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金黄色葡萄球菌临床分离株分泌的葡萄球菌肠毒素样K(SEl-K)的检测与测定。

Detection and measurement of staphylococcal enterotoxin-like K (SEl-K) secretion by Staphylococcus aureus clinical isolates.

作者信息

Aguilar Jorge L, Varshney Avanish K, Wang Xiaobo, Stanford Lindsay, Scharff Matthew, Fries Bettina C

机构信息

Department of Medicine, Albert Einstein College of Medicine, Bronx, New York, USA Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USA.

Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA.

出版信息

J Clin Microbiol. 2014 Jul;52(7):2536-43. doi: 10.1128/JCM.00387-14. Epub 2014 May 7.

Abstract

Staphylococcal enterotoxin-like K (SEl-K) is a potent mitogen that elicits T-cell proliferation and cytokine production at very low concentrations. However, unlike the classical enterotoxins SEB and toxic shock syndrome toxin 1 (TSST-1), the gene for SEl-K is commonly present in more than half of all Staphylococcus aureus clinical isolates and is present in almost all USA300 community-acquired methicillin-resistant S. aureus (CA-MRSA) isolates. Sequencing of the sel-k gene in over 20 clinical isolates and comparative analysis with all 14 published sel-k sequences indicate that there are at least 6 variants of the sel-k gene, including one that is conserved among all examined USA300 strains. Additionally, we have developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) that specifically detects and measures SEl-K protein in culture supernatants and biological fluids. Quantification of in vitro SEl-K secretion by various S. aureus isolates using this novel capture ELISA revealed detectable amounts of SEl-K secretion by all isolates, with the highest secretion levels being exhibited by MRSA strains that coexpress SEB. In vivo secretion was measured in a murine thigh abscess model, where similar levels of SEl-K accumulation were noted regardless of whether the infecting strain exhibited high or low secretion of SEl-K in vitro. We conclude that SEl-K is commonly expressed in the setting of staphylococcal infection, in significant amounts. SEl-K should be further explored as a target for passive immunotherapy against complicated S. aureus infection.

摘要

葡萄球菌肠毒素样K(SEl-K)是一种强效促细胞分裂剂,在极低浓度下就能引发T细胞增殖和细胞因子产生。然而,与经典肠毒素SEB和中毒性休克综合征毒素1(TSST-1)不同,SEl-K基因在超过半数的金黄色葡萄球菌临床分离株中普遍存在,并且几乎在所有USA300社区获得性耐甲氧西林金黄色葡萄球菌(CA-MRSA)分离株中都存在。对20多个临床分离株中的sel-k基因进行测序,并与所有已发表的14个sel-k序列进行比较分析,结果表明sel-k基因至少有6种变体,其中一种在所有检测的USA300菌株中是保守的。此外,我们开发了一种高度灵敏的酶联免疫吸附测定(ELISA),可特异性检测和测量培养上清液和生物体液中的SEl-K蛋白。使用这种新型捕获ELISA对各种金黄色葡萄球菌分离株的体外SEl-K分泌进行定量分析,结果显示所有分离株均可检测到SEl-K分泌,其中共表达SEB的MRSA菌株分泌水平最高。在小鼠大腿脓肿模型中测量了体内分泌情况,结果发现,无论感染菌株在体外SEl-K分泌水平高或低,SEl-K的积累水平相似。我们得出结论,SEl-K在葡萄球菌感染的情况下通常大量表达。SEl-K应作为针对复杂性金黄色葡萄球菌感染的被动免疫治疗靶点进行进一步研究。

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