A new in vitro model for studying human T cell differentiation: T H1/T H2 induction following activation by superantigens

Abstract

A new T H1/T H2 in vitro model for mechanistic studies and drug screening in human T cells was established working with ficoll-separated PBMCs or elutriated lymphocytes cultured in serum-free medium. Human T cells could be kept viable and reactive in this medium for several months. In this model, superantigens (SAs) were used to activate exclusively those T cell clones which were known to express specifically SA-binding Vβ-chains of the T cell receptor. It was possible to identify the activated SA-specific T cells among the whole T cell population by using monoclonal antibodies against these Vβ-chains and to follow responses involving receptor regulation and cytokine expression. The flow cytometric analysis revealed, that SA exposure caused an upregulation of the IL-2 receptor selectively in the SA-specific subpopulation. Only the T cells of this subpopulation could be shifted towards T H1 or T H2 differentiation, which was determined by the distribution of IFN-γ and IL-4 positive cells. Regulation of IFN-γ could be detected by flow cytometry after 18 h and that of IL-4 on the third day of cell culture. The differentiation status could be influenced by various measures: T H1 shifts were achieved in the presence of IL-12 and anti-IL-4, whereas, T H2 shifts were induced more slowly with monocyte-reduced elutriated lymphocytes in the presence of IL-4, IL-6, anti-IL-12, 1α,25-dihydroxy-vitamin-D 3 or combinations thereof. It was found that sialidase stimulated whereas TGF-β and pentoxifylline suppressed both kinds of T cell response. The T H1/T H2 differentiation persisted for several weeks after primary activation if cells were expanded in IL-2 containing serum-free culture medium. Therefore, this human T H1/T H2 in vitro model should be ideal for studying early and late events of infection, allergy, and autoimmunity as well as for investigating the cellular interactions involved. In addition, the early detection of the response pattern makes this model potentially useful for drug screening.