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反应性关节炎/赖特综合征患者滑膜中的沙眼衣原体具有活力,但呈现异常基因表达。

Synovial Chlamydia trachomatis in patients with reactive arthritis/Reiter's syndrome are viable but show aberrant gene expression.

作者信息

Gérard H C, Branigan P J, Schumacher H R, Hudson A P

机构信息

Medical Research Service, Department of Veterans Affairs Medical Center, Allegheny University of the Health Sciences, MCP-Hahnemann School of Medicine, USA.

出版信息

J Rheumatol. 1998 Apr;25(4):734-42.

PMID:9558178
Abstract

OBJECTIVE

We used reverse transcription-polymerase chain reaction (RT-PCR) assays to assess expression of genes from Chlamydia trachomatis in synovial tissues of patients with reactive arthritis (ReA)/Reiter's syndrome (RS) to determine viability/metabolic activity of the bacterium in joints of infected patients.

METHODS

Synovial biopsies were obtained from 18 patients with ReA, RS, or other arthritides; nucleic acids from 16 samples were PCR positive for chlamydial chromosomal DNA. RT-PCR assays targeting primary transcripts from C. trachomatis rRNA operons, and mRNA from the bacterial omp1, hsp60, glyQS, and r-protein S5 and L5 genes, were used to characterize viability/metabolic activity. Host actin mRNA was assessed as control in each sample preparation.

RESULTS

RT-PCR of host cell actin mRNA in the 18 patient samples confirmed the quality of all RNA preparations. RNA from 14/16 PCR positive samples was positive by RT-PCR for chlamydial rRNA primary transcripts. Each of these same 14 samples was also RT-PCR positive in assays targeting glyQS, r-protein S5 and L5, and hsp60 mRNA. However, none of the 14 samples showing chlamydial rRNA and mRNA was positive for omp1 transcripts.

CONCLUSION

Synovial chlamydia are viable/metabolically active, since primary rRNA transcripts and mRNA from chlamydial genes specifying components of the bacterial protein synthetic system were present in most patient samples assayed. Expression of omp1, encoding the major outer membrane protein, is strongly attenuated in persistently infecting synovial chlamydia, while that of hsp60, specifying a highly immunogenic heat shock protein of the organism, is not downregulated.

摘要

目的

我们采用逆转录-聚合酶链反应(RT-PCR)检测法,评估反应性关节炎(ReA)/赖特综合征(RS)患者滑膜组织中沙眼衣原体基因的表达情况,以确定感染患者关节中细菌的生存能力/代谢活性。

方法

对18例患有ReA、RS或其他关节炎的患者进行滑膜活检;16份样本的核酸经PCR检测显示沙眼衣原体染色体DNA呈阳性。采用针对沙眼衣原体rRNA操纵子初级转录本以及细菌omp1、hsp60、glyQS和核糖体蛋白S5及L5基因mRNA的RT-PCR检测法,来表征生存能力/代谢活性。在每份样本制备过程中,以宿主肌动蛋白mRNA作为对照进行评估。

结果

对18例患者样本中宿主细胞肌动蛋白mRNA进行RT-PCR检测,证实了所有RNA制备物的质量。14/16份PCR阳性样本的RNA经RT-PCR检测,沙眼衣原体rRNA初级转录本呈阳性。在针对glyQS、核糖体蛋白S5及L5和hsp60 mRNA的检测中,这14份相同样本的RT-PCR结果也均为阳性。然而,在这14份显示沙眼衣原体rRNA和mRNA阳性的样本中,没有一份omp1转录本呈阳性。

结论

滑膜中的衣原体具有生存能力/代谢活性,因为在大多数检测的患者样本中,存在来自指定细菌蛋白质合成系统成分的衣原体基因的初级rRNA转录本和mRNA。编码主要外膜蛋白的omp1的表达在持续感染的滑膜衣原体中大幅减弱,而指定该生物体一种高度免疫原性热休克蛋白的hsp60的表达并未下调。

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