摘要
目的探讨艾滋病(AIDS)患者高效抗逆转录病毒治疗(HAART)前后T淋巴细胞表面某些归巢分子CD49d、CCR9、CD62L和CD4+T细胞表面辅助受体CCR5的表达变化情况及其意义。
方法采用流式细胞术检测42例艾滋病患者和18例HIV阴性正常健康对照的外周血单核细胞(PBMC),分别观察CD3+、CD4+、CD8+T细胞表面CD49d、CD62L、CCR9和CD4+T细胞上CCR5的表达,并对CD4+CCR9+和CD4+CCR5+ T细胞进一步进行CD45RO表型分析,采用BD FACSDiva软件分析计算各组细胞表达的百分率,分析上述细胞群在治疗前后的变化及意义。
结果AIDS患者未治疗组外周血CD3+CD49d+、CD3+CCR9+、CD3+CD62L+、CD4+CD49d+、CD4+CCR9+、CD4+CD62L+、CD8+CD49d+、CD8+CD62L+、CD4+CCR9+CD45RO+、CD4+CCR5+CD45RO+较治疗组明显降低;CD4+CCR9+CD45RO-、CD4+CCR5+、CD4+CCR5+CD45RO-明显升高,差异均有统计学意义。AIDS患者未治疗组外周血CD3+CD49d+、CD3+CCR9+、CD3+CD62L+、CD3+CD4+、CD4+CD49d+、CD4+CCR9+、CD4+CD62L+、CD4+CCR9+CD45RO+、CD4+CCR5+CD45RO+较对照组明显下降;CD4+CCR9+CD45RO-、CD4+CCR5+、CD4+CCR5+CD45RO-较对照明显升高,差异均有统计学意义。AIDS患者治疗组CD3+CD4+、CD3+CCR9+、CD4+CD62L+、CD8+CCR9+、CD8+CD62L+较对照组降低;CD3+CD8+、CD4+CCR5+较对照组升高,差异均有统计学意义。
结论AIDS患者外周血T淋巴细胞表面肠道归巢分子CD49d、CCR9,淋巴结归巢分子CD62L,辅助受体CCR5出现异常改变;HAART可以逆转以上部分免疫病理改变;肠道归巢分子CD49d、CCR9和淋巴结归巢分子CD62L可建议作为评价艾滋病疾病进展和HAART后机体免疫重建情况的指标之一。本课题受到广东省科技厅科技计划项目(2006B36030016)资助。
Objective To evaluate the expression of CD49d, CCR9, CD62L on T lymphocytes and co-receptor CCR5 on CD4+ T lymphocytes in AIDS patients before and after HAART,and furthermore to explore its clinical significance.
Methods The study was performed in 42 cases of AIDS patients and 18 cases of healthy controls. The expression of CD49d, CCR9, CD62L on CD3+, CD4+ , CD8+T lymphocytes and CCR5 on CD4+ T lymphocytes, and CD45RO on CD4+CCCR9+ and CD4+CCR5+ T lymphocytes in AIDS patients and healthy controls were analysed by Flow cytometry. Software BD FACSDiva was used to calculate the percentage of expression,then study the changes and significance between untreated group and therapy group .
Results Compared with therapy group , the frequency of CD3+CD49d+, CD3+CCR9+,CD3+CD62L+, CD4+CD49d+, CD4+CCR9+,CD4+CD62L+, CD8+CD49d+, CD8+CD62L+, CD4+CCR9+CD45RO+,CD4+CCR5+CD45RO+ T lymphocytes in untreated group were significantly decreased, while the frequency of CD4+CCR9+CD45RO - , CD4 + CCR5+, CD4+CCR5+CD45RO - T lymphocytes significantly increased; compared with controls, the frequency of CD3+CD49d+, CD3+CCR9+, CD3+CD62L+, CD3+CD4+ , CD4+CD49d+, CD4+CCR9+, CD4+CD62L+ , CD4+CCR9+CD45RO+, CD4+CCR5+CD45RO+ T lymphocytes in untreated group were significantly decreased, while the frequency of CD4+CCR9+CD45RO-, CD4+CCR5+, CD4+CCR5+CD45RO-T lymphocytes significantly increased; compared with controls, the frequency of CD3+CD4+,CD3+CCR9+, CD4+CD62L+, CD8+CCR9+, CD8+CD62L+ T cells in therapy group were significantly decreased,while the frequency of CD3+CD8+,CD4+CCR5+ T lymphocytes significantly increased.
Conclusion The expression of gut homing receptor molecule CD49d, CCR9 and lymph node homing molecule CD62L, co-receptor molecule CCR5 in AIDS patients on T lymphocytes was impaired; HAART can partially reverse this immunologic pathological phenomena; CD49d, CCR9 and CD62L may be suggested to indicate the progression of AIDS and immunologic reconstitution after HAART.
引文
[1] Picker LT,Watkins DI. HIV pathogenesis: the first cut is the deepest[J].Nat Immunol,2005,6(5):430-432.
[2] Von Andrian UH, Mackay CR. T-cell function and migration: two sides of the same coin[J]. N Engl J Med,2000,343(14): 1020-1034.
[3] Raul Elgueta, Fernando E Sepulveda, Felipe Vilches, et al. Imprinting of CCR9 on CD4 T Cells Requires IL-4 Signaling on Mesenteric Lymph Node Dendritic Cells[J]. The Journal of Immunology[J].2008,180(10):6501-6507.
[4] Svensson, Marcus Marsal, Jan Ericsson , et al. CCL25 mediates the localization of recently activated CD8 alphabeta(+)lymphocytes to the small-intestinal mucosa[J]. J Clin Invest, 2002,110(8):1113-1121.
[5] Quentin Sattentau . HIV’s gut feeling[J]. Nat immunol, 2008 , 9(3):225-226.
[6] Berlin C, Beg E J, Briskin M J, et al.α4β7 integrin mediates lymphocyte binding to themucosal vascular addressin MAdCAM-1[J]. Cell, 1993,74(5):185-195.
[7] Graham KL, Franco MA, Angel J, et al. Rotaviruses interact with alpha4beta7 and alpha4beta1 integrins by binding the same integrin domains as natural ligands[J].J Gen Virol,2005,86 (12) 3397-408.
[8] Arthos J,Cicala C, Martinelli E, et al. HIV-1 envelope protein binds to and signals through integrin alpha4beta7, the gut mucosal homing receptor for peripheral T cells[J]. Nat Immunol.2008,9(3):301-309.
[9] Smit-McBride Z, Mattapallil- JJ, McCheseney M, et al. Gastrointestinal T lymphocyts retain high potential for cytokine responses but have severe CD4(+)T-cell depletion at all stages of simian immunodefiency virus infection compared to peripheral lymphocytes[J]. J Virol,1998,72(8):6646-6656.
[10] Rosen SD. Ligands for L-selectin: homing, inflammation,and beyond. [J]. Annu Rev Immunol, 2004,(22):129–156.
[11] Bobardt MD, Saphire AC, Hung, HC, et a1. Syndecan captures, protects,and transmits HIV to T lymphocytes[J]. Immunity,2003,18(1):27–39.
[12] Bastiani L, Laal S, Kim M, et a1. Host cell dependent alterations in envelopecomponents of human immunodeficiency virus type 1 virions[J]. J Virol,1997, 71 (12): 3444–3450.
[13] Rosenberg ES, Altfeld M, Poon SH, et al. Immune control of HIV-1 after early treatment of acute infection[J]. Nature, 2000,407(6803):523–526.
[14] Damle NK, Aruffo A. Vascular cell adhesion molecule 1 induces T-cell antigen receptor-dependent activation of CD4+T lymphocytes[J].Proc Natl Acad Sci,1991,88(15), 6403-7.
[15] Youn BS, Kim YG, Mantel C, et a1. Blockingofc-FIIP(L)-independent cycloheximide-induced apoptosis by the CC chemokine receptor 9/TECK interaction[J].Blood,2001,98(4):925-933.
[16] Eun JP, Mora JR, Carman CV, et al. Aberrant activation of integrin alpha 4beta 7 (integrinα4β7) suppresses lymphocyte migration to the gut[J]. Clin Invest ,2007,117(9):2526–2538.
[17] Kanse SM, Matz RL, Preissner KT, et al. Promotion of leukocyte adhesion by a novel interaction between vitronectin and the beta2 integrin Mac-1 (alphaMbeta2, CD11b/CD18)[J]. Arterioscler Thromb Vasc Bio,2004,24(12):2251-2256.
[18] Carramolino L, Zaballos A, Kremer L, et a1.Expression of CCR9 13-chemokine receptor is modulated in thymo-cyte differentiation and is selectively maintained in CD8 T cell from secondary lymphoid organs[J].Blood, 2001,26 (94):850-857.
[19] Hengel, RL Jones, BM Kennedy, et al.CD4+ T Cells Programmed to Traffic to Lymph Nodes Account for Increases in Numbers of CD4+ T Cells Up to 1 Year after the Initiation of Highly Active Antiretroviral Therapy for Human Immunodeficiency Virus Type 1 Infection[J].JID,2001,184(1):93-97.
[20] Issekutz TB. Inhibition of in vivo lymphocyte migration to inflammation and homing to lymphoid tissues by the TA-2 monoclonal antibody. A likely role for VLA-4 in vivo[J]. J Immunol,1991,147(12), 4178-84 .
[21] Damle NK, Klussman K, Leytze G, et al. Costimulation of T lymphocytes with integrin ligands intercellular adhesion molecule-1 or vascular cell adhesion molecule-1 induces functional expression of CTLA-4, a second receptor for B7[J]. J Immunol,1994,152(6),2686-97.
[22] Bargatze RF, Jutila MA, Butcher EC, et al. Distinct roles of L-selectin and integrinsα4β7 andLFA-1 in lymphocyte homing to Peyer’s patch-HEV in situ: the multistep model confirmed and refined[J].Immunity ,1995,13(8):99-108.
[23] Yang Y, Cardarelli PM, Lehnert K, et al. LPAM-1 (integrinalpha 4 beta 7)-ligand binding: overlapping binding sites recognizing VCAM-1, MAdCAM-1 and CS-1 are blocked by fibrinogen, a fibronectin-likepolymer and RGD-like cyclic peptides[J]. Eur J Immunol,1998,28(3):995-1004.
[24] Plana M, Ferrer E, Martinez C, et a1.Immune restoration in HIV-positive antiretrovira1 naive patients after 1 year of zidovudine/lamivudine plus nelilnavir or nevirapine[J]. Antivir Ther,2004,9(2):197-204.
[25] HamannA, Andrew DP, Jablonski-Westrich D, et al. Role ofα4-integrins in lymphocyte homing to mucosal tissues in vivo [J].J Immunol,1994,152(6):3282-3293.
[26] Shimizu YV, Seventer GA, Horgan KJ, et al. Costimulation of proliferative responses of resting CD4+ T cells by the interaction of VLA-4 and VLA-5 with fibronectin or VLA-6 with laminin[J]. J Immunol,1990,145(1), 59-67.
[27] ZimmermanC, Brduscha-Riem K, Blaser C,et al.Visualization, characterization, and turnover of CD8_ memory T cells in virusinfected hosts[J]. J Exp Med,1996, 183(123): 1367–1375.
[28] Papagno L, Spina CA, Marchant A, et al. Immune activation and CD8_ T-cell differentiation towards senescence in HIV-1 infection[J]. PLoS Biol,2004,(13)120-125.
[29] Champagne P, Ogg GS, King KS, et al. Skewed maturation of memory HIV-specific CD8 T lymphocytes[J]. Nature,2001,410(6824):106–111.
[30] Caruso M . Mutation in the VP1-LDV Motif of the Murine Polyomavirus Affects Viral Infectivity and Conditions Virus Tissue Tropism in vivo[J].J Mol Biol,2007,367(13) 54-64.
[31] Oxenius AD, Price A, Easterbrook PJ, et al. Early highly active antiretroviral therapy for acute HIV-1 infection preserves immune function of CD8_ and CD4_ T lymphocytes[J]. Proc Natl Acad,Sci,2000,97(12):3382–3387.
[32] De Martinis M, Modesti M, Profeta VF, et a1.CD50 and CD62L adhesion receptor expression on naive (CD45RA+) and memory(CD45RO+)T lymphocytes in the elderly[J].Pathobiology,2000,68(6):245-250.
[33] Chen BJ, Cui XS, Empowski GD, et a1. Transfer of allogeneic CD62L-memory T cells without graft-versus-host disease[J].Blood,2004,103(4):1534-1541.
[34] Amend B, Doster H, Lange C, et a1. Induction of autoimmunity by expansion of autoreactive CD4+CD62L low cells in vivo [J]. J Immunol,2006, 177(19): 4384-4390.
[35] Savage ND, Harris SH, Rossi AG, et a1.Inhibition of TCR-mediated shedding of L-selectin(CD62L)on human and mouse CD4 T cells by metalloproteinase inhibition:an alysis of the regulation of Th1/Th2 function[J].Eur J Immunol, 2002,32(10):2905-2914.
[36] Paul ME, Shearer WT, Kozinetz CA .Comparision of CD8(+) T-cells subsets in HIV-infected rapid progressor children versus non-rapid progressor children[J].J Allergy Clin Immunol,2001,108(2):258-64.
[37] Kwong PD, Wyatt R, Robinson J, et al. Structure of an HIV gp120 envelope glycoprotein in complex with the CD4 receptor and a neutralizing human antibody[J].Nature,1998, 393(14), 648-59.
[38] Thibault, Sandra R Tardif, Melanie R Gilbert, et al. Virus-associated host CD62L increases attachment of human immunodeficiency virus type 1 to endothelial cells and enhances trans infection of CD4+ T lymphocytes[J].Journal of General Virology 2007, 88(9):2568-73.
[1] Picker LT,Watkins DI. HIV pathogenesis:the first cut is the deepest[J].Nat Immunology,2005,6(5):430-432.
[2] Veazey RS, DeMaria M, Chalifoux LV, et al. Gastrointestinal tract as a major site of CD4+T cell depletion and viral replication in SIV infection[J]. cience,1998, 80(5362):427-431.
[3] Smit-McBride Z, Mattapallil-JJ, McCheseney M, et al.Gastrointestinal T lymphocytes retain high potential for cytokine responses but have severe CD4(+)T-cell depletion at all stages of simian immunodefiency virus infection compared to peripheral lymphocytes[J].J Virol,1998,72(8):6646-6656.
[4] Picker LJ, Hagen SI, Lum R, et al.Insufficient production and tissue delivery of CD4+ memory T cells in rapidly progressive simian immunodefiency virus infection[J].J Exp Med,2004,200(10):1299-1314.
[5] Brenchley JM,Schacker TW,Ruff LE,et al.CD4+T cell depletion during all stages of HIVdisease occurs predominantly in the gastrointestinal tract[J]. J Exp Med,2004,200(6):749-759.
[6] Mehandru S, Poles MA, Tenner-Racz K, et al. Primary HIV-1 infection is associated with preferential depletion of CD4+T lymphocytes. from effector sites in the gastrointestinal tract[J]. J Exp Med,2004,200(6):761-770.
[7] Mowat AM. Anatomal basis of tolerance and immunity to intestinal antigens[J].Nat Rev Immunol,2003,3(4):331-341.
[8] Johansson-Lindbom B, Svensson M, Wurbel MA, et al. Selective generation of gut tropic T cells in gut-associated lymphoid tissue (GALT):requiremengt for GALT dendritic cells and adjuvant[J]. J Exp Med ,2003,198(6):963-969.
[9] Dogan A, Du m, Koulis A, et al. Expression of lymphocyte homing recepters and vascular addressions in low grade gastric B-cell lymphomas of mucosa-associated lymphoidtissue[J].J AmJ Pathol,1997,151(5):1361-1397.
[10] Johansson-Lindbom B, Agace WW. Vitamin A helps gut T cells find their way in the dark[J].Nat Med,2004,10(12):1300-1301.
[11] Marcus Svensson, Jan Marsal, Heli Uronen-Hansson, et al. Involvement of CCR9 at multiple stages of adult T lymphopoiesis[J].Journal of Leukocyte Biology, 008,83(1):156-164.
[12] Jesus, Rivera N, Johnson H, et al. Antibody Blockade of CCL25/CCR9 Ameliorates Early but not Late Chronic Murine Iieitis[J]. astroenterolory 2006, 131(5):1518-1529.
[13] Butcher EC, Picker LJ. lymphocyte homing and homeostasis [J].Science, 1996, 72(5258):60-66.
[14] Sankaran S, George M, Giorgos A, et al. HIV-1 infected Long-term Non-progressors Display Normal CD4+T cell levels and Reduced Immune Activation and Inflammation within the Intestinal Mucosa[C].13thConference on Retrovirus and Opportunistic Infection,2006,Abstract number 288.
[15] Grossman Z, Meizer-Schellersheim M, Paul WE, et al. Pathogenesis of HIV infection :what the virus apsre is as important as what it destroys[J].Nat Med,2006,12(3):289-295.
[16] Arthos J, Cicala C, Martinelli E, et al. HIV-1 envelope protein binds to and signals through integrin alpha4beta7, the gut mucosal homing receptor for peripheral T cells[J]. Nat Immunol,2008,9(3):301-309.
[17] Quentin Sattentau . HIV’s gut feeling[J].Nat Imu,2008 , 9(3):225-226.
[18] Moraima Guadalupe, Elizabeth Reay, Sumathi Sankaran, et al. Severe CD4_ T-Cell Depletion in Gut Lymphoid Tissue during Primary Human Immunodeficiency Virus Type 1 Infection and Substantial Delay in Restoration following Highly Active Antiretroviral Therapy[J].J Virol, 2003, 77(21) :11708–11717.
[19] Pau AK, Tavel JA. Therapeutic use of interleukin-2 in HIV-infected patients[J]. Curr Opin Pharmaco,2002 ,2(4):433-439.
[20] Levy Y. Cytokine-based modulation of immune resonstitution following highly active antiretroviral therapy:can adjuvant tnterleukin-2 fill the gap? [J].JAntimicrob chemother, 2005,55(5):401-409.
[21] Marchetti G, Meroni L, Molteni C, et al. IL-7/IL-7receptor system regulation following IL-2 immunotherpy in HIV-infected patients[J].Antivir Ther ,2004,9(11):447-452.
[22] Vento S, Cainelli F, Temasgen Z. Interleukin-2 therpy and CD4+Tcells in HIV-1 infection[J].Lancet,2006,367(9505):93-95.
[23] Muthukumar A, Wozniakowaki A, Gauduin MC, et al. Elevated interleukin-7 levels not sufficient to maitain T-cell homeostasis during simian immunodeficiency virus-induced disease progression[J].Blood,2004,103(3): 73-979.
[24] Miller CJ, Li Q, Abel K, et al. Propagation and dissemination of infection vaginal transmission of simian immunodeficiency virus[J].Virol,2005,79(14): 217-9227.
[25] Pialoux G, Hocini H, Pérusat S, et al. Phase I study of a candidate vaccine based on recombinant HIV-1 gp160 (MN/LAI) administered by the mucosal route to HIV-seronegative volunteers: the ANRS VAC14 study[J].Vaccine, 2008,26(21):2657-2666.
[26] Li Z, Zhang M, Zhou C, et al. Novel vaccination protocol with two live mucosal vectors elicits strong cell-mediated immunity in the vagina and protects against vaginal virus challenge[J].Immunol,2008,180(4):2504-2513.
[27] Johnson RP. How HIV guts the immune system[J].N Engl J Med,2008, 58(21):2287-2289.
[28] Matsuzaki K, Tsuzuki Y, Matsunaga H, Iet al. In vivo demonstration of T lymphocyte migration and amelioration of ileitis in intestinal mucosa of SAMP1/Yit mice by the inhibition of MAdCAM-1[J]. Clin Exp Immunol, 2005, 140(1): 22-31.
[29] Teramoto K, Miura S, Tsuzuki Y, et al. Increased lymphocyte trafficking to colonic microvessels is dependent on MAdCAM-1 and C-C chemokine mLARC/CCL20 in DSS-induced mice colitis[J].Clin Exp Immunol, 2005,139(14): 421-428.