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用多杀性巴氏杆菌重组黏附蛋白疫苗接种对鸡进行禽霍乱防护。

Protection of chickens from fowl cholera by vaccination with recombinant adhesive protein of Pasteurella multocida.

作者信息

Sthitmatee Nattawooti, Numee Sureerat, Kawamoto Eiichi, Sasaki Hiraku, Yamashita Kaoru, Takahashi Naoyuki, Kataoka Yasushi, Sawada Takuo

机构信息

Laboratory of Veterinary Microbiology, Nippon Veterinary and Life Science University, Musashino, Tokyo 180-8602, Japan.

出版信息

Vaccine. 2008 May 2;26(19):2398-407. doi: 10.1016/j.vaccine.2008.02.051. Epub 2008 Mar 14.

Abstract

The recombinant adhesive protein (rCp39) of Pasteurella multocida strain P-1059 (serovar A:3) was prepared and purified with a hybrid condition of affinity chromatography. The rCp39 was highly protective for chickens from fowl cholera by challenge-exposure with parental strain P-1059 or heterologous strain X-73 (serovar A:1) compared to various kind of vaccines. Sixteen groups of ten chickens each were subcutaneously inoculated twice with 100, 200 or 400 microg proteins of rCp39, native Cp39, native outer membrane protein H (OmpH) or recombinant OmpH, or 100 microg proteins of crude capsular extract (CCE) of strains P-1059 or X-73 at 2 weeks interval. Five chickens of each group were challenge-exposed with each strain 2 weeks after the second inoculation. As the results, 60-100% protections were demonstrated in the chickens against both strains. Fisher's exact test indicated no significant differences (P<0.05) in vaccine types and dosages. ELISA and Western blot analysis indicated that the chicken anti-rCp39 sera reacted to whole-cell lysate of parental or heterologous strains. In conclusion, rCp39 is a cross-protective recombinant adhesive antigen of P. multocida capsular serogroup A strains. Moreover, a hybrid condition of affinity chromatography was successfully demonstrated and protected the immunogenicity of recombinant protein.

摘要

制备并通过亲和层析混合条件纯化了多杀性巴氏杆菌P-1059株(血清型A:3)的重组黏附蛋白(rCp39)。与各种疫苗相比,通过用亲本菌株P-1059或异源菌株X-73(血清型A:1)进行攻毒暴露试验,rCp39对鸡霍乱具有高度保护作用。将16组每组10只鸡,每隔2周皮下接种两次100、200或400微克的rCp39、天然Cp39、天然外膜蛋白H(OmpH)或重组OmpH蛋白,或100微克P-1059或X-73菌株的粗荚膜提取物(CCE)蛋白。每组5只鸡在第二次接种后2周用每种菌株进行攻毒暴露试验。结果显示,鸡对两种菌株的保护率为60%-100%。Fisher精确检验表明疫苗类型和剂量无显著差异(P<0.05)。ELISA和Western blot分析表明,鸡抗rCp39血清与亲本或异源菌株的全细胞裂解物发生反应。总之,rCp39是多杀性巴氏杆菌荚膜血清群A菌株的一种交叉保护性重组黏附抗原。此外,成功展示了亲和层析的混合条件并保留了重组蛋白的免疫原性。

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