Corbeil L B, Arthur J E, Widders P R, Smith J W, Barbet A F
Infect Immun. 1987 Jun;55(6):1381-6. doi: 10.1128/iai.55.6.1381-1386.1987.
The antigens of Haemophilus somnus recognized by convalescent bovine serum were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with a protein A-peroxidase conjugate. The same two 76K and 40K antigens were predominant in whole-bacterium preparations and in outer-membrane-enriched, Triton X-100-insoluble fractions. The surface location of these two antigens was confirmed by absorbing antiserum with whole, live bacteria. Absorption with H. somnus removed antibody reactivity for the 76K antigen and reduced reactivity for the 40K antigen. Absorption with Pasteurella multocida, Actinobacillus equuli, or Escherichia coli did not reduce reactivity, and results with Pasteurella haemolytica were equivocal. The two immunodominant antigens detected in this study were conserved in isolates of H. somnus from thromboembolic meningoencephalitis, pneumonia, reproductive failure, or asymptomatic carriers. Convalescent sera from nearly all 17 cattle studied recognized these two antigens. Other antigens were recognized less consistently. Although other antigens may also be involved, the 76K and 40K surface antigens of H. somnus appear to be important candidates for a subunit vaccine or an immunodiagnostic assay.
利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及与蛋白A-过氧化物酶结合物的Western印迹法,研究了恢复期牛血清识别的睡眠嗜血杆菌抗原。在全菌制剂以及富含外膜的、不溶于Triton X-100的组分中,同样的两种76K和40K抗原占主导地位。通过用完整的活细菌吸收抗血清,证实了这两种抗原的表面定位。用睡眠嗜血杆菌吸收可消除针对76K抗原的抗体反应性,并降低针对40K抗原的反应性。用多杀巴斯德菌、马驹放线杆菌或大肠杆菌吸收不会降低反应性,而溶血巴斯德菌的结果不明确。在本研究中检测到的两种免疫显性抗原在来自血栓栓塞性脑膜脑炎、肺炎、生殖障碍或无症状携带者的睡眠嗜血杆菌分离株中是保守的。几乎所有研究的17头牛的恢复期血清都识别这两种抗原。对其他抗原的识别则不太一致。尽管可能还涉及其他抗原,但睡眠嗜血杆菌的76K和40K表面抗原似乎是亚单位疫苗或免疫诊断检测的重要候选物。