T cells in two patients with Yersinia-induced ReA. N-terminal amino acid sequencing of this protein revealed that it was identical to the 19-kDa urease beta subunit of Y. enterocolitica O:9. This protein has previously been shown to be arthritogenic in preimmunized rats after intra-articular injection. Analysis of the T-cell response to this protein showed that it contains several T-cell epitopes, one of which cross-reacts with other enterobacteria not able to …

Effect of the stable prostacyclin analogue iloprost on water and electrolyte transfer of the rat ileum and colon in vivo.

The effect of iloprost on water and ion transfer was measured simultaneously in tied-off loops of the rat ileum and colon in vivo. (1) In the ileal loops iloprost had no effect on water and ion transfer neither by intraluminal, nor intraaortal or intravenous application. (2) In the colonic loops only intraaortal bolus application of the high dose of 500 micrograms iloprost significantly decreased net water, sodium and chloride absorption, but did not induce net secretion. (3) Inhibition of endogenous prostaglandin synthesis by indomethacin did not change net water and electrolyte transfer in the ileum and colon. (4) Under this pretreatment i.v.-application of 100 micrograms iloprost, ineffe…

Hepatitic C virus infection and autoimmunity : clinical features, diagnostic tools and therapeutical aspects

Summary Since the genom of the hepatitis C virus (HCV) was detected the pathogenesis of a distinct group of chronic liver disease and some non-hepatic disorders of sofar unknown origin could be newly established. A wide variety of clinical features including vasculitis, glomerulonephritis, porphyria and rheumatoid-like symptoms may become apparant separately or in combination. HCV can be precipitated in cryoglobulins and tissue-related HCV was found in liver and kidney specimen, for instance. HCV-related cryoglobulinemia or circulating immune complexes often occur with vasculitis. Hematological changes may be present such as thrombocytopenia. A considerable amount of patients show circulati…

Antibodies to proteinase 3 increase adhesion of neutrophils to human endothelial cells

SUMMARY The detection of anti-neutrophil cytoplasmic antibodies (ANCA), especially those with specificity for proteinase 3, is important in the diagnosis and in monitoring disease activity of Wegener's granulomatosis and related vasculitides. An ubiquitous feature of all ANCA-associated acute vascular injury is lytic necrosis. Adhesion of neutrophils to endothelium is a fundamental early step of the inflammatory response. Recently we were able to show that ANCA recognize their target antigen (proteinase 3) translocated into the membrane of human endothelial cells. The aim of this study was to investigate the effect of ANCA on the adhesion of neutrophils to human endothelial cells. Incubatio…

Biliary adenocarcinoma

Three human cell lines from adenocarcinomas of the extrahepatic biliary tract were established in permanent tissue culture. Mz-ChA-1 and Mz-ChA-2 were cultured from mechanically dissociated gallbladder adenocarcinoma metastases and SK-ChA-1 was grown from malignant ascites of a patient with primary adenocarcinoma of the extrahepatic biliary tree. Cell doubling times in tissue culture are 3-4 days for Mz-ChA-1 and approximately 2 days for Mz-ChA-2 and SK-ChA-1. All three tumour cell lines were successfully transplanted to nude mice, inducing progressive tumour growth. Histologically, nude mouse tumours resembled the original adenocarcinomas. In vitro formation of gland-like structures were r…

Lithotripsy of gallbladder stones in 1992: Improved indications and actual results

ESWL of gallbladder stones and subsequent adjuvant oral dissolution is a safe and comfortable therapy of symptomatic cholecystolithiasis. This method is limited on highly selected 10 to 15% of patients. Recent experience indicates that the ideal indication is given in patients with radiolucent solitary gallstones (diameter ≤2 cm) which are harboured in a gallbladder with unimpaired contractility. 1 year after ESWL 80% of those patients present with a stone free gallbladder. Computed tomography of gallstones may probably improve selection of patients and therapeutic success.

Influence of loperamide and loperamide oxide on the anal sphincter

The objective of this study was to investigate the effects of the opioid loperamide and its recently synthesized pharmacologically inactive prodrug loperamide oxide on the anal sphincter. In a double-blind, placebo-controlled crossover study, anorectal manometry was performed in 12 healthy volunteers five hours after oral bolus application of 10 mg of loperamide, loperamide oxide, or placebo. Loperamide significantly increased the threshold volumes for minimal perception and urgency to defecate (P less than 0.05) and raised the volume required to abolish recovery of the rectoanal inhibitory reflex (P less than 0.05). These findings suggest that loperamide has a specific continence-improving…

Distribution of Ha-ras alleles in patients with colorectal cancer and Crohn's disease.

The allele distribution of the Ha-ras gene on chromosome 11p was analysed by the restriction fragment length polymorphism of the enzymes Mspl/Hpall in 238 individuals. The investigation covered 116 patients with colorectal carcinoma and 122 patients with Crohn's disease, representing two patient populations with the same ethnic origin, one with a malignant and the other a benign disease of the same organ system. A total of 17 different alleles were detected belonging to the common, intermediate, and rare classes according to the original nomenclature of Ha-ras alleles. Patients with Crohn's disease showed no difference in the distribution of Ha-ras alleles when compared with expected freque…

Analysis of antigens recognized on human melanoma cells by A2-restricted cytolytic t lymphocytes (CTL)

We have pursued our analysis of potential tumor-rejection antigens recognized on human melanoma by autologous cytolytic T lymphocytes (CTL). We reported previously that 3 distinct antigens (A,B,C) were recognized on melanoma cell line SK29-MEL in association with HLA-A2. Selection for melanoma-cell variants resistant to anti-A CTL revealed that antigen A consists of at least 2 determinants (Aa, Ab) which can be lost separately. Genetic linkage between Aa and Ab was suggested by concomitant loss of Aa and Ab in an immunoselected tumor-cell variant. This variant was also resistant to an autologous CTL clone restricted by HLA-B45, indicating that this CTL may also recognize a determinant of an…

Zur Vitalitätsprüfung isolierter Rattenleberzellen

Rattenleberzellen wurden mittels extrakorporaler Leberperfusion unter Verwendung eines Hyaluronidase-Kollagenase-haltigen bzw. enzymfreien Perfusionsmediums isoliert. Die isolierten Leberzellen wurden auf ihr Verhalten gegenuber Trypanblau, FDA, in der Phasenkontrastbetrachtung und im Luciferin-Luciferase-Test untersucht. Daruber hinaus wurde der ATP-Gehalt der Zellen sowie die spontane Abgabe von51Cr aus markierten Zellen bestimmt.

Responses to self and non-self intestinal microflora in health and inflammatory bowel disease.

Alternative splicing products of the tenascin gene distinguish rat liver fat storing cells from arterial smooth muscle cells and skin fibroblasts

Abstract Fat storing-(Ito-)cells (FSC) transform into a myofibroblast-like cell type during liver fibrogenesis. A similar development can be observed in cell culture. At the moment, a definite marker to differentiate transformed FSC from smooth muscle cells (SMC) is not available. We recently found that FSC, SMC and skin fibroblasts (SF) synthesize tenascin, a novel matrix protein. As it is reported that various tissues express different tenascin forms by the mechanism of alternative pre-mRNA splicing, we analyzed the tenascin transcripts in these cell types. Total RNA extracted from cultured FSC, SMC and SF, analyzed by Northern blot hybridization, showed a 7.2 kb transcript in FSC, a 8.7 …

Role of sinusoidal endothelial cells of the liver in concanavalin A-induced hepatic injury in mice

CD4+ T lymphocytes have been identified as being responsible for organ damage in the murine model of experimental liver injury induced by intravenous injection of concanavalin A (Con A). Liver sinusoidal endothelial cells (SEC) and Kupffer's cells (KC) are among the first cells that come into contact with lymphocytes in the liver sinusoid. We aimed to investigate the respective role of these cell populations in the initial steps of T-cell-mediated liver injury in Con A-induced hepatitis. By electron microscopy, we could show that intravenously applied Con A bound predominantly to SEC but not to KC. KC depletion by gadolinium chloride treatment of mice did not result in protection from liver…

Untersuchungen zum Verteilungsmuster von Prä S und S-kodierten viralen Hüllproteinen des Hepatitis B Virus (HBV) auf molekularer Ebene im Serum und Lebergewebe von chronischen HBsAg-Trägern

Die biologische Bedeutung der PraS-kodierten Hepatitis B Virus (HBV) — Hullproteine — insbesondere ihre Regulation innerhalb der Leberzelle — ist noch unklar. Daher war es Ziel unserer Untersuchungen, die PraS- und S-kodierten Hullproteine vergleichend im Serum und im Lebergewebe auf molekularer Ebene zu untersuchen.

Lysis of human melanoma cells by autologous cytolytic T cell clones. Identification of human histocompatibility leukocyte antigen A2 as a restriction element for three different antigens.

From the peripheral blood of the melanoma patient (AV), we derived cytolytic T lymphocyte (CTL) clones that lysed the autologous tumor line SK-MEL-29, but not autologous EBV-B cells, K562, and other tumor targets. By immunoselection experiments it was shown that the CTL clones recognized at least three different antigens on the autologous tumor cells. We demonstrate here that these melanoma antigens are presented to the CTL in association with HLA-A2. First, HLA-A2-reactive pregnancy sera as well as an mAb against HLA-A2 inhibited the CTL lysis. Second, immunoselected melanoma subclones that were resistant to lysis by CTL clones against the three antigens described were found to lack expres…

Assoziation einer arteriellen Verschlußkrankheit mit Ciclosporin-Therapie nach Nierentransplantation

Renal transplantation followed by immunosuppression with cyclosporine (whole-blood levels 200-300 micrograms/l) and methylprednisolone (4 mg daily) was performed in a 54-year-old man with chronic glomerulonephritis. Three years later rapidly progressive arterial obstructive disease (peripheral type) developed. Parenteral treatment with prostaglandins, calcium antagonists and nitrates, as well as a lumbar sympathectomy, was unsuccessful so that, in rapid succession, several amputations on upper and lower limbs became necessary. The disease progression was arrested only when azathioprine replaced cyclosporine. Raynaud's phenomenon, present at the time, also disappeared and analgesics were no …

Complete congenital heart block is associated with increased autoantibody titers against calreticulin.

Complete congenital heart block (CCHB) is associated with anti-Ro/SS-A and anti-La/SS-B antibodies. Calreticulin, a calcium-binding, multi-functional protein of the endoplasmic reticulum with C-terminal KDEL-sequence, is not part of the Ro/SS-A ribonucleoprotein complex. In this study anti-calreticulin autoantibody responses in serum samples from 18 infants with CCHB, their mothers and in a control group of 11 anti-Ro/SS-A or anti-La/SS-B positive infants without heart block and their mothers were analysed. Specific enzyme-linked immunosorbent assays were performed. Nine out of 18 sera with CCHB contained IgG anti-calreticulin antibodies. Four sera of those with IgG antibodies also had IgM …

Reactivation of chronic type B hepatitis: the effect on expression of serum HBV-DNA and pre-S encoded proteins.

Hepatitis B markers were studied in seven patients with reactivated liver disease. Reactivation of chronic type B hepatitis, as indicated by the reappearance of hepatitis B e antigen (HBeAg) in the serum, was characterised by the appearance of hepatitis B virus-DNA (HBV-DNA) in the serum. The expression of pre-S 1 encoded protein remained unchanged in five of seven patients, and poly-HSA as a marker for pre-S 2 encoded protein remained detectable in six of seven patients before and after reactivation of chronic hepatitis. The level of serum HBV-DNA correlated well with the level of liver enzymes, which rose from normal to various levels after reactivation of the liver disease. The data sugg…

Soluble E-Selectin Enhances Intercellular Adhesion Molecule-1 (ICAM-1) Expression in Human Tumor Cell Lines

E-selectin mediates neovascularization via its soluble form, while its membrane-bound form initiates binding of tumor cells to vascular endothelium. Therefore, it was studied whether soluble E-selectin regulates further adhesion molecules on tumor cells. In tumor cells but not in related nonmalignant cells, intercellular adhesion molecule (ICAM)-1 expression was strikingly increased from 5 to 68% positive cells by in vitro inoculation of a recombinant E-selectin-IgG1 within 24 h, as analyzed by flow cytometry. The absence of changes in the expression of vascular cell adhesion molecule, integrin ligands (CD11a, CD18, integrin alpha 4), and sialyl-Lewis X indicates a specific effect of solubl…

Cellular cytotoxicity against autologous hepatocytes in alcoholic liver disease.

We tested lymphocyte cytotoxicity against autologous hepatocytes in patients with alcoholic liver disease (ALD). The following cytotoxicity values were found (mean +/- SEM): alcohol-induced steatosis with or without fibrosis 16.5 +/- 2% (n = 29), alcoholic cirrhosis 28 +/- 4% (n = 13), controls with normal liver histology or minimal changes 6 +/- 2% (n = 11). The differences were statistically significant (both forms of ALD versus controls p less than 0.005). T-cell as well as non-T-cell-enriched lymphocyte fractions showed increased cytotoxicity in ALD. We did not observe a correlation between cellular cytotoxicity and the degree of biochemical or histological alterations within the groups…

Salmonella-reactive Synovial Fluid T-cell Clones in a Patient with Post-infectious Salmonella Arthritis

From a patient with reactive arthritis following Salmonella typhimurium enteritis, synovial fluid T-lymphocytes were cloned and expanded in vitro. Seven out of 74 clones showed a marked proliferative response to antigens of heat-killed Salmonella typhimurium with autologous T-cell-depleted peripheral blood mononuclear cells as antigen-presenting cells. The Salmonella-reactive clones were of the CD4+ phenotype, antigen-induced proliferation could be inhibited by a monoclonal antibody to HLA class II. One clone recognized both Salmonella and Campylobacter jejuni antigens in the proliferation assay. The multiclonality of Salmonella-reactive synovial fluid T-cells indicates that the microorgani…

The Effect of Glyceryl Trinitrate on the Intravascular Oesophageal Variceal Pressure in Patients with Cirrhosis and Portal Hypertension

Glyceroltrinitrat (GTN) fuhrt zur Erschlaffung der glatten Muskulatur von Blutgefasen. Es wurde daher fur moglich gehalten, das es den Pfortaderhochdruck bei Patienten mit Leberzirrhose vermindern konnte. Mit der kurzlich beschriebenen endoskopischen Feinnadelpunktion von Osophagusvarizen konnte der Einflus von GTN auf den intravasalen Osophagusvarizendruck (IOVD) gemessen werden. Drei Minuten nach sublingualer Gabe von 2.2 mg GTN fiel der IOVD bei 10 Patienten mit Varizen Grad III von 22.8 ± 2.0 mmHg auf 12.0 ± 0.4 mmHg (p◄0.005) und bei sechs Patienten mit Varizen Grad II von 16.3 ± 0.4 mmHg auf 10.0 ± 0.4 mmHg (p◄0.005). Unsere bisherigen Ergebnisse lassen vermuten, das GTN zur Blutstill…

Autosomal dominant polycystic kidney disease—in vitro culture of cyst-lining epithelial cells

The major form of autosomal dominant polycystic kidney disease (ADPKD) in humans is linked to the PKD1 gene on chromosome 16p. The identity of the gene and the underlying pathogenetic mechanisms are not yet defined. Cyst-lining epithelial cells derived from a polycystic kidney were successfully grown in culture and designated MZ-PKD-1 cells. By linkage analysis, the related pedigree of the nephrectomized patient could be linked to the PKD1 gene on chromosome 16p. Thus, these cells exhibit the genotype of a mutated PKD1 gene and represent an in vitro culture model for ADPKD involving chromosome 16p. The antigenic phenotype was characterized immunohistologically by epithelial differentiation …

Human renal tubular epithelial cells as target cells for antibodies to proteinase 3 (c-ANCA)

Biological standards for hepatitis B virus assays.

Administration of Second-Generation Extracorporeal Shock Waves without Waterbath for Fragmentation of Extra- and Intrahepatic Bile Duct Stones

First-generation extracorporeal shock-wave sources disintegrate 97% of kidney stones [1, 2]. Recently, in selected patients gallbladder and common bile duct stones were also treated. The technique available so far, however, requires immersion of the patient’s body in a tank of degassed water. The procedure is therefore inconvenient, time consuming, and relatively expensive. The high pressure of shocks (up to 1000 bar) generated by underwater spark discharge causes pain, and general anesthesia is necessary in most patients [3, 4].

Lysis of human pancreatic adenocarcinoma cells by autologous HLA-class I-restricted cytolytic T-lymphocyte (CTL) clones.

From the primary site of a pancreatic adenocarcinoma (patient BE) a permanent cell line (MZ-PC-2) was established in tissue culture. In the course of mixed lymphocyte-tumor-cell cultures (MLTC) with autologous blood-derived lymphocytes, we isolated CTL clones that lysed autologous tumor cells but not autologous EBV-transformed B cells (EBV-B) and not K562. Pre-treatment of MZ-PC-2 cells with IFN-gamma was required to obtain significant lysis in 4-hr cytotoxicity assays. IFN-gamma was superior to IFN-alpha in that respect. Among MLTC responder lymphocytes, tumor-reactive CTL proliferated more strongly in response to MZ-PC-2 cells treated with IFN-gamma than to untreated tumor cells. Three CT…

Rare pre-core stop-codon mutant nt. 1897 predominates over wide-spread mutant nt. 1896 in an unusual course of chronic hepatitis B

We present a patient with an unusual course of hepatitis B e antigen (HBeAg)-negative chronic hepatitis B who had repeated reactivations of his disease progressing to cirrhosis with terminal liver failure. Each flare up presented like an acute hepatitis with very high titres of hepatitis B virus (HBV) and high inflammatory activity followed by rapid clearance of viraemia. The pre-core genome of HBV isolated from sera during 5 years of follow up was analysed. Direct sequencing of polymerase chain reaction (PCR) products derived from consecutive sera showed a rare pre-core stop-codon mutation at nucleotide (nt.) 1897 G --> A with an accompanying mutation nt. 1857 C --> T as well as a stop-cod…

Biliary Pancreatitis: Endoscopic Diagnostic and Therapeutic Techniques

Acute pancreatitis constitutes one of the most hazardous emergencies in clinical gastroenterology. Although the diagnosis “biliary pancreatitis” is often used in clinical practice for patients presenting with concrements in the biliary tree or impacted in the papillary orifice [17, 32], the common phrase “biliary pancreatitis” has not so far been exactly defined. Patients with cholecystolithiasis may be included in this group, as well as patients with dysfunction of the sphincter of Oddi. Some authors have reported that juxtapapillary diverticula increase the risk for development of biliary pancreatitis [28].

Recognition of human renal cell carcinoma and melanoma by HLA-A2-restricted cytotoxic T lymphocytes is mediated by shared peptide epitopes and up-regulated by interferon-gamma.

Cytotoxic T lymphocytes (CTL) have previously been isolated from peripheral blood of patients with renal cell carcinoma (RCC). The CD8-positive CTL line MZ1257-CTL-5 (CTL-5) has been shown to lyse autologous cultured RCC cells in an HLA-A2 restricted fashion. Allogeneic, HLA-A2-matched RCC and melanoma cell lines were also lysed by CTL-5, suggesting that melanoma and renal cancer share antigenic determinants. The aim of the study was to determine whether RCC and melanoma share peptide epitopes that are recognized by CTL-5 in the context of HLA-A2 molecules. Peptides were acideulated from various cell lines, separated by reversed phase high performance liquid chromatography (RP-HPLC), and as…

Identification and characterization of autoantibodies against catalase and α-enolase in patients with primary sclerosing cholangitis

SUMMARY Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown aetiology. Recent studies have shown that genetic factors and both cellular and humoral immunological abnormalities are important in the pathogenesis of PSC. The most prominent autoantibodies in PSC are anti-neutrophil cytoplasmic antibodies (ANCA). The autoepitopes of ANCA in PSC are not well defined. The aim of this study was to identify corresponding ANCA autoantigens in patients with PSC. A biochemical approach with enrichment and partial purification of soluble neutrophil proteins, detection of autoantibodies by Western blot and partial amino acid sequencing were used. Two new autoantigen/aut…

Human T helper cells reactive with somatic bacterial antigens belong to the Th1 subset

The aim of this study was to characterize the cytokine secretion patterns of human T helper cells from healthy donors reactive with somatic antigens from various bacteria, the nematode Anisakis and tetanus toxoid. From the peripheral blood of four healthy donors we have established 70 T cell lines reactive with antigens from Yersinia, Salmonella, Morganella, Klebsiella, Serratia, Escherichia, Chlamydia, Shigella, Streptococcus, tetanus toxoid and Anisakis, respectively. Our results show that all T cells reactive with bacteria produce interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), but no interleukin (IL)-4 and no or very little IL-2 and IL-10 and, thus, belong to t…

Prostanoid release of cultured liver sinusoidal endothelial cells in response to endotoxin and tumor necrosis factor

Abstract Vascular endothelial cells release prostanoids, especially prostacyclin, when properly stimulated. In addition to short acting stimuli like thrombin and histamine an increased prostanoid release occurs in the presence of endotoxin, interleukin 1 or tumor necrosis factor (TNF). The response of sinusoidal endothelial liver cells to such stimuli — probably important in hepatic inflammatory disease — is unknown. Sinusoidal endothelial liver cells from the guinea pig were isolated by centrifugal elutriation and investigated as confluent monolayers. Their prostanoid release in response to endotoxin and human recombinant TNF was determined by radioimmunoassays and compared to that obtaine…

Guinea pig Kupffer cells can be activated in vitro to an enhanced superoxide response

Summary In the preceding paper it was shown that Kupffer cells isolated by digestion of the liver and purified by centrifugal elutriation can be activated in vitro by lipopolysaccharide and muramyl dipeptide to an enhanced superoxide response upon zymosan phagocytosis. Lipopolysaccharide and muramyl dipeptide also led to a strongly increased prostaglandin E 2 release during the phagocytosis of zymosan. This activation was accompanied by an increased production of prostaglandin E 2 during the incubation with the stimuli. Prostaglandin E 2 synthesis was inhibited by the cyclooxygenase inhibitor indomethacin, reduced by dexamethasone, but only slightly decreased by the lipoxygenase inhibitor n…

A monoclonal Ro-antibody and the serum of a Ro-positive patient with subacute cutaneous lupus erythematosus (SCLE) react with basal layers of human epidermis.

Skin lesions, especially at areas exposed to sunlight, prove to be a major form of manifestation of diseases related to Ro-antibodies and neonatal-, 'ANA-negative-', and cutaneous types of lupus erythe- matosus. A monoclonal Ro-antibody established by our group reacts with a 60 kD polypeptide in extracts from human spleen, whereas in extracts from human epidermis the monoclonal Ro-antibody and a purified Ro-antibody from a monospecific serum of a patient with subacute cutaneous lupus erythematosus reacted with a 60 kD and a 48 kD protein. Performing immunofluorescence microscopy on HEp2-cells both antibodies showed a nuclear speckled staining pattern and a reaction with cytokeratin filament…

Regulation of cytochrome P450 IID by acute phase mediators in C3H/HeJ mice.

Abstract Cytochrome P450 IID6 is a drug metabolizing enzyme and the major target antigen in LKM-1 antibody positive chronic active hepatitis. The histological hallmark of chronic active hepatitis is a lymphocytic infiltrate in the liver. It is unknown whether and how cytokines produced and secreted by these tissue infiltrating mononuclear cells regulate the cellular expression of cytochrome P450 IID6. To study the effect of interleukin 1, tumor necrosis factor and interleukin 6 on the hepatocellular RNA expression of cytochrome P450 IID, we injected each of the cytokines in C3H HeJ mice. We found a time-dependent suppression of the cytochrome in the liver. Six hours after the intraperitonea…

Semiquantitative assessment of pre-core stop-codon mutant and wildtype hepatitis B virus during the course of chronic hepatitis B using a new PCR-based assay

In most patients with chronic hepatitis B positive for antibodies (anti-HBe) to HBe antigen (HBeAg), a pre-core mutant hepatitis B virus (HBV) with a point-mutation at nt. 1896 can be isolated. Clinical significance of the mutant virus in chronic hepatitis B is not proven yet, and screening of large numbers of sera during different clinical courses of numerous patients is necessary. We therefore aimed to develop a fast and reliable assay, that allows to discriminate wildtype from nt. 1896 G-->A mutant HBV and to determine the ratio of mutant and wildtype HBV in patients' sera. A mutation specific polymerase chain reaction (ms PCR) with new primers served to distinguish nt. 1896 G-->A mutant…

Stimulation of synovial fluid mononuclear cells with the human 65-kD heat shock protein or with live enterobacteria leads to preferential expansion of TCR-γδ+ lymphocytes

SUMMARY T lymphocyte responses to heterologous or self 65-kD heat shock protein (hsp) have been implicated in the pathogenesis of various forms of arthritis. To delineate the relationship of 65-kD hsp to different synovial fluid (SF) T cell subsets, we stimulated synovial fluid (SFMC) and peripheral blood mononuclear cells (PBMC) from patients with different inflammatory rheumatic diseases and from healthy controls with human or mycobacterial 65-kD hsp, tetanus toxoid (TT), heat-killed or live Yersinia enterocotitica. Phenotyping of the resulting T cell lines revealed an increase of up to 97% TCR-γδ+ lymphocytes in the 65-kD hsp-stimulatcd SF-derived lines. This expansion of TCR-γδ+ cells w…

Signal transduction pathways of membrane expression of proteinase 3 (PR‐3) in human endothelial cells

At present, the exact mechanism of the pathogenic effect of anti-PR-3 antibodies remains unknown. Interaction of anti-neutrophil cytoplasmic antibodies (ANCAs) with human umbilical vein endothelial cells (HUVECs) may play a key role. Recently we were able to show that ANCAs recognize their target antigen, PR-3, translocated into the membrane of HUVECs. The objective of this study was to investigate regulation, i.e. signal transduction pathways, of PR-3 expression in endothelial cells. HUVECs were isolated according to the method of Jaffe et al. and cultured under standard conditions. A cyto-enzyme-linked immunosorbent assay (ELISA) with unfixed cells was performed. Membrane-expressed PR-3 w…

Antiglomerular basement membrane antibodies in human sera: Detection by a modified micro-ELISA

An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to human glomerular basement membrane has been developed. Special emphasis has been put on the choice of microtiter plates which were coated with a collagenase digest of human glomerular basement membrane. Results differed markedly between the different microtiter plates. Best results were obtained with a flexible polyvinylchloride microtiter plate with flat wells (Dynatec). This plate exhibited the highest positive/negative ratio and the lowest intraassay standard deviation. Optimal conditions for each step in the ELISA have been determined. The assay proved to be specific, sensitive, and reproducible. Circulating ant…

HCV-associated cryoglobulinaemia presenting with vasculitis, hepatitis, and glomerulonephritis—a therapeutic dilemma

treated by diet. One year prior to presentation the patient suffered a transitory ischaemic brain attack concomitant with the onset of arterial hypertension, which was subsequently treated with calcium antagonists and beta blockers. Purpura-like skin lesions were noted which were reported to have occurred intermittently over a period of 5 years on both legs and occasionally on the arms. Proteinuria and haematuria were noted during periods with skin lesions.

Analysis of radiolucent gallstones by computed tomography for in vivo estimation of stone components.

. Successful oral litholytic and other non-operative therapies of gallstones require exact determination of the stone components. Since computed tomography (CT) provides highly sensitive measurement of density, we performed a study to evaluate whether CT measurement of stone density allows a prediction of the composition of radiolucent gallstones. Twenty-eight patients presenting with 29 radiolucent gallbladder (n= 17) or common bile duct stones (n=12) were included. Prior to operative or endo-scopic therapy the attenuation values (Hounsfield Units, HU) were assessed in vivo by CT under standardized conditions (Somatom II, 125 KV, 130 mAs). After surgical or endoscopic stone removal the con…

Lymphocytes from hepatic inflammatory infiltrate kill rat hepatocytes in primary culture

In the last few years it has become possible in the liver to isolate lymphocytes from inflammatory infiltrates and to culture them in vitro. Most of the lymphocyte clones obtained are CD 8 + cytotoxic cells, but interactions between these lymphocytes and hepatocytes in primary culture have not been analysed previously. In this study, cloned human T lymphocytes from liver biopsies and from the peripheral blood of patients with chronic hepatitis B or primary biliary cirrhosis, after phenotypical and functional characterization into CD 8+ or CD 4+ cytotoxic lymphocytes, were activated in an antigen-independent fashion by adding either anti CD 3 or anti CD 2/R-3 monoclonal antibodies to the cel…

In vivo and in vitro induction of natural killer cells by cloned human tumor necrosis factor

The natural killer (NK) cell activity of mice in the peritoneal cavity is very low or undetectable and testing peritoneal NK cells is a useful model for studying the influence of activating substances upon local injection. Injection of tumor necrosis factor (TNF) at doses of 10-200 ng caused a marked activation of NK cell activity which was maximal after 24 h and declined rapidly on day 2. A similar effect was observed when interferons alpha and beta were injected, and there were additive results when interferon was injected together with TNF. The NK cell nature of the effector cells activated by TNF was substantiated by the finding that previous injection with anti-asialo GM 1 antibody pre…

Liver-infiltrating and circulating CD4+ T cells in chronic hepatitis C: immunodominant epitopes, HLA-restriction and functional significance.

The aim was to assess the specificity and functional significance of liver-infiltrating and peripheral blood T cells in chronic hepatitis C. Peripheral blood mononuclear cells hepatitis C virus from 50 of 58 (86.2%) patients with chronic hepatitis C virus infection and 6 of 28 (21.4%) controls showed a proliferative T cell response to at least one of 16 synthetic peptides covering highly conserved regions of the core, envelope (El) and non-structural regions (NS4) of hepatitis C virus. However, six immunodominant peptides were exclusively recognized by the proliferating blood mononuclear cells from 46 patients with chronic hepatitis C virus infection (79.3%). Fine specificity and HLA-restri…

Immunorecognition of different ganglioside epitopes on human normal and melanoma tissues.

There is increasing evidence that cell-surface gangliosides play a role in tumor growth, progression and metastases. In order to determine the frequency of ganglioside GD3 in patients with metastatic malignant melanoma for further therapeutic trials, GD3 ganglioside expression was determined in 119 tissue samples. Of these melanomas, 93% (111/119) were R-24-positive, which indicates the value of this diagnostic marker for melanoma. To study the structural epitopes of gangliosides, 10 ganglioside antibodies with defined specificities and affinities were tested on over 100 fresh-frozen tissue specimens of human normal and melanoma tissues. All the antibodies tested recognize the ganglioside G…

Relationship of pre-S encoded antigens in liver and clinical manifestations of chronic hepatitis B infection.

Pre-S1 and pre-S2 encoded antigens of hepatitis B virus were localized in liver tissue using monoclonal antibodies. They were found to be exclusively expressed in the cytoplasm of liver cells. Cell bound pre-S1 encoded protein was often detected in patients with chronic liver disease and viremia. Only a small number of the HBsAg positive cells also contained pre-S1 antigen. There was no correlation with nuclear HBcAg. Livers of non-viremic HBsAg carriers contained many HBsAg expressing liver cells, that were frequently also positive for pre-S2 encoded protein but contained no detectable pre-S1 encoded protein at all. It remains open whether cell bound pre-S2 containing proteins of middle si…

Significant autoimmune markers of autoimmune liver disorders: Current status

Candida arthritis: cellular immune responses of synovial fluid and peripheral blood lymphocytes to Candida albicans.

A case of septic Candida albicans arthritis of the knee in a patient with systemic candidiasis is presented. Systemic and intra-articular cellular immune responses to C albicans and various bacterial antigens were monitored for 15 weeks. It is shown that the candida induced blastogenesis of synovial fluid lymphocytes was much more stimulated than that of peripheral blood lymphocytes, and that the proportion of activated cells expressing HLA class II antigens was markedly increased in the synovial fluid. Strong cellular immune responses to Candida albicans could still be shown many weeks after the synovial fluid aspirates had become sterile. For the first time synovial fluid derived, CD4 pos…

ROUTES OF HIV-2 TRANSMISSION IN WESTERN EUROPE

Demonstration of antibodies to the surface (anti-p41) and core proteins (anti-p24) of the human immunodeficiency virus (HIV) in individuals positive for anti-HIV.

Diagnosis of infection with the human immunodeficiency virus (HIV) relies on the demonstration of antibody to this virus. Occasionally, the combined analysis of sera using ELISA and western blot reveals false-positive results. We have compared a newly developed test to detect antibodies to the core (anti-p24) and surface (anti-p41) proteins of HIV with the established tests described above. Anti-p24 and anti-p41 were negative in three individuals positive for anti-HIV by ELISA and immunoblot; they had a low risk to acquire HIV infection and were clinically and immunologically normal and suspected false positive previously. In 62 individuals at risk, anti-p41 was always positive while anti-p…

Tierexperimentelle Untersuchungen über die Wirkung von Azathioprin auf die Bildung zirkulierender Antikörper

Die Wirkung von AP bei peroraler Applikation auf die Ak-Antwort vom Soforttyp wurde im Kaninchenexperiment gepruft. Die primarimmunologische Antwort lies sich durch 4 mg AP/kg/Tag nur teilweise supprimieren. 6 mg waren voll wirksam hinsichtlich der Unterdruckung der Primarreaktion. Die Sekundarreaktion wurde von dieser AP-Dosis nicht wesentlich beeinflust. Dies gelang erst mit toxischen Dosen von 15 mg AP/kg/Tag. Die Wirksamkeit von AP unterhalb von 6 mg/kg/Tag wachst mit dem Antigenstimulus.

No primary association between LMP2 polymorphisms and extraspinal manifestations in spondyloarthropathies

OBJECTIVE—To investigate the potential role of the HLA-linked LMP2 (low molecular weight protein) gene polymorphisms in conjunction with DR4 and DR7 on extraspinal disease manifestations in HLA-B27 positive patients with spondyloarthropathy.
METHODS—172 patients with spondyloarthropathy, 46 healthy, HLA-B27 positive blood donors, and 99 unrelated controls were typed for HLA-class I and II antigens. LMP2 alleles were determined by polymerase chain reaction and subsequent restriction enzyme digestion.
RESULTS—There were statistically non-significant increases of DR4 and DR7 in spondyloarthropathy subjects. However these differences did not relate to specific extraspinal manifestations. There …

Zur Induktion einer aktiven chronischen Hepatitis durch heterologe l�sliche Leberproteine

Die Untersuchungen gehen von dem fruher gefuhrten Nachweis eines im immunologischen Sinne organspezifischen und weitgehend speciesexclusiven Proteins der Leber aus. Entsprechende Proteinpraparationen aus menschlichen Lebern wurden zur Sensibilisierung von Kaninchen eingesetzt. 1. Bei langdauernder Sensibilisierung (uber 1 Jahr) mit kleinen Dosen gelang beim erwachsenen Kaninchen die Induktion einer Immuntoleranz, die durch Applikation von hitzeund sulfanilsaurealteriertem Protein durchbrochen werden konnte; gleichzeitig konnte eine Hypersensibilitat vom verzogerten Typ induziert werden. Histologisch fand sich bei allen Tieren eine aktive chronische Hepatitis; die ubrigen Organe (Herz, Lunge…

Das »Nußknacker-Syndrom« der Vena renalis (Arteria-mesenterica-superior-Syndrom) als Ursache gastrointestinaler Beschwerden

History and clinical findings Since the age of 19 a now 22-year-old man had complained of intermittent abdominal pain, irregular stools and paroxysmal tachycardia. The only preceding illness had been a single episode of iron-deficiency anemia. A laparoscopy, done 8 months after the onset of symptoms, had revealed an inflamed Meckel's diverticulum which was surgically removed. After transient improvement the symptoms recurred 5 months postoperatively. On admission to clarify the cause of the symptoms he had discrete abdominal pain on pressure, but physical examination was otherwise unremarkable. Investigations Routine biochemical tests and endoscopy were normal. Abdominal computed tomography…

Enormous hemangiosarcoma of the heart

This report describes a 26-year-old patient with hemangiosarcoma of the heart and summarizes the clinicopathological features in previous reports of patients with cardiac angiosarcoma. The patient was admitted to our hospital because of a syncope and one episode of nocturnal dyspnea and hemoptysis. In his history he complained of progressive weakness and loss of weight over the past 2 months. Echocardiography and computed tomography of the chest showed inhomogeneous masses in the pericardial cavity completely surrounding the heart and involving the ascending aorta and the superior vena cava. Histological examination of the tissue obtained from the mass by fine needle technique revealed a po…

The T Cell Receptor (TCR) in HLA-B27-Restricted T Cell Responses - an Introduction

Recent data indicate that cytotoxic T lymphocytes (CTL) are involved in the pathogenesis of HLA-B27-associated spondylarthropathies. In the absence of clearly defined "arthritogenic" bacterial or self peptides that are presented by HLA-B27 and recognized by such CD8+CTL, one approach has been to investigate the T cell repertoire of lesional cellular infiltrates by determining T cell receptor (TCR) variable (V) gene segment frequencies. Furthermore, the TCR V alpha and V beta chains of HLA-B27-restricted CTL clones, notably the putative peptide-contacting CDR3-regions of these TCRs, have been sequenced. This article will give a short review of the current literature on the topology of the TC…

Ergebnisse verschiedener Methoden zur Trennung spezifischer und unspezifischer Antistreptolysin-O-Titer in menschlichen Seren

Im Jahre 1964 wurden in unserem Labor bei 1090 Patienten 1239 Antistreptolysin-O-Reaktionen ausgefuhrt. Hiervon wurden 688 bei ebenso vielen Patienten im Rahmen der klinischen Routinediagnostik angewandt. Bei 175 der Falle wurde ein erhohter Antistreptolysintiter (AST) von 400 E oder hoher festgestellt. Nur bei sieben von diesen konnte die AST-Erhohung mit Sicherheit als Folge einer Infektion mitβ-hamolytischen Streptokokken der Gruppe A gedeutet werden.

Correlation of anti-cytoskeleton antibody activities in synovial fluid with interleukin-6 in patients with osteoarthritis and inflammatory joint disease

Synovial fluids and sera from patients with rheumatoid arthritis, psoriatic arthritis, yersinia arthritis, Behçet's syndrome, Crohn's disease, and osteoarthritis were tested for antinuclear antibodies and antibodies to five cytoskeletal components in sensitive enzyme-linked immunosorbent assay (ELISA) systems and for IL-6 concentrations in a proliferation assay (IL-6 dependent hybridoma cell line B13.29, subclone B9). Statistically significant correlations between antibody activities and IL-6 levels were found for vimentin antibodies (r = 0.56; p less than 0.05) and actin antibodies (r = 0.44; p less than 0.05). In patients with chronic and active disease like rheumatoid arthritis and psori…

Expression of intercellular adhesion molecule 1 (ICAM-1, CD54) in colonic epithelial cells.

The expression of intercellular adhesion molecule-1 (ICAM-1, CD54) was examined in 16 surgically removed colonic tumours and two colonic carcinoma cell lines. Immunohistochemistry showed a varying percentage of ICAM-1 positive colonic carcinoma cells in 9/16 tissue specimens, while normal colonic tissue (apart from a slight reactivity of endothelial cells) was not stained. The presence of the ICAM-1 molecule on the cell surface and the expression of ICAM-1 mRNA were investigated for two colonic carcinoma cell lines. It was possible to enhance the expression of ICAM-1 considerably by incubating the cells in the presence of inflammatory cytokines in HT-29 and CaCo-2 cells. The responsiveness …

Pulmonary aspergilloma in a patient with AIDS.

Aspergillus infections are rare in the course of AIDS. They mostly occur as invasive destructive disease in patients with severe CD4 cell depletion. An unusual case of a homosexual AIDS patient who developed a pulmonary aspergilloma is presented.

Tumor associated antigens in human renal cell carcinoma: MHC restricted recognition by cytotoxic T lymphocytes.

Based on previous studies in human melanoma leading to the molecular cloning of genes encoding peptide antigens recognized by MHC-restricted cytotoxic T lymphocytes (CTL) we extended our efforts to renal cancer systems established in tissue culture. In two patients we obtained stable CD8+ CTL clones with high cytolytic activity for the corresponding autologous tumor cell line in vitro. Neither autologous EBV-transformed B lymphocytes or PHA-activated PBL nor natural killer targets K562 were lysed by these CTL clones. MZ1257-RCC CTL5-30 lysed autologous tumor cells as well as normal kidney cell cultures in an HLA-A2 restricted fashion. Further specificity analysis showed cross reactivity wit…

Expression of Ia-antigens on guinea pig Kupffer cells

Summary The expression of the Ia-antigen on guinea pig Kupffer cells was studied employing two monoclonal antibodies against two different determinants of the Ia-molecule. The study was performed in situ on liver sections and on isolated highly purified Kupffer cells kept in culture up to 6 days. The influence of guinea pig hepatocyte culture supernatant and of supernatants of phytohemoagglutinin (PHA)-stimulated human peripheral blood lymphocytes (PBL) on the Ia expression was measured. Immunofluorescence staining of cryostat sections revealed that the monoclonal antibodies used are able to detect Ia-antigens on liver macrophages in situ. The in vitro studies strongly suggest that all Kupf…

Induction of an Anti-Vaccine Response by T Cell Vaccination in Non-human Primates and Humans

Abstract Experimental and spontaneous autoimmune disease in animals can effectively be prevented and treated by application of pathogenic autoreactive T cells in an attenuated form. This approach has become known as T cell vaccination, T cell vaccination exploits specifically the ability of the immune system to regulate its autoreactive T cells by mechanisms of network control. The success of T cell vaccination in a variety of rodent animal models has raised hopes for its use as an effective and specific therapy in human autoimmune disease. The aim of this study was to induce an anti-T cell response by T cell vaccination in humans and primates as a pre-clinical study into the feasibility an…

Cytokine-mediated regulation of monocyte/macrophage cytotoxicity in human immunodeficiency virus-1 infection.

Monocyte/macrophage-mediated tumor cytotoxicity was studied in patients infected with human immunodeficiency virus-1 (HIV-1) at various stages [Center for disease control (CDC) classification] of the disease. using the P-815 tumor cell line as target cells, the results demonstrated reduced monocyte/macrophage cytotoxicity early in HIV-1-related disease (CDCIII, P0.01). This cellular dysfunction sustained during the progression of the disease. Evidence could be presented that neither exogenous application of macrophage-stimulating cytokines (e.g. interferons) nor their endogenous induction in vitro restored monocyte/macrophage cytotoxicity. However, enhanced tumor necrosis factor (TNF)-alpha…

Alpha- and gamma-interferon (IFNα, IFNγ) but not interleukin-1 (IL-1) modulate synthesis and secretion of β2-microglobulin by hepatocytes

Soluble serum beta 2-microglobulin has been thought to result from membrane shedding by activated T-lymphocytes. This hypothesis could explain the increase of beta 2-microglobulin serum levels during virally induced mononucleosis, but not elevated levels as observed in other virally induced and in malignant diseases. In this paper we demonstrate that beta 2-microglobulin is a true secretory protein, and that its synthesis in hepatocytes is modulated by IFNs but not by IL-1. While the 45,000 MW HLA antigen can be found only in cell lysates, beta 2-microglobulin is shown to be secreted also into the culture medium like other secretory proteins (e.g. albumin-factor B-complement C3). Furthermor…

Bacteria-specific cytotoxic CD8+ T cells: a missing link in the pathogenesis of the HLA-B27-associated spondylarthropathies.

The term seronegative spondylarthropathies is used for an entity of rheumatic syndromes of peripheral joints and the spine (ankylosing spondylitis, reactive arthritis, Reiter's syndrome, arthritis in psoriasis and in inflammatory bowel disease) which are strongly associated with the MHC class I molecule HLA-B27. However, the mechanisms whereby HLA-B27 confers disease susceptibility have so far remained unknown. There is strong evidence that gut inflammation and infection with gram-negative bacteria play a role in the induction of B27-associated disease. HLA-B27, like other MHC class I molecules, physiologically binds antigenic peptides in its binding groove and presents them to CD8+ T lymph…

Definition of discrete signals involved in human T-cell activation

Abstract Mitogenic activities of monoclonal antibodies directed at denned receptor structures expressed on the surface of mature human T lymphocytes were employed to study, in detail, signals involved in primary T-cell activation. Based on differential requirements for stimulation, two discrete pathways of human T-cell activation can be defined: the antigen-induced mode of activation initiated through the Ti-T3 antigen-receptor complex and an alternative pathway which can be triggered by monoclonal antibodies directed at the T11 glycoprotein. Perhaps more importantly, the approach taken here allows the definition of stable intermediate cellular stages within the activation cascade and, thus…

Monoklonale Antikörper und Konjugate in der Krebsdiagnostik und Therapie

Bereits um die Jahrhundertwende betonte Paul Ehrlich die mogliche Bedeutung von Antikorpern als Mittel gegen Krebs. Ihre zytotoxische Aktivitat und ihre Spezifitat machen sie theoretisch zu idealen Werkzeugen, um abnorme Zellen zu erkennen und zu zerstoren und die umliegenden normalen Zellen zu verschonen. Leider waren bis vor kurzem die technischen Voraussetzungen nicht gegeben, spezifische Antikorper mit zytotoxischer Aktivitat in groser Menge herzustellen. Durch die von Kohler u. Milstein im Jahre 1975 neu etablierte Technik, monoklonale Antikorper zu erzeugen [9], anderte sich die Situation schlagartig. Diese Methodik erlaubt die Immortalisierung eines einzigen B-Lymphozyten, der schlie…