一种39千道尔顿的蛋白质介导禽多杀性巴氏杆菌与鸡胚成纤维细胞的黏附。

A 39 kDa protein mediates adhesion of avian Pasteurella multocida to chicken embryo fibroblast cells.

作者信息

Borrathybay Entomack, Sawada Takuo, Kataoka Yasushi, Ohtsu Naoko, Takagi Masami, Nakamura Shigeyuki, Kawamoto Eiichi

机构信息

Department of Veterinary Microbiology, Nippon Veterinary and Animal Science University, Musashino, Tokyo 180-8602, Japan.

出版信息

Vet Microbiol. 2003 Dec 30;97(3-4):229-43. doi: 10.1016/j.vetmic.2003.09.010.

DOI:10.1016/j.vetmic.2003.09.010

PMID:14654293

Abstract

To clarify the role of avian Pasteurella multocida capsule in pathogenesis, adhesion of capsulated strains P-1059, X-73 and Pm-18, and noncapsulated strains P-1059B, Pm-1 and Pm-3 to chicken embryo fibroblast (CEF) cells was compared. Number of adherent organisms of the capsulated strains to CEF cells were approximately three times as much as noncapsulated strains indicating that adhesive properties were enhanced by the presence of bacterial capsule. Pretreatments of the bacterial cells with heat, trypsin, or with antiserum caused a marked decrease in adhesion of capsulated strain P-1059 and its noncapsulated variant P-1059B. However, depolymerization of capsular hyaluronic acid with high dose of hyaluronidase enhanced adhesion of these strains. Combined treatments of the bacterial cells with both hyaluronidase and trypsin significantly (P < 0.05) inhibited the adherence of strain P-1059 as compared to the treatment only with trypsin, but strain P-1059B was not affected. SDS-PAGE profiles of crude capsular extract (CCE) prepared from capsulated strain P-1059 and its noncapsulated variant P-1059B grown on dextrose starch agar (DSA) plates by heating at 56 degrees C in a 2.5% NaCl solution demonstrated eight protein bands of 28, 34, 36, 39, 52, 56, 63 and 93 kDa. The 28, 34 and 36 kDa proteins were commonly major for both strains, and the 39 kDa protein was major only for strain P-1059 but poor in strain P-1059B. Outer membrane protein (OMP) profiles were identical with a major protein at 34 kDa and four minor proteins between the two strains. The adhesion of strain P-1059 and strain P-1059B to CEF cells was inhibited significantly (P < 0.01) by treatment with rabbit antisera against P-1059, P-1059B, CCE or 39 kDa protein of strain P-1059 as compared to the treatment with either PBS or with normal rabbit serum. These results indicated that an antigenic 39 kDa protein in the capsule may be responsible for adhesion of avian P. multocida type A strains to CEF cells as a virulence factor.

摘要

为阐明禽多杀性巴氏杆菌荚膜在发病机制中的作用，比较了荚膜菌株P - 1059、X - 73和Pm - 18以及非荚膜菌株P - 1059B、Pm - 1和Pm - 3对鸡胚成纤维细胞（CEF）的黏附情况。荚膜菌株对CEF细胞的黏附菌数约为非荚膜菌株的三倍，表明细菌荚膜的存在增强了黏附特性。用热、胰蛋白酶或抗血清预处理细菌细胞，导致荚膜菌株P - 1059及其非荚膜变体P - 1059B的黏附显著降低。然而，用高剂量透明质酸酶使荚膜透明质酸解聚增强了这些菌株的黏附。与仅用胰蛋白酶处理相比，用透明质酸酶和胰蛋白酶联合处理细菌细胞显著（P < 0.05）抑制了菌株P - 1059的黏附，但菌株P - 1059B不受影响。通过在2.5% NaCl溶液中56℃加热，从在葡萄糖淀粉琼脂（DSA）平板上生长的荚膜菌株P - 1059及其非荚膜变体P - 1059B制备的粗荚膜提取物（CCE）的SDS - PAGE图谱显示有28、34、36、39、52、56、63和93 kDa的八条蛋白带。28、34和36 kDa的蛋白是两株菌的共同主要蛋白，39 kDa的蛋白仅在菌株P - 1059中为主，而在菌株P - 1059B中含量较少。两株菌的外膜蛋白（OMP）图谱相同，有一个34 kDa的主要蛋白和四个次要蛋白。与用PBS或正常兔血清处理相比，用抗P - 1059、P - 1059B、CCE或菌株P - 1059的39 kDa蛋白的兔抗血清处理显著（P < 0.01）抑制了菌株P - 1059和菌株P - 1059B对CEF细胞的黏附。这些结果表明，荚膜中的一种39 kDa抗原性蛋白可能作为毒力因子，介导禽A型多杀性巴氏杆菌菌株对CEF细胞的黏附。