HIV-infection of Kupffer cells results in a dysregulated response to LPS despite effective Anti-Retroviral Therapy
Arevik Mosoian1, Feng Hong1, Yedidya Saiman1, Adeeb Rahman1, Andrea D. Branch1, Sasan Roayaie1, Sander Florman1, Francesc Cunyat2, Mario Stevenson2, Meena Bansal1
1lcahn School of Medicine at Mount Sinai, New York, NY; 2University of Miami Leonard M. Miller School of Medicine, Miami, FL
HIV infection causes CD4+ lymphocyte depletion in the gut and has been linked to the disruption of gut epithelial integrity and increased microbial translocation. Microbial translocation has been linked to liver disease progression in HIV/HCV coinfected patients and to systemic immune activation in HIVinfected patients. Since Kupffer cells (KCs) are known targets of HIV in vivo, are responsible for the uptake of translocated microbial products entering the portal circulation, and are tolerant to effects of LPS, we hypothesized that HIV infection of KCs may alter their response to LPS and postulated that despite effective systemic anti-retroviral therapy, their response to LPS may be persistently dysregulated Methods: KCs were isolated by density centrifugation from livers of patients with and without HIV infection who underwent resection or transplant (3HIV/HCV and 1-HIV/HBV/HCV). All HIV patients had undetectable plasma HIV for a minimum of one year. Purity of KC preps was assessed by RT-qPCR for CD68, CD14, TLR4, CD3, and CD31.HIV-BaL (R5 tropic) and HIV-IIIB (X4 tropic) were used to infect primary KCs. Cytokine levels in response to LPS were examined by RT-qPCR and ELISA in both the in vitro infected KCs and the KCs derived from HIV-infected patients. Evidence of latent HIV infection in KCs was assessed by RTqPCR for HIV multiply-spliced RNA products and by qPCR for HIV DNA. Results: R5 tropic HIV-Bal productively infected KCs and resulted in large amounts of viral production for over one month in culture suggesting that infection is not cytopathic. KCs challenged with LPS showed a minimal inflammatory response. However, infection of KCs with R5 or X4 tropic HIV resulted in a significantly enhanced production of pro-inflammatory cytokines (IL-8, TNFα, IL-6) and the pro-fibrogenic cytokine (TGFβ1) in response to LPS but not to poly-IC suggesting a specific effect on TLR4 ligands. Most notably, we detected HIV DNA by qPCR and multiply-spliced RNA by RT-qPCR in KCs derived from the livers of all aviremic HIV patients and these cells maintained a dysregulated pro-inflammatory response to LPS. Conclusions: HIV-1 infection of Kupffer cells in vitro results in an augmented pro-inflammatory and pro-fibrogenic response to LPS but not poly-IC. Despite undetectable plasma HIV RNA in patients on ART, KCs contain HIV DNA and multiply-spliced RNA and maintain a dysregulated pro-inflammatory response to LPS. These findings suggest that HIV infection of Kupffer cells may contribute to hepatic inflammation and fibrosis by altering response to translocated microbial products.
Andrea D. Branch - Grant/Research Support: Kadmon, Gilead, Janssen
The following people have nothing to disclose: Arevik Mosoian, Feng Hong, Yedidya Saiman, Adeeb Rahman, Sasan Roayaie, Sander Florman, Francesc Cunyat, Mario Stevenson, Meena Bansal