Rodriguez F, Kennedy S, Bryson T D, Fernandez A, Rodriguez J L, Ball H J
Department of Agriculture for Northern Ireland, Veterinary Sciences Division, Belfast.
Zentralbl Veterinarmed B. 1996 Sep;43(7):429-38. doi: 10.1111/j.1439-0450.1996.tb00335.x.
Lung samples from pneumonic lesions in cattle and goats, naturally or experimentally infected with strains of the Mycoplasma mycoides cluster, were fixed in formalin and embedded in paraffin. An immunohistochemical technique using monoclonal or polyclonal antibodies was performed on tissue sections in order to detect Mycoplasma antigens. Four monoclonal antibodies (MAbs), one (2A3) raised against M. mycoides ssp. mycoides small colony (SC) and large colony (LC), two (1D3 and 5E5) against M. mycoides ssp. capri, and one (5A10) against M. bovis, were used. A range of polyclonal antibodies, raised to the individual subspecies of the M. mycoides cluster, and one to Pasteurella haemolytica, was also used. The MAb 2A3 showed positive immunostaining in lung sections from cattle and goats naturally and experimentally infected with M. mycoides ssp. mycoides SC and LC, but not with pneumonic lesions of cattle and goats due to other members of the M. mycoides cluster, M. bovis or Pasteurella spp. The MAb 1D3 showed immunostaining in lung sections from goats naturally and experimentally infected with M. mycoides ssp. capri, but again not with pneumonic lesions caused by other members of the M. mycoides cluster, M. bovis or Pasteurella spp. The MAb 5E5 immunoreacted in sections from pneumonic lesions from all animals infected with one of the three M. mycoides cluster subspecies used in the study, but not with M. bovis or Pasteurella infected tissue. Immunoreaction was mainly found in the cell debris around necrotic areas, as well as in macrophages, neutrophils and epithelial cells. The localization of antigens of the M. mycoides cluster using polyclonal antisera followed basically the same pattern as that obtained with the monoclonals. However, a wide cross reactivity was found between different antisera and relatively high background immunostaining was also seen, especially in necrotic areas. The results suggest that immunohistochemical methods using monoclonal antibodies are useful tools for the diagnosis and study of the pathogenesis of pneumonia caused by the Mycoplasmas of the M. mycoides cluster.
采集自然感染或实验感染丝状支原体簇菌株的牛和山羊肺部肺炎病变样本，用福尔马林固定并石蜡包埋。对组织切片进行免疫组化技术，使用单克隆或多克隆抗体以检测支原体抗原。使用了四种单克隆抗体（MAb），一种（2A3）针对丝状支原体丝状亚种小菌落（SC）和大菌落（LC）产生，两种（1D3和5E5）针对丝状支原体山羊亚种产生，一种（5A10）针对牛支原体产生。还使用了一系列针对丝状支原体簇各个亚种产生的多克隆抗体，以及一种针对溶血巴斯德菌产生的多克隆抗体。单克隆抗体2A3在自然感染和实验感染丝状支原体丝状亚种SC和LC的牛和山羊肺部切片中显示出阳性免疫染色，但在由丝状支原体簇其他成员、牛支原体或巴斯德菌属引起的牛和山羊肺炎病变中未显示阳性。单克隆抗体1D3在自然感染和实验感染丝状支原体山羊亚种的山羊肺部切片中显示出免疫染色，但同样在由丝状支原体簇其他成员、牛支原体或巴斯德菌属引起的肺炎病变中未显示阳性。单克隆抗体5E5在感染本研究中使用的三种丝状支原体簇亚种之一的所有动物的肺炎病变切片中产生免疫反应，但在牛支原体或巴斯德菌感染的组织中未产生免疫反应。免疫反应主要见于坏死区域周围的细胞碎片以及巨噬细胞、中性粒细胞和上皮细胞中。使用多克隆抗血清对丝状支原体簇抗原的定位基本遵循与单克隆抗体相同的模式。然而，在不同抗血清之间发现了广泛的交叉反应，并且还观察到相对较高的背景免疫染色，尤其是在坏死区域。结果表明，使用单克隆抗体的免疫组化方法是诊断和研究由丝状支原体簇支原体引起的肺炎发病机制的有用工具。
