Memory CD4+ T lymphocytes in peripheral blood that express integrins α4ß7 preferentially recirculate through gut-associated lymphoid tissue (GALT), a proposed site of significant HIV-1 replication. Tregs and activated CD4+ T cells in GALT could also be particularly susceptible to infection. We therefore hypothesized that infection of these subsets of memory CD4+ T cells may contribute disproportionately to the HIV-1 reservoir. A cross-sectional study of CD4+ T cell subsets of memory CD45RO+ cells in peripheral blood mononuclear cells (PBMCs) was conducted using leukapheresis from eight subjects with untreated chronic HIV-1 infection. Real-time polymerase chain reaction (PCR) was used to quantify total and integrated HIV-1 DNA levels from memory CD4+ T cells sorted into integrin β7+ vs. β7−, CD25+CD127low Treg vs. CD127high, and activated CD38+ vs. CD38−. More than 80% of total HIV-1 DNA was found to reside in the integrin β7-negative non-gut-homing subset of CD45RO+ memory CD4+ T cells. Less than 10% was found in highly purified Tregs or CD38+ activated memory cells. Similarly, integrated HIV-1 DNA copies were found to be more abundant in resting non-gut-homing memory CD4+ T cells (76%) than in their activated counterparts (23%). Our investigations showed that the majority of both total and integrated HIV-1 DNA was found within non-gut-homing resting CD4+ T cells.
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References
1.
BlanksonJN, PersaudD, SilicianoRF. The challenge of viral reservoirs in HIV-1 infection. Annu Rev Med, 2002; 53:557–593.
2.
GeeraertL, KrausG, PomerantzRJ. Hide-and-seek: The challenge of viral persistence in HIV-1 infection. Annu Rev Med, 2008; 59:487–501.
3.
DaveyRTJr, BhatN, YoderCet al.HIV-1 and T cell dynamics after interruption of highly active antiretroviral therapy (HAART) in patients with a history of sustained viral suppression. Proc Natl Acad Sci USA, 1999; 96,26:15109–15114.
4.
JoosB, FischerM, KusterHet al.HIV rebounds from latently infected cells, rather than from continuing low-level replication. Proc Natl Acad Sci USA, 2008; 105,43:16725–16730.
5.
AgostoLM, YuJJ, DaiJet al.HIV-1 integrates into resting CD4+ T cells even at low inoculums as demonstrated with an improved assay for HIV-1 integration. Virology, 2007; 368:60–72.
6.
CameronPU, SalehS, SallmannGet al.Establishment of HIV-1 latency in resting CD4+ T cells depends on chemokine-induced changes in the actin cytoskeleton. Proc Natl Acad Sci USA, 2010; 107,39:16934–16939.
7.
ChunTW, FinziD, MargolickJet al.In vivo fate of HIV-1-infected T cells: Quantitative analysis of the transition to stable latency. Nat Med, 1995; 1,12:1284–1290.
8.
ChunTW, EngelD, BerreyMMet al.Early establishment of a pool of latently infected, resting CD4(+) T cells during primary HIV-1 infection. Proc Natl Acad Sci USA, 1998; 95,15:8869–8873.
9.
ZaundersJJ, CunninghamPH, KelleherADet al.Potent antiretroviral therapy of primary human immunodeficiency virus type 1 (HIV-1) infection: Partial normalization of T lymphocyte subsets and limited reduction of HIV-1 DNA despite clearance of plasma viremia. J Infect Dis, 1999; 180,2:320–329.
10.
KoelschKK, BoeseckeC, McBrideKet al.Impact of treatment with raltegravir during primary or chronic HIV infection on RNA decay characteristics and the HIV viral reservoir. AIDS, 2011; 25,17:2069–2078.
11.
StrainMC, LittleSJ, DaarESet al.Effect of treatment, during primary infection, on establishment and clearance of cellular reservoirs of HIV-1. J Infect Dis, 2005; 191:1414–1418.
12.
HaaseAT. Perils at mucosal front lines for HIV and SIV and their hosts. Nat Rev Immunol, 2005; 5,10:783–792.
13.
BrenchleyJM, SchackerTW, RuffLEet al.CD4+ T cell depletion during all stages of HIV disease occurs predominantly in the gastrointestinal tract. J Exp Med, 2004; 200,6:749–759.
14.
MehandruS, PolesMA, Tenner-RaczKet al.Mechanisms of gastrointestinal CD4+ T-cell depletion during acute and early human immunodeficiency virus type 1 infection. J Virol, 2007; 81,2:599–612.
15.
BrenchleyJM, PriceDAet al.HIV disease: Fallout from a mucosal catastrophe?Nat Immunol, 2006; 7:235–239.
16.
ReddAD, GrayRH, QuinnTC. Is microbial translocation a cause or consequence of HIV disease progression?J Infect Dis, 2011; 203,5:744–745.
17.
ChunT-W, NickleDC, JustementJSet al.Persistence of HIV in gut-associated lymphoid tissue despite long-term antiretroviral therapy. J Infect Dis, 2008; 197:714–720.
18.
YuklSA, ShergillAK, McQuaidKet al.Effect of raltegravir-containing intensification on HIV burden and T-cell activation in multiple gut sites of HIV-positive adults on suppressive antiretroviral therapy. AIDS, 2010; 24,16:2451–2460.
19.
SchweighofferT, YoshiyaT, TidswellMet al.Selective expression of integrin α4β7 on a subset of human CD4+ memory T cells with hallmarks of gut-tropism. J Immunol, 1993; 151,2:717–729.
20.
von AndrianUH, MackayCR. T-cell function and migration. Two sides of the same coin. N Engl J Med, 2000; 343,14:1020–1034.
21.
ChanBM, ElicesMJ, MurphyEet al.Adhesion to vascular cell adhesion molecule 1 and fibronectin. Comparison of alpha 4 beta 1 (VLA-4) and alpha 4 beta 7 on the human B cell line JY. J Biol Chem, 1992; 267,12:8366–8370.
22.
HolzmannB, WeissmanIL. Peyer's patch-specific lymphocyte homing receptors consist of a VLA-4-like α chain associated with either of two integrin β chains, one of which is novel. EMBO J, 1989; 8,6:1735–1741.
23.
RueggC, PostigoA, SikorskiEet al.Role of integrin alpha 4 beta 7/alpha 4 beta P in lymphocyte adherence to fibronectin and VCAM-1 and in homotypic cell clustering. J Cell Biol, 1992; 117,1:179–189.
24.
FarstadIN, HalstensenTS, KvaleDet al.Topographic distribution of homing receptors on B and T cells in human gut-associated lymphoid tissue: Relation of L-selectin and integrin α4β7 to naive and memory phenotypes. Am J Pathol, 1997; 150,1:187–199.
25.
MonteiroP, GosselinA, WaclecheVSet al.Memory CCR6+CD4+ T cells are preferential targets for productive HIV type 1 infection regardless of their expression of integrin β7. J Immunol, 2011; 186,8:4618–4630.
26.
ErleDJ, BriskinMJ, ButcherECet al.Expression and function of the MAdCAM-1 receptor, integrin alpha 4 beta 7, on human leukocytes. J Immunol, 1994; 153,2:517–528.
27.
BerlinC, BergEL, BriskinMJet al.α4β7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell, 1993; 74,1:185–195.
28.
BensonMJ, Pino-LagosK, RosemblattMet al.All-trans retinoic acid mediates enhanced Treg cell growth, differentiation, and gut homing in the face of high levels of co-stimulation. J Exp Med, 2007; 204,8:1765–1774.
29.
SinghB, ReadS, AssemanCet al.Control of intestinal inflammation by regulatory T cells. Immunol Rev, 2001; 182:190–200.
30.
AnderssonJ, BoassoA, NilssonJet al.The prevalence of regulatory T cells in lymphoid tissue is correlated with viral load in HIV-infected patients. J Immunol, 2005; 174,6:3143–3147.
31.
EppleHJ, LoddenkemperC, KunkelDet al.Mucosal but not peripheral FOXP3+ regulatory T cells are highly increased in untreated HIV infection and normalize after suppressive HAART. Blood, 2006; 108,9:3072–3078.
32.
Moreno-FernandezME, ZapataW, BlackardJTet al.Human regulatory T cells are targets for human immunodeficiency virus (HIV) infection, and their susceptibility differs depending on the HIV type 1 strain. J Virol, 2009; 83,24:12925–12933.
33.
SeddikiN, SassonSC, Santner-NananBet al.Proliferation of weakly suppressive regulatory CD4+ T cells is associated with over-active CD4+ T-cell responses in HIV-positive patients with mycobacterial immune restoration disease. Eur J Immunol, 2009; 39,2:391–403.
34.
ChenX, OppenheimJJ, Winkler-PickettRTet al.Glucocorticoid amplifies IL-2-dependent expansion of functional FoxP3+CD4+CD25+ T regulatory cells in vivo and enhances their capacity to suppress EAE. Eur J Immunol, 2006; 36,8:2139–2149.
35.
DouekDC, BettsMR, HillBJet al.Evidence for increased T cell turnover and decreased thymic output in HIV infection. J Immunol, 2001; 167,11:6663–6668.
36.
SeddikiN, Santner-NananB, TangyeSGet al.Persistence of naive CD45RA+ regulatory T cells in adult life. Blood, 2006; 107,7:2830–2838.
37.
AandahlEM, MichaëlssonJ, MorettoWJet al.Human CD4+CD25+ regulatory T cells control T-cell responses to human immunodeficiency virus and cytomegalovirus antigens. J Virol, 2004; 78,5:2454–2459.
38.
AnderssonJ, BoassoA, NilssonJet al.Cutting edge: The prevalence of regulatory T cells in lymphoid tissues is correlated with viral load in HIV-infected patients. J Immunol, 2005; 174,6:3143–3147.
39.
KinterAL, HennesseyM, BellAet al.CD25+CD4+ regulatory T cells from the peripheral blood of asymptomatic HIV-infected individuals regulate CD4+ and CD8+ HIV-specific T cell immune responses in vitro and are associated with favorable clinical markers of disease status. J Exp Med, 2004; 200,3:331–343.
40.
SakaguchiS. Regulatory T cells: Mediating compromises between host and parasite. Nat Immunol, 2003; 4:10–11.
41.
WeissL, Donkova-PetriniV, CaccavelliLet al.Human immunodeficiency virus-driven expansion of CD4+CD25+ regulatory T cells, which suppress HIV-specific CD4 T-cell responses in HIV-infected patients. Blood, 2004; 104,10:3249–3256.
42.
NilssonJ, BoassoA, VelillaPAet al.HIV-1-driven regulatory T-cell accumulation in lymphoid tissues is associated with disease progression in HIV/AIDS. Blood, 2006; 108,12:3808–3817.
43.
SeddikiN, Santner-NananB, MartinsonJet al.Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J Exp Med, 2006; 203,7:1693–1700.
44.
TrickettA, DwyerJ, TedlaNet al.Safety and feasibility of harvesting cells for adoptive immunotherapy from patients with asymptomatic HIV infection. J Acquir Immune Defic Syndr Hum Retrovirol, 1996; 12,5:523–524.
45.
ZaundersJJ, IpS, MunierMLet al.Infection of CD127+ (interleukin-7 receptor+) CD4+ cells and overexpression of CTLA-4 are linked to loss of antigen-specific CD4 T cells during primary human immunodeficiency virus type 1 infection. J Virol, 2006; 80,20:10162–10172.
46.
MartinA, SmithDE, CarrAet al.Reversibility of lipoatrophy in HIV-infected patients 2 years after switching from a thymidine analogue to abacavir: The MITOX Extension Study. AIDS, 2004; 18,7:1029–1036.
47.
SuzukiK, ShijuukuT, FukamachiTet al.Prolonged transcriptional silencing and CpG methylation induced by siRNAs targeted to the HIV-1 promoter region. J RNAi Gene Silencing, 2005; 1,2:66–78.
48.
BrenchleyJM, HillBJ, AmbrozakDRet al.T-cell subsets that harbor human immunodeficiency virus (HIV) in vivo: Implications for HIV pathogenesis. J Virol, 2004; 78:1160–1168.
49.
ChomontN, El-FarM, AncutaPet al.HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. Nat Med, 2009; 15:893–900.
50.
ZaundersJ, CarrA, McNallyLet al.Effects of primary HIV-1 infection on subsets of CD4+ and CD8+ T lymphocytes. AIDS, 1995; 9:561–566.
51.
ChunTW, NickleDC, JustementJSet al.HIV-infected individuals receiving effective antiviral therapy for extended periods of time continually replenish their viral reservoir. J Clin Invest, 2005; 115,11:3250–3255.
52.
MeditzAL, HaasMK, FolkvordJMet al.HLA-DR+CD38+CD4+ T lymphocytes have elevated CCR5 expression and produce the majority of R5-tropic HIV-1 RNA in vivo. J Virol, 2011; 85,19:10189–10200.
53.
LiuZ, CumberlandWG, HultinLEet al.Elevated CD38 antigen expression on CD8+ T cells is a stronger marker for the risk of chronic HIV disease progression to AIDS and death in the Multicenter AIDS Cohort Study than CD4+ cell count, soluble immune activation markers, or combinations of HLA-DR and CD38 expression. J Acquir Immune Defic Syndr Hum Retrovirol, 1997; 16,2:83–92.
54.
MehandruS, DandekarS. Role of the gastrointestinal tract in establishing infection in primates and humans. Curr Opin HIV AIDS, 2008; 3,1:22–27.
55.
KlonowskiKD, WilliamsKJ, MarzoALet al.Dynamics of blood-borne CD8 memory T cell migration in vivo. Immunity, 2004; 20,5:551–562.
56.
LiQ, DuanL, EstesJDet al.Peak SIV replication in resting memory CD4+ T cells depletes gut lamina propria CD4+ T cells. Nature, 2005; 434,7037:1148–1152.
57.
NishimuraY, SadjadpourR, MattapallilJJet al.High frequencies of resting CD4+ T cells containing integrated viral DNA are found in rhesus macaques during acute lentivirus infections. Proc Natl Acad Sci USA, 2009; 106,19:8015–8020.
58.
MaenetjeP, RiouC, CasazzaJPet al.A steady state of CD4+ T cell memory maturation and activation is established during primary subtype C HIV-1 infection. J Immunol, 2010; 184,9:4926–4935.
59.
BrenchleyJM, KarandikarNJ, BettsMRet al.Expression of CD57 defines replicative senescence and antigen-induced apoptotic death of CD8+ T cells. Blood, 2003; 101,7:2711–2720.
60.
BucyRP, HockettRD, DerdeynCAet al.Initial increase in blood CD4(+) lymphocytes after HIV antiretroviral therapy reflects redistribution from lymphoid tissues. J Clin Invest, 1999; 103,10:1391–1398.
61.
ArthosJ, CicalaC, MartinelliEet al.HIV-1 envelope protein binds to and signals through integrin alpha4beta7, the gut mucosal homing receptor for peripheral T cells. Nat Immunol, 2008; 9,3:301–309.
62.
ParrishNF, WilenCB, BanksLBet al.Transmitted/founder and chronic subtype C HIV-1 use CD4 and CCR5 receptors with equal efficiency and are not inhibited by blocking the integrin α4β7. PLoS Pathog, 2012; 8,5:e1002686.
63.
YuklSA, GianellaS, SinclairEet al.Differences in HIV burden and immune activation within the gut of HIV-positive patients receiving suppressive antiretroviral therapy. J Infect Dis, 2010; 202,10:1553–1561.
64.
TedlaN, DwyerJ, TruskettPet al.Phenotypic and functional characterization of lymphocytes derived from normal and HIV-1-infected human lymph nodes. Clin Exp Immunol, 1999; 117,1:92–99.
65.
PantaleoG, GraziosiC, ButiniLet al.Lymphoid organs function as major reservoirs for human immunodeficiency virus. Proc Natl Acad Sci USA, 1991; 88,21:9838–9842.
66.
SeddikiN, KelleherAD. Regulatory T cells in HIV infection: Who's suppressing what?Curr Infect Dis Rep, 2008; 10,3:252–258.
67.
LernerP, GuadalupeM, DonovanRet al.The gut mucosal viral reservoir in HIV-infected patients is not the major source of rebound plasma viremia following interruption of highly active antiretroviral therapy. J Virol, 2011; 85,10:4772–4782.
68.
GanusovVV, De BoerRJ. Do most lymphocytes in humans really reside in the gut?Trends Immunol, 2007; 28,12:514–518.
69.
MacalM, SankaranS, ChunT-Wet al.Effective CD4+ T-cell restoration in gut-associated lymphoid tissue of HIV-infected patients is associated with enhanced Th17 cells and polyfunctional HIV-specific T-cell responses. Mucosal Immunol, 2008; 1,6:475–488.
70.
SchneiderT, JahnH-U, SchmidtWet al.Loss of CD4 T lymphocytes in patients infected with human immunodeficiency virus type 1 is more pronounced in the duodenl mucosa than in the peripheral blood. Gut, 1995; 37:524–529.
71.
AntonPA, MitsuyasuRT, DeeksSGet al.Multiple measures of HIV burden in blood and tissue are correlated with each other but not with clinical parameters in aviremic subjects. AIDS, 2003; 17,1:53–63.
72.
BuzonM, SeissK, LiCet al.T memory stem cells: A long-term reservoir for HIV-1. IDWeek 2012 Meeting. Vol Paper #594, October 17–21, San Diego, California, 2012.
73.
HoY-C, ShanL, WangJet al.Characterization of non-induced HIV-1 proviruses dampens the hope for HIV-1 eradication. CROI—Conference on Retroviruses and Opportunistic Infections. Vol Paper #43Atlanta, Georgia, 2013.
74.
HaaseAT. Population biology of HIV-1 infection: Viral and CD4+ T cell demographics and dynamics in lymphatic tissues. Annu Rev Immunol, 1999; 17:625–656.
75.
McIlroyD, AutranB, CheynierRet al.Infection frequency of dendritic cells and CD4+ T lymphocytes in spleens of human immunodeficiency virus-positive patients. J Virol, 1995; 69,8:4737–4745.
76.
GavinM, RudenskyA. Control of immune homeostasis by naturally arising regulatory CD4+ T cells. Curr Opin Immunol, 2003; 15:690–696.
SassonSC, ZaundersJJ, KelleherAD. The IL-7/IL-7 receptor axis: Understanding its central role in T-cell homeostasis and the challenges facing its utilization as a novel therapy. Curr Drug Targets, 2006; 7,12:1571–1582.
