CAJ | 학술논문

目的探讨肠道病毒71型( EV71)感染患儿免疫活性细胞模式识别受体(pattern recognition receptor，PRR)及细胞因子水平的变化。方法 EV71感染患儿71例，其中轻症EV71感染组20例，重症EV71感染组25例，危重症EV71感染组26例，同年龄正常对照组20例。采用real -time PCR检测外周血单个核细胞维甲酸蛋白Ⅰ(retinoic acidinducible gene Ⅰ，RIG-Ⅰ)、黑色素瘤分化相关基因5（melanoma differentitation-associated gene 5，MDA5）及细胞因子(IL-12、IFN-α)表达；流式细胞术检测外周血单核/巨噬细胞( monocyte/macrophages，MC)、髓样树突状细胞(myloid dentritic cells,mDC)及浆样树突状细胞（plasmacytoid dentritic cells，pDC）Toll样受体(TLRs)表达率；ELISA检测细胞因子IL-12及IFN-α的变化。结果 (1)轻症患儿仅TLR7升高，重症EV71感染患儿外周血MC、mDC、pDCTLR7表达明显升高(P＜0.05)，MC、mDC高表达TLR2、3、4，危重症患儿呈下降趋势。(2)EV71感染患儿胞内模式识别受体RIG- Ⅰ/MDA5 mRNA表达明显增加；(3)轻症患儿DC相关细胞因子有上升趋势，重症患儿DC相关细胞因子IL-12、IFN-α明显增高(P＜0.05)，轻症及危重症患儿明显降低(P＜0.05)。结论 TLR7可能是免疫活性细胞识别EV71的主要模式识别受体；RIG-I/MDA5也可能参与识别EV71；合并细菌感染或细胞破坏释放的内源性配体导致TLR2或TLR4活化，诱导炎症反应。
목적 탐토장도병독71형( EV71)감염환인면역활성세포모식식별수체(pattern recognition receptor，PRR)급세포인자수평적변화。방법 EV71감염환인71례，기중경증EV71감염조20례，중증EV71감염조25례，위중증EV71감염조26례，동년령정상대조조20례。채용real -time PCR검측외주혈단개핵세포유갑산단백Ⅰ(retinoic acidinducible gene Ⅰ，RIG-Ⅰ)、흑색소류분화상관기인5（melanoma differentitation-associated gene 5，MDA5）급세포인자(IL-12、IFN-α)표체；류식세포술검측외주혈단핵/거서세포( monocyte/macrophages，MC)、수양수돌상세포(myloid dentritic cells,mDC)급장양수돌상세포（plasmacytoid dentritic cells，pDC）Toll양수체(TLRs)표체솔；ELISA검측세포인자IL-12급IFN-α적변화。결과 (1)경증환인부TLR7승고，중증EV71감염환인외주혈MC、mDC、pDCTLR7표체명현승고(P＜0.05)，MC、mDC고표체TLR2、3、4，위중증환인정하강추세。(2)EV71감염환인포내모식식별수체RIG- Ⅰ/MDA5 mRNA표체명현증가；(3)경증환인DC상관세포인자유상승추세，중증환인DC상관세포인자IL-12、IFN-α명현증고(P＜0.05)，경증급위중증환인명현강저(P＜0.05)。결론 TLR7가능시면역활성세포식별EV71적주요모식식별수체；RIG-I/MDA5야가능삼여식별EV71；합병세균감염혹세포파배석방적내원성배체도치TLR2혹TLR4활화，유도염증반응。
Objective To investigate the expression of pattern recognition receptor (PRR)of immune cells from the patients with enterovirus 71 ( EV 71 ) infection and the changes of cytokines. Methods Seventy-one patients and 20 age-matched healthy children were enrolled in the study. The patients were divided into three groups according to the critical degree: 20 mild patients, 25 severe patients and 26 critical patients. Real-time PCR were used to evaluate the levels of retinoic acidinducible gene Ⅰ ( RIG- Ⅰ ), melanoma differentitation-associated gene 5 ( MDA5 ) and cytokines IL-12, IFN-α mRNA expression in peripheral blood mononuclear cells (PBMC). The proportions of Toll like receptors(TLRs) on monocytes/macrophages (MC) , myloiddentritic cells (mDC) and plasmacytoiddentritic cells (pDC) were analyzed by flow cytometry. The concentrations of plasma cytokines( IL-12, IFN-α) were measured by ELISA. Results ( 1 ) The proportion of TLR7 is the unique TLR which is increased in mild patients and it was significantly elevated in MC, mDC and pDC in severe group (P＜0.05), TLR2, TLR3 and TLR4 also had an enhanced expression in MC and mDC. The enhanced expression of TLRs mentioned above had a decreased trend in critical patients. (2)Transcription levels of RIG- Ⅰ and MDA5 was significantly elevated in EV71 infected children.(3)The expression levels of DC-associated cytokines gene( IL-12 and IFN-α) have an increased trend in mild cases and these cytokines were remarkably increased in severe patients (P＜0.05), whereas decreased in critical cases (P＜0.05). Conclusion TLR7 might be the main PRR recognizing EV71 in immune cells and RIG-Ⅰ/MDA5 might also take part in the recognition of EV71. The exogenous or endogenous ligands released from bacterial infection and cell destruction could result in the activation of TLR2 or TLR4, inducing the inflammatory response.