Gogolewski R P, Kania S A, Inzana T J, Widders P R, Liggitt H D, Corbeil L B
Infect Immun. 1987 Jun;55(6):1403-11. doi: 10.1128/iai.55.6.1403-1411.1987.
The ability of convalescent serum to passively protect calves against Haemophilus somnus-induced pneumonia was studied. Preimmune and convalescent serum were obtained from calves before or after recovery from experimental chronic H. somnus pneumonia. Passive protection was assessed in another group of calves by intrabronchial inoculation of H. somnus that had been incubated with preimmune or convalescent serum. Each calf was inoculated with each treatment in alternating caudal lung lobes. Twenty-four hours after inoculation almost no pneumonia was present in lungs inoculated with bacteria incubated with convalescent serum, whereas severe pneumonia was present in lungs inoculated with bacteria incubated with preimmune serum. Quantitation of calf pneumonia in both treatment groups indicated a significantly different protective capacity between convalescent serum and preimmune serum (P less than 0.0005). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western blotting of purified H. somnus lipopolysaccharide resulted in intense reactivity with convalescent serum, but no reactivity was detected with preimmune serum. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis of H. somnus outer membrane-enriched fractions, Western blots with convalescent serum gave intense reactions against H. somnus outer membrane antigens with apparent molecular masses of 78 and 40 kilodaltons and weaker reactions with 60-, 34-, 31-, 29-, 18-, and 15-kilodalton outer membrane antigens. No reactivity was detected with preimmune serum. Antibodies eluted from H. somnus after adsorption of convalescent serum reacted almost identically to unadsorbed convalescent serum in Western blots against bacterial outer membrane-enriched fractions. Thus, most of the antigens recognized by convalescent serum are likely to be on the bacterial surface and accessible to antibody. Surface antigens recognized by protective convalescent serum are candidate antigens for a subunit vaccine against H. somnus pneumonia.
研究了恢复期血清被动保护犊牛免受睡眠嗜血杆菌所致肺炎的能力。在实验性慢性睡眠嗜血杆菌肺炎恢复之前或之后，从犊牛获取免疫前血清和恢复期血清。通过将睡眠嗜血杆菌与免疫前或恢复期血清一起孵育后经支气管接种到另一组犊牛中，评估被动保护作用。每头犊牛在交替的尾侧肺叶接种每种处理。接种后24小时，接种与恢复期血清一起孵育的细菌的肺中几乎没有肺炎，而接种与免疫前血清一起孵育的细菌的肺中有严重肺炎。两个处理组犊牛肺炎的定量分析表明，恢复期血清和免疫前血清之间的保护能力有显著差异（P小于0.0005）。对纯化的睡眠嗜血杆菌脂多糖进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，随后进行蛋白质印迹分析，结果显示与恢复期血清有强烈反应，但未检测到与免疫前血清的反应。对富含睡眠嗜血杆菌外膜的组分进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，用恢复期血清进行蛋白质印迹分析，对表观分子量为78和40千道尔顿的睡眠嗜血杆菌外膜抗原有强烈反应，对60、34、31、29、18和15千道尔顿的外膜抗原有较弱反应。未检测到与免疫前血清的反应。恢复期血清吸附后从睡眠嗜血杆菌洗脱的抗体在针对富含细菌外膜的组分的蛋白质印迹分析中，与未吸附的恢复期血清反应几乎相同。因此，恢复期血清识别的大多数抗原可能位于细菌表面且可被抗体识别。保护性恢复期血清识别的表面抗原是针对睡眠嗜血杆菌肺炎的亚单位疫苗的候选抗原。
