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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Analysis of the Humoral Immune Response toChlamydia pneumoniae by Immunoblotting and Immunoprecipitation
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Analysis of the Humoral Immune Response toChlamydia pneumoniae by Immunoblotting and Immunoprecipitation

机译：免疫印迹和免疫沉淀分析对肺炎衣原体的体液免疫反应

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Chlamydia pneumoniae is a widely spread agent of respiratory tract infections in humans. A reliable serodiagnosis of the disease is hampered by the poor knowledge about immunodominant antigens in C. pneumoniae infections. We applied a novel strategy to identify immunogenic proteins of C. pneumoniae TW183 combining metabolic radiolabeling of de novo-synthesized chlamydial antigens with immunoprecipitation. By this technique C. pneumoniae antigens of approximately 160, 97 to 99, 60 to 62, 40, 27, and 15 kDa were detected in the vast majority of sera from patients with a current C. pneumoniae infection. By immunoblotting purified elementary bodies of C. pneumoniae TW183 with the same sera, only the 60- to 62-kDa antigen could be detected consistently. Sequential immunoprecipitation performed at different stages of the chlamydial developmental cycle revealed that the 60- to 62-kDa antigen is strongly upregulated after 24 to 48 h of host cell infection and is presented as a major immunogen in both C. pneumoniae-infected patients and mice. We conclude that, due to its high sensitivity and concurrent preservation of conformational epitopes, metabolic radiolabeling of chlamydial antigens combined with immunoprecipitation may be a useful method to reveal important immunogens in respiratory C. pneumoniae infection which might have been missed by immunoblot analysis.
机译：肺炎衣原体是人类呼吸道感染的一种广泛传播媒介。对 C的免疫优势抗原的了解不足，阻碍了该疾病的可靠血清学诊断。肺炎感染。我们应用了一种新颖的策略来鉴定 C的免疫原性蛋白。肺炎TW183结合了从头合成的衣原体抗原的代谢放射标记和免疫沉淀。通过这种技术， C。在目前患有 C病的患者的绝大多数血清中检测到约160、97至99、60至62、40、27和15 kDa的肺炎抗原。肺炎感染。通过免疫印迹纯化的 C基本体。具有相同血清的肺炎链球菌TW183，只能一致地检测到60-62kDa的抗原。在衣原体发育周期的不同阶段进行的顺序免疫沉淀显示，宿主细胞感染24至48小时后，60至62 kDa抗原被强烈上调，并且在这两个C中均被视为主要的免疫原。感染肺炎的患者和小鼠。我们得出结论，由于衣原体抗原的代谢放射标记与免疫沉淀相结合，具有很高的敏感性并同时保留了构象表位，可能是揭示呼吸道C中重要免疫原的有用方法。免疫印迹分析可能遗漏了肺炎感染。

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