Innate Sensing of HIV-Infected Cells

Abstract

Cell-free HIV-1 virions are poor stimulators of type I interferon (IFN) production. We examined here how HIV-infected cells are recognized by plasmacytoid dendritic cells (pDCs) and by other cells. We show that infected lymphocytes are more potent inducers of IFN than virions. There are target cell-type differences in the recognition of infected lymphocytes. In primary pDCs and pDC-like cells, recognition occurs in large part through TLR7, as demonstrated by the use of inhibitors and by TLR7 silencing. Donor cells expressing replication-defective viruses, carrying mutated reverse transcriptase, integrase or nucleocapsid proteins induced IFN production by target cells as potently as wild-type virus. In contrast, Env-deleted or fusion defective HIV-1 mutants were less efficient, suggesting that in addition to TLR7, cytoplasmic cellular sensors may also mediate sensing of infected cells. Furthermore, in a model of TLR7-negative cells, we demonstrate that the IRF3 pathway, through a process requiring access of incoming viral material to the cytoplasm, allows sensing of HIV-infected lymphocytes. Therefore, detection of HIV-infected lymphocytes occurs through both endosomal and cytoplasmic pathways. Characterization of the mechanisms of innate recognition of HIV-infected cells allows a better understanding of the pathogenic and exacerbated immunologic events associated with HIV infection.

Author Summary

AIDS is characterized by a hyperactivation of the immune system. Innate and inflammatory responses, associated with an exacerbated production of cytokines like type I interferons (IFN) and of chemokines, deregulate the normal functioning of T lymphocytes and other cells. The events that trigger this inappropriate activation remain poorly understood. Plasmacytoid dendritic cells (pDCs) normally produce IFN when they encounter viruses. Here we examined how HIV-infected cells are recognized by pDCs, as well as by other immune and non-immune cells. We show that viruses transmitted via cell-to-cell contacts are more potent inducers of IFN than cell-free viral particles. In pDCs, recognition occurs in large part through TLR7, a cellular receptor detecting viral genetic materials after capture in intracellular vesicles. Donor cells expressing replication-defective viruses are also able to trigger IFN production by target cells. We further show that in TLR7-negative, non-hematopoietic cells an additional cytoplasmic pathway allows sensing of HIV-infected lymphocytes. Therefore, detection of HIV-infected lymphocytes occurs at different intracellular localizations, and does not require ongoing viral replication. Characterization of the mechanisms of innate HIV-1 recognition allows a better understanding of the pathology of HIV infection, and has consequences for the design of vaccine strategies.