CAJ | 학술논문

目的分析人类白细胞抗原(HLA)-A0201限制性的特异性CTL,研究急性肝炎急性期和慢性乙型肝炎活动期患者T淋巴细胞对特异性抗原表位免疫应答的差异.方法收集HLA-A0201阳性的5例急性肝炎急性期和6例慢性乙型肝炎活动期患者的外周血单个核细胞(PBMC),酶联免疫斑点技术(ELISPOT)测定针对HBV聚合酶区(Pol575-583)、包膜区(Env348-357)和核心区(Core18-27)3个CD8+T淋巴细胞表位肽特异性CTL的数量和功能.数据采用t检验.结果经Pol575-583、Env348-357和Core18-27三条抗原肽刺激,急性乙型肝炎急性期患者组斑点形成细胞数(SFC)分别为110±13、165±17和185±20;慢性乙型肝炎活动期患者组SFC分别为22±4、23±5和30±5,两组差异有统计学意义(t值分别为10.9、15.2和8.0,均P<0.05).急性乙型肝炎急性期患者各抗原肽特异性CTL的应答能力Pol575-583<Env348-357<Core18-27,其中Pol575-583和Core18-27比较,差异有统计学意义(t=4.0,P<0.05),而Env348-357和Core18-27比较,差异无统计学意义(P>0.05).非特异性HLA-2402限制性Core117-125刺激也出现SFC增加,但与阴性对照组比较,差异无统计学意义(P>0.05).结论急性感染者HBV特异性CTL应答水平显著高于慢性HBV感染者,慢性乙型肝炎患者体内的多克隆CTL数量和功能低下.
목적 분석인류백세포항원(HLA)-A0201한제성적특이성CTL,연구급성간염급성기화만성을형간염활동기환자T림파세포대특이성항원표위면역응답적차이.방법 수집HLA-A0201양성적5례급성간염급성기화6례만성을형간염활동기환자적외주혈단개핵세포(PBMC),매련면역반점기술(ELISPOT)측정침대HBV취합매구(Pol575-583)、포막구(Env348-357)화핵심구(Core18-27)3개CD8+T림파세포표위태특이성CTL적수량화공능.수거채용t검험.결과 경Pol575-583、Env348-357화Core18-27삼조항원태자격,급성을형간염급성기환자조반점형성세포수(SFC)분별위110±13、165±17화185±20;만성을형간염활동기환자조SFC분별위22±4、23±5화30±5,량조차이유통계학의의(t치분별위10.9、15.2화8.0,균P<0.05).급성을형간염급성기환자각항원태특이성CTL적응답능력Pol575-583<Env348-357<Core18-27,기중Pol575-583화Core18-27비교,차이유통계학의의(t=4.0,P<0.05),이Env348-357화Core18-27비교,차이무통계학의의(P>0.05).비특이성HLA-2402한제성Core117-125자격야출현SFC증가,단여음성대조조비교,차이무통계학의의(P>0.05).결론 급성감염자HBV특이성CTL응답수평현저고우만성HBV감염자,만성을형간염환자체내적다극륭CTL수량화공능저하.
Objective To analyze human leucocyte antigen (HLA)-A0201 restricted antigen-specific cytotoxic lymphocytes (CTL), and to investigate the difference of T cell response to specific antigen epitopes between patients with acute phase of acute hepatitis B and active phase of chronic hepatitis B. Methods Peripheral blood mononuclear cells (PBMC) from 5 patients with acute phase of acute hepatitis B and 6 patients with active phase of chronic hepatitis B were isolated. The numbers and functions of CD8+ T-lymphocyte epitope peptide specific CTL were detected using enzyme-linked immunosorbent spot (ELISPOT) assay, and the 3 peptides were from HBV polymerase region (Pol575-583), envelope region (Env348-357) and core region (Core18-27), respectively. The data were analyzed using t test. Results The spot formation cell counts (SFC) of Pol575-583, Env348-357 and Core18-27 stimulations in patients with acute phase of acute hepatitis B were 110±13, 165±17 and 185±20, respectively; and those in patients with active phase of chronic hepatitis B were 22±4, 23±5 and 30±5, respectively; the differences were all significant (t=10.9, 15.2 and 8.0, respectively, all P<0.05). The CTL responses to the three peptides in patients with acute phase of acute hepatitis B were Pol575-583<Env348-357<Core18-27; and the difference between responses to Pol575-583 and Core18-27 was significant (t=4.0, P<0.05), while there was no statistical difference between CTL responses to Env348-357 and Core18-27 (P>0.05). The SFC were increased upon non-antigen specific HLA-A2404 restricted epitope (Core117-125), but the difference was not significant compared with negative control group (P>0.05). Conclusions Hepatitis B virus-specific CTL responses in patients with acute hepatitis B are significantly higher than those in patients with chronic hepatitis B. The number and function of polyclonal CTL are both impaired in patients with chronic hepatitis B.