0000000000422105
AUTHOR
Peter Scheurich
Frequency-Analysis of Precursors of Cytotoxic T Lymphocytes in Radiation Chimeras: Enumeration of Antigenspecific CTL-P Restricted to Thymic MHC- and Bone Marrow-MHC-Determinants
The mechanisms controlling the acquisition of T cell restriction specificity and immunocompetence are, despite of numerous investigations, not well understood. From studies of the CTL-immune responsiveness in thymus- and bone marrow-grafted chimeric mice, it became apparent, that it is the thymus which is crucial not only for the maturation or T cells, but also for the specificity repertoire of the T cells (1,2). From these data it was suggested, that during intra-thymic maturation both mutational events and positive selection mechanisms influence the repertoire such that only T cells restricted to thymic epithelial cell MHC determinants mature and will be exported to the peripheral lymphoi…
Postnatal development of functional T cell subsets in the mouse: a frequency analysis of mitogen reactive precursors of proliferating, of cytotoxic and of IL 2 producing T cells
In order to study the postnatal development of functional T cell subsets in the mouse, a mitogen-driven limiting dilution culture system was used for a precursor frequency analysis of proliferating, of cytolytic and of IL 2-producing T cells, respectively, present in spleen and thymus of mice from neonatal to adult age. In adult mice, the majority (up to 100%) of splenic T cells was capable to respond to Concanavalin A. In contrast, an up to tenfold lower frequency of mitogen-reactive precursors was found within positively selected Thy-1+ spleen cells of neonatal mice. Within this fraction of Con A reactive neonatal T cells, there was an apparent imbalance in the CTLp/PTLp ratio within the …
Tumor necrosis factor-receptor 2 is up-regulated on lamina propria T cells in Crohn's disease and promotes experimental colitis in vivo
Tumor necrosis factor (TNF) plays a pivotal role in the pathogenesis of Crohn's disease (CD). However, little is known about the role of TNF receptors (TNF-R) in this disease. Here, we found that TNF-R2 (in contrast to TNF-R1) was significantly up-regulated on lamina propria and peripheral blood T cells in CD compared to control patients. To directly test the functional role of TNF-R2 in Th1-mediated experimental colitis in vivo, we took advantage of transgenic animals overexpressing TNF-R2 in T cells. Reconstitution of SCID mice with CD4+ CD62L+ T cells from TNF-R2 transgenic mice led to an earlier wasting syndrome, a more severe colitis and augmented Th1 cytokine production than reconstit…
8-Azido-adenosine 5'-triphosphate as a Photoaffinity Label for Bacterial F1 ATPase
1. 8-Azido-adenosine 5'-triphosphate (n83ATP) is a suitable photoaffinity label for F1 ATPase from Micrococcus luteus. The nucleotide is a substrate in the presence of bivalent cations and inhibits the enzyme irreversibly upon irradiation with ultraviolet light above 300 nm. 2. More than 80% of the label is covalently bound to the beta subunits in the presence of bivalent cations. Labeling and inactivation is decreased by protection with ADP, ATP or adenyl-5'-yl imidodiphosphate. To a much smaller degree the alpha subunits also become labeled. 3. n83AMP does not specifically bind to the beta subunits upon irradiation. Like n83ATP and n83ADP, it also labels the alpha subunits to a small exte…
Quantitative representation of all T cells committed to develop into cytotoxic effector cells and/or interleukin 2 activity-producing helper cells within murine T lymphocyte subsets
A limiting dilution culture system based on stimulation with concanavalin A (Con A) has been used to study the quantitative distribution of helper and of cytotoxic precursor cells in Lyt-2-defined subpopulations of murine T cells. Virtually all of the selected Lyt-2+ and Lyt-2-T cells grow and expand to large clonal colonies within an 8-9-day culture period. Our data show that upon stimulation with Con A, 90% of the Lyt-2-T cells were capable to produce interleukin 2 (IL 2) activity. In addition, IL 2 activity is produced by 8-10% of Lyt-2+ T cells. However, at the clonal level, the average of the IL2 activity produced by Lyt-2+ T cells is about 8-fold less as compared to Lyt-2-T cells. Pre…
Photoaffinity labeling of the coupling factor 1 from the thermophilic bacterum PS3 by 8-azido ATP
AbstractTo localize the nucleotide binding sites of the F1ATPase (TF1) from the thermophilic bacterium PS3 we have used 14C-labeled 8-azido ATP (8-N3ATP) as photoaffmity label. 8-N3ATP is hydrolyzed by the F,ATPase in the absence of ultraviolet light. Irradiation by ultraviolet light of the enzyme in the presence of 8-N3ATP results in reduction of ATPase activity and in preferential nucleotide specific labeling of the α subunits (0.8–0.9 mol 8-N3ATP/TF1,α:β = 4:1). Inactivation and labeling do not depend on the presence of Mg2+. Both effects decrease upon addition of various nucleotide di- or triphosphates.
Eine einfache Darstellung von 8-Azidoadenosin-5′-triphosphat; ein Agens zur Photoaffinitätsmarkierung von ATP-bindenden Proteinen
Eine vereinfachte Synthese von 8-Azido-ATP (3) wird beschrieben. Die neue Methode ist kurzer und fuhrt zu wesentlich besseren Ausbeuten als die bekannte Methode. Neben 3 entstehen dabei 8-Azido-ADP und 8-Azido-AMP. Es wurden mit einem Reaktionsansatz alle drei 8-Azidonucleotide gleichzeitig erhalten. 8-Azido-ATP (3) wurde durch Elementaranalyse, Dunnschichtchromatographie, Infrarotspektroskopie und UV-Absorptionsspektroskopie charakterisiert. Die Photolyse bei verschiedenen pH-Werten wurde untersucht. Durch F1ATPase wird 3 umgesetzt. A Facile Synthesis of 8-Azidoadenosine 5′-Triphosphate; a Photoaffinity Label for ATP-Binding Proteins A simple procedure for the synthesis of 8-azido-ATP (3) …