Proteome and antigen profiling of Coxiella burnetii developmental forms
Proteome and antigen profiling of Coxiella burnetii developmental forms
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DOI：
10.1128/iai.00883-06
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发表时间：
2007-01-01
影响因子：
3.1
通讯作者：
Heinzen, Robert A.
中科院分区：
文献类型：
--
作者：
Coleman, Sherry A.;Fischer, Elizabeth R.;Heinzen, Robert A.
高耐药小细胞变异体（SCV）从大细胞变异体（LCV）产生的双相发育周期被认为是人类Q热病原体贝氏柯克斯体毒力的基础。本研究对C.进行贝氏体发育形式的研究，以深入了解其独特的生物学和免疫学特性。通过二维（2-D）凝胶电泳分离的SCV和LCV裂解物的银染凝胶在两种发育形式中分辨出超过675种蛋白质。48种蛋白质在LCV中的丰度比在SCV中的丰度高两倍以上，6种蛋白质在SCV中的丰度比在LCV中的丰度高两倍以上。通过质谱法分别鉴定了SCV和LCV的4种和15种上调蛋白，并且它们的预测功能作用与代谢活性LCV和结构抗性SCV一致。一维和2-D免疫印迹的细胞形式裂解物探测血清感染/接种疫苗的豚鼠和恢复期血清从人类患者从急性Q热恢复，分别揭示了独特的SCV/LCV抗原和共同的SCV/LCV抗原，往往差异合成。通过质谱鉴定了在人类感染期间识别的抗原，包括先前描述的贝氏隐孢子虫的免疫显性蛋白和在临床Q热的病理生理学和/或诱导保护性免疫中可能重要的新型免疫原性蛋白。
A biphasic developmental cycle whereby highly resistant small-cell variants (SCVs) are generated from large-cell variants (LCVs) is considered fundamental to the virulence of Coxiella burnetti, the causative agent of human Q fever. In this study a proteome analysis of C. burnetii developmental forms was conducted to provide insight into their unique biological and immunological properties. Silver-stained gels of SCV and LCV lysates separated by two-dimensional (2-D) gel electrophoresis resolved over 675 proteins in both developmental forms. Forty-eight proteins were greater than twofold more abundant in LCVs than in SCVs, with six proteins greater than twofold more abundant in SCVs than in LCVs. Four and 15 upregulated proteins of SCVs and LCVs, respectively, were identified by mass spectrometry, and their predicted functional roles are consistent with a metabolically active LCV and a structurally resistant SCV. One-dimensional and 2-D immunoblots of cell form lysates probed with sera from infected/vaccinated guinea pigs and convalescent-phase serum from human patients who had recovered from acute Q fever, respectively, revealed both unique SCV/LCV antigens and common SCV/LCV antigens that were often differentially synthesized. Antigens recognized during human infection were identified by mass spectroscopy and included both previously described immunodominant proteins of C burnetii and novel immunogenic proteins that may be important in the pathophysiology of clinical Q fever and/or the induction of protective immunity.