A Novel Monoclonal Antibody Against Rat LFA-1: Blockade of LFA-1 and CD4 Augments Class II MHC Expression on T Cells

Abstract

Previously, we have shown that myelin basic protein (MBP)-specific Lewis rat GP2.E5/R1 (R1) T cells cultured with antigen and irradiated syngeneic splenocytes (IrrSPL) in the presence of anti-CD4 and LRTC1 monoclonal antibodies (MAbs) become highly effective antigen presenting cells (APC). The purpose of these studies was to identify the ligand for the LRTC1 MAb and to determine whether this MAb affected MBP-stimulated IL-2 production and expression of MHC class II molecules by T cells. In the current studies, we show that the LRTC1 MAb specifically immunoprecipitated molecular species of ∼95,150, and 180 kD. Commercially available anti-CD18 (β2 integrin, β-chain of LFA-1, MAC-1, and pl50, 95) and LRTC1 MAb immunoprecipitated proteins with identical mobilities on 1-D and 2-D SDS-PAGE gels. Moreover, anti-CD18 and LRTC1 immunoprecipitates also showed identical mobilities on 1-D gels after enzymatic cleavage of N-linked oligosaccharides and thereby had the same patterns of differential glycosylation. Anti-CD4 MAb W3/25 and LRTC1 MAb synergistically inhibited T-cell IL-2 mRNA and IL-2 bioactivity, but augmented antigenstimulated surface I-A on R1 T cells. In conclusion, these studies describe the characteristics of a novel anti-LFA-1 MAb, LRTC1, which should prove useful in studying costimulatory and adhesion pathways among rat leukocytes.