Localization and characterization of two putative TMH family proteins in Chlamydia psittaci.
Localization and characterization of two putative TMH family proteins in Chlamydia psittaci.
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DOI：
10.1016/j.micres.2015.11.005
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发表时间：
2016-02
影响因子：
6.7
通讯作者：
Haiying Wu;Chuan Wang;Chuanhao Jiang;Yafeng Xie;Liangzhuan Liu;Yin Song;Xiaohua Ma;Yimou Wu
中科院分区：
文献类型：
--
作者：
Haiying Wu;Chuan Wang;Chuanhao Jiang;Yafeng Xie;Liangzhuan Liu;Yin Song;Xiaohua Ma;Yimou Wu
鹦鹉热衣原体（C.鹦鹉热）是鹦鹉热病的一种专性细胞内因子，在人类中引起非典型肺炎。C.鹦鹉热，pupeli属于inc家庭，大概发挥类似的作用。CPSIT_0844和CPSIT_0846是C.鹦鹉为了鉴定这两种蛋白，用原核表达的融合蛋白制备了抗血清。免疫荧光法显示CPSIT_0844和CPSIT_0846定位于包涵体膜上。鹦鹉热感染的细胞通过RT-PCR和western blot分析检测时间表达，CPSIT_0844和CPSIT_0846早在感染后（p.i.）和6 h p.i.，免疫荧光法检测分泌物中，p.i.并在整个生长周期中保持在包涵体膜中。CPSIT_0844和CPSIT_0846能被C.但与沙眼衣原体和衣原体的抗血清无反应，这与它们在C.沙眼和C.肺炎。这些结果表明CPSIT_0844和CPSIT_0846可能是C.鹦鹉热可用于诊断C.鹦鹉此外，CPSIT_0844和CPSIT_0846可诱导THP-1细胞表达炎性细胞因子IL-1β、IL-6和TNF-α，这可能与衣原体诱导的炎症病理有关。
Chlamydia psittaci(C. psittaci), an obligate intracellular agent of psittacosis, causes an atypical pneumonia in humans. The transmembrane head proteins (TMH) ofC. psittaci, putatively belong to the Inc family and presumably play similar roles. CPSIT_0844 and CPSIT_0846 were the putative TMH proteins ofC. psittaci. To identify these two proteins, antisera were raised with fusion proteins which were prokaryotic expressed inEscherichia coliand purified. By immunofluorescence assay, CPSIT_0844 and CPSIT_0846 were localized in the inclusion membrane ofC. psittaci-infected cells. By RT-PCR and western blot analysis to detect the temporal expression, CPSIT_0844 and CPSIT_0846 were detected as early as 12 h post-infection (p.i.) and 6 h p.i., separately; meanwhile, in secretions monitored with immunofluorescence assay, these proteins were observed in the inclusion membrane at 18 h p.i. and remained in the inclusion membrane throughout the growth cycle. CPSIT_0844 and CPSIT_0846 could specifically be recognized by the antiserum ofC. psittacibut failed to react with the antiserums ofChlamydiatrachomatisandChlamydiapneumoniae, which is consistent with the fact that they had no significant orthologs inC. trachomatisandC. pneumoniae. These results revealed that CPSIT_0844 and CPSIT_0846, the putative TMH family proteins, might be unique toC. psittaciand could be used to diagnose the infection caused byC. psittaci. Moreover, CPSIT_0844 and CPSIT_0846 could induce the expression of the inflammatory cytokines IL-1β, IL-6 and TNF-α in THP-1 cells, which might contribute to chlamydia-induced inflammatory pathologies.