Costimulatory molecule expression on human uveal melanoma cells: Functional analysis of CD40 and B7-H1

Abstract

Costimulatory molecules play important roles in regulating T cell function in tumor immunity. In this study, we investigated costimulatory molecule expression on human uveal melanoma cells (a primary culture, and OCM-1, OMM-1 and 92-1 cell lines) and assessed the functional roles of selected costimulatory molecules. Uveal melanoma cells were incubated in the presence or absence of IFN-γ and expression of costimulatory molecules on the cells was measured by flow cytometry. The costimulatory effect of B7-H1-expressing uveal melanoma cells on cytokine production by purified T cells was studied in uveal melanoma/T cell co-culture experiments using a blocking anti-B7-H1 monoclonal antibody (mAb). The functional role of CD40-mediated interactions in modifying immune responses to uveal melanoma cells was assessed in vitro using recombinant human CD40 ligand (rhCD40L). MHC class I and B7-H1 were consistently detected and further upregulated by IFN-γ stimulation in all human uveal melanoma cell cultures. CD40 was consistently detected and further upregulated by IFN-γ stimulation in primary culture, OCM-1, and OMM-1 but not 92-1. IL-2 production from purified CD3+ T cells co-stimulated with IFN-γ-treated uveal melanoma cells was significantly enhanced by the addition of anti-B7-H1 mAb. Treatment of primary culture, OCM-1, or OMM-1 with rhCD40L induced or enhanced secretion of chemokines IL-8, MCP-1, IP-10 and RANTES. These results suggest that the expression of B7-H1 on IFN-γ-treated uveal melanoma cells contributes to suppression of T cells by decreasing IL-2 production. In contrast, CD40 expressed on uveal melanoma cells plays an important role in augmenting anti-tumor immunity by stimulating chemokine production. The dual effects of CD40 and B7-H1 may contribute to positive or negative regulation of anti-tumor immune responses to human uveal melanoma.