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目的比较细粒棘球蚴重组铁蛋白(rEgferritin)和重组线粒体苹果酸脱氢酶(rEgmMDH)对感染细粒棘球蚴(Echinococcus granulosus)小鼠的免疫差异。方法 30只雌性ICR小鼠随机分为3组(rEgferritin免疫组、rEgmMDH免疫组和对照组)。将rEgferritin、rEgmMDH和PBS与佐剂乳化后,分别于背部皮下多点注射免疫小鼠,每次注射100μl/只(含抗原10μg),共免疫3次,每次间隔2周。末次免疫后2周,3组小鼠分别于腹腔注射0.1 ml原头节悬液(约1 500个)进行攻击感染,22周后剖杀,无菌取脾组织,并记录包囊数量和直径,评价免疫保护力。用流式细胞仪检测脾组织中CD4+T细胞和CD8+T细胞,计算两者比例。结果 rEgferritin免疫组30%(3/10)小鼠有包囊,包囊总数为5个;rEgmMDH免疫组小鼠均有包囊,包囊总数为118个;对照组9只小鼠中7只有包囊,包囊总数为35个。rEgferritin免疫组小鼠获得了84.7%的免疫保护力,而rEgmMDH组无免疫保护力。小鼠脾淋巴细胞CD4+亚群百分比,rEgferritin免疫组[(57.50±7.02)%]显著高于对照组[(43.60±6.09)%](P<0.05)。小鼠脾淋巴细胞CD8+亚群百分比,rEgferritin免疫组[(25.66±6.18)%]和rEgmMDH免疫组[(19.23±5.40)%]与对照组[(21.80±6.12)%]比较,差异均无统计学意义(P>0.05)。小鼠脾淋巴细胞CD4+/CD8+亚群比值,rEgferritin免疫组和rEgmMDH免疫组与对照组比较,差异均无统计学意义(P>0.05)结论 rEgferritin能抑制细粒棘球蚴的生长,而rEgmMDH促进了细粒棘球蚴的生长。
Objective To compare the immunogenicity of rEgferritin and recombinant mitochondrial malate dehydrogenase (rEgmMDH) in mice infected with Echinococcus granulosus. Methods Thirty female ICR mice were randomly divided into 3 groups (rEgferritin immunized group, rEgmMDH immunized group and control group). After emulsifying rEgferritin, rEgmMDH and PBS with adjuvant, mice were immunized subcutaneously with multiple injections of 100 μl / each antigen (10 μg antigen) in the back for 3 weeks for 2 weeks. Two weeks after the last immunization, 3 groups of mice were challenged by intraperitoneal injection of 0.1 ml of the original cephalic suspension (about 1 500) respectively. After 22 weeks, the mice were sacrificed and the spleen tissues were aseptically removed. The number and diameter of the cysts , Evaluation of immune protection. The percentage of CD4 + T cells and CD8 + T cells in spleen tissue were detected by flow cytometry. RESULTS: Thirty (3/10) mice in the rEgferritin-immunized group had cysts and the total number of cysts was 5. The mice in the rEgmMDH immunized group had cysts with a total number of 118 cysts. In the control group, 7 of the 9 mice Cysts, the total number of cysts 35. The mice immunized with rEgferritin achieved 84.7% immunoprotective power, whereas the rEgmMDH group had no immunoprotective power. The percentage of CD4 + subsets in splenic lymphocytes of mice was significantly higher than that in control group [(57.50 ± 7.02)% vs (43.60 ± 6.09)%] (P <0.05). Compared with the control group [(21.80 ± 6.12)%], there was no statistical difference in the percentage of CD8 + subsets of splenic lymphocytes, immunized group of rEgferritin [(25.66 ± 6.18)%] and rEgmMDH immunized group [(19.23 ± 5.40)%] Significance (P> 0.05). The ratio of CD4 + / CD8 + subsets of mice spleen lymphocytes, rEgferritin immunized group and rEgmMDH immunized group were not significantly different from the control group (P> 0.05) .Conclusion rEgferritin can inhibit the growth of Echinococcus granulosus while rEgmMDH promotes Echinococcus granulosus growth.