Frequencies and interactions of regulatory T cells. I. The balance between help and suppression regulates the primary immune response to keyhole limpet hemocyanin in vitro.

Frequencies and efficiencies of regulatory T cells from non-immunized mice were estimated in several assay systems differing from each other in cellular composition and antigen dose (NIP-KLH). The NIP-specific and total IgM responses were quantified. Using 10(4) syngeneic B cells and 50 micrograms/ml NIP-KLH, helper (Th) cells from 5 day immune donors were detected in frequencies of 1:3000-1:4000 in lymphnode and spleen T cells, with an efficiency of 70-90 ng IgM/Th cell in C57B1/6 mice. In non-immune spleen T cells, Th cells were observed in frequencies of 1:16,000-1:38,000, with comparable efficiency, but these Th cells appeared suppressed at increased T cell doses. Polyclonal activation led to the appearance of multiple independently regulated populations of Th cells with similar efficiencies. In the presence of 10(5) syngeneic spleen cells, treated once with anti-Thy-1 antibody and complement and 50 micrograms/ml NIP-KLH, suppressor activity was observed in the same T cell population. Similar to help, suppression fluctuated with increasing T cell numbers. Using 1 x 10(6) spleen cells and 50 micrograms/ml NIP-KLH as assay system, T cells enhanced the responses. Again, several independently regulated populations were observed, with efficiencies slightly higher than those of the above-described Th cells. By maintaining the cellular components of the assay system constant (10(4) B cells) and titrating the antigen, Th cell frequencies showed little variation up to 100 micrograms/ml NIP-KLH and were always suppressed at higher T cell numbers. At 200 micrograms/ml NIP-KLH, the frequency was increased to approximately 1:2000, and not suppressed, i.e., was identical to the frequency observed in mice immunized 5 days previously. Efficiencies increased with increasing doses of antigen. The results strongly indicate that regulatory T cell function shows "plasticity", in the sense that the appearance and the frequencies of helping and suppressing T cell populations highly depend on the micro-environment present in culture.