Lu Y S, Aguila H N, Lai W C, Pakes S P
Department of Pathology, University of Texas, Southwestern Medical Center, Dallas 75235.
Infect Immun. 1991 Apr;59(4):1470-5. doi: 10.1128/iai.59.4.1470-1475.1991.
The role of rabbit antibodies against Pasteurella multocida outer membrane proteins and lipopolysaccharides (LPS) in resistance remains unknown. Pooled immune sera against P. multocida outer membranes were prepared from specific-pathogen-free rabbits immunized with sucrose gradient-purified P. multocida outer membranes. Western immunoblotting showed that purified outer membrane protein antibodies reacted strongly against the outer membrane proteins but not the purified LPS. Affinity-purified LPS antibodies exhibited strong reactivity against purified LPS and very little reactivity against outer membrane vesicles. Mice were inoculated intranasally with immune serum or normal rabbit serum, challenged intranasally with 10(6) CFU of P. multocida, and euthanatized 48 h later to determine the number of P. multocida organisms in the lungs. Mice inoculated with pooled immune serum had a 3,300-fold reduction (P less than 0.001) in the numbers of P. multocida in the lungs as compared with the controls. Similarly, mice inoculated with purified outer membrane protein antibodies had a 201-fold reduction (P less than 0.001) in the numbers of P. multocida. Conversely, mice inoculated with affinity-purified LPS antibodies had a 1.1-fold reduction (P greater than 0.50) in the numbers of P. multocida. These results show that antibodies against the outer membrane proteins but not the LPS are the components of rabbit immune sera which inhibit P. multocida proliferation in mouse lungs.
兔抗多杀性巴氏杆菌外膜蛋白和脂多糖(LPS)抗体在抵抗作用中的角色仍不清楚。用蔗糖梯度纯化的多杀性巴氏杆菌外膜免疫无特定病原体的兔,制备抗多杀性巴氏杆菌外膜的混合免疫血清。蛋白质免疫印迹显示,纯化的外膜蛋白抗体与外膜蛋白反应强烈,但与纯化的LPS无反应。亲和纯化的LPS抗体对纯化的LPS表现出强反应性,对外膜囊泡的反应性非常低。给小鼠鼻内接种免疫血清或正常兔血清,然后鼻内用10(6) CFU多杀性巴氏杆菌攻击,48小时后安乐死以确定肺中多杀性巴氏杆菌的数量。与对照组相比,接种混合免疫血清的小鼠肺中多杀性巴氏杆菌数量减少了3300倍(P<0.001)。同样,接种纯化外膜蛋白抗体的小鼠肺中多杀性巴氏杆菌数量减少了201倍(P<0.001)。相反,接种亲和纯化LPS抗体的小鼠肺中多杀性巴氏杆菌数量减少了1.1倍(P>0.50)。这些结果表明,兔免疫血清中抑制小鼠肺中多杀性巴氏杆菌增殖的成分是抗外膜蛋白抗体而非抗LPS抗体。