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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Immunology I Abstract 13 THE KSHV LYTIC PROTEIN VOX2 AND ITS CELLULAR COUNTERPART, CD200, INHIBIT EPITOPE-SPECIFIC T CELL RESPONSES Rachel Colman, Karen Misstear, Heather Long, Omar Quereshi^, David Sansom^, Andrew Hislop, David J. Blackbourn CRUK Institute for Cancer Studies and ^Division of Immunity and Infection, University of Birmingham, Birmingham B15 2TT, UK Abstract Background & Objectives: The KSHV lytic protein vOX2 shares 36% identity with human cellular CD200. An engineered soluble derivative of vOX2 inhibited neutrophil activity in vitro and had anti-inflammatory activity in vivo, suggesting the protein inhibits innate immune responses. In the present study we determined whether adaptive immune responses were modulated by native, membrane-bound vOX2 and CD200. Methods: Antigen presenting cells (APC) were engineered to express either vOX2 or CD200 on their surface by retroviral transduction and membrane expression was confirmed by flow cytometry and IFA. APC loaded with epitope peptides were incubated with appropriate epitope-specific CD4+ or CD8+ T cell clones and T cell responses were measured by IFN-γ release. Ex-vivo restimulation of epitope-specific CD8+ T cells was performed by coculturing peripheral blood mononuclear cells (PBMC) with autologous APC engineered to express either vOX2 or CD200 on their surface. T cell function was measured by IFN-γ release. Results: Expression of vOX2 or CD200 on the surface of APCs inhibited IFN-γ secretion by co-cultured CD4+ and CD8+ T cell clones up to 50% when compared to vector only transduced APCs. Restimulation of PBMC was similarly inhibited. Neither HLA class I, nor CD80 and CD86 costimulatory molecule expression levels were downregulated on APCs by vOX2 or CD200 expression. Conclusion: These data demonstrate that both KSHV vOX2 and cellular CD200 inhibit antigen-specific T-lymphocyte activity. They suggest vOX2 suppresses adaptive immune responses against KSHV lytically infected cells, facilitating virus replication and dissemination in vivo. Presenting author Email: r.colman@bham.ac.uk 34
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Session 3 abstracts 14-21: Pathogenesis 35
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