B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers

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Abstract

Chronic-phase HIV and simian immunodeficiency virus (SIV) replication is reduced by as much as 10,000-fold in elite controllers (ECs) compared with typical progressors (TPs), but sufficient viral replication persists in EC tissues to allow viral sequence evolution and induce excess immune activation. Here we show that productive SIV infection in rhesus monkey ECs, but not TPs, is markedly restricted to CD4+ follicular helper T (TFH) cells, suggesting that these EC monkeys' highly effective SIV-specific CD8+ T cells can effectively clear productive SIV infection from extrafollicular sites, but their relative exclusion from B cell follicles prevents their elimination of productively infected TFH cells. CD8+ lymphocyte depletion in EC monkeys resulted in a dramatic re-distribution of productive SIV infection to non-TFH cells, with restriction of productive infection to TFH cells resuming upon CD8+ T cell recovery. Thus, B cell follicles constitute 'sanctuaries' for persistent SIV replication in the presence of potent anti-viral CD8+ T cell responses, potentially complicating efforts to cure HIV infection with therapeutic vaccination or T cell immunotherapy.

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Figure 1: The distribution of productive SIV infection within CD4+ memory T cell subsets differs in chronic-phase, but not acute-phase, attenuated versus WT SIVmac239 infection.
Figure 2: The distribution of productive WT SIV infection within CD4+ memory T cells in LN correlates with immune control.
Figure 3: Productive SIV infection in monkeys is anatomically restricted to B cell follicles in EC, but not TP, monkeys.
Figure 4: The restriction of productive SIV infection to CD4+ TFH cells in EC LNs is lost with in vivo CD8+ lymphocyte depletion.
Figure 5: The restriction of productive SIV infection to CD4+ TFH cells in EC LNs is not affected by activation of extrafollicular CD4+ memory T cells with IL-7 administration.
Figure 6: CD4+ TFH cells contain higher levels of cell-associated SIV RNA than non-TFH CD4+ memory T cells in SIV+ monkeys with effective suppression of viral replication.

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Acknowledgements

The authors dedicate this paper to the memory of Michael Piatak, Jr., in recognition of his key enabling contributions to this study and so many others. This work was supported by the US National Institutes of Health; grants 5R37A1054292 (L.J.P.), 5U19AI096109 (L.J.P. and A.A.O.), and 8P51OD01109255, contract HHSN261200800001E (R.S., M.P., J.D.E. and J.D.L.) and Intramural Program of the US National Institute of Allergy and Infectious Diseases, and the Bill and Melinda Gates Foundation (grant no. 41185; L.J.P.). The authors thank R. Wiseman and D. Watkins for MHC typing; Dr. K. Reimann and the US National Institutes of Health Nonhuman Primate Reagent Resource Program for provision of the CD8α-specific monoclonal antibody M-T807R1; D. Hazuda (Merck Research Labs) for providing cART drugs for cohort 1; Cytheris SA (Issy Les Moulineaux, France) for recombinant rhesus IL-7; S. Hansen for animal study management; A. Sylwester, L. Koo, J. Clock, A. Konfe, H.W. Kim, M. Rohankhedkar, M. Reyes, N. Coombes, B. Assaf, K. Oswald, R. Fast, Y. Li, C. Trubey, J. Turner, S. Planer and L. Boshears for technical or administrative assistance.

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L.J.P. and Y.F. conceived the study, designed the experiments and wrote the paper, assisted by A.A.O., H.P. and J.D.L. Y.F. also supervised the experiments, performed immunologic and SIV coculture assays, and analyzed the data, assisted by R.L., J.Y.B. and S.I.H. A.W.L. and M.K.A. managed the animal protocols, assisted by T.S. K.M., V.M.H., C.D., C.L., D.M. and J.D.E. provided tissue-based analysis including immunohistochemistry and RNAscope. M.P., Jr. and J.D.L. planned and performed SIV quantification, assisted by R.S. J.H. and R.G. developed and provided optimized cART regimens (cohort 2). P.T.E. performed the statistical analysis of study data.

Correspondence to Louis J Picker.

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Fukazawa, Y., Lum, R., Okoye, A. et al. B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers. Nat Med 21, 132–139 (2015) doi:10.1038/nm.3781

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