Coexpression and functional cooperation of CTLA-4 and CD28 on activated T lymphocytes.

• DOI: 10.1084/jem.176.6.1595
• Publication Date: 2004-06-24T07:56:10Z
• Authors: P S Linsley; J L Greene; P Tan; J Bradshaw; J A Ledbetter; C Anasetti; N K Damle
• Keywords: Antigens, Differentiation, T-Lymphocyte; Mice, Inbred BALB C; Immunoconjugates; Antibodies, Monoclonal; Antigen-Presenting Cells; CHO Cells; Lymphocyte Activation; Antigens, Differentiation; Abatacept; Kinetics; Mice; 03 medical and health sciences; 0302 clinical medicine; CD28 Antigens; Antigens, CD; Cricetinae; CD4 Antigens; Cell Adhesion; Animals; CTLA-4 Antigen; Cells, Cultured; Plasmids

T cell costimulation by molecules on the antigen presenting cell (APC) is required for optimal T cell proliferation. The B7 molecule on APC binds the T lymphocyte receptor CD28, triggering increased interleukin 2 (IL-2) production and subsequent T cell proliferation. CTLA-4 is a predicted T cell membrane receptor homologous to CD28, which also binds the B7 counter receptor, but whose distribution and function are unknown. Here we have developed monoclonal antibodies (mAbs) specific for CTLA-4 and have investigated these questions. mAbs were produced that bound CTLA-4 but not CD28, and that blocked binding of CTLA-4 to B7. CTLA-4 expression as measured by these mAbs was virtually undetectable on resting T cells, but was increased several hundred-fold during T cell activation. On activated lymphocytes, CTLA-4 was expressed equally on CD4+ and CD8+ T cell subsets and was coexpressed with CD25, CD28, and CD45RO. CTLA-4 expression was lower than that of CD28, reaching a maximum of approximately 1/30-50 the level of CD28. Despite its lower expression, CTLA-4 was responsible for much of the B7 binding by large activated T cells. Anti-CTLA-4 mAb 11D4 and anti-CD28 mAb 9.3 acted cooperatively to inhibit T cell adhesion to B7, and to block T cell proliferation in primary mixed lymphocyte culture. When coimmobilized with anti T cell receptor (TCR) mAb, anti-CTLA-4 mAbs were less effective than anti-CD28 mAb 9.3 at costimulating proliferation of resting or activated T cells. However, coimmobilized combinations of anti-CD28 and anti-CTLA-4 were synergistic in their ability to augment anti-TCR-induced proliferation of preactivated CD4+ T cells. These results indicate that CTLA-4 is coexpressed with CD28 on activated T lymphocytes and cooperatively regulates T cell adhesion and activation by B7.