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目的通过检测W135群脑膜炎球菌结合物(简称W135群结合物)小鼠免疫血清中IgG抗体亚类及IgG抗体亲合力,判断W135群结合物的免疫应答特征。方法 (1)免疫血清的制备:将NIH雌性小鼠随机分为3组(A组、B组、对照组),每组30只。A组采用含2.5μg多糖的PSW135CNBr-TT分别于0、2、4、8周腹股沟皮下注射小鼠;B组采用含2.5μg多糖的PSW135CNBr-TT分别于0、4、8周腹股沟皮下注射小鼠;对照组采用含2.5μg多糖的PSW135分别于0、2、4周腹股沟皮下注射小鼠。分别于每次免疫7 d后,经小鼠眼眶静脉采血,分离血清备用;(2)IgG抗体亚类和亲合力的检测:采用间接ELISA分析各组免疫血清中IgG抗体及IgG抗体亚类含量;采用单一浓度的硫氰酸钾溶液作为洗脱液,用间接ELISA测定免疫血清中IgG相对亲合力。结果 A组和B组获得的小鼠免疫血清,IgG抗体均以IgG1为主(占总IgG抗体的80%以上),IgG抗体及其亚类抗体含量均呈典型的剂次加强效应;对照组各针次免疫血清中的IgG3抗体含量所占比例最高,未见免疫剂次加强效应。A组IgG抗体的相对亲合力分别是46.6%、25.4%、17.6%和47.6%,B组IgG抗体的相对亲合力分别是46.6%、23.6%和48.3%,对照组IgG抗体的相对亲合力分别是92.4%、81.9%和81.0%。结论 W135群结合物小鼠免疫血清呈现典型的胸腺依赖性(thymus dependent,TD)抗原的免疫应答特征。
OBJECTIVE To determine the immune response characteristics of W135 colonies by detecting the affinity of IgG antibodies in the immune sera of mice of W135 meningococcal conjugate (W135 conjugate for short) and the affinity of IgG antibodies. Methods (1) Preparation of immune serum: NIH female mice were randomly divided into 3 groups (group A, group B, control group), 30 in each group. In group A, PSW135CNBr-TT containing 2.5μg polysaccharide was used to subcutaneously inoculate mice in groins at 0, 2, 4 and 8 weeks respectively. In group B, PSW135CNBr-TT containing 2.5μg polysaccharide was injected subcutaneously at 0, Mice in the control group were injected with PSW135 containing 2.5μg polysaccharide subcutaneously at 0, 2 and 4 weeks respectively. Blood samples were collected from the orbital vein of mice by 7 days after immunization, respectively. (2) Detection of subclasses and avidity of IgG: The contents of IgG and IgG antibodies in each group were analyzed by indirect ELISA. Using a single concentration of potassium thiocyanate solution as an eluent, indirect ELISA was used to determine the relative affinity of IgG in the immune serum. Results The immune serum and IgG antibodies of mice in groups A and B were mainly IgG1 (accounting for more than 80% of the total IgG antibodies), and the levels of IgG antibodies and their subclasses were typical dose-escalation effects. The control group The immunoglobulin IgG3 antibody content of each needle in the highest proportion, no immune agents to strengthen the effect. The relative affinity of IgG antibodies in group A were 46.6%, 25.4%, 17.6% and 47.6%, respectively. The relative affinity of IgG antibodies in group B was 46.6%, 23.6% and 48.3% respectively. The relative affinity of IgG antibodies in control group was Are 92.4%, 81.9% and 81.0%. Conclusion The immune serum of W135 group conjugate mice shows the immune response characteristic of typical thymus dependent (TD) antigen.