Reconstitution of Peripheral T Cells by Tissue-Derived CCR4+ Central Memory Cells Following HIV-1 Antiretroviral Therapy

Main Article Content

Yolanda D. Mahnke
Kipper Fletez-Brant
Irini Sereti
Mario Roederer


Background. Highly active antiretroviral therapy induces clinical benefits to HIV-1 infected individuals, which can be striking in those with progressive disease. Improved survival and decreased incidence of opportunistic infections go hand in hand with a suppression of the plasma viral load, an increase in peripheral CD4+ T-cell counts, as well as a reduction in the activation status of both CD4+ and CD8+ T cells.

Methods. We investigated T-cell dynamics during ART by polychromatic flow cytometry in total as well as in HIV-1-specific CD4+ and CD8+ T cells. We also measured gene expression by single cell transcriptomics to assess functional state.

Results. The cytokine pattern of HIV-specific CD8+ T cells was not altered after ART, though their magnitude decreased significantly as the plasma viral load was suppressed to undetectable levels. Importantly, while CD4+ T cell numbers increased substantially during the first year, the population did not normalize: the increases were largely due to expansion of mucosal-derived CCR4+ CD4+ TCM; transcriptomic analysis revealed that these are not classical Th2-type cells.

Conclusion. The apparent long-term normalization of CD4+ T-cell numbers following ART does not comprise a normal balance of functionally distinct cells, but results in a dramatic Th2 shift of the reconstituting immune system.


Download data is not yet available.

Article Details

How to Cite
Mahnke YD, Fletez-Brant K, Sereti I, Roederer M. Reconstitution of Peripheral T Cells by Tissue-Derived CCR4+ Central Memory Cells Following HIV-1 Antiretroviral Therapy. PAI [Internet]. 2016 Oct. 13 [cited 2023 Sep. 25];1(2):260-9. Available from:


1. Autran B, Carcelain G, Li TS, Blanc C, Mathez D, Tubiana R, Katlama C, Debre P, Leibowitch J. Positive effects of combined antiretroviral therapy on CD4+ T cell homeostasis and function in advanced HIV disease. Science. 1997;277(5322):112-6. PubMed PMID: 9204894.

2. Guihot A, Bourgarit A, Carcelain G, Autran B. Immune reconstitution after a decade of combined antiretroviral therapies for human immunodeficiency virus. Trends Immunol. 2011;32(3):131-7. PubMed PMID: 21317040. doi: 10.1016/

3. Pakker NG, Notermans DW, de Boer RJ, Roos MT, de Wolf F, Hill A, Leonard JM, Danner SA, Miedema F, Schellekens PT. Biphasic kinetics of peripheral blood T cells after triple combination therapy in HIV-1 infection: a composite of redistribution and proliferation. Nat Med. 1998;4(2):208-14. PubMed PMID: 9461195.

4. Bucy RP, Hockett RD, Derdeyn CA, Saag MS, Squires K, Sillers M, Mitsuyasu RT, Kilby JM. Initial increase in blood CD4(+) lymphocytes after HIV antiretroviral therapy reflects redistribution from lymphoid tissues. J Clin Invest. 1999;103(10):1391-8. PubMed PMID: 10330421. Pubmed Central PMCID: 408455. doi: 10.1172/JCI5863

5. Steffens CM, Smith KY, Landay A, Shott S, Truckenbrod A, Russert M, Al-Harthi L. T cell receptor excision circle (TREC) content following maximum HIV suppression is equivalent in HIV-infected and HIV-uninfected individuals. AIDS. 2001;15(14):1757-64. PubMed PMID: 11579236.

6. Kovacs JA, Lempicki RA, Sidorov IA, Adelsberger JW, Herpin B, Metcalf JA, Sereti I, Polis MA, Davey RT, Tavel J, Falloon J, Stevens R, Lambert L, Dewar R, Schwartzentruber DJ, Anver MR, Baseler MW, Masur H, Dimitrov DS, Lane HC. Identification of dynamically distinct subpopulations of T lymphocytes that are differentially affected by HIV. J Exp Med. 2001;194(12):1731-41. PubMed PMID: 11748275. Pubmed Central PMCID: 2193579.

7. Mohri H, Perelson AS, Tung K, Ribeiro RM, Ramratnam B, Markowitz M, Kost R, Hurley A, Weinberger L, Cesar D, Hellerstein MK, Ho DD. Increased turnover of T lymphocytes in HIV-1 infection and its reduction by antiretroviral therapy. J Exp Med. 2001;194(9):1277-87. PubMed PMID: 11696593. Pubmed Central PMCID: 2195973.

8. Hazenberg MD, Stuart JW, Otto SA, Borleffs JC, Boucher CA, de Boer RJ, Miedema F, Hamann D. T-cell division in human immunodeficiency virus (HIV)-1 infection is mainly due to immune activation: a longitudinal analysis in patients before and during highly active antiretroviral therapy (HAART). Blood. 2000;95(1):249-55. PubMed PMID: 10607709.

9. Lepej SZ, Begovac J, Vince A. Changes in T-cell subpopulations during four years of suppression of HIV-1 replication in patients with advanced disease. FEMS Immunol Med Microbiol. 2006;46(3):351-9. PubMed PMID: 16553807. 10.1111/j.1574-695X.2005.00034.x

10. Autran B. Effects of antiretroviral therapy on immune reconstitution. Antivir Ther. 1999;4 Suppl 3:3-6. PubMed PMID: 16021864.

11. Barbour JD, Ndhlovu LC, Xuan Tan Q, Ho T, Epling L, Bredt BM, Levy JA, Hecht FM, Sinclair E. High CD8+ T cell activation marks a less differentiated HIV-1 specific CD8+ T cell response that is not altered by suppression of viral replication. PLoS One. 2009;4(2):e4408. PubMed PMID: 19198651. Pubmed Central PMCID: 2634967. 10.1371/journal.pone.0004408

12. Lim A, Tan D, Price P, Kamarulzaman A, Tan HY, James I, French MA. Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high on antiretroviral therapy. AIDS. 2007;21(12):1525-34. PubMed PMID: 17630546. doi: 10.1097/QAD.0b013e32825eab8b

13. Almeida M, Cordero M, Almeida J, Orfao A. Relationship between CD38 expression on peripheral blood T-cells and monocytes, and response to antiretroviral therapy: a one-year longitudinal study of a cohort of chronically infected ART-naive HIV-1+ patients. Cytometry B Clin Cytom. 2007;72(1):22-33. PubMed PMID: 17051525. doi: 10.1002/cyto.b.20144

14. Glencross DK, Janossy G, Coetzee LM, Lawrie D, Scott LE, Sanne I, McIntyre JA, Stevens W. CD8/CD38 activation yields important clinical information of effective antiretroviral therapy: findings from the first year of the CIPRA-SA cohort. Cytometry B Clin Cytom. 2008;74 Suppl 1:S131-40. PubMed PMID: 18228566. doi: 10.1002/cyto.b.20391

15. Casazza JP, Betts MR, Picker LJ, Koup RA. Decay kinetics of human immunodeficiency virus-specific CD8+ T cells in peripheral blood after initiation of highly active antiretroviral therapy. J Virol. 2001;75(14):6508-16. PubMed PMID: 11413318. Pubmed Central PMCID: 114374. doi: 10.1128/JVI.75.14.6508-6516.2001

16. Lopez M, Soriano V, Rallon N, Cascajero A, Gonzalez-Lahoz J, Benito JM. Suppression of viral replication with highly active antiretroviral therapy has no impact on the functional profile of HIV-specific CD8(+) T cells. Eur J Immunol. 2008;38(6):1548-58. PubMed PMID: 18421792. doi: 10.1002/eji.200738054

17. Mahnke YD, Greenwald JH, DerSimonian R, Roby G, Antonelli LR, Sher A, Roederer M, Sereti I. Selective expansion of polyfunctional pathogen-specific CD4(+) T cells in HIV-1-infected patients with immune reconstitution inflammatory syndrome. Blood. 2012;119(13):3105-12. PubMed PMID: 22219223. Pubmed Central PMCID: 3321870. doi: 10.1182/blood-2011-09-380840

18. Antonelli LR, Mahnke Y, Hodge JN, Porter BO, Barber DL, DerSimonian R, Greenwald JH, Roby G, Mican J, Sher A, Roederer M, Sereti I. Elevated frequencies of highly activated CD4+ T cells in HIV+ patients developing immune reconstitution inflammatory syndrome. Blood. 2010;116(19):3818-27. PubMed PMID: 20660788. Pubmed Central PMCID: 2981537. doi: 10.1182/blood-2010-05-285080

19. Dominguez MH, Chattopadhyay PK, Ma S, Lamoreaux L, McDavid A, Finak G, Gottardo R, Koup RA, Roederer M. Highly multiplexed quantitation of gene expression on single cells. J Immunol Methods. 2013;391(1-2):133-45. PubMed PMID: 23500781. doi: 10.1016/j.jim.2013.03.002

20. Roederer M, Nozzi JL, Nason MC. SPICE: exploration and analysis of post-cytometric complex multivariate datasets. Cytometry A. 2011;79(2):167-74. PubMed PMID: 21265010. Pubmed Central PMCID: 3072288. doi: 10.1002/cyto.a.21015

21. Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JY, Zhang J. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 2004;5(10):R80. PubMed PMID: 15461798. Pubmed Central PMCID: 545600. doi: 10.1186/gb-2004-5-10-r80

22. Lin H, Lin C, Weng R. A note on Platt’s probabilistic outputs for support vector machines machine learning. Mach Learn. 2003;68:267-76.

23. Platt J. Probablistic outputs for support vector machines and comparisons to regularized likelihood methods. In: Smola A, Bartlett P, Scholkopf B, Schuurmans D, editors. Advances in Large Margin Classifiers. Cambridge, MA: MIT Press; 1999. p. 61-74.

24. Cortes C, Vapnik V. Support-Vector Networks. Machine Learning. 1995;20:273-97.

25. Meyer D, Dimitriadou E, Hornik K, Weingessel A, Leisch F. e1071: Misc Functions of the Department of Statistics (e1071), TU Wien. R package version 1.6-4. 2014. Available from:

26. Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, Smyth GK. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Research. 2015;43. doi: 10.1093/nar/gkv007.

27. Smyth G, Michaud J, Scott H. The use of within-array replicate spots for assessing differential expression in microarray experiments. Bioinformatics. 2005;21:2067-75.

28. Smyth GK. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol. 2004;3:Article3. PubMed PMID: 16646809. doi: 10.2202/1544-6115.1027

29. Huber P. Robust Statistics: Wiley; 1981.

30. Venables WN, Ripley BD. Modern Applied Statistics with S, Fourth Edition. New York: Springer; 2002.

31. Benjamini Y, Hochnerg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society. 1995;Series B 57 (1):289-300.

32. Mahnke YD, Song K, Sauer MM, Nason MC, Giret MT, Carvalho KI, Costa PR, Roederer M, Kallas EG. Early immunologic and virologic predictors of clinical HIV-1 disease progression. AIDS. 2012. PubMed PMID: 23211771. doi: 10.1097/QAD.0b013e32835ce2e9

33. Liu Z, Hultin LE, Cumberland WG, Hultin P, Schmid I, Matud JL, Detels R, Giorgi JV. Elevated relative fluorescence intensity of CD38 antigen expression on CD8+ T cells is a marker of poor prognosis in HIV infection: results of 6 years of follow-up. Cytometry. 1996;26(1):1-7. PubMed PMID: 8809474. 10.1002/(SICI)1097-0320(19960315)26:1<1::AID-CYTO1>3.0.CO;2-L

34. Bonecchi R, Bianchi G, Bordignon PP, D'Ambrosio D, Lang R, Borsatti A, Sozzani S, Allavena P, Gray PA, Mantovani A, Sinigaglia F. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. J Exp Med. 1998;187(1):129-34. PubMed PMID: 9419219. Pubmed Central PMCID: 2199181.

35. Campbell JD, HayGlass KT. T cell chemokine receptor expression in human Th1- and Th2-associated diseases. Arch Immunol Ther Exp (Warsz). 2000;48(6):451-6. PubMed PMID: 11197598.

36. Rivino L, Messi M, Jarrossay D, Lanzavecchia A, Sallusto F, Geginat J. Chemokine receptor expression identifies Pre-T helper (Th)1, Pre-Th2, and nonpolarized cells among human CD4+ central memory T cells. J Exp Med. 2004;200(6):725-35. PubMed PMID: 15381728. Pubmed Central PMCID: 2211963. doi: 10.1084/jem.20040774

37. Gosselin A, Monteiro P, Chomont N, Diaz-Griffero F, Said EA, Fonseca S, Wacleche V, El-Far M, Boulassel MR, Routy JP, Sekaly RP, Ancuta P. Peripheral blood CCR4+CCR6+ and CXCR3+CCR6+CD4+ T cells are highly permissive to HIV-1 infection. J Immunol. 2010;184(3):1604-16. PubMed PMID: 20042588. doi: 10.4049/jimmunol.0903058

38. Acosta-Rodriguez EV, Rivino L, Geginat J, Jarrossay D, Gattorno M, Lanzavecchia A, Sallusto F, Napolitani G. Surface phenotype and antigenic specificity of human interleukin 17-producing T helper memory cells. Nat Immunol. 2007;8(6):639-46. PubMed PMID: 17486092. doi: 10.1038/ni1467

39. Duhen T, Geiger R, Jarrossay D, Lanzavecchia A, Sallusto F. Production of interleukin 22 but not interleukin 17 by a subset of human skin-homing memory T cells. Nat Immunol. 2009;10(8):857-63. PubMed PMID: 19578369. doi: 10.1038/ni.1767

40. Annunziato F, Galli G, Cosmi L, Romagnani P, Manetti R, Maggi E, Romagnani S. Molecules associated with human Th1 or Th2 cells. Eur Cytokine Netw. 1998;9(3 Suppl):12-6. PubMed PMID: 9831180.

41. Arno A, Ruiz L, Juan M, Zayat MK, Puig T, Balague M, Romeu J, Pujol R, O'Brien WA, Clotet B. Impact on the immune system of undetectable plasma HIV-1 RNA for more than 2 years. AIDS. 1998;12(7):697-704. PubMed PMID: 9619800.

42. Kim CJ, Nazli A, Rojas OL, Chege D, Alidina Z, Huibner S, Mujib S, Benko E, Kovacs C, Shin LY, Grin A, Kandel G, Loutfy M, Ostrowski M, Gommerman JL, Kaushic C, Kaul R. A role for mucosal IL-22 production and Th22 cells in HIV-associated mucosal immunopathogenesis. Mucosal Immunol. 2012;5(6):670-80. PubMed PMID: 22854709. doi: 10.1038/mi.2012.72

43. Rutz S, Eidenschenk C, Ouyang W. IL-22, not simply a Th17 cytokine. Immunol Rev. 2013;252(1):116-32. PubMed PMID: 23405899. doi: 10.1111/imr.12027

44. Eyerich S, Eyerich K, Pennino D, Carbone T, Nasorri F, Pallotta S, Cianfarani F, Odorisio T, Traidl-Hoffmann C, Behrendt H, Durham SR, Schmidt-Weber CB, Cavani A. Th22 cells represent a distinct human T cell subset involved in epidermal immunity and remodeling. J Clin Invest. 2009;119(12):3573-85. PubMed PMID: 19920355. Pubmed Central PMCID: 2786807. doi: 10.1172/JCI40202

45. Becker Y. The changes in the T helper 1 (Th1) and T helper 2 (Th2) cytokine balance during HIV-1 infection are indicative of an allergic response to viral proteins that may be reversed by Th2 cytokine inhibitors and immune response modifiers--a review and hypothesis. Virus Genes. 2004;28(1):5-18. PubMed PMID: 14739648. doi: 10.1023/B:VIRU.0000012260.32578.72

46. Klein SA, Dobmeyer JM, Dobmeyer TS, Pape M, Ottmann OG, Helm EB, Hoelzer D, Rossol R. Demonstration of the Th1 to Th2 cytokine shift during the course of HIV-1 infection using cytoplasmic cytokine detection on single cell level by flow cytometry. AIDS. 1997;11(9):1111-8. PubMed PMID: 9233457.

47. Levy JA. HIV pathogenesis and long-term survival. AIDS. 1993;7(11):1401-10. PubMed PMID: 8280406.

48. Soriano A, Lozano F, Oliva H, Garcia F, Nomdedeu M, De Lazzari E, Rodriguez C, Barrasa A, Lorenzo JI, Del Romero J, Plana M, Miro JM, Gatell JM, Vives J, Gallart T. Polymorphisms in the interleukin-4 receptor alpha chain gene influence susceptibility to HIV-1 infection and its progression to AIDS. Immunogenetics. 2005;57(9):644-54. PubMed PMID: 16189667. doi: 10.1007/s00251-005-0041-x

49. Spellberg B, Edwards JE, Jr. Type 1/Type 2 immunity in infectious diseases. Clin Infect Dis. 2001;32(1):76-102. PubMed PMID: 11118387. doi: 10.1086/317537

50. Chun TW, Nickle DC, Justement JS, Meyers JH, Roby G, Hallahan CW, Kottilil S, Moir S, Mican JM, Mullins JI, Ward DJ, Kovacs JA, Mannon PJ, Fauci AS. Persistence of HIV in gut-associated lymphoid tissue despite long-term antiretroviral therapy. J Infect Dis. 2008;197(5):714-20. PubMed PMID: 18260759. doi: 10.1086/527324

51. Moir S, Chun TW, Fauci AS. Pathogenic mechanisms of HIV disease. Annu Rev Pathol. 2011;6:223-48. PubMed PMID: 21034222. 10.1146/annurev-pathol-011110-130254

52. Cerf-Bensussan N, Jarry A, Brousse N, Lisowska-Grospierre B, Guy-Grand D, Griscelli C. A monoclonal antibody (HML-1) defining a novel membrane molecule present on human intestinal lymphocytes. Eur J Immunol. 1987;17(9):1279-85. PubMed PMID: 3498635. doi: 10.1002/eji.1830170910

53. Rendon JL, Choudhry MA. Th17 cells: critical mediators of host responses to burn injury and sepsis. J Leukoc Biol. 2012;92(3):529-38. PubMed PMID: 22753950. Pubmed Central PMCID: 3427614. doi: 10.1189/jlb.0212083

54. Baekkevold ES, Wurbel MA, Kivisakk P, Wain CM, Power CA, Haraldsen G, Campbell JJ. A role for CCR4 in development of mature circulating cutaneous T helper memory cell populations. J Exp Med. 2005;201(7):1045-51. PubMed PMID: 15795234. Pubmed Central PMCID: PMC2213118. doi: 10.1084/jem.20041059

55. Guadalupe M, Sankaran S, George MD, Reay E, Verhoeven D, Shacklett BL, Flamm J, Wegelin J, Prindiville T, Dandekar S. Viral suppression and immune restoration in the gastrointestinal mucosa of human immunodeficiency virus type 1-infected patients initiating therapy during primary or chronic infection. J Virol. 2006;80(16):8236-47. PubMed PMID: 16873279. Pubmed Central PMCID: 1563811. doi: 10.1128/JVI.00120-06

56. Mehandru S, Poles MA, Tenner-Racz K, Jean-Pierre P, Manuelli V, Lopez P, Shet A, Low A, Mohri H, Boden D, Racz P, Markowitz M. Lack of mucosal immune reconstitution during prolonged treatment of acute and early HIV-1 infection. PLoS Med. 2006;3(12):e484. PubMed PMID: 17147468. Pubmed Central PMCID: 1762085. doi: 10.1371/journal.pmed.0030484

57. Mahnke YD, Beddall MH, Roederer M. OMIP-017: human CD4(+) helper T-cell subsets including follicular helper cells. Cytometry Part A : the journal of the International Society for Analytical Cytology. 2013;83(5):439-40. doi: 10.1002/cyto.a.22269

Most read articles by the same author(s)