流感嗜血杆菌D蛋白作为A群脑膜炎奈瑟菌多糖蛋白结合疫苗蛋白载体的免疫效果评价  

Evaluation on immune effect of meningococcal polysaccharide conjugate vaccine using protein D of Haemophilus influenzae as a carrier

作　　者：王丽婵 谭亚军[1] 卫辰[1] 张庶民[1] 张华捷[1] 马霄[1] WANG Li-chan;TAN Ya-jun;WEI Chen;ZHANG Shu-min;ZHANG Hua-jie;MA Xiao(Division of Diphtheria,Tetanus and Pertussis Vaccine and Toxins,National Institutes of Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院百白破疫苗与毒素室卫生部生物技术产品检定方法及其标准化重点实验室,北京102629

出　　处：《中国生物制品学杂志》2021年第9期1023-1027,1032,共6页Chinese Journal of Biologicals

基　　金：国家科技重大专项重大新药创制(2018ZX09738005)。

摘　　要：目的对流感嗜血杆菌D蛋白(protein D,PD)作为A群脑膜炎奈瑟菌多糖蛋白结合疫苗蛋白载体的免疫效果进行评价。方法采用CDAP活化法,将PD、破伤风类毒素(tetanus toxin,TT)分别与A群脑膜炎奈瑟菌多糖结合制备结合物;将结合物分别免疫BALB/c小鼠,腹部皮下免疫3次,每次加强免疫前及末次免疫后7 d经眼眶采血,分离血清,间接ELISA法检测抗体水平及亚类,同时在末次免疫后7 d取小鼠脾脏,检测细胞免疫水平。结果 PD与A群多糖结合后能增强抗-多糖抗体的IgG水平,第3次免疫后抗体水平达1∶12 800,显著高于单独多糖组(1∶400)(P<0.05);其脾细胞经多糖及多糖蛋白混合物刺激后,IFNγ和IL-4的斑点形成细胞数量与多糖组比较均升高(P<0.05);但Th1/Th2亚群比值与单独多糖组比较并未增高(P>0.05),反而显著低于GAMP-TT组和单独PD组(P<0.05)。结论重组PD与A群多糖结合后能够增强多糖的免疫原性,既可提高体液免疫反应能力,又可激发细胞免疫反应,提示PD具备作为A群脑膜炎奈瑟菌多糖蛋白载体的潜力。Objective To evaluate the immune effect of group A meningococcal polysaccharide conjugate vaccine using recombinant protein D(PD) of Haemophilus influenae as a carrier.Methods PD and tetanus toxoid(TT) were conjugated to group A meningococcal capsular polysaccharide by CDAP activation method respectively.BALB/c mice were immunized s.c.with the conjugates respectively for 3 times,of which the serum samples were collected before each booster immunization and 7 d after the last immunization,and determined for antibody level and subclass by indirect ELISA.The spleens of mice were collected 7 d after the last immunization and determined for cellular immunity.Results The conjugation of PD to group A meningococcal polysaccharide increased the IgG level against the polysaccharide.The antibody level after the third immunization with the conjugate was 1∶12 800,which was significantly higher than that with polysaccharide alone(1 ∶ 400)(P<0.05).The number of spleen cells with spot formation by IFNγ and IL-4 after stimulation with polysaccharide and polysaccharide-protein conjugate in the mice immunization with polysaccharide-PD conjugate increased as compared with that with polysaccharide alone(P<0.05),while the ratio of Th1/Th2 subsets showed no significant increase(P > 0.05)and was significantly lower than that with GAMP-TT and with PD alone(P<0.05).Conclusion The conjugation of recombinant PD and group A meningococcal polysaccharide enhanced the immu-nogenicity of polysaccharide,increased the humoral immune response and stimulated the cellular immune response,indicating the potential of PD as a protein carrier of polysaccharide of group A Neisseria meningitidis.

关 键 词：流感嗜血杆菌 D蛋白 A群脑膜炎奈瑟菌 荚膜多糖 载体 体液免疫 细胞免疫 

分 类 号：K392.33[历史地理—历史学]