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目的鉴定美洲大蠊过敏原Per a 7表达,研究其对辅助性T细胞17(Th17细胞)相关基因和蛋白表达的影响。方法用含编码Per a 7基因的大肠杆菌诱导表达并纯化出Per a 7蛋白。将磁珠分离后CD4+T淋巴细胞与Per a 7激发活化的树突状细胞共培养。采用实时荧光定量PCR检测IL-17、RORγt和Foxp3mRNA表达,ELISA检测上清液中IL-17含量。结果成功克隆分子量约为33kD的重组蛋白Per a 7。重组Per a 7激发后树突状细胞与CD4+T淋巴细胞共培养后,IL-17、RORγt mRNA表达以及上清液中IL-17含量增加(P<0.05),而Foxp3mRNA表达无明显变化(P>0.05)。结论美洲大蠊过敏原Per a 7能激发树突状细胞诱导T淋巴细胞向Th17细胞方向分化,在过敏性疾病发生中起了重要的作用。
Objective To identify the expression of Per a 7 in Periplaneta americana and investigate its effect on the expression of related genes and proteins in T helper 17 (Th17) cells. Methods Per a 7 protein was induced and expressed in Escherichia coli containing Per a 7 gene. CD4 + T lymphocytes isolated from the magnetic beads were co-cultured with Per a 7-activated dendritic cells. The expression of IL-17, RORγt and Foxp3 mRNA were detected by real-time fluorescence quantitative PCR. The content of IL-17 in supernatant was detected by ELISA. Results The recombinant protein Per a 7 with a molecular weight of about 33 kD was successfully cloned. After co-cultured with CD4 + T lymphocytes, the expression of IL-17 and RORγt mRNA and the content of IL-17 in the supernatant increased (P <0.05), while the expression of Foxp3 mRNA did not change significantly P> 0.05). Conclusion Per a 7, an American cockroach allergen, can stimulate dendritic cells to induce T lymphocytes to differentiate into Th17 cells and play an important role in the development of allergic diseases.