Abstracts (complete list) - Wissenschaft Online

Maria Diedrichs-Möhring, Georges M.G.M. Verjans, Seerp G. Baarsma, Gerhild
Wildner
Comparison of antigen-specific cytokine secretion and
proliferation of T lymphocytes in vitro
Response of T lymphocytes following stimulation with antigen or mitogen can be
determined either by proliferation or by secretion of distinct cytokines. Due to limited
material it is often not possible to establish cultures for early and lately secreted
cytokines as well as to detect proliferation. Therefore, we combined both methods.
Lymphocytes were stimulated in 96-well-plates with different antigens or mitogen.
Culture supernatants were collected after 24 h, 48 h, 72 h and 96 h and frozen until
use. The remaining cultures were used to determine proliferation by 3H-thymidine
uptake. Supernatants from the various time points were pooled to analyze the secreted
cytokines by multiplex bead immunoassay. To verify this approach peripheral blood
lymphocytes from two donors were stimulated in the same way and cytokine
concentrations were determined from samples of single time points (24 h, 48 h, 72 h
and 96 h) as well as from pooled supernatants.
Using this approach we were able to detect early as well as lately secreted cytokines in
the supernatants of stimulated T lymphocytes. Interestingly, the peak of distinct
cytokine secretion in response to a certain antigen varied between different individuals.
Comparing cytokine secretion and proliferation of T lymphocytes we found no
correlation: high amounts of TNF-α and / or IFN-γ were found in weakly proliferating
cultures and vice versa. These results indicate that cytokine secretion and proliferation
are two different aspects of lymphocyte response following stimulation and these
parameters are not replaceable by each other.
Supported by Deutsche Forschungsgemeinschaft, SFB 571