西咪替丁伍用弓形虫ROP2蛋白免疫小鼠诱导免疫反应的初步研究

发布时间：2019-05-06 18:08

【摘要】：目的观察西咪替丁伍用弓形虫棒状体蛋白2(ROP2)免疫小鼠诱导的免疫反应。方法 80只BALB/c小鼠随机分为A组、B组、C组和D组,各组分别经皮下注射PBS、弓形虫ROP2蛋白+PBS、弓形虫ROP2蛋白+福氏佐剂和弓形虫ROP2蛋白+西咪替丁[200 mg/(kg.d)]免疫BALB/c小鼠,每次ROP2蛋白免疫剂量为100μg/只(200μl),共免疫3次,每次间隔2周。于首次免疫前和每次免疫后2周,采血分离血清。末次免疫后2周,每组小鼠随机处死8只,无菌分离脾细胞,CCK-8法测定淋巴细胞增殖活性。流式细胞术检测小鼠全血T细胞亚群。ELISA法检测小鼠血清IgG抗体和γ干扰素(IFN-γ)水平。各组剩余12只小鼠经腹腔感染弓形虫RH株速殖子(5×104个/鼠),观察攻击感染后小鼠生存时间。结果末次免疫后2周,A、B、C和D组小鼠的血清IFN-γ水平分别为(659.750±239.962)、(872.750±197.011)、(1 600.750±480.680)和(1 494.375±451.655)pg/ml,血清特异性IgG抗体水平分别为0.504±0.078、0.592±0.160、1.068±0.111和1.046±0.147,脾细胞增殖活性分别为0.504±0.078、0.592±0.160、0.831±0.130和0.762±0.089,外周血CD4+/CD8+比值分别为0.504±0.078、0.592±0.160、0.831±0.130和0.762±0.089,C和D组均显著高于A和B组(前3个指标,均P0.01;后者,均P0.05),D组与C组的差异均无统计学意义(P0.05)。各免疫组弓形虫攻击感染后,A、B、C和D组小鼠的生存时间中位数分别为96、108、132和132 h,C组和D组小鼠平均存活时间显著长于A组和B组(P0.05),D组与C组的相比,差异无统计学意义(P0.05)。结论西咪替丁可增强ROP2蛋白诱导的细胞免疫和体液免疫反应。
[Abstract]:Objective to observe the immune response induced by cimetidine combined with Toxoplasma gondii rod protein 2 (ROP2) immunization in mice. Methods 80 BALB/c mice were randomly divided into three groups: group A, group B, group C and group D. ROP2 protein PBS, of Toxoplasma gondii was subcutaneously injected into each group. Toxoplasma gondii ROP2 protein adjuvant and Toxoplasma gondii ROP2 protein cimetidine [200 mg/ (kg.d)] were used to immunize BALB/c mice at a dose of 100 渭 g / mouse (200 渭 l) per ROP2 protein for 3 times at an interval of 2 weeks. Blood samples were collected before the first immunization and 2 weeks after each immunization. Two weeks after the last immunization, 8 mice in each group were killed at random, spleen cells were isolated aseptically, and lymphocyte proliferation activity was measured by CCK- 8 method. Flow cytometry was used to detect T lymphocyte subsets and Elisa was used to detect serum IgG antibody and interferon-纬 (IFN- 纬) levels in mice. The remaining 12 mice in each group were infected with Toxoplasma gondii RH strain Tachyzoites (5 脳 10 4 / mouse) through abdominal cavity to observe the survival time after infection. Results the serum IFN- 纬 levels in group A, B, C and D were (659.750 卤239.962), (872.750 卤197.011), (1 600.750 卤480.680) and (1 494.375 卤451.655) pg/ml, respectively at 2 weeks after the last immunization. The serum specific IgG antibody levels were 0.504 卤0.078, 0.592 卤0.160, 1.068 卤0.111 and 1.046 卤0.147, respectively. The proliferative activities of splenocytes were 0.504 卤0.078, 0.592 卤0.160, 0.831 卤0.130 and 0.762 卤0.089, respectively. The ratio of CD4 / CD8 in peripheral blood was 0.504 卤0.078, 0.592 卤0.160, 0.831 卤0.130 and 0.762 卤0.089, respectively. The ratios in C and D groups were significantly higher than those in A and B groups (P 0.01). In the latter group, there was no significant difference between), D group and C group (P0.05). The median survival time of mice in groups A, B, C and D were 96108132 and 132h, respectively. The average survival time of mice in groups C and D was significantly longer than that in groups A and B (P0.05), D group compared with group C, P 0.05, P < 0.01), and the median survival time of mice in group C and D was significantly longer than that in group A and group B (P0.05). There was no significant difference (P0.05). Conclusion cimetidine can enhance cellular and humoral immune responses induced by ROP2 protein.
【作者单位】：蚌埠医学院病原生物学教研室安徽省感染与免疫重点实验室;安徽病原生物学省级实验室人兽共患病安徽省重点实验室安徽医科大学病原生物学教研室;
【基金】：国家重点基础研究发展规划项目(973计划)(No.2010CB530001) 安徽省教育厅重点资助项目(No.KJ2012A201)~~
【分类号】：R392

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