Molecular cloning, expression and characterization of the functional domain of CTLA4 from the rhesus monkey, Macaca mulatta

Cytotoxic T lymphocyte-associated antigen 4 (CTLA4) is a potent inhibitor of T cell activation. The genes encoding the membrane and soluble forms of Macaca mulatta CTLA4 (mmCTLA4) were isolated from peripheral blood mononuclear cells (PBMCs). The predicted mmCTLA4 protein is nearly identical to human CTLA4 (hCTLA4), with the exception of a serine instead of an asparagine residue at position 49 and a leucine instead of a methionine residue at position 141 of its extracellular domain. The fusion protein mmCTLA4Ig, containing the extracellular domain of mmCTLA4 and the constant region of the human IgG1 antibody, was expressed in Pichia pastoris. The mmCTLA4Ig produced by P. pastoris exhibited specific binding to human B7-positive Raji cells that could be inhibited by competitive binding of hCTLA4Ig. MmCTLA4Ig and hCTLA4Ig could comparably suppress the proliferation of lymphocytes derived from rhesus monkeys, humans, or mice that had been stimulated either by concanavalin A (Con A) or allogeneic cells. These results suggest that mmCTLA4 is a negative regulator of T cell activation and that mmCTLA4Ig may be useful for immunotherapy of immunologic diseases in the rhesus monkey.