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Kinetics and extent of protein tyrosine kinase activation in individual T cells upon antigenic stimulation by Müller; Demotz; Bulliard; Valitutti is a Immunology and Microbiology article available to read on EtoBox.

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Using human CD4^+^ T‐cell clones and peptide‐pulsed antigen‐presenting cells (APC) we measured, at the single cell level, different steps in the T‐cell activation cascade. Simultaneous analysis of T‐cell antigen receptor (TCR) down‐regulation and interferon‐γ (IFN‐γ) production shows that both the level of TCR occupancy and the amount of IFN‐γ produced by single T cells increase in an antigen dose‐dependent fashion. Conversely, commitment of T cells to IFN‐γ production does not occur as soon as a defined number of TCR have been engaged, but requires the same duration of sustained signalling at low as well as at high antigen concentrations. Measurement of phosphotyrosine levels by flow cytometry reveals that, upon conjugation with APC, individual T cells undergo an antigen dose‐dependent activation of protein tyrosine kinases (PTK), which parallels the level of TCR occupancy. In antigen‐stimulated T cells the increased phosphotyrosine staining is localized in the area of contact with APC, as shown by confocal microscopy. PTK activation is sustained for at least 2 hr after conjugation, and is required to maintain a sustained increase in intracellular Ca^2+^ concentration. Our results

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Author

Müller; Demotz; Bulliard; Valitutti

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0019-2805)

Published

1999

Language

EN

Field

Immunology and Microbiology (Life Sciences)