Hensel A, Stockhofe-Zurwieden N, Petzoldt K, Lubitz W
Institute of Microbiology and Genetics, Biocentre, University of Vienna, Austria.
Infect Immun. 1995 Aug;63(8):3048-53. doi: 10.1128/iai.63.8.3048-3053.1995.
A dose-defined aerosol infection of pigs was used to study the immunogenic and protective potentials of oral immunization with dead or live Actinobacillus pleuropneumoniae serotype 9 reference strain CVI 13261 against an aerogenic challenge. Pigs were vaccinated with a single dose of 10(11) CFU of viable (n = 8) or inactivated (n = 8) A. pleuropneumoniae given orally in a gelatin capsule. After 3 weeks, vaccinated pigs and nonvaccinated controls were challenged aerogenically with a dose of 10(8) CFU of A. pleuropneumoniae CVI 13261. The protective efficacy of oral immunization was evaluated by clinical and postmortem examinations. Bronchoalveolar lavage in pigs was performed during the experiment to obtain lavage samples for assessment of local antibodies. Isotype-specific antibody responses in sera and in bronchoalveolar lavage fluids were determined by enzyme-linked immunosorbent assays based on whole-cell antigen. Oral immunization did not induce clinical side effects. After aerosol challenge, two animals of both vaccinated groups (25% in each case) showed a moderate fever for 2 days, whereas all four pigs (100%) of the nonvaccinated control group developed severe fever. In contrast to the controls, which developed severe pleuropneumonia, the vaccinated pigs had only mild pulmonary lesions. Three weeks after challenge, 13 of 16 vaccinated pigs (81%) were found to be free of pathomorphological changes of the lungs. From two of these pigs immunized with live bacteria we were able to reisolate A. pleuropneumoniae. A significant systemic and pulmonary increase in the concentrations of immunoglobulin A (IgA), IgM, and IgG antibodies reactive with A. pleuropneumoniae was detectable after aerosol challenge in both vaccinated groups. Immunization with viable bacteria was found to induce significantly higher concentrations of each Ig isotype in bronchoalveolar lavage fluids and sera than immunization with inactivated A. pleuropneumoniae. These serological findings were not reflected in the reduction in clinical disease after challenge in comparison to the case for the pigs vaccinated with inactivated bacteria. We concluded that a single oral administration of A. pleuropneumoniae provides partial clinical protection against aerosol challenge infection in the respiratory tract.
采用剂量明确的猪气溶胶感染模型,研究口服灭活或活的胸膜肺炎放线杆菌9型参考菌株CVI 13261对气溶胶攻击的免疫原性和保护潜力。给猪口服一粒明胶胶囊,其中含有10(11) CFU的活(n = 8)或灭活(n = 8)胸膜肺炎放线杆菌单剂量疫苗。3周后,给接种疫苗的猪和未接种疫苗的对照猪进行10(8) CFU胸膜肺炎放线杆菌CVI 13261的气溶胶攻击。通过临床和尸检评估口服免疫的保护效果。在实验期间对猪进行支气管肺泡灌洗以获取灌洗样本,用于评估局部抗体。基于全细胞抗原,通过酶联免疫吸附测定法测定血清和支气管肺泡灌洗液中的同型特异性抗体反应。口服免疫未引起临床副作用。气溶胶攻击后,两个接种组各有两只动物(各占25%)出现中度发热2天,而未接种疫苗的对照组的所有四只猪(100%)均出现高热。与出现严重胸膜肺炎的对照组相比,接种疫苗的猪仅有轻度肺部病变。攻击3周后,16只接种疫苗的猪中有13只(81%)肺部无病理形态学变化。从其中两只接种活菌的猪中,我们能够重新分离出胸膜肺炎放线杆菌。在两个接种组中,气溶胶攻击后均可检测到与胸膜肺炎放线杆菌反应的免疫球蛋白A(IgA)、IgM和IgG抗体浓度在全身和肺部显著增加。发现用活菌免疫比用灭活胸膜肺炎放线杆菌免疫在支气管肺泡灌洗液和血清中诱导的每种Ig同型浓度显著更高。与接种灭活菌的猪相比,这些血清学结果并未反映在攻击后临床疾病的减轻上。我们得出结论,单次口服胸膜肺炎放线杆菌可为呼吸道气溶胶攻击感染提供部分临床保护。