Abstract
Whether the human brain is a robust reservoir for HIV-1 subtype C has yet to be established. We aimed to determine whether HIV-1 subtype C infection can be detected in the brain tissue of a viremic individual at post-mortem and whether the viral burden was differential between different brain regions. This study reports a 38-year-old Zambian female decedent with severe wasting who was on Atripla for antiretroviral therapy. The cause of death was determined to be HIV/AIDS end-stage disease. The QuantStudio 3 Real-Time PCR System analyzed formalin-fixed paraffin-embedded tissue DNA from a systematic sampling of the entire left-brain hemisphere. Plasma and cerebral spinal fluid HIV-1 RNA loads were 576,123 and 14,962 copies/mL, respectively. The lymph node DNA viral load was 2316 copies per 106 cells. Two hundred and six (96.3%) tissue blocks had amplifiable DNA. HIV-1 viral DNA was detected in 35.9% of the blocks, the highest in the basal ganglia (66.7%) and the frontal lobe (46%). Overall, HIV detection was random, with low viral copies detected by quantitative polymerase chain reaction (qPCR); the lowest was observed in the occipital (median, IQR, range) 0.0 [0.0–0.0], 0.0–31.3, and the highest in the basal ganglia (mean ± SD, range, 125.1149.5, 0.0–350.0). Significant differences in HIV-1 DNA distribution were observed between the occipital versus parietal (p = 0.049), occipital versus frontal (p = 0.019), occipital versus basal ganglia (p = 0.005), cerebellum versus frontal (p = 0.021), cerebellum versus basal ganglia (p = 0.007), and temporal versus frontal (p = 0.034).
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References
Amornkul PN, Karita E, Kamali A, Rida WN, Sanders EJ, Lakhi S, Price MA, Kilembe W, Cormier E, Anzala O, Latka MH, Bekker LG, Allen SA, Gilmour J, Fast PE (2013) Disease progression by infecting HIV-1 subtype in a seroconverter cohort in sub-Saharan Africa. AIDS 27(17):2775–2786. https://doi.org/10.1097/QAD.0000000000000012
Archibald SL, Masliah E, Fennema-Notestine C, Marcotte TD, Ellis RJ, McCutchan JA, Heaton RK, Grant I, Mallory M, Miller A, Jernigan TL (2004) Correlation of in vivo neuroimaging abnormalities with postmortem human immunodeficiency virus encephalitis and dendritic loss. Arch Neurol 61(3):369–376. https://doi.org/10.1001/ARCHNEUR.61.3.369
Avettand-Fènoë V, Hocqueloux L, Ghosn J, Cheret A, Frange P, Melard A, Viard JP, Rouzioux C (2016) Total HIV-1 DNA, a marker of viral reservoir dynamics with clinical implications. Clin Microbiol Rev 29(4):859–880. https://doi.org/10.1128/CMR.00015-16
Bachiller S, Jiménez-Ferrer I, Paulus A, Yang Y, Swanberg M, Deierborg T, Boza-Serrano A (2018) Microglia in neurological diseases: a road map to brain-disease dependent-inflammatory response. Front Cell Neurosci 12. Front Media S.A. https://doi.org/10.3389/fncel.2018.00488
Banerjee A, Zhang X, Manda KR, Banks WA, Ercal N (2010) HIV proteins (gp120 and Tat) and methamphetamine in oxidative stress-induced damage in the brain: potential role of the thiol antioxidant N-acetylcysteine amide. Free Radical Biol Med 48(10):1388–1398. https://doi.org/10.1016/J.FREERADBIOMED.2010.02.023
Bozzi G, Simonetti FR, Watters SA, Anderson EM, Gouzoulis M, Kearney MF, Rote P, Lange C, Shao W, Gorelick R, Fullmer B, Kumar S, Wank S, Hewitt S, Kleiner DE, Hattori J, Bale MJ, Hill S, Bell J, Maldarelli F (2019) No evidence of ongoing HIV replication or compartmentalization in tissues during combination antiretroviral therapy: implications for HIV eradication. Sci Adv 5(9). https://doi.org/10.1126/SCIADV.AAV2045/SUPPL_FILE/AAV2045_SM.PDF
Carr JM, Cheney KM, Coolen C, Davis A, Shaw D, Ferguson W, Chang G, Higgins G, Burrell C, Li P (2007) Development of methods for coordinate measurement of total cell-associated and integrated human immunodeficiency virus type 1 (HIV-1) DNA forms in routine clinical samples: levels are not associated with clinical parameters, but low levels of integrated HIV-1 DNA may be prognostic for continued successful therapy. J Clin Microbiol 45(4):1288–1297. https://doi.org/10.1128/JCM.01926-06/ASSET/B3BAA430-30BF-40F4-BEA0-DCC2AC2F3A77/ASSETS/GRAPHIC/ZJM0040772260004.JPEG
Carroll E, Sanchez-ramos J (2011) Hyperkinetic movement disorders associated with HIV and other viral infections. Handb Clin Neurol 100:323–334. https://doi.org/10.1016/B978-0-444-52014-2.00025-2
Chéret A, Bacchus-Souffan C, Avettand-Fenoël V, Mélard A, Nembot G, Blanc C, Samri A, Sáez-Cirión A, Hocqueloux L, Lascoux-Combe C, Allavena C, Goujard C, Valantin MA, Leplatois A, Meyer L, Rouzioux C, Autran B, Hoen B, Bourdeaux C, Abel S (2015) Combined ART started during acute HIV infection protects central memory CD4+ T cells and can induce remission. J Antimicrob Chemother 70(7):2108–2120. https://doi.org/10.1093/JAC/DKV084
Clifford DB, Ances BM (2013) HIV-associated neurocognitive disorder. Lancet Infect Dis 13(11):976–986. https://doi.org/10.1016/S1473-3099(13)70269-X
Cohn LB, Chomont N, Deeks SG (2020) The biology of the HIV-1 latent reservoir and implications for cure strategies. Cell Host Microbe 27(4):519. https://doi.org/10.1016/J.CHOM.2020.03.014
Fukazawa Y, Lum R, Okoye AA, Park H, Matsuda K, Bae JY, Hagen SI, Shoemaker R, Deleage C, Lucero C, Morcock D, Swanson T, Legasse AW, Axthelm MK, Hesselgesser J, Geleziunas R, Hirsch VM, Edlefsen PT, Piatak M, Picker LJ (2015) B cell follicle sanctuary permits persistent productive simian immunodeficiency virus infection in elite controllers. Nat Med 21(2):132–139. https://doi.org/10.1038/nm.3781
Ganasen KA, Fincham D, Smit J, Seedat S, Stein D (2008) Utility of the HIV Dementia Scale (HDS) in identifying HIV dementia in a South African sample. J Neurol Sci 269(1–2):62–64. https://doi.org/10.1016/j.jns.2007.12.027
Gartner MJ, Roche M, Churchill MJ, Gorry PR, Flynn JK (2020) Understanding the mechanisms driving the spread of subtype C HIV-1. EBioMedicine. https://doi.org/10.1016/j.ebiom.2020.102682
Gendelman HE, Persidsky Y, Ghorpade A, Stins M, Fiala M, Morrisett R (1997) The neuropathogenesis of the AIDS dementia complex. AIDS 11:35–45. https://pubmed.ncbi.nlm.nih.gov/9451964/
Graf EH, Mexas AM, Yu JJ, Shaheen F, Liszewski MK, di Mascio M, Migueles SA, Connors M, O’Doherty U (2011) Elite suppressors harbor low levels of integrated HIV DNA and high levels of 2-LTR circular HIV DNA compared to HIV+ patients on and off HAART. PLoS Pathogens 7(2). https://doi.org/10.1371/JOURNAL.PPAT.1001300
Hosmane NN, Kwon KJ, Bruner KM, Capoferri AA, Beg S, Rosenbloom DIS, Keele BF, Ho YC, Siliciano JD, Siliciano RF (2017) Proliferation of latently infected CD4+ T cells carrying replication-competent HIV-1: potential role in latent reservoir dynamics. J Exp Med 214(4):959–972. https://doi.org/10.1084/JEM.20170193
Ivey NS, MacLean AG, Lackner AA (2009) Acquired immunodeficiency syndrome and the blood-brain barrier. J Neurovirol 15(2):111–122. https://doi.org/10.1080/13550280902769764
Ketzler S, Weis S, Haug H, Budka H (1990) Loss of neurons in the frontal cortex in AIDS brains. Acta Neuropathol 80(1):92–94. https://doi.org/10.1007/BF00294228
Lamers SL, Gray RR, Salemi M, Huysentruyt LC, McGrath MS (2011) HIV-1 phylogenetic analysis shows HIV-1 transits through the meninges to brain and peripheral tissues. Infect Genet Evol 11(1):31–37. https://doi.org/10.1016/J.MEEGID.2010.10.016
López-Villegas D, Lenkinski RE, Frank I (1997) Biochemical changes in the frontal lobe of HIV-infected individuals detected by magnetic resonance spectroscopy. Proc Natl Acad Sci USA 94(18):9854–9859. https://doi.org/10.1073/PNAS.94.18.9854
Martrus G, Altfeld M (2016) Immunological strategies to target HIV persistence. Curr Opin HIV AIDS 11(4):402–408. https://doi.org/10.1097/COH.0000000000000289
McRae MP (2016) HIV and viral protein effects on the blood brain barrier. Tissue Barr 4(1). https://doi.org/10.1080/21688370.2016.1143543
Mishra M, Varghese RK, Verma A, Das S, Aguiar RS, Tanuri A, Mahadevan A, Shankar SK, Satishchandra P, Ranga U (2015) Genetic diversity and proviral DNA load in different neural compartments of HIV-1 subtype C infection. J NeuroVirol 21(4):399–414. https://doi.org/10.1007/S13365-015-0328-0
Naif HM (2013) Pathogenesis of HIV infection. Infectious Disease Reports 5(Suppl 1):e6–e6. https://doi.org/10.4081/idr.2013.s1.e6
Nir TM, Fouche JP, Ananworanich J, Ances BM, Boban J, Brew BJ, Chaganti JR, Chang L, Ching CRK, Cysique LA, Ernst T, Faskowitz J, Gupta V, Harezlak J, Heaps-Woodruff JM, Hinkin CH, Hoare J, Joska JA, Kallianpur KJ, Jahanshad N (2021) Association of immunosuppression and viral load with subcortical brain volume in an international sample of people living with HIV. JAMA Netw Open 4(1):e2031190–e2031190. https://doi.org/10.1001/JAMANETWORKOPEN.2020.31190
Novitsky V, Woldegabriel E, Kebaabetswe L, Rossenkhan R, Mlotshwa B, Bonney C, Finucane M, Musonda R, Moyo S, Wester C, van Widenfelt E, Makhema J, Lagakos S, Essex M (2009) Viral load and CD4+ T-cell dynamics in primary HIV-1 subtype C infection. J Acquir Immune Defic Syndr 50(1):65–76. https://doi.org/10.1097/QAI.0B013E3181900141
Pasternak AO, Adema KW, Bakker M, Jurriaans S, Berkhout B, Cornelissen M, Lukashov VV (2008) Highly sensitive methods based on seminested real-time reverse transcription-PCR for quantitation of human immunodeficiency virus type 1 unspliced and multiply spliced RNA and proviral DNA. J Clin Microbiol 46(7):2206–2211. https://doi.org/10.1128/JCM.00055-08
Prevedel L, Ruel N, Castellano P, Smith C, Malik S, Villeux C, Bomsel M, Morgello S, Eugenin EA (2019) Identification, localization, and quantification of HIV reservoirs using microscopy. Curr Protoc Cell Biol 82(1):e64. https://doi.org/10.1002/CPCB.64
Rao VR, Ruiz AP, Prasad VR (2014) Viral and cellular factors underlying neuropathogenesis in HIV associated neurocognitive disorders (HAND). AIDS Res Ther 11(1):13. https://doi.org/10.1186/1742-6405-11-13
Re MC, Vitone F, Biagetti C, Schiavone P, Alessandrini F, Bon I, de Crignis E, Gibellini D (2010) HIV-1 DNA proviral load in treated and untreated HIV-1 seropositive patients. Clin Microbiol Infect 16(6):640–646. https://doi.org/10.1111/j.1469-0691.2009.02826.x
Rotta I, de Almeida SM (2011) Genotypical diversity of HIV clades and central nervous system impairment. Arq Neuropsiquiatr 69(6):964–972. https://doi.org/10.1590/S0004-282X2011000700023
Scutari R, Alteri C, Perno CF, Svicher V, Aquaro S (2017) The role of HIV infection in neurologic injury. Brain Sci 7(4). https://doi.org/10.3390/BRAINSCI7040038
Stevenson M, Haggerty S, Lamonica CA, Meier CM, Welch SK, Wasiak AJ (1990) Integration is not necessary for expression of human immunodeficiency virus type 1 protein products. J Virol 64(5):2421–2425. https://doi.org/10.1128/JVI.64.5.2421-2425.1990
Tavazzi E, Morrison D, Sullivan P, Morgello S, Fischer T (2014) Brain inflammation is a common feature of HIV-infected patients without HIV encephalitis or productive brain infection. Curr HIV Res 12(2):97–110. https://doi.org/10.2174/1570162X12666140526114956
Taylor BS, Sobieszczyk ME, McCutchan FE, Hammer SM (2008) The challenge of HIV-1 subtype diversity. N Engl J Med 358(15):1590–1602. https://doi.org/10.1056/NEJMra0706737
Trautmann L (2016) Kill: boosting HIV-specific immune responses. Curr Opin HIV AIDS 11(4):409–416. https://doi.org/10.1097/COH.0000000000000286
Tso FY, Kang G, Kwon EH, Julius P, Li Q, West JT, Wood C (2018) Brain is a potential sanctuary for subtype C HIV-1 irrespective of ART treatment outcome. PLoS ONE 13(7):e0201325. https://doi.org/10.1371/journal.pone.0201325
UNAIDS (2021) UNAIDS data 2021. https://www.unaids.org/en/resources/documents/2021/2021_unaids_data
Veenstra M, León-Rivera R, Li M, Gama L, Clements JE, Berman JW (2017) Mechanisms of CNS viral seeding by HIV+ CD14+ CD16+ monocytes: establishment and reseeding of viral reservoirs contributing to HIV-associated neurocognitive disorders. MBio 8(5). https://doi.org/10.1128/MBIO.01280-17/ASSET/4AF9A5CB-2074-42C2-ACCC-DC4589846558/ASSETS/GRAPHIC/MBO0051735590004.JPEG
Wallet C, de Rovere M, van Assche J, Daouad F, de Wit S, Gautier V, Mallon PWG, Marcello A, van Lint C, Rohr O, Schwartz C (2019) Microglial cells: the main HIV-1 reservoir in the brain. Front Cell Infect Microbiol 9. https://www.frontiersin.org/article/10.3389/fcimb.2019.00362
Wiley CA, Soontornniyomkij V, Radhakrishnan L, Masliah E, Mellors J, Hermann SA, Dailey P, Achim CL (2006) Distribution of brain HIV load in AIDS. Brain Pathol 8(2):277–284. https://doi.org/10.1111/j.1750-3639.1998.tb00153.x
Williams ME, Zulu SS, Stein DJ, Joska JA, Naudé PJW (2020) Signatures of HIV-1 subtype B and C Tat proteins and their effects in the neuropathogenesis of HIV-associated neurocognitive impairments. Neurobiol Dis 136:104701. https://doi.org/10.1016/J.NBD.2019.104701
Wong JK, Yukl SA (2016) Tissue reservoirs of HIV. Curr Opin HIV AIDS 11(4):362–370. https://doi.org/10.1097/COH.0000000000000293
Zayyad Z, Spudich S (2015) Neuropathogenesis of HIV: from initial neuroinvasion to HIV-associated neurocognitive disorder (HAND). Curr HIV/AIDS Rep 12(1):16–24. https://doi.org/10.1007/S11904-014-0255-3
Zazzi M, Romano L, Catucci M, Venturi G, de Milito A, Almi P, Gonnelli A, Rubino M, Occhini U, Valensin PE (1997) Evaluation of the presence of 2-LTR HIV-1 unintegrated DNA as a simple molecular predictor of disease progression. J Med Virol 52(1):20–25. https://doi.org/10.1002/(SICI)1096-9071(199705)52:1%3c20::AID-JMV4%3e3.0.CO;2-T
Acknowledgements
We thank Dr. Chibamba Mumba and members of the ZAMDAPP, the University Teaching Hospitals (UTH)-Department of Pathology and Microbiology for recruitment, and the Wood Laboratory for the support and critical discussions of the work. We thank Dr. Luchenga Mucheleng'anga and Dr. Cordelia Himwaze for doing the autopsy and getting samples for testing.
Funding
The Public Health Service Awards partly funded this study: National Institute of Drug Abuse R01 DA044920; Fogarty International D43 TW010354; National Cancer Institute U54CA221204 through CW. JM, PJ, and SNS are Fogarty Fellows.
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JM managed the research study at the UTH and UNL, performed the histopathology and molecular work, and drafted the manuscript; PJ provided pathology expertise, helped with conceptualization, and helped write and edit the manuscript; SNS and DY performed the statistical analysis; GK helped perform histopathology work; SM assisted with the conceptualization of the proposal; CW and JTW provided mentorship, analyzed the results, and helped write and edit the manuscript.
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Musumali, J., Julius, P., Siyumbwa, S.N. et al. Systematic post-mortem analysis of brain tissue from an HIV-1 subtype C viremic decedent revealed a paucity of infection and pathology. J. Neurovirol. 28, 527–536 (2022). https://doi.org/10.1007/s13365-022-01099-8
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DOI: https://doi.org/10.1007/s13365-022-01099-8