|Detection of CD28 in CD3+ Mouse Splenocytes by Flow Cytometry. CD3+ mouse splenocytes were stained with Rat Anti-Mouse CD28 APC‑conjugated Monoclonal Antibody (Catalog # FAB4832A, filled histogram) or isotype control antibody (Catalog # IC005A, open histogram). View our protocol for Staining Membrane-associated Proteins.|
CD28 and CTLA-4, together with their ligands B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T and B cell responses. CD28 and CTLA-4 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily. Both CD28 and CTLA-4 are composed of a single Ig V‑like extracellular domain, a transmembrane domain and an intracellular domain. CD28 and CTLA-4 are both expressed on the cell surface as disulfide-linked homodimers or as monomers. The genes encoding these two molecules are closely linked on human chromosome 2 and mouse chromosome 1. Mouse CD28 is expressed constitutively on virtually 100% of mouse T cells and on developing thymocytes. Cell surface expression of mouse CD28 is down-regulated upon ligation of CD28 in the presence of PMA or PHA. In contrast, CTLA-4 is not expressed constitutively but is upregulated rapidly following T cell activation and CD28 ligation. Cell surface expression of CTLA-4 peaks approximately 48 hours after activation. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7‑1 and B7‑2 with a 20-100 fold higher affinity than CD28. CD28/B7 interaction has been shown to prevent apoptosis of activated T cells via the up-regulation of Bcl‑xL. CD28 ligation has also been shown to regulate Th1/Th2 differentiation. Agonist activity has been reported using MAB4831 (4,5).