Primary B lymphocytes infected with KSHV can be expanded in vitro and are recognized by LANA-specific CD4+ T cells

Samantha Nicol, Shereen Sabbah, Jae Jung, Kevin Brulios, Andrew Bell, Andrew Hislop

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Kaposi sarcoma-associated herpesvirus (KSHV) has a tropism for B lymphocytes in which it establishes latency and can also cause lymphoproliferative disorders of these cells manifesting as primary effusion lymphoma (PEL) and multi-centric Castleman disease (MCD). T cell immunity is vital for the control of KSHV infection and disease however few models of B lymphocyte infection exist to study immune recognition of such cells. Here we developed a model of B lymphocyte infection with KSHV where infected tonsillar B lymphocytes were expanded by providing mitogenic stimuli and these challenged with KSHV-specific CD4+ T cells. Infected cells expressed viral proteins found in PELs namely LANA and vIRF3, albeit at lower levels, with similar patterns of gene expression for the major latency, vIL-6 and vIRF3 transcripts. Despite low level expression of ORF50, transcripts for the immune evasion genes K3 and K5 were detected, with some downregulation of cell surface expressed CD86 and ICAM. The vast majority of infected lymphocytes expressed IgM and the Ig light chain, recapitulating these features seen in infected cells in MCD. We assessed the ability of the infected lymphocytes to be targeted by a panel of MHC class II matched CD4+ T cells and found that LANA-specific T cells restricted to different epitopes recognized these infected cells. Given that at least some KSHV latent antigens are thought to be poor targets for CD8+ T cells, we suggest that CD4+ T cells are potentially important effectors for the in vivo control of KSHV infected B lymphocytes.
Original languageEnglish
JournalJournal of virology
Early online date27 Jan 2016
Publication statusE-pub ahead of print - 27 Jan 2016


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