间日疟原虫重组蛋白Pvs25和Pvs28免疫小鼠应答机制观察
李冬梅;刘军;刘英杰;郑丽;冯辉;陈光;曹雅明;
中国医科大学基础医学院免疫学教研室,中国医科大学基础医学院免疫学教研室,中国医科大学基础学院病原生物学教研室,中国医科大学基础医学院免疫学教研室,中国医科大学基础医学院免疫学教研室,中国医科大学基础医学院免疫学教研室,中国医科大学基础医学院免疫学教研室 沈阳110001,沈阳110001,沈阳110001,沈阳110001,沈阳110001,沈阳110001,沈阳110001
Mechanism of murine immune response to Plasmodium vivax Pvs25 and Pvs28 recombinant proteins
摘要 目的观察间日疟传播阻断疫苗候选抗原Pvs25和Pvs28的免疫应答机制。方法分别用与弗氏佐剂乳化后的Pvs25和Pvs28重组蛋白皮下免疫DBA/2小鼠;采用ELISA分别对各组小鼠脾细胞培养上清的IFN-γ、IL-4、IL-10动态检测;以ELISPOT方法检测小鼠脾脏IFN-γ+和IL-4+细胞数量。结果经重组蛋白免疫的两组小鼠脾细胞培养上清IFN-γ的含量于初次免疫后第14d升高,与对照组相比差异显著(P<0.01),但随后降至对照组水平;IL-4和IL-10均在初次免疫后第28d出现有意义的升高(P<0.01),后升高更为明显,分别直至免疫后期70d、56d出现下降,但直至112d仍明显高于免疫前的水平(P<0.01)。初次免疫后第70d?98d小鼠脾脏IFN-γ+和IL-4+细胞数量,与对照组相比IFN-γ+无明显变化,而IL-4+则明显增多(P<0.01)。结论Pvs25和Pvs28重组蛋白可诱导小鼠产生以Th2反应为主的免疫应答。
关键词 :
间日疟原虫 ,
传播阻断疫苗 ,
Pvs25 ,
Pvs28
Abstract :The mechanism of murine immune response to Pvs25 and Pvs28 recombinant protein (rPvs25 and rPvs28), the Plasmodium vivax malaria transmission blocking vaccine candidates, was investigated. Mice were injected subcutaneouly with rPvs25 or rPvs28 emulsified in Freund’s adjuvant. Level of IFN-γ, IL-10 and IL-4 in spleen cell culture supernatants derived from mice at different times before or after vaccination was assayed by ELISA kits. Numbers of IL-4+ and IFN-γ+ cells in murine spleen lymphocytes were detected by enzyme-linked immunospot assay (ELISPOT) on day 70,day 98 after first vaccination. It was found that level of IFN-γ elevated sharply at day 14 (P<0.01) and then dropped to the level of control group. Levels of IL-4 and IL-10 increased from day 28 after first vaccination and got further higher henceforth. Despite of its later decrease, the levels kept higher till day 112 compared with the control (P<0.01). Number of IL-4+ cells from mice spleen elevated on day 70 and day 98 (P<0.01), while that of IFN-γ+ cells didn’t change, compared with the control. It is suggested that rPvs25 and rPvs28 can induce murine immune response, predominantly the Th2 response.
Key words :
Plasmodium vivax
transmission-blocking vaccine
Pvs25
Pvs28
cytokines
收稿日期: 2007-04-20
[1]Tsuboi T,Tachibana M,Kaneko O,et al.Transmission-bloc-king vaccine of vivax malaria[J].Parasitol Int,2003,52:1-11.
[2]Tomas AM,Margos G,Di mopoulos G,et al.P25and P28pro-teins of malaria ookinte surface have multiple and partially redun-dant functions[J].EMBOJ,2001,20(15):3975-3983.
[3]Tsuboi T,Kaslow DC,Gozar MM,et al.Sequence polymor-phismin two novelplasmodiumvivaxookinete surface proteins,Pvs25and Pvs28,that are malatia transmission-blocking vaccine candidates[J],Mol Med,1998,4:772-782.
[4]Sattabongkot J,Tsuboi T,Hisaeda H,et al.Blocking of trans-mission to mosquitoes by antibody toPlasmodiumvivaxmalaria vaccine candidates Pvs25and Pvs28despite antigenic polymor-phismin fieldisolates[J].AmJ Trop Med Hyg,2003,69(5):536-541.
[5]刘军,冯辉,郑丽,等.我国分离株间日疟原虫传播阻断疫苗候选抗原Pvs25基因多态性分析[J].中国人兽共患病学报,2005,21(6):15-18.
[6]Hisaeda H,Stowers AW,Tsuboi T,et al.Antibodies to malari-a vaccine candidates Pvs25and Pvs28completely blockthe ability ofPlasmodium vivaxto infect mosquitoes[J].Infect I mmun,2000,68(12):6618-6623.
[7]Arakawa T,Tsuboi T,Kishi moto A,et al.Serumantibodiesin-duced by intranasal i mmunization of mice withPlasmodium vivaxPvs25co-administered with cholera toxin completely block parasite transmission to mosquitoes[J].Vaccine,2003,21:3143-3148.
[8]Coban C,Ishii KJ,Stowers AW,et al.Effect of CpG Oligode-oxynucleotides on the i mmunogenicity of Pfs25,aPlasmodium falciparumtransmission-Blocking vaccine Antigen[J].Infect I mmun,2004,72:584-588.
[1]
魏海潮, 王琳, 赵艳, 张学星, 胡钰冰, 张杨明慧, 王庆辉, 曹雅明. 疫苗候选抗原Pvs 48/45在中缅边境的遗传多样性特点 [J]. 中国人兽共患病学报, 2020, 36(1): 7-11.
[2]
徐馨,刘怀鄂,刘永华,杨照青. 间日疟原虫抗药性研究进展 [J]. 中国人兽共患病学报, 2015, 31(1): 74-76.
[3]
李欢,孙菲,文岚,王晓春. 多重PCR快速检测恶性疟和间日疟的研究 [J]. 中国人兽共患病学报, 2014, 30(3): 288-291.
[4]
李军伟 周水森 黄芳. 西藏林芝地区间日疟原虫PvMSP-3α多态性研究 [J]. 中国人兽共患病学报, 2012, 28(6): 536-538.
[5]
方强;夏惠;王雪梅;齐文娟;常雪莲;高琪;. 间日疟原虫乳酸脱氢酶编码区全长基因的克隆、序列分析及线性B细胞表位预测 [J]. 中国人兽共患病学报, 2010, 26(06): 519-523.
[6]
胡守锋;陶志勇;夏惠;. 间日疟原虫和恶性疟原虫可溶性抗原免疫反应性比较 [J]. 中国人兽共患病学报, 2009, 25(10): 988-990.
[7]
李冬梅;刘英杰;刘军;王庆辉;郭生玉;曹雅明;. 间日疟传播阻断疫苗Pvs25和Pvs28重组蛋白免疫活性的实验观察 [J]. 中国人兽共患病学报, 2007, 23(06): 598-600.
[8]
肖方震;张山鹰;许龙善;黄江宏;谢汉国;欧阳榕;. 间日疟原虫裂殖子表面蛋白3α基因多态性研究 [J]. 中国人兽共患病学报, 2006, 22(09): 813-816.
[9]
刘军;冯辉;郑丽;凌静;朱晓彤;曹雅明;. 我国间日疟原虫传播阻断疫苗候选抗原Pvs48基因特点分析 [J]. 中国人兽共患病学报, 2006, 22(06): 558-560.
[10]
冯辉;郑丽;刘军;杨毅梅;曹雅明;. 间日疟原虫云南分离株传播阻断疫苗候选抗原Pvs28基因多态性分析 [J]. 中国人兽共患病学报, 2006, 22(04): 360-362.
[11]
刘军,冯辉,郑丽,杨毅梅,金行一,曹雅明. 我国云南分离株间日疟原虫传播阻断疫苗候选抗原pvs25基因多态性分析 [J]. 中国人兽共患病学报, 2005, 21(06): 458-460.
[12]
刘军,冯辉,金行一,曹雅明. 我国分离株间日疟原虫与红细胞黏附关键区DBPⅡ区基因多态性分析 [J]. 中国人兽共患病学报, 2005, 21(04): 323-325.
[13]
张山鹰,许龙善,张莹珍,谢汉国,高琪,陆惠民. 间日疟原虫不同MSP-1等位基因型形态学观察 [J]. 中国人兽共患病学报, 2005, 21(03): 234-236.
[14]
胡锡敏,林世干,陈冬燕,崔宜庆,童重锦,王善青. 逆转录聚合酶链反应检测和鉴定疟原虫的研究 [J]. 中国人兽共患病学报, 2004, 20(10): 894-896.
[15]
张山鹰,陆惠民,许龙善,高琪,沈毓祖,黄伟达. 我国不同疟区间日疟原虫裂殖子表面蛋白1(PvMSP-1)基因多态性研究 [J]. 中国人兽共患病学报, 2004, 20(01): 26-30.