Borrathybay Entomack, Peng Qingzhong, Yan Fang, He Cui, Nazierbieke Wulumuhan
College of Biology and Environmental Sciences, Jishou University, Jishou 416000, China.
Wei Sheng Wu Xue Bao. 2010 Mar;50(3):360-6.
We evaluated the cross-protective effect of a 39-kDa adhesive protein (Cp39), recombinant adhesive protein (rCp39), and capsular proteins from in vitro and in vivo grown avian P. multocida in mice.
The Cp39 was purified by electroelution from capsular proteins of in vivo grown strain C48-3. The rCp39 was expressed in E. coli B21 as a soluble protein by IPTG inducing, and purified with pMAL protein fusion and purification system. Mice of each group were subcutaneously immunized twice with 100 microg of rCp39, Cp39 or capsular proteins from in vivo or in vitro grown strain C48-3 with in complete or incomplete Freund adjuvant at 2-week intervals. Five mice of each group were challenged with 100 LD50 of avian P. multocida strain C48-3 or rabbit P. multocida strain C51-3 two weeks after the second immunization, while the antibody response specific for rCp39 was determined by ELISA.
SDS-PAGE indicated that the Cp39 protein was expressed in vitro and in vivo grown P. multocida. Mice immunized with rCp39, native Cp39 or capsular proteins from in vivo and in vitro grown strain C48-3 were protected completely against the challenge with the homologous strain C48-3, while 60 - 80% protection was demonstrated in the mice against the challenge with the heterologous strain C51-3.
In conclusion, rCp39 is a cross immunoprotective antigen of P. multocida capsular serogroup A strains, and might be a useful vaccine candidate against fowl cholera.
我们评估了39 kDa黏附蛋白（Cp39）、重组黏附蛋白（rCp39）以及体内和体外培养的禽多杀性巴氏杆菌荚膜蛋白在小鼠体内的交叉保护作用。
通过电洗脱从体内培养菌株C48 - 3的荚膜蛋白中纯化出Cp39。rCp39通过IPTG诱导在大肠杆菌B21中表达为可溶性蛋白，并使用pMAL蛋白融合与纯化系统进行纯化。每组小鼠每隔2周皮下免疫两次，每次100 μg的rCp39、Cp39或体内或体外培养的菌株C48 - 3的荚膜蛋白，使用不完全或完全弗氏佐剂。每组5只小鼠在第二次免疫后两周用100 LD50的禽多杀性巴氏杆菌菌株C48 - 3或兔多杀性巴氏杆菌菌株C51 - 3进行攻毒，同时通过ELISA测定针对rCp39的抗体反应。
SDS - PAGE表明Cp39蛋白在体内和体外培养的多杀性巴氏杆菌中均有表达。用rCp39、天然Cp39或体内和体外培养的菌株C48 - 3的荚膜蛋白免疫的小鼠对同源菌株C48 - 3的攻毒具有完全保护作用，而对异源菌株C51 - 3的攻毒，小鼠的保护率为60 - 80%。
总之，rCp39是多杀性巴氏杆菌荚膜A血清群菌株的交叉免疫保护抗原，可能是一种用于预防禽霍乱的有效疫苗候选物。
