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目的观察猪带绦虫TSO45W-4B-TSOL18重组蛋白疫苗免疫小鼠诱导产生的体液和细胞免疫应答。方法将SPF级昆明小鼠60只随机分为3组,每组20只,分别为重组蛋白疫苗组和重组蛋白疫苗加弗氏佐剂组和生理盐水对照组。采用多点皮下注射法免疫3次,间隔2周。于免疫后0、2、4、6、8周摘眼球取血,分离血清,采用ELISA法检测小鼠血清IgG与IgG2a水平;无菌取脾,制备脾细胞悬液,采用四甲基偶氮唑盐比色法(MTT法)检测脾细胞增殖水平,采用ELISA法检测小鼠脾细胞经特异性抗原刺激后产生IL-2和IL-4的能力。结果与0周时及生理盐水对照组相比,免疫小鼠血清IgG水平在免疫后2~8周升高,F值分别为20.108、235.886、209.012、49.058,6周时达较高水平,IgG2a水平在免疫后2~8周升高,F值分别为80.214、76.540、152.314、82.561,6周时达较高水平,脾细胞增殖水平在免疫后2~8周升高,F值分别为21.643、56.982、225.880、151.682,6周时达较高水平,小鼠脾细胞经特异性抗原刺激后产生的IL-2水平在免疫后2~8周升高,F值分别为16.699、25.967、77.133、34.662,6周时达较高水平,IL-4水平在免疫后2~8周升高,F值分别为26.087、147.353、130.654、62.807,6周时达较高水平,与0周时及生理盐水对照组相比差异均有统计学意义(P<0.05);免疫后2~8周重组蛋白疫苗加弗氏佐剂组与重组蛋白疫苗组比较上述指标差异均有统计学意义(P<0.05)。结论猪带绦虫TSO45W-4B-TSOL18重组蛋白疫苗免疫可诱导小鼠产生体液和细胞免疫应答,免疫效果以重组蛋白疫苗加弗氏佐剂组较好。
Objective To observe the humoral and cellular immune responses induced by the TSO45W-4B-TSOL18 recombinant protein vaccine induced by Taenia solium in mice. Methods Sixty SPF Kunming mice were randomly divided into three groups (n = 20 in each group): recombinant protein vaccine and recombinant protein vaccine plus Freund’s adjuvant and saline control group respectively. Multi-point subcutaneous injection immunization 3 times, 2 weeks apart. The blood samples were collected at 0, 2, 4, 6, and 8 weeks after immunization, the serum was separated and the serum IgG and IgG2a levels were detected by ELISA. Spleen cells were prepared by splenectomy. The proliferation of spleen cells was detected by azole-salt colorimetric assay (MTT). The ability of splenocytes to produce IL-2 and IL-4 after stimulation with specific antigens was detected by ELISA. Results Compared with the 0 week group and the saline control group, the serum IgG levels of the immunized mice increased at 2 to 8 weeks after immunization, the F values were 20.108, 235.886, 209.012 and 49.058, respectively. The level of IgG2a was higher at After 2 to 8 weeks of immunization, the F value increased to 80.214, 76.540, 152.314 and 82.561, respectively. The levels of splenocyte proliferation increased from 2 to 8 weeks after immunization with F values of 21.643, 56.982 and 225.880 , 151.682 at 6 weeks, the level of IL-2 in mouse splenocytes stimulated by specific antigen increased at 2 ~ 8 weeks after immunization, F values were 16.699, 25.967, 77.133, 34.662, respectively IL-4 levels increased at 2 ~ 8 weeks after immunization, F values were 26.087,147.353,130.654,62.807 respectively, reaching a high level at 6 weeks, compared with those at 0 week and saline control group (P <0.05). There was significant difference between the recombinant protein vaccine plus Freund’s adjuvant group and the recombinant protein vaccine group at 2 ~ 8 weeks after immunization (P <0.05). Conclusion The vaccine of TSO45W-4B-TSOL18 recombinant protein of Taenia solium can induce humoral and cellular immune responses in mice, and the immunogenicity of the recombinant protein vaccine plus Freund’s adjuvant is better.