Expression of properdin in human monocytes

Properdin is the only known positive regulator of the alternative pathway of complement activation. Northern blot analysis of cell lines derived from fibroblasts, B-cells, hepatoma cells, and cells of the monocyte-macrophage lineage revealed properdin expression only in the myelomonocytic cell line HL-60, in the monoblastic cell line U-937 and in the monocytic line Mono Mac 6. Culture of Mono Mac 6 cells for 24 h with phorbol 12-myristate 13-acetate, bacterial lipopolysaccharide and the cytokines interleukin-1 beta and tumour necrosis factor-alpha enhanced mRNA abundance, with the strongest effect (tenfold) being observed with the lipopolysaccharide. In contrast, recombinant interferon-gamm…

Essential role of surface-bound chemoattractant in leukocyte migration

MANY chemotactic factors, usually proteins or peptides, have been isolated and studied, but little is known about the basic mechanism of leukocyte migration. This movement is termed chemotaxis if its direction is determined by substances in the cells' environment1. The chemotactic agent is assumed to convey information to the leukocytes by interaction with receptors. The subsequent sequence of events thus triggered in the cells is unknown but metabolic changes such as activation of an esterase have been reported as occurring as the cells move forward (for review see ref. 2). A role for surface-bound chemoattractant in cell locomotion was suggested by the observation that mouse fibroblasts m…

Complement Receptor Analogous Factors in Human Serum: I. Isolation of a Molecule Inhibitory for Complement Dependent Rosette Formation, its Identification as α1-Antitrypsin and its Functional Characterization

Abstract A glycoprotein was isolated from human plasma which partially inhibited C3 carrying erythrocytes from binding to complement receptor cells (CR + C). Based on its physicochemical characteristics and its antigenicity this glycoprotein was identified as aI-antitrypsin (α 1 -AT). The activity of α 1 -AT towards-C3 and its fragments was unaffected by heating but it was destroyed by periodic acid. The isolated carbohydrate moiety of α 1 -AT showed the same effect as the intact molecule. Using F(ab) 2 of IgG-anti-α 1 -AT, α 1 -AT could be demonstrated on Raji cells and human erythrocytes. Treatment of these CR + C with IgG-anti-α 1 -AT resulted in a blockade of their C3 receptor activity.…

Ability of the T cell-replacing polyanion dextran sulfate to trigger the alternate pathway of complement activation.

Dextran sulfate (DS) consumed C3 in C4 deficient guinea pig serum. This temperature-dependent reaction required Mg++ ions and could therefore be blocked by EDTA. Isolated C3 was not influenced by DS, but serum factors were required for C3 consumption. The C3 proactivator as well as C3 were converted to their activated state by DS in guinea pig and human serum, as revealed by immunoelectrophoretical analysis. DS generated anaphylatoxin activity in serum. It is concluded that DS activates C3 via the alternate pathway of complement activation. This potency of the polyanion might serve as a tentative explanation for its T cell-replacing effect in an antibody-forming system, which was reported b…

Complement receptors on lymphocytes

Formation and function of a complement-activating enzyme generated from factors of guinea pig serum and cobra venom

An enzymatic complex can be formed by factors from guinea pig serum and cobra venom, which is able to activate C3 bypassing C1, C4 and C2. Formation and action of the enzyme are described. The action on C3 results in an activation of the terminal complement components and in membrane destruction provided suitable membrane receptors are available.

Detection of proteolytic (C 3-cleaving) activity on mouse mastocytoma (P815) cells and other mouse cell lines by formation of cell contact with C 3-carrying mouse lymphocytes

Mouse mastocytoma cells (P 815) formed rosettes with normal mouse spleen lymphocytes which had been coated with uncleaved human C 3; this interaction was clearly dependent on the amount of C 3. Lymphocytes treated with C 3 b or buffer alone were ineffective. Formation of cell contact could be inhibited by the presence of protease inhibitors such as diisopropyl fluorophosphate, phenyl methyl sulfonyl fluoride and tosyllysyl chloromethyl ketone. Seve n out of 13 different cell lines behaved like P 815 cells. The results strongly suggested that a proteolytic activity on mouse tumor cells led to a cooperation with uncleaved C 3 on a carrier cell to connect these two cells. We interpreted these …

Physicochemical characterization of the fifth (C5), sixth (C6), seventh (C7), eighth (C8) and ninth (C9) component of guinea pig complement.

A physicochemical characterization of the purified guinea pig complement components C5 to C9 is given. For this purpose the sedimentation rate, the diffusion coefficient, the molecular weight and the isoelectric point were determined and compared with the values already known for the guinea pig and human complement system. For the determination of the physicochemical parameters gel filtration on Sephadex G-200, ultracentrifugation applying a sucrose density gradient and thin-layer isoelectric focusing were used. By comparing the values of the human and guinea pig complement a remarkable similarity is shown.

Importance of Factors H and I for the Adherence of C3b-Coated Erythrocytes to Cells

Abstract The role of cell membrane-associated human factor H for the binding of cell-bound Cab to complement receptor-carrying (CR + ) cells was investigated. Pretreatment of CR + cells with antibodies to factor H inhibited the adherence of Cab-coated red cells to human tonsil lymphocytes (TL) and peripheral blood monocytes (Mo). The Cab receptor reactivity of human polymorphonuclear leucocytes (PMN) was not influenced and the one of Raji lymphoblastoid cells only slightly influenced; iC3b and Cad receptor reactivity was in no case affected. When diisopropylfluorophosphate (DFP) in a concentration of 0.1 mM was present during pretreatment of the CR + cells with anti H, the antibodies gained…

Role of β1H for the binding of C3b-coated particles to human lymphoid and phagocytic cells

Coating of EAC14oxy23b with highly purified human serum beta 1H globulin (beta 1H) led to acceleration of rosette formation with human peripheral blood lymphocytes (PBL), tonsil lymphocytes, B lymphoblastoid (Raji) cells, granulocytes and monocytes. This reaction was discernible from C3bi-dependent rosette formation. Enhancement of rosette formation of C3b cells by beta 1H was most effective at limiting amounts of C3 per EAC14oxy23b. The beta 1H effect was not due to trace contamination with C3b inactivator. beta 1H-dependent rosette formation with the various lymphoid and phagocytic cells could be suppressed by the F(ab')2 fragment of anti-beta 1H suggesting beta 1H-mediated binding of bet…

COMPLEMENT-DEPENDENT B-CELL ACTIVATION BY COBRA VENOM FACTOR AND OTHER MITOGENS?

It has been proposed that two distinct signals are required for the triggering of the precursors of antibody-forming bone marrow-derived cells (B cells): (a) the binding of antigen or of a mitogen to the corresponding receptor sites on B-cell membranes and (b) the interaction of activated C3 with the C3 receptor of B lymphocytes. There is growing evidence that B-cell mitogens and T (thymus-derived cell)-independent antigens are capable of activating the alternate pathway of the complement system (bypass). Therefore, the effect of another potent bypass inducer was investigated with regard to B-cell activation and the role of C3. Purified, pyrogen-free cobra venom factor was mitogenic for bot…

Description of a Simple, Specific, and Sensitive Test for the Detection of Detergent-Solubilized C3b Receptor

Abstract The C3b receptor was isolated from detergent-solubilized human erythrocyte membranes by a previously described technique (1). The receptor glycoprotein was shown to enhance EAC14oxy23b rosette formation with Raji lymphoblastoid cells. This provided a specific and sensitive test to detect the solubilized C3b receptor either in crude or highly purified form. The property of the C3b receptor tested by this assay appears to be analogous to properties of β1H.

Multiple sclerosis patients show an increased spontaneous activity of their peripheral blood monocytes as measured by chemiluminescence

I has been reported that myelin basic protein (BP) reacts extremely sensitively to peroxide, which is formed when monocytes/macrophages are stimulated to produce a "respiratory burst" (RB). We measured the RB activity by means of chemiluminescence in peripheral blood monocytes/macrophages (MO) of 17 MS patients, 5 patients with a viral infection of the CNS, and 14 control persons. The median of the spontaneous RB activity of MS patients compared with the median of our control group showed a highly significant increase (P = 0.0002). All MS patients examined possessed a clearly increased MO activity. The highest values, however, were found in MS patients in a bout (means = 315%, means = 296%)…

Alpha-1-antitrypsin-induced inhibition of complement-dependent phagocytosis.

Abstract In a previous investigation, inhibition of complement-dependent rosette formation by alpha1-antitrypsin (α1-AT) was observed, and it was demonstrated that α1-AT interacts through its carbohydrate portion with C3 and its fragments. In the present study, the effect of α1-AT on the complement-receptor-mediated phagocytosis by human peripheral blood monocytes was examined. Purified α1-AT inhibited in a dose-dependent manner phagocytosis of C3-carrying yeast particles. Inhibition was selective, concerned only C3-receptor-mediated phagocytosis, neither Fc-receptor-mediated phagocytosis nor uptake of untreated yeast particles was blocked by α1-AT. It was demonstrated that α1-AT exerted it…

Migration of Leukocytes into Filters Coated Homogeneously with Immune Complexes, Antigens, Lectins or Tripeptides

Cellulose nitrate filters were incubated in solutions of albumin, a chemotactically active tripeptide (f-Met-Leu-Phe), immune complexes or lectins and afterwards washed with buffer. They showed a dose-dependent increased leukocyte migration, when tested in typical Boyden chambers in comparison to filters treated only with buffer. The tripeptide, the immune complexes and the lectins were stimulatory at very low concentrations and acted inhibitory at high concentrations. Treating filters with formaldehyde or glutardialdehyde had no clear stimulatory effect. These findings extend earlier observations obtained with casein. They show that cells move very effectively on solid substrata in the abs…

Expression of Histocompatibility Antigens during the Growth Cycle of Cultured Lymphoid Cells

Histocompatibility antigens are genetically determined markers which are located on plasma membranes of tissue cells of each member of a species. HL-A antigens are the gene products of the major histocompatibility locus in man and represent the human counterparts of the H-2, Ag-B, ChL-A and DL-A systems in mice, rats, chimpanzees and dogs, respectively (Palm, 1964; Snell and Stimpfling, 1966; Rapaport et al., 1970; Balner et al., 1971; Klein and Shreffler, 1971). The great interest in the serologic, genetic, chemical and immunological characterization of histocompatibility antigens is attributable to the fact they provide cell surface markers useful in selecting transplant donors and recipi…