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首页> 美国卫生研究院文献>Iranian Journal of Veterinary Research >Evaluation of immunogenicity and protective efficacy of recombinant ptfA of avian Pasteurella multocida
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Evaluation of immunogenicity and protective efficacy of recombinant ptfA of avian Pasteurella multocida

机译：禽多杀性巴氏杆菌重组ptfA的免疫原性和保护功效的评价

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摘要

Avian Pasteurella multocida is the causative agent of fowl cholera, a disease much affecting the poultry industry. In order to study the efficacy of the recombinant subunit vaccine constructed with ptfA gene of avian P. multocida, the ptfA gene fragment amplified by PCR from avian P. multocida was cloned into the prokaryotic expression vector pET32a and the recombinant plasmid pET32a-ptfA was obtained. The pET32a-ptfA was expressed in Escherichia coli BL21(DE3) and the target protein rPtfA was purified. The purified protein was then mixed with Freund’s adjuvant and the recombinant subunit vaccine was obtained. Three groups of chickens labeled as rPtfA, attenuated live vaccine and PBS were vaccinated with the recombinant subunit vaccine, attenuated live vaccine and PBS, respectively. Serum antibodies, peripheral blood lymphocyte proliferation (PBLP) and interferon-γ (IFN-γ) level secreted by peripheral blood lymphocyte were tested. The immunized chickens were finally challenged with virulent avian P. multocida and the protection rate was counted. Indirect ELISA showed the levels of antibodies in rPtfA and attenuated vaccine groups were most significantly higher than the other groups (P<0.01), and the former was slightly lower than the latter. Peripheral blood lymphocyte proliferation experiments and IFN-γ experiments indicated that SI value and the levels of IFN-γ induced by ConA in the two vaccine groups were significantly higher than those of the PBS groups (P<0.01), and that the attenuated vaccine group was higher than the rPtfA group. The protection rates of rPtfA and attenuated live vaccines were 45% and 75%, respectively. The results indicated that the PtfA recombinant subunit vaccine was capable of improving the immunity level and inducing a protective effect for the vaccinated chickens, but it was barely satisfactory.
机译：禽多杀巴斯德氏菌是禽霍乱的病原体，该病对家禽业影响很大。为了研究用禽多杀性疟原虫的ptfA基因构建的重组亚单位疫苗的功效，通过PCR从禽多杀性疟原虫中扩增得到的ptfA基因片段被克隆到原核表达载体pET32a中，得到重组质粒pET32a-ptfA 。 pET32a-ptfA在大肠杆菌BL21（DE3）中表达，并纯化目标蛋白rPtfA。然后将纯化的蛋白质与弗氏佐剂混合，获得重组亚单位疫苗。将三组分别标记为rPtfA，减毒活疫苗和PBS的鸡分别接种重组亚单位疫苗，减毒活疫苗和PBS。检测外周血淋巴细胞分泌的血清抗体，外周血淋巴细胞增殖（PBLP）和干扰素-γ（IFN-γ）水平。最后，用强毒的禽多杀性巴氏杆菌攻击免疫的鸡，并计算保护率。间接ELISA显示rPtfA和减毒疫苗组中的抗体水平最显着高于其他组（P <0.01），前者略低于后者。外周血淋巴细胞增殖实验和IFN-γ实验表明，两个疫苗组中ConA诱导的SI值和IFN-γ水平明显高于PBS组（P <0.01），减毒疫苗组高于rPtfA组。 rPtfA和减毒活疫苗的保护率分别为45％和75％。结果表明，PtfA重组亚单位疫苗能够提高免疫水平并诱导对接种鸡的保护作用，但效果不理想。

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