HIV Productively Infects Highly Differentiated and Exhausted CD4+ T Cells During AIDS
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Abstract
Background: Throughout HIV infection, productively infected cells generate billions of viral particles and are thus responsible for body-wide HIV dissemination, but their phenotype during AIDS is unknown. As AIDS is associated with immunological changes, analyzing the phenotype of productively infected cells can help understand HIV production during this terminal stage.
Methods: Blood samples from 15 untreated viremic participants (recent infection, n=5; long-term infection, n=5; active opportunistic AIDS-defining disease, n=5) and 5 participants virologically controlled on antiretroviral therapy (ART) enrolled in the Analysis of the Persistence, Reservoir and HIV Latency (APRIL) study (NCT05752318) were analyzed. Cells expressing the capsid protein p24 (p24+ cells) after 18 hours of resting or 24 hours of stimulation (HIV-Flow) revealed productively infected cells from viremic participants or translation-competent reservoir cells from treated participants, respectively.
Results: The frequency of productively infected cells tended to be higher during AIDS in comparison with recent and long-term infections (median, 340, 72, and 32/million CD4+ T cells, respectively) and correlated with the plasma viral load at all stages of infection. Altogether, these cells were more frequently CD4low, HLA-ABClow, CD45RA-, Ki67+, PD-1+, with a non-negligible contribution from pTfh (CXCR5+PD-1+) cells, and were not significantly enriched in HIV coreceptors CCR5 nor CXCR4 expression. The comparison markers expression between stages showed that productively infected cells during AIDS were enriched in memory and exhausted cells. In contrast, the frequencies of infected pTfh were lower during AIDS compared to non-AIDS stages. A UMAP analysis revealed that total CD4+ T cells were grouped in 7 clusters and that productive p24+ cells were skewed to given clusters throughout the course of infection. Overall, the preferential targets of HIV during the latest stages seemed to be more frequently highly differentiated (memory, TTD-like) and exhausted cells and less frequently pTfh-like cells. In contrast, translation-competent reservoir cells were less frequent (5/million CD4+ T cells) and expressed more frequently HLA-ABC and less frequently PD-1.
Conclusions: In long-term infection and AIDS, productively infected cells were differentiated and exhausted. This could indicate that cells with these given features are responsible for HIV production and dissemination in an immune dysfunction environment occurring during the last stages of infection.
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References
1. Brenchley JM, Hill BJ, Ambrozak DR, Price DA, Guenaga FJ, Casazza JP, Kuruppu J, Yazdani J, Migueles SA, Connors M, Roederer M, Douek DC, Koup RA. T-cell subsets that harbor human immunodeficiency virus (HIV) in vivo: implications for HIV pathogenesis. J Virol. 2004;78(3):1160-8. doi: 10.1128/jvi.78.3.1160-1168.2004. PubMed PMID: 14722271; PMCID: PMC321406.
2. Chomont N, El-Far M, Ancuta P, Trautmann L, Procopio FA, Yassine-Diab B, Boucher G, Boulassel MR, Ghattas G, Brenchley JM, Schacker TW, Hill BJ, Douek DC, Routy JP, Haddad EK, Sekaly RP. HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. Nature Medicine. 2009;15(8):893-900. doi: 10.1038/nm.1972. PubMed PMID: 19543283; PMCID: 2859814.
3. Banga R, Procopio FA, Noto A, Pollakis G, Cavassini M, Ohmiti K, Corpataux JM, de Leval L, Pantaleo G, Perreau M. PD-1(+) and follicular helper T cells are responsible for persistent HIV-1 transcription in treated aviremic individuals. Nature Medicine. 2016;22(7):754-61. doi: 10.1038/nm.4113. PubMed PMID: 27239760.
4. Fromentin R, Bakeman W, Lawani MB, Khoury G, Hartogensis W, DaFonseca S, Killian M, Epling L, Hoh R, Sinclair E, Hecht FM, Bacchetti P, Deeks SG, Lewin SR, Sekaly RP, Chomont N. CD4+ T Cells Expressing PD-1, TIGIT and LAG-3 Contribute to HIV Persistence during ART. PLoS Pathog. 2016;12(7):e1005761. doi: 10.1371/journal.ppat.1005761. PubMed PMID: 27415008; PMCID: PMC4944956.
5. Gosselin A, Wiche Salinas TR, Planas D, Wacleche VS, Zhang Y, Fromentin R, Chomont N, Cohen EA, Shacklett B, Mehraj V, Ghali MP, Routy JP, Ancuta P. HIV persists in CCR6+CD4+ T cells from colon and blood during antiretroviral therapy. AIDS. 2017;31(1):35-48. doi: 10.1097/QAD.0000000000001309. PubMed PMID: 27835617; PMCID: PMC5131694.
6. Pardons M, Baxter AE, Massanella M, Pagliuzza A, Fromentin R, Dufour C, Leyre L, Routy JP, Kaufmann DE, Chomont N. Single-cell characterization and quantification of translation-competent viral reservoirs in treated and untreated HIV infection. PLoS Pathog. 2019;15(2):e1007619. doi: 10.1371/journal.ppat.1007619. PubMed PMID: 30811499; PMCID: PMC6411230.
7. Bruner KM, Murray AJ, Pollack RA, Soliman MG, Laskey SB, Capoferri AA, Lai J, Strain MC, Lada SM, Hoh R, Ho YC, Richman DD, Deeks SG, Siliciano JD, Siliciano RF. Defective proviruses rapidly accumulate during acute HIV-1 infection. Nature Medicine. 2016;22(9):1043-9. doi: 10.1038/nm.4156. PubMed PMID: 27500724; PMCID: 5014606.
8. Hiener B, Horsburgh BA, Eden JS, Barton K, Schlub TE, Lee E, von Stockenstrom S, Odevall L, Milush JM, Liegler T, Sinclair E, Hoh R, Boritz EA, Douek D, Fromentin R, Chomont N, Deeks SG, Hecht FM, Palmer S. Identification of Genetically Intact HIV-1 Proviruses in Specific CD4(+) T Cells from Effectively Treated Participants. Cell Reports. 2017;21(3):813-22. doi: 10.1016/j.celrep.2017.09.081. PubMed PMID: 29045846; PMCID: 5960642.
9. Wagner TA, McLaughlin S, Garg K, Cheung CY, Larsen BB, Styrchak S, Huang HC, Edlefsen PT, Mullins JI, Frenkel LM. HIV latency. Proliferation of cells with HIV integrated into cancer genes contributes to persistent infection. Science. 2014;345(6196):570-3. doi: 10.1126/science.1256304. PubMed PMID: 25011556; PMCID: 4230336.
10. Lee GQ, Orlova-Fink N, Einkauf K, Chowdhury FZ, Sun X, Harrington S, Kuo HH, Hua S, Chen HR, Ouyang Z, Reddy K, Dong K, Ndung’u T, Walker BD, Rosenberg ES, Yu XG, Lichterfeld M. Clonal expansion of genome-intact HIV-1 in functionally polarized Th1 CD4+ T cells. The Journal of Clinical Investigation. 2017;127(7):2689-96. doi: 10.1172/JCI93289. PubMed PMID: 28628034; PMCID: 5490740.
11. Einkauf KB, Lee GQ, Gao C, Sharaf R, Sun X, Hua S, Chen SM, Jiang C, Lian X, Chowdhury FZ, Rosenberg ES, Chun TW, Li JZ, Yu XG, Lichterfeld M. Intact HIV-1 proviruses accumulate at distinct chromosomal positions during prolonged antiretroviral therapy. The Journal of Clinical Investigation. 2019;129(3):988-98. doi: 10.1172/JCI124291. PubMed PMID: 30688658; PMCID: 6391088.
12. Cole B, Lambrechts L, Gantner P, Noppe Y, Bonine N, Witkowski W, Chen L, Palmer S, Mullins JI, Chomont N, Pardons M, Vandekerckhove L. In-depth single-cell analysis of translation-competent HIV-1 reservoirs identifies cellular sources of plasma viremia. Nat Commun. 2021;12(1):3727. doi: 10.1038/s41467-021-24080-1. PubMed PMID: 34140517; PMCID: PMC8211655.
13. Siliciano JD, Kajdas J, Finzi D, Quinn TC, Chadwick K, Margolick JB, Kovacs C, Gange SJ, Siliciano RF. Long-term follow-up studies confirm the stability of the latent reservoir for HIV-1 in resting CD4+ T cells. Nature Medicine. 2003;9(6):727-8. doi: 10.1038/nm880. PubMed PMID: 12754504.
14. Baxter AE, Niessl J, Fromentin R, Richard J, Porichis F, Charlebois R, Massanella M, Brassard N, Alsahafi N, Delgado GG, Routy JP, Walker BD, Finzi A, Chomont N, Kaufmann DE. Single-Cell Characterization of Viral Translation-Competent Reservoirs in HIV-Infected Individuals. Cell Host Microbe. 2016;20(3):368-80. doi: 10.1016/j.chom.2016.07.015. PubMed PMID: 27545045; PMCID: PMC5025389.
15. Yukl SA, Kaiser P, Kim P, Telwatte S, Joshi SK, Vu M, Lampiris H, Wong JK. HIV latency in isolated patient CD4(+) T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing. Sci Transl Med. 2018;10(430). doi: 10.1126/scitranslmed.aap9927. PubMed PMID: 29491188; PMCID: PMC5959841.
16. Sannier G, Dube M, Dufour C, Richard C, Brassard N, Delgado GG, Pagliuzza A, Baxter AE, Niessl J, Brunet-Ratnasingham E, Charlebois R, Routy B, Routy JP, Fromentin R, Chomont N, Kaufmann DE. Combined single-cell transcriptional, translational, and genomic profiling reveals HIV-1 reservoir diversity. Cell Reports. 2021;36(9):109643. doi: 10.1016/j.celrep.2021.109643. PubMed PMID: 34469719.
17. Whitney JB, Hill AL, Sanisetty S, Penaloza-MacMaster P, Liu J, Shetty M, Parenteau L, Cabral C, Shields J, Blackmore S, Smith JY, Brinkman AL, Peter LE, Mathew SI, Smith KM, Borducchi EN, Rosenbloom DI, Lewis MG, Hattersley J, Li B, Hesselgesser J, Geleziunas R, Robb ML, Kim JH, Michael NL, Barouch DH. Rapid seeding of the viral reservoir prior to SIV viraemia in rhesus monkeys. Nature. 2014;512(7512):74-7. doi: 10.1038/nature13594. PubMed PMID: 25042999; PMCID: PMC4126858.
18. Colby DJ, Trautmann L, Pinyakorn S, Leyre L, Pagliuzza A, Kroon E, Rolland M, Takata H, Buranapraditkun S, Intasan J, Chomchey N, Muir R, Haddad EK, Tovanabutra S, Ubolyam S, Bolton DL, Fullmer BA, Gorelick RJ, Fox L, Crowell TA, Trichavaroj R, O’Connell R, Chomont N, Kim JH, Michael NL, Robb ML, Phanuphak N, Ananworanich J, group RVs. Rapid HIV RNA rebound after antiretroviral treatment interruption in persons durably suppressed in Fiebig I acute HIV infection. Nature Medicine. 2018;24(7):923-6. doi: 10.1038/s41591-018-0026-6. PubMed PMID: 29892063; PMCID: PMC6092240.
19. Wei X, Ghosh SK, Taylor ME, Johnson VA, Emini EA, Deutsch P, Lifson JD, Bonhoeffer S, Nowak MA, Hahn BH, et al. Viral dynamics in human immunodeficiency virus type 1 infection. Nature. 1995;373(6510):117-22. doi: 10.1038/373117a0. PubMed PMID: 7529365.
20. Perelson AS, Neumann AU, Markowitz M, Leonard JM, Ho DD. HIV-1 dynamics in vivo: virion clearance rate, infected cell life-span, and viral generation time. Science. 1996;271(5255):1582-6. doi: 10.1126/science.271.5255.1582. PubMed PMID: 8599114.
21. Gantner P, Buranapraditkun S, Pagliuzza A, Dufour C, Pardons M, Mitchell JL, Kroon E, Sacdalan C, Tulmethakaan N, Pinyakorn S, Robb ML, Phanuphak N, Ananworanich J, Hsu D, Vasan S, Trautmann L, Fromentin R, Chomont N. HIV rapidly targets a diverse pool of CD4(+) T cells to establish productive and latent infections. Immunity. 2023. doi: 10.1016/j.immuni.2023.01.030. PubMed PMID: 36804957.
22. Pantaleo G, Cohen OJ, Schacker T, Vaccarezza M, Graziosi C, Rizzardi GP, Kahn J, Fox CH, Schnittman SM, Schwartz DH, Corey L, Fauci AS. Evolutionary pattern of human immunodeficiency virus (HIV) replication and distribution in lymph nodes following primary infection: implications for antiviral therapy. Nature Medicine. 1998;4(3):341-5. doi: 10.1038/nm0398-341. PubMed PMID: 9500610.
23. Pantaleo G, Graziosi C, Demarest JF, Butini L, Montroni M, Fox CH, Orenstein JM, Kotler DP, Fauci AS. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease. Nature. 1993;362(6418):355-8. doi: 10.1038/362355a0. PubMed PMID: 8455722.
24. Fox CH, Tenner-Racz K, Racz P, Firpo A, Pizzo PA, Fauci AS. Lymphoid germinal centers are reservoirs of human immunodeficiency virus type 1 RNA. J Infect Dis. 1991;164(6):1051-7. doi: 10.1093/infdis/164.6.1051. PubMed PMID: 1955708.
25. Xu H, Wang X, Malam N, Lackner AA, Veazey RS. Persistent Simian Immunodeficiency Virus Infection Causes Ultimate Depletion of Follicular Th Cells in AIDS. J Immunol. 2015;195(9):4351-7. doi: 10.4049/jimmunol.1501273. PubMed PMID: 26408660; PMCID: PMC4610871.
26. Buranapraditkun S, Pissani F, Teigler JE, Schultz BT, Alter G, Marovich M, Robb ML, Eller MA, Martin J, Deeks S, Michael NL, Streeck H. Preservation of Peripheral T Follicular Helper Cell Function in HIV Controllers. J Virol. 2017;91(14). doi: 10.1128/JVI.00497-17. PubMed PMID: 28468877; PMCID: PMC5487582.
27. Locci M, Havenar-Daughton C, Landais E, Wu J, Kroenke MA, Arlehamn CL, Su LF, Cubas R, Davis MM, Sette A, Haddad EK, International AVIPCPI, Poignard P, Crotty S. Human circulating PD-1+CXCR3-CXCR5+ memory Tfh cells are highly functional and correlate with broadly neutralizing HIV antibody responses. Immunity. 2013;39(4):758-69. doi: 10.1016/j.immuni.2013.08.031. PubMed PMID: 24035365; PMCID: PMC3996844.
28. Boswell KL, Paris R, Boritz E, Ambrozak D, Yamamoto T, Darko S, Wloka K, Wheatley A, Narpala S, McDermott A, Roederer M, Haubrich R, Connors M, Ake J, Douek DC, Kim J, Petrovas C, Koup RA. Loss of circulating CD4 T cells with B cell helper function during chronic HIV infection. PLoS Pathog. 2014;10(1):e1003853. doi: 10.1371/journal.ppat.1003853. PubMed PMID: 24497824; PMCID: PMC3911819.
29. Perreau M, Savoye AL, De Crignis E, Corpataux JM, Cubas R, Haddad EK, De Leval L, Graziosi C, Pantaleo G. Follicular helper T cells serve as the major CD4 T cell compartment for HIV-1 infection, replication, and production. J Exp Med. 2013;210(1):143-56. doi: 10.1084/jem.20121932. PubMed PMID: 23254284; PMCID: PMC3549706.
30. Fenwick C, Joo V, Jacquier P, Noto A, Banga R, Perreau M, Pantaleo G. T-cell exhaustion in HIV infection. Immunol Rev. 2019;292(1):149-63. doi: 10.1111/imr.12823. PubMed PMID: 31883174; PMCID: PMC7003858.
31. Cockerham LR, Jain V, Sinclair E, Glidden DV, Hartogenesis W, Hatano H, Hunt PW, Martin JN, Pilcher CD, Sekaly R, McCune JM, Hecht FM, Deeks SG. Programmed death-1 expression on CD4(+) and CD8(+) T cells in treated and untreated HIV disease. AIDS. 2014;28(12):1749-58. doi: 10.1097/QAD.0000000000000314. PubMed PMID: 24871455; PMCID: PMC4206412.
32. Chew GM, Fujita T, Webb GM, Burwitz BJ, Wu HL, Reed JS, Hammond KB, Clayton KL, Ishii N, Abdel-Mohsen M, Liegler T, Mitchell BI, Hecht FM, Ostrowski M, Shikuma CM, Hansen SG, Maurer M, Korman AJ, Deeks SG, Sacha JB, Ndhlovu LC. TIGIT Marks Exhausted T Cells, Correlates with Disease Progression, and Serves as a Target for Immune Restoration in HIV and SIV Infection. PLoS Pathog. 2016;12(1):e1005349. doi: 10.1371/journal.ppat.1005349. PubMed PMID: 26741490; PMCID: PMC4704737.
33. Sun L, Yang K, Zhang L, Qi LM, Chen JM, Li P, Xiao J, Zhao HX, Wang P. Programmed death-1 expression and regulatory T cells increase in the Intestinal mucosa of cytomegalovirus colitis in patients with HIV/AIDS. AIDS Res Ther. 2020;17(1):54. doi: 10.1186/s12981-020-00315-x. PubMed PMID: 32891157; PMCID: PMC7487894.
34. Iannetta M, Savinelli S, Rossi R, Mascia C, Marocco R, Vita S, Zuccala P, Zingaropoli MA, Mengoni F, Massetti AP, Falciano M, d’Ettorre G, Ciardi MR, Mastroianni CM, Vullo V, Lichtner M. Myeloid and lymphoid activation markers in AIDS and non-AIDS presenters. Immunobiology. 2019;224(2):231-41. doi: 10.1016/j.imbio.2018.11.011. PubMed PMID: 30522891.
35. Bernal-Fernandez G, Hermida C, Espinosa-Cueto P, Cubilla-Tejeda AC, Salazar-Gonzalez JF, Ortiz-Ortiz L, Leyva-Meza R, Diaz-Silvestre H, Mancilla R. Impact of opportunistic Mycobacterium tuberculosis infection on the phenotype of peripheral blood T cells of AIDS patients. J Clin Lab Anal. 2006;20(3):80-6. doi: 10.1002/jcla.20105. PubMed PMID: 16721821; PMCID: PMC6807506.
36. Castagna A, Monno L, Carta S, Galli L, Carrara S, Fedele V, Punzi G, Fanti I, Caramello P, Lepri AC, De Luca A, Ceccherini-Silberstein F, Monforte AD, Group IFS. Switch of predicted HIV-1 tropism in treated subjects and its association with disease progression. Medicine (Baltimore). 2016;95(44):e5222. doi: 10.1097/MD.0000000000005222. PubMed PMID: 27858869; PMCID: PMC5591117.
37. Shang H, Zhang Z, Jiang Y, Han X, Wang Y, Zhang M, Ye X, Liu Y, Diao Y, Dai D, Geng W. Activation and coreceptor expression of T lymphocytes in HIV/AIDS patients of China. J Clin Immunol. 2005;25(1):68-72. doi: 10.1007/s10875-005-0359-2. PubMed PMID: 15742159.
38. Sallusto F, Lenig D, Forster R, Lipp M, Lanzavecchia A. Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature. 1999;401(6754):708-12. doi: 10.1038/44385. PubMed PMID: 10537110.
39. Marichannegowda MH, Zemil M, Wieczorek L, Sanders-Buell E, Bose M, O’Sullivan AM, King D, Francisco L, Diaz-Mendez F, Setua S, Chomont N, Phanuphak N, Ananworanich J, Hsu D, Vasan S, Michael NL, Eller LA, Tovanabutra S, Tagaya Y, Robb ML, Polonis VR, Song H. Tracking coreceptor switch of the transmitted/founder HIV-1 identifies co-evolution of HIV-1 antigenicity, coreceptor usage and CD4 subset targeting. bioRxiv. 2023. doi: 10.1101/2023.01.21.525033. PubMed PMID: 36712089; PMCID: PMC9882280.
40. Becht E, McInnes L, Healy J, Dutertre CA, Kwok IWH, Ng LG, Ginhoux F, Newell EW. Dimensionality reduction for visualizing single-cell data using UMAP. Nat Biotechnol. 2018. doi: 10.1038/nbt.4314. PubMed PMID: 30531897.
41. Van Gassen S, Callebaut B, Van Helden MJ, Lambrecht BN, Demeester P, Dhaene T, Saeys Y. FlowSOM: Using self-organizing maps for visualization and interpretation of cytometry data. Cytometry A. 2015;87(7):636-45. doi: 10.1002/cyto.a.22625. PubMed PMID: 25573116.
42. Dobrowolski C, Valadkhan S, Graham AC, Shukla M, Ciuffi A, Telenti A, Karn J. Entry of Polarized Effector Cells into Quiescence Forces HIV Latency. mBio. 2019;10(2). doi: 10.1128/mBio.00337-19. PubMed PMID: 30914509; PMCID: PMC6437053.
43. Shan L, Deng K, Gao H, Xing S, Capoferri AA, Durand CM, Rabi SA, Laird GM, Kim M, Hosmane NN, Yang HC, Zhang H, Margolick JB, Li L, Cai W, Ke R, Flavell RA, Siliciano JD, Siliciano RF. Transcriptional Reprogramming during Effector-to-Memory Transition Renders CD4(+) T Cells Permissive for Latent HIV-1 Infection. Immunity. 2017;47(4):766-75 e3. doi: 10.1016/j.immuni.2017.09.014. PubMed PMID: 29045905; PMCID: PMC5948104.
44. Dirk BS, Pawlak EN, Johnson AL, Van Nynatten LR, Jacob RA, Heit B, Dikeakos JD. HIV-1 Nef sequesters MHC-I intracellularly by targeting early stages of endocytosis and recycling. Sci Rep. 2016;6:37021. doi: 10.1038/srep37021. PubMed PMID: 27841315; PMCID: PMC5107982.
45. Lama J, Mangasarian A, Trono D. Cell-surface expression of CD4 reduces HIV-1 infectivity by blocking Env incorporation in a Nef- and Vpu-inhibitable manner. Curr Biol. 1999;9(12):622-31. doi: 10.1016/s0960-9822(99)80284-x. PubMed PMID: 10375528.
46. Carl S, Greenough TC, Krumbiegel M, Greenberg M, Skowronski J, Sullivan JL, Kirchhoff F. Modulation of different human immunodeficiency virus type 1 Nef functions during progression to AIDS. J Virol. 2001;75(8):3657-65. doi: 10.1128/JVI.75.8.3657-3665.2001. PubMed PMID: 11264355; PMCID: PMC114857.
47. Mann JK, Chopera D, Omarjee S, Kuang XT, Le AQ, Anmole G, Danroth R, Mwimanzi P, Reddy T, Carlson J, Radebe M, Goulder PJR, Walker BD, Abdool Karim S, Novitsky V, Williamson C, Brockman MA, Brumme ZL, Ndung’u T. Nef-mediated down-regulation of CD4 and HLA class I in HIV-1 subtype C infection: association with disease progression and influence of immune pressure. Virology. 2014;468-470:214-25. doi: 10.1016/j.virol.2014.08.009. PubMed PMID: 25193656; PMCID: PMC4252354.
48. Ross TM, Oran AE, Cullen BR. Inhibition of HIV-1 progeny virion release by cell-surface CD4 is relieved by expression of the viral Nef protein. Curr Biol. 1999;9(12):613-21. doi: 10.1016/s0960-9822(99)80283-8. PubMed PMID: 10375525.
49. Schnittman SM, Lane HC, Greenhouse J, Justement JS, Baseler M, Fauci AS. Preferential infection of CD4+ memory T cells by human immunodeficiency virus type 1: evidence for a role in the selective T-cell functional defects observed in infected individuals. Proc Natl Acad Sci U S A. 1990;87(16):6058-62. doi: 10.1073/pnas.87.16.6058. PubMed PMID: 2385584; PMCID: PMC54471.
50. Cohen Stuart JW, Hazebergh MD, Hamann D, Otto SA, Borleffs JC, Miedema F, Boucher CA, de Boer RJ. The dominant source of CD4+ and CD8+ T-cell activation in HIV infection is antigenic stimulation. J Acquir Immune Defic Syndr. 2000;25(3):203-11. doi: 10.1097/00126334-200011010-00001. PubMed PMID: 11115950.
51. Khoury G, Rajasuriar R, Cameron PU, Lewin SR. The role of naive T-cells in HIV-1 pathogenesis: an emerging key player. Clin Immunol. 2011;141(3):253-67. doi: 10.1016/j.clim.2011.09.002. PubMed PMID: 21996455.
52. Wu VH, Nobles CL, Kuri-Cervantes L, McCormick K, Everett JK, Nguyen S, Del Rio Estrada PM, Gonzalez-Navarro M, Torres-Ruiz F, Avila-Rios S, Reyes-Teran G, Bushman FD, Betts MR. Assessment of HIV-1 integration in tissues and subsets across infection stages. JCI Insight. 2020;5(20). doi: 10.1172/jci.insight.139783. PubMed PMID: 32970634; PMCID: PMC7605534.
53. Harper J, Gordon S, Chan CN, Wang H, Lindemuth E, Galardi C, Falcinelli SD, Raines SLM, Read JL, Nguyen K, McGary CS, Nekorchuk M, Busman-Sahay K, Schawalder J, King C, Pino M, Micci L, Cervasi B, Jean S, Sanderson A, Johns B, Koblansky AA, Amrine-Madsen H, Lifson J, Margolis DM, Silvestri G, Bar KJ, Favre D, Estes JD, Paiardini M. CTLA-4 and PD-1 dual blockade induces SIV reactivation without control of rebound after antiretroviral therapy interruption. Nature Medicine. 2020;26(4):519-28. doi: 10.1038/s41591-020-0782-y. PubMed PMID: 32284611; PMCID: PMC7790171.