Activation of T cells by cross-linking an anti-CD3 antibody with a second anti-T cell antibody: mechanism and subset-specific activation.

Binding of anti-CD3 antibody BMA030-F(ab')2 to T cells is not able to induce T cell proliferation even after additional cross-linking by plastic-bound goat anti-mouse Ig antibodies (panning) and addition of either interleukin (IL) 1 or IL2. In search for agents able to complement the signals provided by BMA030-F(ab')2, several antibodies directed against CD2, CD4, CD5, CD6 and CD8 were used. Neither of these caused T cell proliferation either by themselves or when simply added together with BMA030-F(ab')2. However, if BMA030-F(ab')2 and any other of these anti-CD2, CD4, CD5, CD6 or CD8 antibodies were cross-linked, then proliferation of T cells occurred. The mitogenic effect of cross-linking BMA030-F(ab')2 and a second anti-T cell antibody is dependent on the presence of monocytes or exogenously added IL2. The enhancing effect of monocytes could not be replaced by addition of IL1, suggesting that other soluble factors or monocyte contact might be involved in induction of activation signals. The mechanism leading to this mitogenic effect is dependent on combining cross-linking of BMA030 with the second anti-T cell antibody, whereby the second antibody appears to act as an "anchor" for the CD3 antibody, controlling and/or changing the signal transduction via the CD3 structure. This "anchor" hypothesis could be substantiated by the following observations: the best stimulatory signals were obtained at a BMA030/anti-T cell antibody ratio of 1:1; the effect is independent of the antibody isotype and the epitope recognized by the second antibody; the binding of BMA030 and separate cross-linking of the anti-T cell antibody was not sufficient to trigger T cell mitogenesis; immobilization by direct binding of the CD3 antibody alone or of the CD3 and second membrane structure/antibody complex separately was not sufficient to stimulate T cell proliferation. A further interesting aspect of this finding is the fact that a selective T cell stimulation is possible, since cross-linking BMA030-F(ab')2 plus anti-CD4 antibodies induced a selective stimulation of T4 cells, while cross-linking BMA030-F(ab')2 plus anti-CD8 activated solely T8 cells. It is thus possible to selectively activate T cell subsets in vitro.