Cytokine profiles and regulatory function of GPIIb/IIIa-reactive CD4+ T-cell lines treated twice with anti-CD154 mAb.
GPIIb/IIIa-reactive CD4+ T-cell lines SiM4 and WY9 were cultured twice with autologous lymphoblastoid B-cell line, antigenic recombinant fragment, and IL-2 in the presence of anti-CD154 or isotype-matched control mAb. (A) The T-cell lines treated with anti-CD154 or isotype-matched control mAb (2 × 105) were stimulated with phorbol myristate acetate (1 μg/mL) and ionomycin (25 ng/mL) for 48 hours, and the levels of IFN-γ, IL-2, IL-4, IL-6, and IL-10 in the culture supernatants were measured by ELISA. The results shown are the means ± SDs of triplicate values. Asterisks indicate significant differences in cytokine levels between the anti-CD154 mAb–treated and isotype-matched control mAb–treated T-cell lines. The results of 1 of 2 experiments with similar results are shown. (B) Line WY9 (3 × 105) was cultured with autologous B cells (3 × 105), IIIa22-262 (5 μg/mL), and pokeweed mitogen (1 μg/mL) for 10 days in the presence or absence of serially diluted WY9-treated twice with anti-CD154 mAb (0.5 to 2 × 105) or treated with isotype-matched control mAb (2 × 105). Anti–IL-10 mAb (10 μg/mL) was added at the start of some cultures. IgG anti-GPIIb/IIIa antibodies in culture supernatants were measured by ELISA. The results shown are the means ± SDs of duplicate values. Asterisks indicate significant difference between anti-GPIIb/IIIa antibody levels in cultures with and without the anti-CD154 mAb–treated WY9 cells. The results of 1 of 2 experiments with similar results are shown.
