【摘要】 目的研制预防沙门菌属A组甲型副伤寒沙门菌感染的多糖蛋白结合疫苗。方法通过对甲型副伤寒沙门菌尼泊尔株NTP-6的发酵培养,热酚水法提取脂多糖(LPS),酸水解脱毒,纯化出有效的型特异O-SP抗原；然后用CDAP对O-SP活化、ADH衍生后,在EDAC的缩合作用下,结合到破伤风类毒素TT上,制备出甲型副伤寒结合疫苗。用含2．5μg多糖甲型副伤寒结合疫苗免疫小鼠,以LPS为包被抗原,用间接ELISA法测定血清抗甲型副伤寒LPS IgG抗体；并测定血清多糖抗体的补体介导杀菌活性；在小鼠和豚鼠体内观察甲型副伤寒结合疫苗的安全性。结果甲型副伤寒结合疫苗第2次免疫NIH后,小鼠血清抗甲型副伤寒LPS IgG抗体平均几何滴度(GMT)均较第1次有4倍以上升高,第3次免疫后,有明显的加强效应；抗体的补体介导杀菌活性效价达1：1280以上；小鼠和豚鼠体内甲型副伤寒结合疫苗安全性良好。结论成功建立了甲型副伤寒结合疫苗的制备工艺；制备的甲型副伤寒结合疫苗有良好的免疫原性,并具有明显的加强效应；刺激的血清多糖抗体有较高的补体介导杀菌活性效价；安全性达到人用生物制品相关要求。更多还原

【Abstract】 Objective To develop polysaccharide-protein conjugate vaccine for proventing Salmonella paratyphi A infection.Methods Salmonella paratyphi A NTP-6 strain was fermented,then LPS was extracted with bot-phenyl method and detoxificated with 1% acetic acid at 100℃for 1.5 h; the O-SP mixture was purified with Sephadex G-75,and the first and second peak were collected as effective polysaceharide antigen.O-SP was activated with CDAP,bound to TT with ADH as a spacer,and condensed with EDAC.Solutions of 2.5μg of sac- charide,alone or as conjugate,were injected subcutaneous into young mice.Antibodies against LPS in serum of the mice were measured by ELISA.Complement-mediated bactericidal activity was also assayed.The safety of con- jugate vaccine was evaluated in mice and guinea pig.Results After the second injection,the mean geometric titer (GMT)of anfi-LPS IgG increased by mere than 4 times,and the third injection showed significantly booster re- sponse.In the complement-mediated bactericidal activity test,the titers of antiserum were above 1:1280.In mice and guinea pig,conjugate vaccine had not shown any harmful effect.Conclusion A Salmonella paratyphi A con- jugate vaccine preparation procedure was successfully constructed.The TI antigen of O-SP was effectively converted into TD antigen; Clinical evaluation of S.paratyphi A conjugate vaccine is planned.更多还原