Corrected: Author correction ARTICLE https://doi.org/10.1038/s41467-019-12987-9 OPEN Macrophage-associated wound healing contributes to African green monkey SIV pathogenesis control Fredrik Barrenas1,2,16, Kevin Raehtz3,4,16, Cuiling Xu5, Lynn Law6,7, Richard R. Green6,7, Guido Silvestri8,9, Steven E. Bosinger8,9, Andrew Nishida1, Qingsheng Li 10, Wuxun Lu10, Jianshui Zhang10, Matthew J. Thomas6,11, Jean Chang6,7, Elise Smith6,7, Jeffrey M. Weiss1, Reem A. Dawoud8, George H. Richter5, Anita Trichel12, Dongzhu Ma13, Xinxia Peng 14, Jan Komorowski 2,15, Cristian Apetrei3,4, Ivona Pandrea4,5 & Michael Gale Jr 6,7,11* 1234567890():,; Natural hosts of simian immunodeficiency virus (SIV) avoid AIDS despite lifelong infection. Here, we examined how this outcome is achieved by comparing a natural SIV host, African green monkey (AGM) to an AIDS susceptible species, rhesus macaque (RM). To asses gene expression profiles from acutely SIV infected AGMs and RMs, we developed a systems biology approach termed Conserved Gene Signature Analysis (CGSA), which compared RNA sequencing data from rectal AGM and RM tissues to various other species. We found that AGMs rapidly activate, and then maintain, evolutionarily conserved regenerative wound healing mechanisms in mucosal tissue. The wound healing protein fibronectin shows distinct tissue distribution and abundance kinetics in AGMs. Furthermore, AGM monocytes exhibit an embryonic development and repair/regeneration signature featuring TGF-β and con- comitant reduced expression of inflammatory genes compared to RMs. This regenerative wound healing process likely preserves mucosal integrity and prevents inflammatory insults that underlie immune exhaustion in RMs. 1 Department of Microbiology, University of Washington, Seattle, WA, USA. 2 Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden. 3 Division of Infectious Diseases, Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. 4 Department of Microbiology and Molecular Genetics, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. 5 Department of Pathology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. 6 Department of Immunology, University of Washington, Seattle, WA, USA. 7 Center for Innate Immunity and Immune Diseases, University of Washington, Seattle, WA, USA. 8 Department of Pathology & Laboratory Medicine, Emory University, Atlanta, GA, USA. 9 Division of Microbiology & Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA. 10 Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE, USA. 11 Washington National Primate Research Center, University of Washington, Seattle, WA, USA. 12 Divison of Laboratory Animal Resources, University of Pittsburgh, Pittsburgh, PA, USA. 13 Department of Orthopedic Surgery, University of Pittsburgh, Pittsburgh, PA, USA. 14 Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, NC, USA. 15 Institute of Computer Science, PAN, Warsaw, Poland. 16 These authors contributed equally: Fredrik Barrenas, Kevin Raehtz. *email: [email protected] NATURE COMMUNICATIONS | (2019) 10:5101 | https://doi.org/10.1038/s41467-019-12987-9 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-019-12987-9 urbing the HIV pandemic remains a global health priority, acute SIV infection, total RNA-seq, bioinformatic, virologic, and but significant scientific hurdles prevent an effective vac- histologic analyses are performed. We report highly dissimilar C 1–3 fi cine or therapy . To this end, de ning the molecular expression dynamics between the two SIV hosts. To fully inter- HIV/SIV immunopathogenic mechanisms remains critical. The rogate these differences, we develop a systems biology approach, most useful model systems to this end are possibly the African referred to as Conserved Gene Signature Analysis (CGSA), based non-human primate (NHP) species that carry SIV in the wild, on correlating expression signatures in acute SIV infection across termed natural SIV reservoir hosts, e.g. the African green monkey informative transcriptomic datasets from the public domain, (AGM—Chlorocebus genus), sooty mangabey (SM—Cercocebus including across diverse species. These datasets entail relevant atys) and mandrill (Mandrillus sphinx). Remarkably, these species tissues (e.g. mucosal tissue, epithelial cells) and conditions (e.g. remain disease-free after SIV infection despite high levels of tissue damage, cytokine stimulation), such that CGSA enables viremia and major mucosal CD4+ T cell depletion4–6. data-driven, unbiased discovery of pathogenic or protective In contrast, SIV-infected rhesus macaques (RMs) and other mechanisms of SIV disease progression. Asian NHPs exhibit sequelae typical of AIDS including CD4+ T Using CGSA, we identify a strong wound healing signature in cell loss, lymphoid tissue fibrosis, hypercoagulation, intestinal the AGM model, including monocyte regulated tissue remodeling epithelium breakdown, and opportunistic infections7,8. Com- similar to that of the axolotl, a neotenic salamander species with parative studies of natural and pathogenic SIV hosts have deli- remarkable wound healing abilities. In contrast, RMs induce neated several immunological and genomic features as novel microbial pattern recognition receptor and inflammatory signal- treatment targets, aimed at reducing harmful inflammation dur- ing. Our observations implicate monocyte-directed mucosal ing HIV infection5,9. immune programming in AGM SIV responses, that con- The defining trait of the natural SIV host phenotype is lack of comitantly maintain intestinal integrity and avoid microbial protracted immune activation10–13. In infected humans and activation. macaques, markers of immune activation are strong indicators of Thus, maintenance of mucosal integrity through control of mortality14. Immunological mechanisms linked to the avoidance monocyte immune responses, could be a valuable therapeutic of pathogenic immune activation in natural hosts include: (1) strategy to avoid systemic immune activation that underlies SIV/ alterations in sequence or expression of CD415 or CCR516,17 on HIV disease progression and AIDS. target cells, restricting infection in vulnerable lymphocyte popu- lations; (2) resolution of interferon signaling and innate immune Results activation;10,18,19 (3) maintenance of mucosal integrity7,15,18,20,21. SIV inoculation and rectal tissue transcriptomic profiling.To Additionally, (4) macrophages, which are prevalent at SIV/HIV identify the host responses that determine pathogenic or natural infection sites22, are less susceptible to SIV in natural hosts5,23. infection outcomes, 28 AGMs and 24 RMs underwent a single Crucial pathogenic events occur within days of SIV infection, high-dose intrarectal inoculation with two genetically diverse including seeding of viral reservoirs3 and systemic immune swarm stocks (SIV 92081 and SIV 251) previously shown to activation24. In RMs, the intestinal tract undergoes dramatic loss sab mac have comparable infection kinetics11,31. As several SIVs replicate of immune cells and tight junction protein expression during the poorly in non-adapted hosts10,32, we selected viruses to ensure acute infection25, partly due to macrophage SIV infection26. This comparable viral loads, and natural or pathogenic infection out- allows microbial components to penetrate the epithelial barrier, comes in AGMs and RMs, respectively. aggravating inflammatory responses7. Transcriptomic analyses of Tissue samples were collected upon necropsy at specific time RMs acutely infected with SIV found activation of genes asso- points over an acute infection course: pre-viremic (D1–4), early ciated with NK cells, monocytes and T cells in rectal tissues24, viremic (D5–6), viral peak (D9–12) and viral set point (D50 or underscoring the prevalent hypothesis that AIDS starts with a D84) (Fig. 1a). Pre-inoculation baseline samples were collected by massive insult to the mucosal immune structures, leading to pinch biopsy. In collected blood samples, plasma was separated to persistent inflammation and systemic immune defects20. This measure viral load. In both infection models, peak viral load hypothesis describes the first days after SIV/HIV infection as occurred at D9–12 (1.39 × 107 in AGMs, 2.48 × 107 in RMs, critical for the immune programming driving infection outcome. Fig. 1b). At set point, viral load in AGMs was approximately one In natural SIV hosts, transcriptional studies of acute SIV log lower than RMs (6.95 × 104 and 5.69 × 105, respectively, p = infection have utilized intravenous inoculation and focused on 0.023 by two-sided T-test). Viral loads in AGM and RM rectal peripheral blood immune cells. In SIV-infected AGMs, CD4+ tissues were comparable (Supplementary Fig. 1). cells mount interferon responses earlier than in RMs, but resolve them after the acute phase, exemplifying how early innate immune programming could potentially drive nonpathogenic Contrasting responses to acute SIV infection in AGMs and infection outcome18,19,27. Similarly, both colon and lymph nodes RMs. Rectal tissues collected during necropsy and pre- show strong but transient immune activation, Th1 polarization inoculation underwent total RNA-seq profiling. The resulting and progenitor cell regeneration during acute infection in sequence reads were mapped to the genomes of the respective AGMs27. Intestinal damage in SIV-infected AGMs, recapitulates