摘要
目的制备伤寒Vi结合疫苗并研究其特性。方法载体蛋白质重组铜绿假单胞菌外毒素A(rEPA)经ADH衍生后,与伤寒Vi多糖结合制备结合物,并对结合物进行化学特性和生物学特性的研究,尤其是研究结合物对小鼠的免疫原性。结果制备的伤寒Vi结合物相对分子质量(Mr)明显大于游离的伤寒Vi多糖和载体蛋白质,其多糖和蛋白质比值为1．2,结合物含量测定证明至少有70％的多糖是以结合状态存在,HPLC(TSK5000)的结果显示伤寒Vi结合物中Kd(partition coefficient)≤0．2占94％,内毒素含量、过敏原性、豚鼠和小鼠的动物毒性试验等都证明结合物安全,免疫双扩散试验证明结合物的多糖和蛋白质组分的抗原性都完好保持,在小鼠上的免疫原性试验证明结合物具有与单纯Vi多糖完全不同的免疫学特性,可以产生更高的特异性免疫应答,并具有明显的加强免疫效果。结论按试验设计方案制备的伤寒Ⅵ结合疫苗,经化学和生物学检测,证明结合物确以多糖蛋白质结合状态存在,并证实该结合物具有较好的生物安全性和免疫学效果。
Objective To prepare a typhoid Vi conjugate vaccine and investigate its characteristics. Methods Recombinant exoprotein A（rEPA） of Pseudomonas aeruginosa was derived with adipic acid dihydrazide （ADH） and bound to the capsular polysaccharide of Salmonella typhi （Vi）. The conjugates were analyzed with proper chemical and biological methods. Immunogenicity of the conjugates was tested in particular in mice model. Results The conjugates were much larger in size than polysaccharide and cartier protein alone. The Vi/protein ratio （wt/wt） was 1.2 and more than 70% of Vi stayed in conjugate form. Analysis of HPLC（TSK5000） showed that the conjugates with Kd（ partition coefficient） ≤ 0.2 occupied 94%. Endotoxin content, anaphylactoid test, abnormal toxicity test in guinea pigs and mice gave evidence of the safety of the conjugates. Double-immunodiffusion demonstrated that both Vi and rEPA in the conjugates kept intact immunogerticity. Immunogerticity test in mice showed that the conjugates could induce significant responses, and elicit boosted responses while Vi not. Conclusion Vi conjugate vaccine was successsfully developed. It was safe and got strong immunngenicity in mice.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第7期665-668,共4页
Chinese Journal of Microbiology and Immunology
关键词
伤寒
结合疫苗
特性
Typhoid
Conjugates vaccine
Characteristic
