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目的探讨重组的全长日本血吸虫副肌球蛋白(rSj97)的免疫保护作用以及作为疫苗的适用佐剂组合。方法重组副肌球蛋白分别与弗氏佐剂(rSj97-FA)、ISA206(rSj97-ISA206)、CpGODN(rSj97-CpGODN-IFA)联合免疫C57BL/6小鼠,在0、2、4周共进行3次免疫注射,剂量为每只每次25μgrSj97。第3次免疫后2周,经腹部贴片感染日本血吸虫尾蚴(40±2)条。第12周剖杀冲虫,计算减虫率与减卵率。同时,在0、2、6、8、12周对各组小鼠采血,检测血清中特异性IgG1与IgG2a抗体。结果相对于未免疫组,rSj97-FA组的减虫率为27.6%、减卵率为-2.3%,rSj97-ISA206组的减虫率为45.3%(P<0.01)、减卵率为35.4%(P<0.05),rSj97-CpGODN-IFA组的减虫率为7.3%、减卵率为6.4%,对照组(仅注射CpGODN)减虫率为13%、减卵率为3%。rSj97-FA、rSj97-ISA206与rSj97-CpGODN-IFA组在首次免疫后2周,rSj97特异性IgG1与IgG2a水平较免疫前显著升高(P<0.01),攻击感染后仍维持在同一水平;对照组与未免疫对照组,其特异性IgG1水平至感染后6周方显著升高(P<0.01),但仍为较低水平(A值<0.1),而IgG2a一直保持在基线水平,无显著变化。结论重组副肌球蛋白分别与3种佐剂联合使用均能诱导小鼠产生特异性IgG1和IgG2a,但仅ISA206佐剂组诱导C57BL/6小鼠产生显著的抗日本血吸虫保护力,rSj97-ISA206是潜在用于大动物免疫的疫苗组合。
Objective To investigate the immunoprotective effect of recombinant full-length Schistosoma japonicum para-myosin (rSj97) and its adjuvant-based adjuvant. Methods C57BL / 6 mice were co-immunized with rSj97-FA, ISA206 (rSj97-ISA206) and CpG ODN (rSj97-CpG ODN-IFA) Three immunizations at a dose of 25 μg rSj97 each. Two weeks after the third immunization, Schistosoma japonicum cercariae (40 ± 2) were infected by abdominal patch. The twelfth week to kill the insects, calculate the worm reduction rate and egg reduction rate. At the same time, blood samples were taken at 0, 2, 6, 8 and 12 weeks to detect the specific IgG1 and IgG2a antibodies in serum. Results Compared with the non-immunized group, the rSj97-FA group had a worm reduction rate of 27.6% and an egg reduction rate of -2.3%. The worm reduction rate of rSj97-ISA206 group was 45.3% (P <0.01) (P <0.05). The worm reduction rate of rSj97-CpGODN-IFA group was 7.3%, and the rate of oviposition rate was 6.4%. In the control group (CpGODN only), the worm reduction rate was 13% and the egg reduction rate was 3%. rSj97-ISA, rSj97-ISA206 and rSj97-CpG ODN-IFA group, rSj97-specific IgG1 and IgG2a levels were significantly higher than those before immunization (P <0.01) two weeks after the first immunization. The levels of specific IgG1 in group and non-immunized group were significantly increased (P <0.01) at 6 weeks after infection, but remained low (A value <0.1), while IgG2a remained at baseline level, with no significant difference Variety. CONCLUSION: Recombinant paramyosin can induce specific IgG1 and IgG2a production in mice with the combination of three adjuvants respectively. However, only the ISA206 adjuvant induced significant protection against Schistosoma japonicum in C57BL / 6 mice, and rSj97-ISA206 Vaccine combinations that are potentially useful for immunization of large animals.