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鸡胚培养的多杀性巴氏杆菌在鸡体内的交叉保护作用及抗原表达

Cross-protection and antigen expression by chicken embryo-grown Pasteurella multocida in chickens.

作者信息

Ibrahim R S, Sawada T, Shahata M, Ibrahim A

机构信息

Department of Poultry Diseases, Assiut University, Assiut, Egypt.

出版信息

J Comp Pathol. 2000 Nov;123(4):278-84. doi: 10.1053/jcpa.2000.0419.

Abstract

The growth of Pasteurella multocida strain X-73 (serotype 1) and P-1059 (serotype 3) in vivo instead of in vitro resulted in the expression of additional antigens, as revealed by SDS-PAGE profile and western blotting of the outer membrane detergent-insoluble fraction (DIF-OM). Minor protein bands ranging from 58-138 kDa were expressed by both strains; major bands ranging from 35-64 and 35-43 kDa were expressed by X-73 and P-1059, respectively. Antigens at 35-39 kDa were dense and dominant. Treatment of DIF-OM from both strains with sodium periodate and proteinase K, but not with trypsin, reduced ELISA reactivity; this suggested that DIF-OM was composed of glycoprotein. Western blotting with heterologous antisera demonstrated antigenic cross-reactivity between strains X-73 and P-1059 grown in vivo. Inactivated vaccine prepared from X-73 grown in vivo protected chickens against challenge with X-73, P-1059 and strain P-1662 (serotype 4); a similar vaccine prepared from in-vitro culture, however, protected only against the homologous serotype. Moreover, antiserum against X-73 grown in vivo but not against the same strain grown in vitro gave considerable passive protection against challenge with the heterologous strains P-1059 and P-1662.

摘要

多杀性巴氏杆菌X-73株(血清型1)和P-1059株(血清型3)在体内而非体外生长时,会表达额外的抗原,这通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱和外膜去污剂不溶性组分(DIF-OM)的蛋白质印迹法得以揭示。两株菌均表达了范围在58 - 138 kDa的次要蛋白条带;X-73株和P-1059株分别表达了范围在35 - 64 kDa和35 - 43 kDa的主要条带。35 - 39 kDa的抗原密集且占主导地位。用高碘酸钠和蛋白酶K而非胰蛋白酶处理两株菌的DIF-OM,降低了酶联免疫吸附测定反应性;这表明DIF-OM由糖蛋白组成。用异源抗血清进行的蛋白质印迹法显示,体内生长的X-73株和P-1059株之间存在抗原交叉反应性。由体内生长的X-73株制备的灭活疫苗可保护鸡免受X-73株、P-1059株和P-1662株(血清型4)的攻击;然而,由体外培养物制备的类似疫苗仅能保护免受同源血清型的攻击。此外,针对体内生长的X-73株而非体外生长的同一菌株的抗血清,对异源菌株P-1059株和P-1662株的攻击给予了相当程度的被动保护。

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