摘要
目的:探探讨CD4^+CD25^+调节性T细胞(regulatory T Cell,Treg)在慢性乙型肝炎(chronic hepatitis B,CHB)患者免疫发病机制中的作用以及其可能在治疗中的应用前景.方法:收集未经抗病毒治疗的CHB患者34例和健康对照18例外周血单个核细胞(peripheral blood mononuclear cell,PBMC)标本,以三色/四色流式分析法对PBMC中CD4^+CD25^+ Treg的频率及表面分子表达进行分析,并同时通过磁珠分选去除CHB患者PBMC中的CD4^+CD25^+Treg,分别以MHC-肽-五聚体法和酶联斑点计数法(enzyme-linked immunospot assay,Elispot)检测HBV core18-27抗原肽刺激的对HBV特异性的CTL(cytotoxic T lymphocyte)频率的升高以及IFN-γ的分泌.结果:CHB患者外周血中CD4^+CD25^+ CD45RO^+CTLA4^+T细胞群以及CD4^+CD127^(lo)CD25^(hi-int)T细胞群所占CD4^+T细胞群的比例与健康对照相比均明显上升(3.78%±1.87%.4.40%±2.11%vs 1.58%±0.76%,2.11%±1.26%;t=4.86,t=5.96;P<0.01)去除CHB患者中CD4^+CD25^+Treg后,特异性CTL的频率以及其分泌IFN-γ的频数与未去除组比出现显著上调(0.94%±0.38%,26±13 vs 0.20%±0.18%,119±30;t=5.25,t= 9.886;P<0.01).结论:CHB患者循环中增多的Treg可能参与抑制抗HBV的免疫应答抑制,去除Treg以及联合病毒抗原肽刺激的进一步研究可能为CHB的免疫治疗提供新的思路.
AIM： To evaluate the proportion of CD4^＋CD25^＋ regulatory T cells （Tregs） in the immunopathogenesis of patients with chronic hepatitis B （CHB）, and to evaluate their potential application in CHB immunotherapy. METHODS： Peripheral blood mononuclear cells （PBMCs） of 34 patients with CHB and 18 healthy donors were evaluated for the CD4^＋CD25^＋ Treg phenotype and the proportion of CD4^＋CD25^＋ Tregs as a percentage of the total number of CD4^＋ cells using flow cytometry with triple-col- or/fourth-color staining. CD4^＋CD25^＋ Tregs were isolated from CHB patients by magnetic beads. Then PBMCs of Treg-depleted and non-Treg-depleted CHB patients were stimulated with HBV core18-27 peptide. The percentage of HBV core18-27-specific cytotoxic lymphocytes （CTLs） and interferon （IFN）-γ-secreting CTLs was de- termined in PBMCs of Treg-depleted and non- Treg-depleted CHB patients by HLA-pentamer and Elispot （enzyme-linked immunospot assay） analysis. RESULTS： The percentage of CD4＋CD25＋ Tregs in CHB patients was reflected by detecting the proportion of CD4＋CD12^loCD25^hi-int Tregs （4.40%± 2.11%） and CD4＋CD25＋CD45RO＋CTLA4＋ Tregs （3.78% ±1.87%）, which were significantly higher than the proportion of CD4＋CD12^loCD25^hi-int Tregs （2.11% ± 1.26%） and CD4＋CD25＋CD45RO＋CTLA4＋ Tregs （1.58 % ± 0.76%） in healthy donors （P 〈 0.01）. The frequen- cy and IFN-γ-secreting spots of NY-ESO-1- specific CD8＋ T cells was higher in CD4＋CD25＋- depleted PBMCs from CHB patients than in the total PBMC population of all patients tested （0.94% ＋ 0.38%, 26 ±13 vs 0.20% ± 0.18%, 119＋30, t =5.25, t = 9.886, P 〈 0.01）. CONCLUSION： The proportion of CD4^＋CD25^＋ Tregs in CHB patients seems to be greater than that in healthy donors. The elimination of Tregs can significantly improve the anti-viral CTL responses in CHB patients, compared with stimulation with HBV core18-27 peptide alone. This may contribute to immunotherapy in CHB patients.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第20期2225-2230,共6页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.30640091
北京大学人民医院研究发展基金
No.RDB2006-01
关键词
调节性T细胞
乙型肝炎
慢性
Regulatory T-Lymphocytes
Hepatitis B
Chronic
作者简介
张恒辉，2005年北京大学内科学硕士研究生毕业．研究实习员主要研究方向肝炎及肝癌的免疫治疗
通讯作者：陈红松．100044．北京市西直门南大街11号．北京大学人民医院肝病研究所．chen2999@sohu.com电诂：010-88325726传真：010--68318386
