CAJ | 학술논문

目的：探讨乙型肝炎患者外周血单个核细胞（PBMC）中Toll样受体2（TLR2）与Th17细胞的相关性，为阐述HBV感染诱导炎症应答机制提供理论和实验依据。方法选取2012年7月-2013年7月唐都医院感染科门诊和住院的34例乙型肝炎初治患者，其中24例慢性乙型肝炎和10例急性乙型肝炎；另外选取健康对照者10例，分离PBMC，利用HBV C基因型Envelope区肽段（特异性）或佛波酯联合伊屋诺霉素（非特异性）刺激，流式细胞术检测TLR2表达及Th17细胞百分比。进一步用TLR2的激动剂刺激PBMC，检测Th17细胞变化情况。组间比较采用Kruskal-Wallis H检验。结果在非特异性刺激条件下，Th17细胞在慢性乙型肝炎患者体内的百分比（4．08±1．78）％显著高于急性乙型肝炎患者（1．85±1．28）％及健康对照者（2.09±0．53）％（P＝0.0009、0．0004），而TLR2＋及IL-17A＋TLR2＋的表达在急、慢性乙型肝炎患者与健康人外周血中差异均无统计学意义（P均＞0.05）。在特异性刺激条件下Th17及TLR2的表达在慢性乙型肝炎患者体内的表达显著高于急性乙型肝炎组[（5．45±1．61）％ vs （3.20±1.13）％；（5．19±3．18）％vs （1．88±1．30）％]，差异具有统计学意义（P＝0．0006、0．0006）。加入TLR2激动剂后急、慢性乙型肝炎患者体内Th17细胞的比例均显著升高，但在急性乙型肝炎患者中，刺激前后差异无统计学意义（P＞0.05）。结论 TLR2可以直接影响Th 17细胞的应答，从而促进乙型肝炎中炎症应答反应。
목적：탐토을형간염환자외주혈단개핵세포（PBMC）중Toll양수체2（TLR2）여Th17세포적상관성，위천술HBV감염유도염증응답궤제제공이론화실험의거。방법선취2012년7월-2013년7월당도의원감염과문진화주원적34례을형간염초치환자，기중24례만성을형간염화10례급성을형간염；령외선취건강대조자10례，분리PBMC，이용HBV C기인형Envelope구태단（특이성）혹불파지연합이옥낙매소（비특이성）자격，류식세포술검측TLR2표체급Th17세포백분비。진일보용TLR2적격동제자격PBMC，검측Th17세포변화정황。조간비교채용Kruskal-Wallis H검험。결과재비특이성자격조건하，Th17세포재만성을형간염환자체내적백분비（4．08±1．78）％현저고우급성을형간염환자（1．85±1．28）％급건강대조자（2.09±0．53）％（P＝0.0009、0．0004），이TLR2＋급IL-17A＋TLR2＋적표체재급、만성을형간염환자여건강인외주혈중차이균무통계학의의（P균＞0.05）。재특이성자격조건하Th17급TLR2적표체재만성을형간염환자체내적표체현저고우급성을형간염조[（5．45±1．61）％ vs （3.20±1.13）％；（5．19±3．18）％vs （1．88±1．30）％]，차이구유통계학의의（P＝0．0006、0．0006）。가입TLR2격동제후급、만성을형간염환자체내Th17세포적비례균현저승고，단재급성을형간염환자중，자격전후차이무통계학의의（P＞0.05）。결론 TLR2가이직접영향Th 17세포적응답，종이촉진을형간염중염증응답반응。
Objective To investigate the relationship between Toll-like receptor 2 (TLR2 )in peripheral blood mononuclear cells (PB-MCs)and T helper 17 (Th17)cells among hepatitis B patients and to provide a theoretical and experimental basis for the study on the mech-anism of inflammatory response to hepatitis B virus (HBV)infection.Methods Thirty-four previously untreated patients with hepatitis B (24 cases of chronic hepatitis B and 10 cases of acute hepatitis B)who visited or were hospitalized in the Center of Infectious Diseases, Tangdu Hospital,from July 2012 to July 2013,as well as ten healthy controls,were enrolled in this study.PBMCs were isolated and stimu-lated by HBV (genotype C)envelope peptides (specific)or propylene glycol monomethyl ether acetate (PMA)plus ionomycin (nonspecif-ic).Flow cytometry was performed to measure the expression of TLR2 and the percentage of Th17 cells.PBMCs were further stimulated by TLR2 agonist Pam3Csk4,and the changes in percentage of Th17 cells were evaluated.Comparison between groups was made by Kruskal-Wallis H test.Results When stimulated by PMA plus ionomycin,patients with chronic hepatitis B had a significantly higher percentage of Th17 cells than patients with acute hepatitis B and healthy controls [(4.08 ±1.78)% vs (1.85 ±1.28)%,P=0.0009;(4.08 ± 1.78)% vs (2.09 ±0.53)%,P=0.0004],while the percentages of TLR2 +and IL-17A+TLR2 +T cells in peripheral blood CD3 +CD4 +T cells showed no significant differences between patients with acute hepatitis B,patients with chronic hepatitis B,and healthy con-trols (P>0.05 for all).When stimulated by HBV envelope peptides,patients with chronic hepatitis B had significantly higher percentages of IL-17A+T cells and TLR2 +T cells than patients with acute hepatitis B [(5.45 ±1.61)%vs (3.20 ±1.13)%,P=0.0006;(5.19 ±3.18)% vs (1.88 ±1.30)%,P=0.0006].After the addition of Pam3Csk4,patients with chronic hepatitis B had a significantly in-creased percentage of Th17 cells,while patients with acute hepatitis B had a nonsignificantly increased percentage of Th17 cells.Conclusion TLR2 could directly influence Th17 cell responses,thereby playing a proinflammatory role in the immune response to HBV infection.