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目的研究弓形虫亲环蛋白(Cyclophilin,CyP)亚单位疫苗的免疫保护性。方法 30只BALB/c小鼠随机分为3组:实验组、佐剂组和PBS对照组,分别肌注pET-28a-TgCyP重组蛋白100μg/只、佐剂100 ul/只和PBS 100 ul/只,共免疫3次,每次间隔2周。末次免疫后2周,各组分别取4只小鼠处死,取脾脏,检测CD4+、CD8+及细胞因子。同时,采用阿尔玛蓝细胞增殖与细胞毒性检测试剂测定小鼠脾淋巴细胞的增殖应答。其余小鼠腹腔攻击感染Rh株弓形虫速殖子500个,观察其存活情况。结果 pET-28a-TgCyP重组蛋白亚单位疫苗能诱发小鼠产生细胞免疫和体液免疫反应,小鼠的CD4+T细胞(62.59±4.17)%、CD8+T细胞(8.53±0.46)%与PBS及佐剂对照组比较显著增殖(P<0.01),其脾细胞培养液白细胞介素-2(IL-2)、γ干扰素(IFN-γ)显著升高,小鼠脾淋巴细胞增殖应答显著增强(P<0.01)。弓形虫攻击感染216 h后,实验组小鼠免疫保护率为33.3%,佐剂对照组及PBS对照组小鼠均全部死亡。结论 pET-28a-TgCyP重组蛋白亚单位疫苗具有较强的免疫原性,能产生良好的免疫保护作用。
Objective To study the immunoprotection of cyclophilin (CyP) subunit vaccine. Methods Thirty BALB / c mice were randomly divided into three groups: experimental group, adjuvant group and PBS control group. The mice were intramuscularly injected with pET-28a-TgCyP recombinant protein 100 μg / adjuvant 100 ul / PBS and 100 ul / Only, co-immunization 3 times, each interval of 2 weeks. Two weeks after the last immunization, four mice in each group were sacrificed and their spleens were taken for detection of CD4 +, CD8 + and cytokines. At the same time, the proliferative response of mouse splenic lymphocytes was measured by using Almar’s blue cell proliferation and cytotoxicity detection reagent. The remaining mice were challenged intraperitoneally with 500 tachyzoites infected with Rh strain Toxoplasma gondii and their survival was observed. Results The pET-28a-TgCyP recombinant protein subunit vaccine induced cellular and humoral immune responses in mice. The levels of CD4 + T cells (62.59 ± 4.17)%, CD8 + T cells (8.53 ± 0.46)% and PBS The adjuvant control group showed significant proliferation (P <0.01), and the spleen cell culture medium IL-2 and IFN-γ were significantly increased, and the splenic lymphocyte proliferation response was significantly increased (P <0.01). After 216 h of Toxoplasma gondii infection, the immunoprotection rate of the mice in the experimental group was 33.3%, and all the mice in the adjuvant control group and the PBS control group all died. Conclusion pET-28a-TgCyP recombinant protein subunit vaccine has strong immunogenicity, can produce good immune protection.