SjFer与GM-CSF共表达核酸疫苗对小鼠免疫保护效果的研究

【摘要】：目的 探讨粒细胞集落刺激因子(GM-CSF)对日本血吸虫铁蛋白核酸疫苗抗攻击感染免疫保护力的影响。 方法 以pGEX-5x-3/SjFer为模板，PCR扩增SjFer，以小鼠总RNA为模板RT-PCR扩增GM-CSF。常规方法构建SjFer/pc、GM-CSF-SjFer/pc及GM-CSF/pc。85只雌性供试小鼠随机分为5组，即生理盐水、pcDNA3.0、GM-CSF/pc、SjFer/pc、GM-CSF-SjFer/pc组，每组小鼠17只。于0，2，4周注射纯化质粒50 ug/次到每只小鼠左后肢股四头肌。每次免疫前尾静脉采血收集血清，ELISA检测血清内特异性IgG水平。末次免疫后一周每组随机选5只小鼠无菌取脾，MTT法检测淋巴细胞增殖情况。末次免疫后两周每只经腹部感染(40±1)条尾蚴，45 天后宰杀小鼠，以观察减虫率和减卵率。结果 SjFer/pc和GM-CSF-SjFer/pc均可诱导小鼠血清内特异性IgG水平。SjFer/pc和GM-CSF-SjFer/pc诱导小鼠产生的减虫率分别为36.27 ％和39.22 ％，减卵率分别为36.10 ％和49.04 ％。结论 GM-CSF对SjFer/pc的免疫保护力有增强作用。

Partial immunoprotection elicited by immunization of DNA vaccine co-encoding SjFer and GM-CSF in mice

Abstract: To study immunoprotection against Schistosoma japonicum(Sj) elicited by immunization of DNA vaccine co-encoding SjFerritin and GM-CSF. Methods SjFer were amplified by PCR from pGEX-5x-3/SjFer temple. GM-CSF were amplified by RT-PCR from totle mice RNA. Reconstructed plasmids were constructed by routine methods. Eighty-five female mice were randomly divided into 5 groups, 17 each .Group A, B and C are controls, immunized with monmal saline, pcDNA3.0 and GM-CSF/pc respectively, and group D, E were immunized with SjFer/pc and GM-CSF-SjFer/pc at weeks 0,2,4. Sera were collected before immunization and challenge infection . Serum lgG were detected by ELISA, Spleen lymphocyte cells of mice were collected before challenge infection.Proliferation activity of spleen lymphocytes was detected by MTT assay. Mice were infected with (40±1) Sj cercariae per mouse 2 weeks after final immunization. Forty-five days later, adult worms and eggs were counted. Results Level of IgG in sera increased after immunization. And proliferation activity of spleen lymphocytes were increased compared with group A, B and C. The worm reduction rate of groups D and E were 36.27 % and 39.22 %. The egg reduction rate of them were 36.10 % and 49.04 %, respectively. Inclusion GM-CSF increased immunoprotection against challenge infection of Sj.