CAJ | 학술논문

目的探讨HBV转基因(Tg)小鼠脾脏树突状细胞(DC)及肝脏B7-H1表达水平与HBV免疫耐受的相关性. 方法制备小鼠脾脏DC,混合淋巴细胞反应检测其同种抗原刺激能力,流式细胞仪检测DC表面主要组织相容性复合物-Ⅱ(MHC-Ⅱ)及CD80、CD86、B7-H1等共刺激分子表达水平,酶联免疫吸附法检测白细胞介素-2(IL-2)、干扰素γ、IL-10水平,逆转录聚合酶链反应法和Western blot法检测肝组织B7-H1表达水平.计量数据组间比较采用f检验.结果 DC和T淋巴细胞比例分别为1:1、1:10、1:100时,HBV Tg小鼠脾脏DC刺激同种小鼠T淋巴细胞增殖数量(每分钟放射细胞数)分别为(865.4±39.3)个、(680.2±34.8)个和(320.0±12.7)个,正常小鼠刺激同种小鼠T淋巴细胞增殖数量分别为(22 385.6±99.7)个,(17 850.6±79.4)个和(760.0±32.1)个,HBV Tg小鼠脾脏DC刺激同种小鼠T淋巴细胞增殖能力明显均弱于正常小鼠DC,t值分别为16.674、19.674和21.712,P值均<0.01,差异有统计学意义.同时,HBV Tg小鼠MHC-Ⅱ,CD80表达下调,而CD86、B7-H1表达差异无统计学意义.HBV Tg小鼠分泌IL-2、干扰素γ、IL-10水平均降低,而HBV Tg小鼠和正常小鼠肝组织表达B7-H1的差异无统计学意义. 结论 HBV免疫耐受与MHC-Ⅱ、CD80下调表达导致的HBV Tg小鼠脾脏DC功能缺陷有关,而与负性共刺激分子B7-H1表达无关.
목적 탐토HBV전기인(Tg)소서비장수돌상세포(DC)급간장B7-H1표체수평여HBV면역내수적상관성. 방법 제비소서비장DC,혼합림파세포반응검측기동충항원자격능력,류식세포의검측DC표면주요조직상용성복합물-Ⅱ(MHC-Ⅱ)급CD80、CD86、B7-H1등공자격분자표체수평,매련면역흡부법검측백세포개소-2(IL-2)、간우소γ、IL-10수평,역전록취합매련반응법화Western blot법검측간조직B7-H1표체수평.계량수거조간비교채용f검험.결과 DC화T림파세포비례분별위1:1、1:10、1:100시,HBV Tg소서비장DC자격동충소서T림파세포증식수량(매분종방사세포수)분별위(865.4±39.3)개、(680.2±34.8)개화(320.0±12.7)개,정상소서자격동충소서T림파세포증식수량분별위(22 385.6±99.7)개,(17 850.6±79.4)개화(760.0±32.1)개,HBV Tg소서비장DC자격동충소서T림파세포증식능력명현균약우정상소서DC,t치분별위16.674、19.674화21.712,P치균<0.01,차이유통계학의의.동시,HBV Tg소서MHC-Ⅱ,CD80표체하조,이CD86、B7-H1표체차이무통계학의의.HBV Tg소서분비IL-2、간우소γ、IL-10수평균강저,이HBV Tg소서화정상소서간조직표체B7-H1적차이무통계학의의. 결론 HBV면역내수여MHC-Ⅱ、CD80하조표체도치적HBV Tg소서비장DC공능결함유관,이여부성공자격분자B7-H1표체무관.
Objective To investigate whether there is an association between the expression of B7-H1 in HBV transgenic mice and the immune tolerance to HBV. Methods T cells stimulatory capacity of DC was analyzed using mixed lymphocyte reaction. Expression of MHC-Ⅱ, CD80, CD86, B7-H1 on DC was detected by Flow Cytometry. IL-2, IFN γ, IL-10 production of T cells were determined by using ELISA. B7-H1 mRNA and protein expression in liver tissue were detected by RT-PCR and Western blotting respectively. Results The ability of DC cells from HBV transgenic mice to stimulate T cell proliferation was significantly impaired compared with DC cells from control mice (t=16.674, 19.674, 21.712, P<0.01). Expression of MHC-Ⅱ, CDS0 on DC was markedly decreased in transgenic mice (t=7.910, 6.413, P<0.05). Meanwhile, the expression of CD86 and B7-H1 on DC cells in HBV transgenic mice were not significantly different from that in control mice. The levels of IL-2, IFN γ, IL-10 in supernantant of T cells was significantly lower compared with controls (t=18.712, 18.712 and 11.683, P<0.05). There was no significant difference in B7-H1 expression at mRNA and protein levels in liver tissue compared with controls. Conclusions Functional defeaet of DC, partly due to decreased expression of MHC-Ⅱ, CD80, but not related to B7-H1 expression, is the cause for immune tolerance to HBV in HBV transgenic mice.