- Open Access
Dynamic analysis of CD127 expression on memory CD8 T cells from patients with chronic hepatitis B during telbivudine treatment
- Guocai Lv†1,
- Linjung Ying†1,
- Wen-Jiang Ma1,
- Xi Jin1,
- Lin Zheng1,
- Lanjuan Li1 and
- Yida Yang1Email author
© Lv et al; licensee BioMed Central Ltd. 2010
- Received: 21 May 2010
- Accepted: 31 August 2010
- Published: 31 August 2010
Accumulating evidence supports the theory that expression of CD127 on CD8 T cells during the process of antiviral immune response indicates a subset of effect CD8 T cells that successfully develop into fully protective memory. CD8 T cells expression of CD127 may be used as a predictor to evaluate disease status in chronic viral infection. The aim of this study was to investigate the CD127 expression level on different subsets of CD8 T cell and explore the relationship between CD127 expression on CD8 memory T cells and serum hepatitis B virus (HBV) DNA and hepatitis B e antigen (HBeAg) levels in patients with chronic hepatitis B (CHB). We also aimed to investigate the CD127 expression pattern on CD8 memory T cells of CHB patients who were treated with Telbivudine.
Twenty HBeAg-positive CHB patients were selected and treated with telbivudine 600 mg/day for 48 weeks. The memory CD8 T cells were characterized by expression of CD45RA and CD27 markers. CD127 expression on the CD8 T-cell surface was measured by four-colour flow cytometry. Our results showed that CD127 expression on memory CD8 T cells was reduced in CHB patients. There was a strong negative correlation between CD127 expression on memory CD8 T cells and serum HBV DNA and HBeAg levels in CHB patients. Moreover, successful antiviral therapy increased CD127 expression on CD8 memory T cells as well as on HBV-specific CD8 T cells in CHB patients.
These results suggest that diminished CD127 expression on CD8 memory T cells of CHB patients is a potential mechanism explaining cellular immune function impairment in CHB infection, and that CD127 expression on CD8 memory T cells is a useful indicator for evaluating the effects of anti-HBV therapy.
- CD127 Expression
- HBeAg Level
- Telbivudine Treatment
- Successful Antiviral Therapy
Chronic hepatitis B virus (HBV) infection remains a serious global health problem. It affects approximately 350 million people worldwide and more than 130 million in Chian. It is widely accepted that the adaptive immune responses, particularly the cellular immune response, mediate clearance of HBV. Unfortunately, in most patients, chronic HBV infection leads to severe abnormalities of CD8 T-cell function, as shown by a low level of antiviral cytokines and impaired cytotoxic T-lymphocyte (CTL) activity .
Naive CD8 T cells that encounter their cognate antigen undergo a complex process of maturation and differentiation that ultimately leads to the generation of long-lived memory CD8 T cells, which mediate immune production from subsequent challenge with the same antigen . Memory CD8 T cells are characterized by their abilities to survive homeostatically in the absence of antigen and proliferate vigorously upon antigenic re-encounter. Memory CD8 T cells are easily activated upon antigen rechallenge, in which situation they quickly produce antiviral cytokines or cytotoxic molecular [4, 5].
Interleukin (IL)-7 signalling is essential to CD8 T-cell proliferation and function. The IL-7 receptor (IL-7R), a heterodimer, is composed of a unique α chain (CD127) and a common γ chain (CD132) . During viral infection, CD127 expression on CD8 T cells occurs only when the antigen load is contained and sufficient CD4 T-cell help is available . Persistent viral antigen load suppresses CD127 expression on primed T cells and correlates with exhaustion of a previously stable primed T-cell population . Studies on patients with acute HBV infection showed that CD127 expression on HBV-specific CD8 T cells increased markedly after viral clearance .
In the present study, we demonstrated that CD127 expression on CD8 memory T cells was reduced in patients with chronic hepatitis B (CHB). There was a strong negative correlation between CD127 expression on CD8 memory T cells and serum HBV DNA and hepatitis B e antigen (HBeAg) levels in these patients. Moreover, successful antiviral therapy increased CD127 expression on CD8 memory T cells as well as on HBV-specific CD8+ T cells in patients with CHB. These results suggest that CD127 expression is a potential indicator for evaluating the effects of anti-HBV therapy.
This study was approved by the Ethics Review Committee of the First Affiliated Hospital, School of Medicine, Zhejiang University (Hangzhou, Zhejiang, China). The diagnosis of CHB was made according to the diagnostic standards from the National Program for Prevention and Treatment of Viral Hepatitis. A total of 20 HLA-A2+ patients with CHB (8 women and 12 men; mean age 27 years) were enrolled in the study. Human leucocyte antigen (HLA) typing was performed using polymerase chain reaction (PCR) amplification with sequence-specific primers, and it was confirmed by flow cytometry.
Hepatitis B surface antigen (HBsAg), HBeAg, anti-HBc, anti-HBe and anti-HBs were quantified by radioimmunoassay (Abbott Laboratories, Abbott Park, IL, USA). HBV DNA was measured using the Amplicor HBV test (Roche Diagnostics, Basel, Switzerland) with a detection limit of 300 copies/mL. All patients were HBeAg positive and had never received anti-HBV therapy before.
At baseline, the average serum HBV DNA of the 20 patients was 7.8 ± 0.09 log10 copies/mL [median: (7.9 5.0-9.8) log10 copies/mL], and the serum alanine aminotransferase (ALT) was 174.6 ± 7.78 IU/L [median: 113 (99-567) IU/L]. All patients received telbivudine 600 mg/day for 48 weeks. The serum ALT level, HBsAg, HBeAg, anti-HBc, anti-HBe, anti-HBs and HBV DNA were tested every 12 weeks during the telbivudine therapy. Healthy donors (n = 10) were included as controls.
Peripheral blood mononuclear cells (PBMC) were isolated from ethylenediaminetetraacetic acid (EDTA) anticoagulated blood samples on a Ficoll-Histopaque density gradient. After isolation, cells were washed twice in phosphate-buffered saline (PBS) and studied immediately. CD127, CD8, CD27 and CD45RA expression on the PBMC was detected by direct staining.
CD127 expression on HBV-specific CD8+ T cells was performed as described previously . Briefly, PBMC were stained with surface PC5-anti-CD8 (BD Pharmingen, San Diego, CA, USA) and pentamer+ CD8 T cells were detected by staining with phycoerythrin (PE)-labelled pentameric peptide-HLA2 complex (ProImmune, Oxford, UK) containing HBV Core 18-27 (FLPSDFFPSV) and HBV Core 18-27 (FLPSDFFPSI). Gated on CD8 T cells, CD127 expression on HBV-specific CD8 T cells was analyzed by fluorescein isothiocynate (FITC)-anti-CD127 and PE-labelled pentamers. Cells were washed three times with PBS, and 1 × 106 events in the lymphogate were collected by flow cytometry (EPICSXL; Coulter, Fullerton, CA, USA). Data were analyzed using CellQuest software (Coulter).
The Wilcoxon matched pairs test and the Mann-Whitney test of SPSS version 12.0 were used to assess differences among groups. Spearman correlation analysis was performed between CD127 expression and serum HBV DNA and HBeAg levels. P-values less than 0.05 were considered statistically significant.
CD127 expression on memory CD8 T cells was reduced in patients with chronic hepatitis B
Relationship between CD127 expression on memory CD8 T cells and serum HBV DNA and HBeAg levels in patients with chronic hepatitis B
Antiviral therapy increased CD127 expression on CD8 memory T cells in patients with chronic hepatitis B
Emergence of CD127 HBV-specific CD8+ T cells after successful antiviral treatment
In our study, we demonstrated that CD127 expression on memory CD8 T cells was reduced in patients with CHB. There was a strong negative correlation between CD127 expression on memory CD8 T cells and serum HBV DNA and HBeAg levels in CHB patients. Moreover, successful antiviral therapy with telbivudine increased CD127 expression on CD8 memory T cells as well as on HBV-specific CD8 T cells in CHB patients.
It is widely accepted that CD8 T cells play an essential role in the immune response to viral infection. In successful responses to acute HBV, hepatitis C virus (HCV) and lymphocytic choriomeningitis virus (LCMV) infection, the up-regulation of CD127 expression on CD8 T cells is closely associated with the downregulation of CD38 and PD-1 and the upregulation of CCR7 expression [9, 11, 12]. All occurred in concert with resolution of disease and containment of viral antigen, supporting the theory that the emergence of CD127 is governed by withdrawal of antigenic stimulation. Colle et al.  reported that human immunodeficiency virus (HIV) infection was associated with a decrease in the proportion of CD127+ cells among memory CD8 T lymphocytes, which resulted in a higher CD127- CD8 T cells count in patients with HIV infection. There was a strong negative correlation between CD127 expression on CD8 T cells and HIV viral load . All of these results support the hypothesis that high CD127 expression on human CD8 T cells is specific for cleared virus [e.g. influenza virus, respiratory syncytial virus (RSV) and acute HBV infection] while low CD127 expression on human CD8 T cells is specific for persisting virus [e.g. HIV, cytomegalovirus (CMV), HCV and HBV] .
Recently some reports have suggested that CD127 might be a useful marker for predicting response to antiviral therapy in HIV- and HCV-infected patients. Badr et al.  reported that during HCV infection, early therapeutic intervention with pegylated (PEG)-interferon (IFN)-α rescued long-lived, polyfunctional memory CD8 T cells expressing high levels of CD127 and Bcl-2 (CD127hiBclhi). In contrast, HCV-specific CD8 T cells in acute infections evolving to chronicity expressed low levels of CD127 and Bcl-2, exhibited diminished proliferation and cytokine production, and eventfully disappeared from the periphery. Colle et al.  also reported that CD127 was increased in memory CD8 T lymphocytes from HAART patients. Our longitudinal study indicated that successful antiviral therapy with telbivudine increased CD127 expression on CD8 memory T cells as well as on HBV-specific CD8 T cells in CHB patients. These consistent results clearly suggest that measurement of CD127 expression might be useful for predicting response to antiviral therapy.
In chronic HBV-infected patients, the frequency and function of circulating and intrahepatic antiviral T-cell responses is inversely proportional to the level of HBV DNA [16, 17]. Nucleoside analogues are known to interfere with viral replication, directly lowering HBV DNA levels, but whether they influence the development of effective memory T-cell differentiation and function has not been proven. Our findings indicate that treatment-induced suppression of HBV replication resulted in upregulation of CD127 expression on memory CD8 T cells in all well responders to telbivudine, but not in non-responders. These comparison results obtained in the responders and non-responders to antiviral therapy support the notion that increased expression of CD127 on memory CD8 T cells is linked to successful inhibition of viraemia. These results indicate measurement of CD127 expression on memory CD8 T cells may be useful to guide antiviral therapy in patients with CHB. However, longitudinal studies are required to draw a clear conclusion on this matter.
Taken together, our results suggest the mechanism linking HBV replication and abnormalities in CD8 T-cell function in patients with CHB. We also demonstrate a strong negative correlation between HBV viraemia and CD127 expression in memory CD8 T cells. Telbivudine-induced inhibition of HBV replication resulted in significant upregulation of CD127 expression in memory CD8 T cells, reducing its negative influence on CD8 T cells' activation and function in CHB patients. Most important, we demonstrate successful antiviral treatment can rescue such a functional signature on memory CD8 T cells, which will indicate to achieve sustained inhibition of HBV replication and resolution of chronic liver disease .
This work was supported by grant no. R20090018 from the China National S&T Major Project to Y. D. Yang, grant no. 2008C23073 from the Department of Science and Technology of Zhejiang Province, China to Y. D. Yang, grant no. 2009C33009 from the Department of Science and Technology of Zhejiang Province, China to L. Zheng and grant no. Y200708441 from Health Department of Zhejiang Province, China to W. J. Ma.
- Lai CL, Ratzin V, Yuen MF, Poynard T: Viral hepatitis B. Lancet 2003, 362: 2089-2094. 10.1016/S0140-6736(03)15108-2PubMedView ArticleGoogle Scholar
- Bertoletti A, Maini M, Williams R: Role of hepatitis B virus specific cytotoxic T cells in liver damage and viral control. Antiviral Res 2003,60(2):61-66. 10.1016/j.antiviral.2003.08.012PubMedView ArticleGoogle Scholar
- Seder RA, Ahmed R: Similarities and differences in CD4 and CD8 effector and memory T cell generation. Nat Immunol 2003, 4: 835-842. 10.1038/ni969PubMedView ArticleGoogle Scholar
- Sallusto F, Geginat J, Lanzavecchia A: Central memory and effector memory T cell subsets: function, generation, and maintenance. Annu Rev Immunol 2004, 22: 745-763. 10.1146/annurev.immunol.22.012703.104702PubMedView ArticleGoogle Scholar
- Kaech SM, Wherry EJ: Heterogeneity and cell-fate decisions in effector and memory CD8 T cell differentiation during viral infection. Immunity 2007,27(3):393-405. 10.1016/j.immuni.2007.08.007PubMedPubMed CentralView ArticleGoogle Scholar
- MacPherson PA, Fex C, Sanchez-Dardon J, Hawley-Foss N, Angel JB: Interleukin-7 receptor expression on CD8 T cells is reduced in HIV infection and partially restored with effective antiretroviral therapy. J Acquir Immune Defic Syndr 2001,28(5):454-457.PubMedView ArticleGoogle Scholar
- Fuller MJ, Hildeman DA, Sabbaj S, et al.: Cutting edge: emergence of CD127 high functionally competent memory T cells is compromised by high viral loads and inadequate T cell help. J Immunol 2005,174(10):5926-5930.PubMedView ArticleGoogle Scholar
- Paiardini M, Cervasi B, Albrecht H, et al.: Loss of CD127 expression defines an expansion of effector CD8 T cells in HIV-infected individuals. J Immunol 2005,174(5):2900-2909.PubMedView ArticleGoogle Scholar
- Boettler T, Panther E, Bengsch B, et al.: Expression of the interleukin-7 receptor alpha chain (CD127) on virus-specific CD8 T cells identifies functionally and phenotypically defined memory T cells during acute resolving hepatitis B virus infection. J Virol 2006,80(7):3532-3540. 10.1128/JVI.80.7.3532-3540.2006PubMedPubMed CentralView ArticleGoogle Scholar
- Wherry EJ, Day CL, Draenert R, et al.: HIV-specific CD8 T cells express low levels of IL-7Ralpha: implications for HIV-specific T cell memory. Virology 2006,353(2):366-373. 10.1016/j.virol.2006.06.017PubMedView ArticleGoogle Scholar
- Badr G, Bédard N, Abdel-Hakeem MS, et al.: Early interferon therapy for hepatitis C virus infection rescues polyfunctional, long-lived CD8 memory T cells. J Virol 2008,82(20):10 017-10 031. 10.1128/JVI.01083-08View ArticleGoogle Scholar
- Kaech SM, Tan JT, Wherry EJ, Konieczny BT, Surh CD, Ahmed R: Selective expression of the interleukin 7 receptor identifies effector CD8 T cells that give rise to long-lived memory cells. Nat Immunol 2003,4(12):1191-11988. 10.1038/ni1009PubMedView ArticleGoogle Scholar
- Colle JH, Moreau JL, Fontanet A, et al.: CD127 expression and regulation are altered in the memory CD8 T cells of HIV-infected patients - reversal by highly active anti-retroviral therapy (HAART). Clin Exp Immunol 2006,143(3):398-403. 10.1111/j.1365-2249.2006.03022.xPubMedPubMed CentralView ArticleGoogle Scholar
- Sharma TS, Hughes J, Murillo A, et al.: CD8 T-cell interleukin-7 receptor alpha expression as a potential indicator of disease status in HIV-infected children. PLoS One 2008,3(12):e3986. 10.1371/journal.pone.0003986PubMedPubMed CentralView ArticleGoogle Scholar
- van Leeuwen EM, de Bree GJ, Remmerswaal EB, et al.: IL-7 receptor alpha chain expression distinguishes functional subsets of virus-specific human CD8 T cells. Blood 2005,106(6):2091-2098. 10.1182/blood-2005-02-0449PubMedView ArticleGoogle Scholar
- Sobao Y, Tomiyama H, Sugi K, et al.: The role of hepatitis B virus-specific memory CD8 T cells in the control of viral replication. J Hepatol 2002,36(1):105-115. 10.1016/S0168-8278(01)00264-1PubMedView ArticleGoogle Scholar
- Webster GJ, Reignat S, Brown D, et al.: Longitudinal analysis of CD8+ T cells specific for structural and nonstructural hepatitis B virus proteins in patients with chronic hepatitis B: implications for immunotherapy. J Virol 2004,78(11):5707-5719. 10.1128/JVI.78.11.5707-5719.2004PubMedPubMed CentralView ArticleGoogle Scholar
- Evans A, Riva A, Cooksley H, et al.: Programmed death 1 expression during antiviral treatment of chronic hepatitis B: Impact of hepatitis B e-antigen seroconversion. Hepatology 2008,48(3):759-769. 10.1002/hep.22419PubMedView ArticleGoogle Scholar
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