Figure 5

Clone 10 treatment reduces T cell proliferation and activation. Extracellular matrix metalloproteinase inducer (EMMPRIN) levels in non-cultured T cells immediately upon isolation ((A); non-cult.), cultured without activation ((B); non-act.), and cultured for 72 h with anti-CD3 and anti-CD28 ((C); act.) are compared using fluorescence-activated cell sorting (FACS) analysis. EMMPRIN levels in T cells from four individual volunteers are displayed (D). Human PBMCs incubated in the absence (non-act.) or presence (act.) of anti-CD3 and anti-CD28 were treated with various concentrations of clone 10, IgM isotype control, a commercial anti-human EMMPRIN antibody (Ancell) and IgG Isotype control. After 48 h cells were incubated with tritiated thymidine and proliferation assessed (E); results are mean ± SEM of quadruplicates; ***P <0.001 compared to act. T cells (one-way analysis of variance (ANOVA) with Tukey’s post hoc comparisons). Similarly, non-activated and activated cells were treated with clone 10 or IgM isotype control and carboxyfluorescein diacetate succinimidyl ester (CFSE) intensity was determined in CD3+ cells by FACS analysis (F) and quantified (G). Gelatin zymography was used to determine matrix metalloproteinase 9 (MMP-9) levels in conditioned media from peripheral blood mononuclear cells (PBMCs) non-activated (non-act.), activated (act.), activated and with clone 10 treatment (act. + C10), and activated with IgM isotype treatment (act. + IgM) (H).