论文部分内容阅读
目的：制备志贺氏Ｉ型菌的Ｏ－ 特异多糖（Ｏ－ ＳＰ） 与破伤风类毒素（ＴＴ） 的结合物Ｏ－ ＳＰ－ ＴＴ，并通过动物实验检测其抗原性和血清中抗Ｏ－ ＳＰＩｇＧ、ＩｇＭ 抗体水平及抗体杀菌活性。方法：提取纯化志贺氏Ｉ型菌的Ｏ－ ＳＰ，以己二酸二肼（ＡＤＨ） 为连接剂，以破伤风类毒素为载体，制备Ｏ－ ＳＰ－ ＴＴ结合物。对提取物、中间产物、结合物的特性进行检测。结果：Ｏ－ ＳＰ、Ｏ－ ＳＰ－ ＡＨ、Ｏ－ ＳＰ－ ＴＴ均可与兔超免疫血清形成清晰的沉淀线。单独使用Ｏ－ ＳＰ免疫小鼠后抗体无显著性升高。Ｏ－ ＳＰ－ ＴＴ免疫注射后第１ 、２ 次间，第２、３ 次间ＩｇＧ抗体及抗ＴＴ抗体水平均有显著性升高；Ｏ－ ＳＰ－ ＴＴ能刺激高水平抗Ｏ－ ＳＰＩｇＧ、ＩｇＭ 抗体，而死菌对ＩｇＧ的作用较差。Ｏ－ ＳＰ－ ＴＴ免疫后血清中抗体可激活补体产生杀菌作用，杀菌效价为１∶１６０ 。结论：本实验采用的化学方法均未破坏抗原的特异性；Ｏ－ ＳＰ单独使用无免疫原性；Ｏ－ ＳＰ－ ＴＴ注射后针对Ｏ－ ＳＰ、ＴＴ 均加强了免疫应答，并较死菌对ＩｇＧ的作用强，其刺激产生的抗体具有杀菌活性；本实验结果为Ｏ－ ＳＰ－ ＴＴ结合物的进一步应用研究提供了一定的基础资料。
OBJECTIVE: To prepare O-SP-TT, a conjugate of O-specific polysaccharide (O-SP) and tetanus toxoid (TT) of Shigella type I bacteria and detect its antigenicity and anti-O- SPIgG, IgM antibody levels and antibody bactericidal activity. Methods: O-SP purified from Shigella flexneri I was collected, and the conjugate of O-SP-TT was prepared by using diphthalic acid dihydrazide (ADH) as linker and tetanus toxoid as carrier. The characteristics of extracts, intermediates and conjugates were tested. Results: O-SP, O-SP-AH and O-SP-TT all formed clear precipitation lines with rabbit hyperimmune serum. There was no significant increase in antibody levels in mice immunized with O-SP alone. The level of IgG and anti-TT antibody between the first and second time and the second and third time after O-SP-TT injection were significantly increased. O-SP-TT stimulated high level of anti-O-IggG and IgM Antibodies, while the effect of dead bacteria on IgG is poor. Antibody in serum after O-SP-TT immunization can activate the complement to produce bactericidal effect, the sterilization titer is 1:160. Conclusion: The chemical methods used in this experiment did not destroy the antigen specificity; O-SP alone without immunogenicity; O-SP-TT injection for the O-SP, TT were to strengthen the immune response, The strong effect of IgG, the antibody stimulated by it has bactericidal activity; The experimental results provide some basic data for the further application of the O-SP-TT conjugate.