Journal of NeuroVirology

, Volume 24, Issue 4, pp 411–419 | Cite as

In vivo characterization of macrophage-tropic simian immunodeficiency virus molecular clones in rhesus macaques

  • Sanjeev Gumber
  • Praveen Kumar Amancha
  • Po-Jen Yen
  • Francois Villinger
  • Dana Gabuzda
  • Siddappa N. Byrareddy


Macrophages are a major target of HIV/SIV infection and play an important role in pathogenesis by serving as viral reservoirs in the central nervous system. Previously, a unique early SIVmac251 envelope (Env) variant, deSIV147 was cloned from blood of a rhesus macaque with rapid disease progression and SIV-associated encephalitis. Here, we show that infectious molecular clone deSIV147 caused systemic infection in rhesus macaques following intravenous or intrarectal exposure. Next, we inoculated deSIV147 into macaques depleted of CD4+ T cells and found that animals were SIV-positive, with high plasma and CSF viral loads. These macaques also showed SIVp17-positive macrophages in brain, lymph nodes, colon, lung, and liver. Furthermore, accumulation of perivascular macrophages, multinucleated giant cells, and microgliosis was detected. These findings suggest that the neurotropic deSIV147 clone will be useful to study macrophage infection in HIV/SIV-associated neurocognitive disorders, gain insights into myeloid cell reservoirs in brain and other anatomical sites, as well as test strategies for eradication.


Macrophage-tropic Rhesus macaques Central nerveous system Myeloid cells Macrophages SIV 



We thank Ms. Stephanie Ehnert and the veterinary/support staff of the Yerkes National Primate Research center for their help in conducting macaque studies.


This study was supported in part by NIH R01AI113883 & R21AI114415 to SNB. DG was supported by NIH R01 MH 97659. Several antibodies and TZm-bl cells were obtained from NIH AIDS Reagent Reference Program.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13365_2018_628_Fig4_ESM.gif (48 kb)
Supplementary Fig.S1

Frequencies of classical monocytes (CD14+16-), intermediate monocytes (CD14+CD16+), and mDCs (CD3-CD8-CD20-CD14-CD16-HLADR+CD11c+) from CD4+ T cells depleted (top panel) and non-depleted (bottom panel) animals are shown. (TIFF 661 kb) (GIF 47 kb)

13365_2018_628_MOESM1_ESM.tif (661 kb)
High resolution image (TIFF 661 kb)
13365_2018_628_Fig5_ESM.gif (19 kb)
Supplementary Fig.S2:

Area under curve data for the CD14+16- (classical monocytes) is calculated from the longitudinal data shown in supplementary Fig S1A. A) is the data on individual animal and B) is the Mean and SD of both the groups. The sample size is too small to perform statistical analysis for the observed difference between the two groups. (TIFF 103 kb) (GIF 18 kb)

13365_2018_628_MOESM2_ESM.tif (103 kb)
high resolution image (TIFF 103 kb)
13365_2018_628_Fig6_ESM.gif (363 kb)
Supplementary Fig.S3:

IHC staining from representative tissue sections from CD4+ T cells depleted SIVmac251desiv147#C4 infected rhesus macaques (RDn15 & RCb15). Cervical lymph node (A), lung (B), small intestine (C), and mesenteric lymph node (D) showing SIVmac251 p17 (green) and CD68 (Fuchsin-red) labeled cells along with SIV/CD68 colocalization. (TIFF 661 kb) (TIFF 15577 kb) (GIF 362 kb)

13365_2018_628_MOESM3_ESM.tif (15.2 mb)
High resolution image (TIFF 15577 kb)


  1. Albright AV, Shieh JT, O’Connor MJ, Gonzalez-Scarano F (2000) Characterization of cultured microglia that can be infected by HIV-1. J Neuro-Oncol 6(Suppl 1):S53–S60Google Scholar
  2. Alexaki A, Liu Y, Wigdahl B (2008) Cellular reservoirs of HIV-1 and their role in viral persistence. Curr HIV Res 6:388–400CrossRefPubMedPubMedCentralGoogle Scholar
  3. Aylward EH, Brettschneider PD, McArthur JC, Harris GJ, Schlaepfer TE, Henderer JD, Barta PE, Tien AY, Pearlson GD (1995) Magnetic resonance imaging measurement of gray matter volume reductions in HIV dementia. Am J Psychiatry 152:987–994CrossRefPubMedGoogle Scholar
  4. Babas T, Munoz D, Mankowski JL, Tarwater PM, Clements JE, Zink MC (2003) Role of microglial cells in selective replication of simian immunodeficiency virus genotypes in the brain. J Virol 77:208–216CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bissel SJ, Wang G, Bonneh-Barkay D, Starkey A, Trichel AM, Murphey-Corb M, Wiley CA (2008) Systemic and brain macrophage infections in relation to the development of simian immunodeficiency virus encephalitis. J Virol 82:5031–5042CrossRefPubMedPubMedCentralGoogle Scholar
  6. Bixby JG, Laur O, Johnson WE, Desrosiers RC (2010) Diversity of envelope genes from an uncloned stock of SIVmac251. AIDS Res Hum Retrovir 26:1115–1131CrossRefPubMedGoogle Scholar
  7. Brown D, Mattapallil JJ (2014) Gastrointestinal tract and the mucosal macrophage reservoir in HIV infection. Clin Vaccine Immunol 21:1469–1473CrossRefPubMedPubMedCentralGoogle Scholar
  8. Burdo TH, Lackner A, Williams KC (2013) Monocyte/macrophages and their role in HIV neuropathogenesis. Immunol Rev 254:102–113CrossRefPubMedPubMedCentralGoogle Scholar
  9. Byrareddy SN, Arthos J, Cicala C, Villinger F, Ortiz KT, Little D, Sidell N, Kane MA, Yu J, Jones JW, Santangelo PJ, Zurla C, McKinnon LR, Arnold KB, Woody CE, Walter L, Roos C, Noll A, Van Ryk D, Jelicic K, Cimbro R, Gumber S, Reid MD, Adsay V, Amancha PK, Mayne AE, Parslow TG, Fauci AS, Ansari AA (2016) Sustained virologic control in SIV+ macaques after antiretroviral and alpha4beta7 antibody therapy. Science 354:197–202CrossRefPubMedPubMedCentralGoogle Scholar
  10. Byrareddy SN, Kallam B, Arthos J, Cicala C, Nawaz F, Hiatt J, Kersh EN, McNicholl JM, Hanson D, Reimann KA, Brameier M, Walter L, Rogers K, Mayne AE, Dunbar P, Villinger T, Little D, Parslow TG, Santangelo PJ, Villinger F, Fauci AS, Ansari AA (2014) Targeting alpha4beta7 integrin reduces mucosal transmission of simian immunodeficiency virus and protects gut-associated lymphoid tissue from infection. Nat Med 20:1397–1400CrossRefPubMedPubMedCentralGoogle Scholar
  11. Byrareddy SN, Little D, Mayne AE, Villinger F, Ansari AA (2015) Phenotypic and functional characterization of monoclonal antibodies with specificity for rhesus macaque CD200, CD200R and Mincle. PLoS One 10:e0140689CrossRefPubMedPubMedCentralGoogle Scholar
  12. Clements JE, Mankowski JL, Gama L, Zink MC (2008) The accelerated simian immunodeficiency virus macaque model of human immunodeficiency virus-associated neurological disease: from mechanism to treatment. J Neuro-Oncol 14:309–317Google Scholar
  13. Cysique LA, Soares JR, Geng G, Scarpetta M, Moffat K, Green M, Brew BJ, Henry RG, Rae C (2017). White matter measures are near normal in controlled HIV infection except in those with cognitive impairment and longer HIV duration. J NeurovirolGoogle Scholar
  14. Daniel MD, Letvin NL, King NW, Kannagi M, Sehgal PK, Hunt RD, Kanki PJ, Essex M, Desrosiers RC (1985) Isolation of T-cell tropic HTLV-III-like retrovirus from macaques. Science 228:1201–1204CrossRefPubMedGoogle Scholar
  15. Del Prete GQ, Scarlotta M, Newman L, Reid C, Parodi LM, Roser JD, Oswald K, Marx PA, Miller CJ, Desrosiers RC, Barouch DH, Pal R, Piatak M, Jr., Chertova E, Giavedoni LD, O'Connor DH, Lifson JD, Keele BF (2013). Comparative characterization of transfection- and infection-derived simian immunodeficiency virus challenge stocks for in vivo nonhuman primate studies. J Virol 87: 4584–4595Google Scholar
  16. Dick AD, Pell M, Brew BJ, Foulcher E, Sedgwick JD (1997) Direct ex vivo flow cytometric analysis of human microglial cell CD4 expression: examination of central nervous system biopsy specimens from HIV-seropositive patients and patients with other neurological disease. AIDS 11:1699–1708CrossRefPubMedGoogle Scholar
  17. Ellis RJ, Hsia K, Spector SA, Nelson JA, Heaton RK, Wallace MR, Abramson I, Atkinson JH, Grant I, McCutchan JA (1997) Cerebrospinal fluid human immunodeficiency virus type 1 RNA levels are elevated in neurocognitively impaired individuals with acquired immunodeficiency syndrome. HIV Neurobehavioral Research Center Group Ann Neurol 42:679–688PubMedGoogle Scholar
  18. Fauci AS, Desrosiers RC (1997). Pathogenesis of HIV and SIV. In: Retroviruses. Coffin JM, Hughes SH, Varmus HE, (eds): Cold Spring Harbor (NY)Google Scholar
  19. Igarashi T, Brown CR, Endo Y, Buckler-White A, Plishka R, Bischofberger N, Hirsch V, Martin MA (2001) Macrophage are the principal reservoir and sustain high virus loads in rhesus macaques after the depletion of CD4+ T cells by a highly pathogenic simian immunodeficiency virus/HIV type 1 chimera (SHIV): implications for HIV-1 infections of humans. Proc Natl Acad Sci U S A 98:658–663CrossRefPubMedPubMedCentralGoogle Scholar
  20. Johnson PR, Hirsch VM (1992) SIV infection of macaques as a model for AIDS pathogenesis. Int Rev Immunol 8:55–63CrossRefPubMedGoogle Scholar
  21. Kanki PJ, McLane MF, King NW, Jr., Letvin NL, Hunt RD, Sehgal P, Daniel MD, Desrosiers RC, Essex M (1985). Serologic identification and characterization of a macaque T-lymphotropic retrovirus closely related to HTLV-III. Science 228: 1199–1201Google Scholar
  22. Kent KA, Gritz L, Stallard G, Cranage MP, Collignon C, Thiriart C, Corcoran T, Silvera P, Stott EJ (1991) Production and of monoclonal antibodies to simian immunodeficiency virus envelope glycoproteins. AIDS 5:829–836CrossRefPubMedGoogle Scholar
  23. Kestler H, Kodama T, Ringler D, Marthas M, Pedersen N, Lackner A, Regier D, Sehgal P, Daniel M, King N et al (1990) Induction of AIDS in rhesus monkeys by molecularly cloned simian immunodeficiency virus. Science 248:1109–1112CrossRefPubMedGoogle Scholar
  24. Kuper M, Rabe K, Esser S, Gizewski ER, Husstedt IW, Maschke M, Obermann M (2011) Structural gray and white matter changes in patients with HIV. J Neurol 258:1066–1075CrossRefPubMedGoogle Scholar
  25. Letvin NL, Daniel MD, Sehgal PK, Desrosiers RC, Hunt RD, Waldron LM, MacKey JJ, Schmidt DK, Chalifoux LV, King NW (1985) Induction of AIDS-like disease in macaque monkeys with T-cell tropic retrovirus STLV-III. Science 230:71–73CrossRefPubMedGoogle Scholar
  26. Maartens G, Celum C, Lewin SR (2014) HIV infection: epidemiology, pathogenesis, treatment, and prevention. Lancet 384:258–271CrossRefPubMedGoogle Scholar
  27. McArthur JC, McClernon DR, Cronin MF, Nance-Sproson TE, Saah AJ, St Clair M, Lanier ER (1997) Relationship between human immunodeficiency virus-associated dementia and viral load in cerebrospinal fluid and brain. Ann Neurol 42:689–698CrossRefPubMedGoogle Scholar
  28. Miller CJ, Marthas M, Greenier J, Lu D, Dailey PJ, Lu Y (1998) In vivo replication capacity rather than in vitro macrophage tropism predicts efficiency of vaginal transmission of simian immunodeficiency virus or simian/human immunodeficiency virus in rhesus macaques. J Virol 72:3248–3258PubMedPubMedCentralGoogle Scholar
  29. Mori K, Ringler DJ, Kodama T, Desrosiers RC (1992) Complex determinants of macrophage tropism in env of simian immunodeficiency virus. J Virol 66:2067–2075PubMedPubMedCentralGoogle Scholar
  30. Nwogu JN, Ma Q, Babalola CP, Adedeji WA, Morse GD, Taiwo B (2016) Pharmacokinetic, pharmacogenetic, and other factors influencing CNS penetration of antiretrovirals. AIDS Res Treat 2016:2587094PubMedPubMedCentralGoogle Scholar
  31. Perelson AS, Neumann AU, Markowitz M, Leonard JM, Ho DD (1996) HIV-1 dynamics in vivo: virion clearance rate, infected cell life-span, and viral generation time. Science 271:1582–1586CrossRefPubMedGoogle Scholar
  32. Sharer LR, Baskin GB, Cho ES, Murphey-Corb M, Blumberg BM, Epstein LG (1988) Comparison of simian immunodeficiency virus and human immunodeficiency virus encephalitides in the immature host. Ann Neurol 23(Suppl):S108–S112CrossRefPubMedGoogle Scholar
  33. Strain MC, Letendre S, Pillai SK, Russell T, Ignacio CC, Gunthard HF, Good B, Smith DM, Wolinsky SM, Furtado M, Marquie-Beck J, Durelle J, Grant I, Richman DD, Marcotte T, McCutchan JA, Ellis RJ, Wong JK (2005) Genetic composition of human immunodeficiency virus type 1 in cerebrospinal fluid and blood without treatment and during failing antiretroviral therapy. J Virol 79:1772–1788CrossRefPubMedPubMedCentralGoogle Scholar
  34. Strickland SL, Gray RR, Lamers SL, Burdo TH, Huenink E, Nolan DJ, Nowlin B, Alvarez X, Midkiff CC, Goodenow MM, Williams K, Salemi M (2011) Significant genetic heterogeneity of the SIVmac251 viral swarm derived from different sources. AIDS Res Hum Retrovir 27:1327–1332CrossRefPubMedGoogle Scholar
  35. Swingler S, Mann AM, Zhou J, Swingler C, Stevenson M (2007) Apoptotic killing of HIV-1-infected macrophages is subverted by the viral envelope glycoprotein. PLoS Pathog 3:1281–1290CrossRefPubMedGoogle Scholar
  36. Takahashi Y, Byrareddy SN, Albrecht C, Brameier M, Walter L, Mayne AE, Dunbar P, Russo R, Little DM, Villinger T, Khowawisetsut L, Pattanapanyasat K, Villinger F, Ansari AA (2014) In vivo administration of a JAK3 inhibitor during acute SIV infection leads to significant increases in viral load during chronic infection. PLoS Pathog 10:e1003929CrossRefPubMedPubMedCentralGoogle Scholar
  37. Valcour V, Sithinamsuwan P, Letendre S, Ances B (2011) Pathogenesis of HIV in the central nervous system. Curr HIV/AIDS Rep 8:54–61CrossRefPubMedGoogle Scholar
  38. von Giesen HJ, Adams O, Koller H, Arendt G (2005) Cerebrospinal fluid HIV viral load in different phases of HIV-associated brain disease. J Neurol 252(7):801–807CrossRefGoogle Scholar
  39. Wang J, Crawford K, Yuan M, Wang H, Gorry PR, Gabuzda D (2002) Regulation of CC chemokine receptor 5 and CD4 expression and human immunodeficiency virus type 1 replication in human macrophages and microglia by T helper type 2 cytokines. J Infect Dis 185:885–897CrossRefPubMedGoogle Scholar
  40. Wiley CA, Schrier RD, Nelson JA, Lampert PW, Oldstone MB (1986) Cellular localization of human immunodeficiency virus infection within the brains of acquired immune deficiency syndrome patients. Proc Natl Acad Sci U S A 83:7089–7093CrossRefPubMedPubMedCentralGoogle Scholar
  41. Williams DW, Engle EL, Shirk EN, Queen SE, Gama L, Mankowski JL, Zink MC, Clements JE (2016) Splenic damage during SIV infection: role of T-Cell depletion and macrophage polarization and infection. Am J Pathol 186:2068–2087CrossRefPubMedPubMedCentralGoogle Scholar
  42. Williams K, Schwartz A, Corey S, Orandle M, Kennedy W, Thompson B, Alvarez X, Brown C, Gartner S, Lackner A (2002) Proliferating cellular nuclear antigen expression as a marker of perivascular macrophages in simian immunodeficiency virus encephalitis. Am J Pathol 161:575–585CrossRefPubMedPubMedCentralGoogle Scholar
  43. Williams KC, Corey S, Westmoreland SV, Pauley D, Knight H, deBakker C, Alvarez X, Lackner AA (2001) Perivascular macrophages are the primary cell type productively infected by simian immunodeficiency virus in the brains of macaques: implications for the neuropathogenesis of AIDS. J Exp Med 193:905–915CrossRefPubMedPubMedCentralGoogle Scholar
  44. Wohlschlaeger J, Wenger E, Mehraein P, Weis S (2009) White matter changes in HIV-1 infected brains: a combined gross anatomical and ultrastructural morphometric investigation of the corpus callosum. Clin Neurol Neurosurg 111:422–429CrossRefPubMedGoogle Scholar
  45. Yen PJ, Mefford ME, Hoxie JA, Williams KC, Desrosiers RC, Gabuzda D (2014) Identification and characterization of a macrophage-tropic SIV envelope glycoprotein variant in blood from early infection in SIVmac251-infected macaques. Virology 458-459:53–68CrossRefPubMedGoogle Scholar
  46. Zink MC, Clements JE (2002) A novel simian immunodeficiency virus model that provides insight into mechanisms of human immunodeficiency virus central nervous system disease. J Neuro-Oncol 8(Suppl 2):42–48Google Scholar

Copyright information

© Journal of NeuroVirology, Inc. 2018

Authors and Affiliations

  • Sanjeev Gumber
    • 1
  • Praveen Kumar Amancha
    • 2
  • Po-Jen Yen
    • 3
  • Francois Villinger
    • 2
  • Dana Gabuzda
    • 3
  • Siddappa N. Byrareddy
    • 4
  1. 1.Division of Pathology, Yerkes National Primate Research CenterEmory UniversityAtlantaGeorgia
  2. 2.New Iberia Research CenterUniversity of Louisiana at LafayetteNew IberiaUSA
  3. 3.Department of Cancer Immunology and VirologyDana-Farber Cancer InstituteBostonUSA
  4. 4.Department of Pharmacology and Experimental NeuroscienceUniversity of Nebraska Medical CenterOmahaUSA

Personalised recommendations