Characterization of antigen-binding and MHC class II-bearing T cells with suppressive activity in response to tolerogenic stimulus

Abstract

Antigen specific non-responsiveness is generally developed through clonal deletion, anergy, and suppression. The term “suppression” is being considered as a functional immune deficit that can be adoptively transferred by regulatory/suppressor T cells. Following tolerance induction protocols the aim of the present study was to characterize the T cells involved in antigen-specific suppression. After defining the immunogenic and tolerogenic protocols in vitro and in vivo, it was shown that CD90+ cells from tolerogenic mice were able to reduce specific antibody production when adaptively transferred to immunized mice. These cells were shown to highly express CD25 and Foxp3, co-localizing with CD4 and MHC class II antigens (MHCII), while a small percentage of these cells (8–14%) was binding free antigen. Further isolation of CD90+MHCII+ and CD90+HSA+ from mice having received the tolerogenic treatment and adaptive transfer to immunized mice showed that CD90+MHCII+ cells were able to suppress antigen-specific response only when transferred along with the second antigenic challenge, while CD90+HSA+ cells were suppressive only when given along with the first antigenic challenge. The suppressive effect of these two sub-populations could also be obtained in in vitro spleen cell proliferation assays in response to the HSA antigenic stimulus. Both in vitro and in vivo tolerogenic treatments were shown to correlate with soluble MHCII production in culture supernatants or serum respectively. Increase of MHCII in the serum could only be detected upon adaptive transfer of CD90+HSA+ cells, whereas transfer of CD90+MHCII+ cells resulted in increased levels of IL-10 and IFN-γ in the serum. These results defined at least two different levels of suppression, one during the onset which was antigen-specific and MHC restricted, and another non-specific at the end of an immune response.