CAJ | 학술논문

目的：观察间日疟原虫裂殖子主要蛋白1（PvMSP1）对树突状细胞（DC）分化成熟和功能的影响，并探讨该蛋白通过Toll样受体（TLR）通路活化DC的机制。方法选择不同剂量的PvMSP1（1.0、10.0、100.0μg/ml）体外刺激人单核细胞来源的DC，采用流式细胞术分析DC成熟性相关分子CD83、CD86、HLA-DR的表达变化；ELISA检测DC培养上清中IL-10、IL-12的表达水平；RT-PCR检测DC TLR4、TLR9 mRNA的表达水平；MTT法检测DC刺激自体淋巴细胞增殖的能力。同时选择未刺激的DC作为阴性对照组，LPS刺激的DC作为阳性对照组。对所得数据进行方差分析和q检验。结果与未刺激组比较，LPS诱导组CD83、CD86、HLA-DR的百分含量均增加，PvMSP1诱导组CD83、CD86、HLA-DR的表达也均升高（P均<0.05）；LPS诱导组IL-10、IL-12的表达量明显增加（P<0.01），PvMSP1诱导组IL-10、IL-12的表达量也均增加（P均<0.05）；LPS组DC TLR4 mRNA的表达增加（P<0.05），TLR9 mRNA的表达无明显变化（P>0.05），PvMSP1诱导组DC TLR4 mRNA的表达增加（P<0.01），TLR9 mRNA无明显变化（P>0.05）；DC能够刺激自体淋巴细胞增殖。结论 PvMSP1具有促进DC分化成熟的作用，且经其诱导成熟的DC具备抗原递呈功能；PvMSP1可能经TLR4通路而非TLR9通路诱导DC成熟。
목적：관찰간일학원충렬식자주요단백1（PvMSP1）대수돌상세포（DC）분화성숙화공능적영향，병탐토해단백통과Toll양수체（TLR）통로활화DC적궤제。방법선택불동제량적PvMSP1（1.0、10.0、100.0μg/ml）체외자격인단핵세포래원적DC，채용류식세포술분석DC성숙성상관분자CD83、CD86、HLA-DR적표체변화；ELISA검측DC배양상청중IL-10、IL-12적표체수평；RT-PCR검측DC TLR4、TLR9 mRNA적표체수평；MTT법검측DC자격자체림파세포증식적능력。동시선택미자격적DC작위음성대조조，LPS자격적DC작위양성대조조。대소득수거진행방차분석화q검험。결과여미자격조비교，LPS유도조CD83、CD86、HLA-DR적백분함량균증가，PvMSP1유도조CD83、CD86、HLA-DR적표체야균승고（P균<0.05）；LPS유도조IL-10、IL-12적표체량명현증가（P<0.01），PvMSP1유도조IL-10、IL-12적표체량야균증가（P균<0.05）；LPS조DC TLR4 mRNA적표체증가（P<0.05），TLR9 mRNA적표체무명현변화（P>0.05），PvMSP1유도조DC TLR4 mRNA적표체증가（P<0.01），TLR9 mRNA무명현변화（P>0.05）；DC능구자격자체림파세포증식。결론 PvMSP1구유촉진DC분화성숙적작용，차경기유도성숙적DC구비항원체정공능；PvMSP1가능경TLR4통로이비TLR9통로유도DC성숙。
Objective To investigate the effects of Plasmodium vivax merozoite surface protein 1(PvMSP1)on differentia-tion,maturation and function of dendritic cells(DC)and the mechanisms of PvMSP1 on the activation of DC via toll like receptors (TLR). Methods DCs were incubated with different doses of PvMSP1(1.0,10.0,100.0μg/ml)in vitro. The changes of CD83, CD86,and HLA-DR on DC were detected by flow cytometry(FCM);the expressions of cytokine IL-10 and IL-12 of DC were mea-sured by ELISA;the expressions of TLR4 and TLR9 mRNA of DC were measured by RT-PCR;the proliferation induction to autol-ogous lymphocytes of DC was measured by MTT. Meanwhile,the untreated DC and LPS inducing DC were as the negative control and positive control,respectively. All the data were analyzed statistically. Results Compared with the untreated DC,the propor-tions of CD83,CD86 and HLA-DR on DC induced by LPS and PvMSP1 increased significantly(all P<0.05);the expressions of IL-10 and IL-12 of DC induced by LPS increased significantly(P<0.01),and those induced by PvMSP1 also increased signifi-cantly(all P<0.05). In the LPS inducing group,the TLR4 mRNA production increased(P<0.05)and the TLR9 mRNA produc-tion had no significantly changes(P>0.05). In the PvMSP1-treated group,the DC TLR4 mRNA production increased(P<0.01) and the TLR9 mRNA production had no significantly changes(P>0.05);DC stimulated the proliferation of autologous lympho-cytes. Conclusion PvMSP1 enhances DC differentiation and maturation,and the mature DC induced by PvMSP1 has the ability of antigen presenting. The route for PvMSP1 inducing DC maturation might be TLR4 pathway rather than TLR9 pathway.