title: Differential functional effects of a humanized anti-CD4 antibody on resting and activated human T cells.
abstract: A fully humanized immunoglobulin G1 (IgG1) anti-CD4 monoclonal antibody is currently being evaluated in phase I/II clinical trials for rheumatoid arthritis. In order to understand the mode of action of this antibody in vivo, we have carried out a detailed functional analysis in vitro of the effects of this antibody on T-cell activation. The anti-CD4 antibody was found to inhibit both antigen-specific responses involving recognition of human leucocyte antigen (HLA) class II and processed antigenic peptides as well as non-class II dependent responses via anti-CD3 antibodies. The antibody did not cause total blockade of T-cell proliferation, but rather induced a shift in the dose-response curve, decreasing the sensitivity of cells to antigen or anti-CD3-mediated stimulation. The antibody appears to allow at least a partial early signal into the T cell as it does not inhibit the increase in tyrosine phosphorylation induced by anti-CD3 antibodies. A comparison of the intact antibody with that of either the F(ab')2 fragment or an engineered non-Fc receptor (FcR) binding form revealed that the intact antibody was the most effective at inhibiting proliferation of resting peripheral blood CD4+ T cells. However, this difference was only apparent when excess antibody was removed from culture prior to antigen or anti-CD3 mediated stimulation. The intact antibody induced both CD4 down-modulation and increases in CD4-associated tyrosine phosphorylation of resting CD4+ T cells, which were not seen with the non-FcR binding versions, which may account for the enhanced potency of the intact antibody at inhibiting T-cell activation. Interestingly, the anti-CD4 antibody induced a differential effect on activated CD4+ T cell clones compared with resting CD4+ T cells with respect to degree of CD4 cross-linking required to induce functional effects in the T cell. Both intact and non-FcR binding antibodies were equally effective at inhibiting T-cell proliferation of activated T-cell clones. In addition CD4 down-modulation and increased CD4-associated tyrosine phosphorylation were observed with T-cell clones in the absence of secondary cross-linking. Such observations may be of relevance when studying the effects of the antibody at sites of inflammation, where there will be CD4+ T cells of differing activation states as well as varying numbers of FcR positive cells.
authors.affiliation: Immunology Unit, Glaxo-Wellcome Medicines Research Centre, Stevenage, UK.
authors.forename: S J
authors.initials: SJ
authors.lastname: Brett
authors.forename: W
authors.initials: W
authors.lastname: Rowan
authors.forename: M
authors.initials: M
authors.lastname: Smith
authors.forename: M
authors.initials: M
authors.lastname: Bartholomew
authors.forename: J P
authors.initials: JP
authors.lastname: Tite
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D015536:Down-Regulation / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology*; D011961:Receptors, Fc / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D017252:CD3 Complex; D015704:CD4 Antigens; D011961:Receptors, Fc