重组弓形虫磷酸甘油酸变位酶2联合IL-2、IFN-γ佐剂鼻内免疫小鼠诱导的小肠黏膜免疫应答  

INDUCTION OF INTESTINE MUCOSAL IMMUNE RESPONSES IN MICE BY INTRANASAL IMMUNIZATION WITH RECOMBINANT TOXOPLASMA GONDII PHOSPHOGLYCERATE MUTASE 2 PROTEIN COMBINED WITH DIFFERENT ADJUVANTS

作　　者：祝建疆[1,2] 李雅清[1,2] 王海龙[2] 孟晓丽[2] 杨建一[1] 殷国荣[2] 

机构地区：[1]山西医科大学细胞生物学与遗传学教研室,太原030001 [2]山西医科大学医学寄生虫学研究所,太原030001

出　　处：《寄生虫与医学昆虫学报》2012年第4期199-203,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家自然科学基金项目（No.81071374）

摘　　要：为探讨IL-2和IFN-γ的佐剂效应，本研究观察了重组弓形虫磷酸甘油酸变位酶2（rTgPGAM2）联合IL-2或IFN-γ滴鼻免疫小鼠诱导小肠黏膜IgA抗体分泌细胞（IgA—ASCs）和小肠冲洗液sIgA的免疫应答。将48只6周龄雌性BALB／c小鼠随机均分为6组，免疫组分别用500IUIL-2、1000UIFN-γ、30μgrTgPGAM2、30μgrTgPGAM2＋500IUIL-2或30μgrTgPGAM24-1000UIFN-1滴鼻免疫小鼠，抗原和佐剂溶于20μLPBS中，对照组用20μLPBS滴鼻。免疫3次，一免和二免间隔2周，二免后1周三免。三免后第14天，颈椎脱臼处死小鼠，免疫组化检测小鼠十二指肠、空肠和回肠黏膜中IgA-ASCs，计算阳性率，ELISA测定小肠冲洗液sIgA水平。结果表明，免疫组十二指肠、空肠和回肠黏膜中IgA—ASCs阳性率高于PBS组，其中rTgPGAM2、rTgPGAM2联合IL-2或IFN-γ佐剂组差异显著（P〈0．05或P〈0．01）。rTgPGAM2联合IL-2或IFN-γ佐剂组小肠冲洗液sIgA水平显著高于PBS组（P〈0．01）。rTgPGAM2、TgPGAM2＋IL-2和TgPGAM2＋IFN-γ滴鼻免疫小鼠均可有效诱导肠黏膜免疫应答，IL-2和IFN-γ可显著增强抗原的免疫效应，IL-2的佐剂效应优于IFN-γ。In order to analyze the adjuvant effect of IL-2 and IFN-γ, we observed the immune responses including IgA antibody secreting cells （IgA-ASCs） in small intestine mucosal and sIgA level in intestine washes which induced by intranasal immunization with recombinant Toxoplasma gondii phosphoglycerate mutase 2 （ rTgPGAM2 ） proteins combined with IL-2 or IFN-γ in mice. Forty-eight 6-week-old female BALB/c mice were randomly divided into 6 groups. The 8 mice of each immunity group were immunized intranasal three times with 500 IU IL-2, 1000 U IFN-γ, 30 μg rTgPGAM2, 30 μg rTgPGAM2 ＋ 500 IU IL-2 or 30 txg rTgPGAM2 ＋ 1000 U IFN-3,, soluble in total volume of 20μL of PBS, respectively. The first and the second immunization were 2 weeks of interval, and there was 1 week of interval between the third immunization and the second one. Mice in control group were treated with 20 txL PBS instead. All mice were sacrificed by cervical dislocation 14 days after the third immunization. The positive rates of IgA-ASCs in mice of duodenum, jejunum, and ileum were detected by immunohistochemistry. The sIgA in intestinal washes were analyzed with ELISA assay. The results showed that IgA-ASCs positive rate in the immunity groups were higher than that of the control group, especially in rTgPGAM2, rTgPGAM2 combined with IL-2 or IFN-γ adjuvant groups （P 〈 0.05 or P 〈 0. 01 ）. There was also a significantly higher level of sIgA in small intestinal washes of rTgPGAM2 combined with IL-2 or IFN-γ adjuvant groups compared with the control group （P 〈 0. 01 ）. We can conclude that intranasal immunization with rTgPGAM2, TgPGAM2 ＋ IL-2 and TgPGAM2 ＋ IFN-γ effectively induced intestinal mucosal immune responses in mice, IL-2 and IFN-γsignificantly enhanced the immune responses to the antigen. As an adjuvant, IL-2 can perform better effect than IFN-γ.

关 键 词：刚地弓形虫 磷酸甘油酸变位酶2 抗体分泌细胞 鼻内免疫 黏膜佐剂 

分 类 号：S852.4[农业科学—基础兽医学]