Absence of Bacteria in the Temporal Arteries of Patients with Giant Cell Arteritis.
Absence of Bacteria in the Temporal Arteries of Patients with Giant Cell Arteritis.
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巨细胞动脉炎患者的颞动脉中没有细菌。
DOI：
10.1097/rhu.0000000000000344
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Katz,BradleyJ
中科院分区：
文献类型：
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作者：
Koening,CurryL;Peterson,SpencerG;Podnecky,NicoleL;Schweizer,HerbertP;Li,DeanY;Katz,BradleyJ
致编辑：巨细胞动脉炎（GCA）是影响50岁以上白色人最常见的血管炎类型。颞浅动脉活检通常用于确诊GCA。虽然GCA的病理生理学已被广泛研究，但其原因仍不清楚。GCA的感染性触发因素已被考虑，但没有生物体最终与GCA有关。2，3在一些受试者的颞动脉壁中存在肉芽肿性炎症和多核巨细胞，这使我们假设细菌感染引起GCA。4灵敏的基因组测序技术现在可以检测细菌和其他传染性有机体，这些生物体曾经无法通过培养或其他分子技术识别。为了确定GCA是否由细菌感染引起，我们在培养基中培养了GCA受试者的颞动脉，并每天分析标本的细菌生长。我们还使用16 S rRNA序列技术来鉴定位于GCA受试者颞动脉壁中的细菌基因组序列。这项研究得到了犹他州大学和湖城军人事务部医学中心的机构审查委员会的批准。根据赫尔辛基宣言原则采集颞浅动脉活检（STAB）样本，所有受试者在参与研究前提供知情同意书。GCA受试者符合1990年美国风湿病学会（ACR）GCA标准，并且STAB的组织病理学特征与GCA一致。5对具有GCA临床特征但病理学家描述为不确定的组织病理学特征的受试者的颞浅动脉活检进行了分析。对照STAB来自因怀疑GCA而接受活检的受试者，但他们的组织病理学动脉正常，随后由医生进行的临床随访确定这些患者没有GCA。颞动脉标本在手术室采用严格的无菌程序采集。将一部分标本固定在福尔马林中，石蜡包埋，并送去进行组织病理学分析。将每份标本的另一部分放入含或不含RNALater（Qiagen，瓦伦西亚，CA）的无菌标本容器中，并在-80 ℃下冷冻。按照标准微生物和无菌技术，在无菌组织培养罩中处理标本。将大约1 - 2 mm的STAB切片置于无菌磷酸盐缓冲盐水中并匀浆。将总计20 μL的均质化混悬液铺板于Luria-Bertani（Teknova，霍利斯特，CA）、脑心灌注液（哈代Diagnostics，Santa Maria，CA）和巧克力培养基（哈代Diagnostics，圣玛利亚，CA）上。将其余悬浮液添加到3 mL Luria-Bertani（Gibco，Grand Island，NV）和脑心输液肉汤（Remel，莱内克萨，KS）中。将所有培养物在37 ℃下振荡孵育长达14天，并每天检查生长情况。分离并表征任何观察到的菌落
To the Editor: Giant cell arteritis (GCA) is the most common type of vasculitis to affect white individuals older than 50 years. 1 Biopsy of a superficial temporal artery is commonly used to confirm the diagnosis of GCA. Although the pathophysiology of GCA has been extensively studied, the cause remains unknown. An infectious trigger for GCA has been considered, but no organism has conclusively been linked to GCA. 2, 3 The presence of granulomatous inflammation and multinucleated giant cells in the temporal artery wall of some subjects led us to hypothesize that a bacterial infection causes GCA. 4 Sensitive genomic sequencing techniques now make it possible to detect bacteria and other infectious organisms that were once unidentifiable by culture or other molecular techniques. To determine if GCA is caused by a bacterial infection, we cultured temporal arteries from subjects with GCA in media and analyzed the specimens daily for growth of bacteria. We also used 16S rRNA sequence technology to identify bacterial genomic sequences located in the temporal artery wall of subjects with GCA. This study was approved by the institutional review boards of the University of Utah and the Department of Veterans Affairs Medical Center, Salt Lake City. Superficial temporal artery biopsy (STAB) samples were collected in accordance with the Declaration of Helsinki principles, and all subjects provided informed consent before participating in the study. Subjects with GCA met the 1990 American College of Rheumatology (ACR) criteria for GCA and had a STAB with histopathologic features consistent with GCA. 5 Superficial temporal artery biopsies from subjects with clinical features of GCA but histopathologic features described by a pathologist as inconclusive were also analyzed. Control STABs came from subjects who underwent biopsy for suspicion of GCA, but they had histopathologically normal arteries, and subsequent clinical follow-up by a physician determined these patients to not have GCA. Temporal artery specimens were collected in the operating room using strict sterile procedures. A portion of the specimen was fixed in formalin, embedded in paraffin, and sent for histopathologic analysis. Another portion of each specimen was placed into a sterile specimen container with or without RNALater (Qiagen, Valencia, CA) and frozen at− 80 C. Specimens were handled in a sterile tissue culture hood following standard microbiological and aseptic techniques. Investigators performing experiments were masked to the affected status of the samples analyzed.Sections of STABs measuring approximately 1 to 2 mm were placed in sterile phosphate-buffered saline and homogenized. A total of 20 μL of the homogenized suspension was plated onto Luria-Bertani (Teknova, Hollister, CA), brain-heart infusion (Hardy Diagnostics, Santa Maria, CA) and chocolate media (Hardy Diagnostics, Santa Maria, CA). The remainder of the suspension was added to 3 mL of each Luria-Bertani (Gibco, Grand Island, NV) and brain-heart infusion broth (Remel, Lenexa, KS). All cultures were incubated at 37 C with shaking for up to 14 days and checked daily for growth. Any observed colonies were isolated and characterized