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Peripheral blood based T cell-containing and T cell-depleted culture systems for human IgE synthesis: the role of T
D J Wheeler1, R A Robins, D I Pritchard
1Department of Immunology, University of Nottingham, UK.
Background:
Comparable T cell-containing and T cell-depleted culture systems for human IgE synthesis are currently not available.
Objective:
This has prompted us to develop peripheral blood mononuclear cell (PBMC) based culture systems for human IgE synthesis in the presence and absence of T cells.
Methods:
In this paper we describe simplified conditions for in vitro synthesis of high levels of IgE by human peripheral blood B cells, both in T cell-containing cultures and in anti-CD40 stimulated T cell-depleted cultures.
Results:
T cell-depleted cultures released approximately 20 times more IgE [range 410-2220 ng/mliter (mean 1270 ng/mliter); based on six experiments using cells from three donors] than did T cell-containing cultures [range 23-105 ng/mliter (mean 58 ng/mliter); based on 15 experiments using cells from three donors]. Reconstitution experiments were performed to investigate the role of T cells on IgE synthesis. Adding T cells back to the anti-CD40 stimulated T cell-depleted cultures resulted in a dose-dependent inhibition of IgE production. In the absence of anti-CD40 low numbers of T cells stimulated, while high numbers suppressed, IgE production: the optimal ratio of T cells to non-T cells for maximal IgE production was found to be 1:1. At this ratio, irradiated (non-replicating) T cells supported a much greater IgE synthesis than did non-irradiated T cells.
Conclusion:
The development of these systems provides directly comparable T cell-containing and T cell-depleted cultures for human IgE synthesis from peripheral blood, allowing further study of the role of T cells in IgE regulation. These systems will also be of use for determining whether potential modulators of IgE synthesis act on the T cells or on other cell types.