论文部分内容阅读
目的:探讨金黄色葡萄球菌超抗原与耳部瘢痕疙瘩形成的相关性。方法:采用病例对照研究方法。2017年6月—2018年3月，取南通大学附属医院收治的10例（女9例、男1例，年龄19~59岁）耳部瘢痕疙瘩患者行耳部瘢痕疙瘩核心切除术后弃用瘢痕组织及3例（均为女性，年龄20~24岁）色素痣患者手术后弃用的正常皮肤组织。取耳部瘢痕疙瘩表面分泌物，培养细菌并进行鉴定。取瘢痕疙瘩和正常皮肤组织，采用蛋白质印记法检测金黄色葡萄球菌肠毒素A+肠毒素B+毒性休克综合征毒素1（TSST-1）的蛋白表达，并根据蛋白表达情况将瘢痕疙瘩分为超抗原阳性组和超抗原阴性组。蛋白质印迹法检测2组瘢痕疙瘩T细胞受体（TCR） Vβ蛋白表达。Masson、苏木精-伊红（HE）染色观察2组瘢痕疙瘩真皮胶原纤维形成和炎症细胞浸润情况。酶联免疫吸附测定法检测超抗原阳性组瘢痕疙瘩人金黄色葡萄球菌肠毒素A、肠毒素B、TSST-1。对数据行配对样本n t检验。n 结果:耳部瘢痕疙瘩表面分泌物培养出细菌，优势菌培养24 h可见细菌周围出现溶血现象，菌落呈白色或金黄色，鉴定为金黄色葡萄球菌。4例患者正常皮肤肠毒素A+肠毒素B+TSST-1阴性，蛋白表达量为0.267±0.016。4例患者瘢痕疙瘩金黄色葡萄球菌肠毒素A+肠毒素B+TSST-1蛋白表达阳性，蛋白表达量为0.472±0.016，纳入超抗原阳性组；6例患者瘢痕疙瘩金黄色葡萄球菌肠毒素A+肠毒素B+TSST-1蛋白表达阴性，蛋白表达量为0.255±0.004，纳入超抗原阴性组。超抗原阳性组瘢痕疙瘩金黄色葡萄球菌肠毒素A+B+TSST-1蛋白表达量明显高于超抗原阴性组和正常皮肤（n t=15.22、8.63，n P＜0.01）。超抗原阳性组瘢痕疙瘩TCR Vβ蛋白表达量为0.389±0.023，明显高于超抗原阴性组的0.169±0.014（n t=8.62，n P＜0.001）。Masson染色显示，2组瘢痕疙瘩真皮内均见大量胶原纤维增生。HE染色显示，超抗原阴性组瘢痕疙瘩真皮可见血管周围少量炎症细胞浸润，超抗原阳性组瘢痕疙瘩血管周围可见大量炎症细胞浸润。4例抗超原阳性瘢痕疙瘩患者中肠毒素A阳性2例、肠毒素B阳性2例，其中1例患者检测出肠毒素A和肠毒素B阳性，未检出TSST-1。n 结论:金黄色葡萄球菌分泌的超抗原是耳部瘢痕疙瘩众多发病原因中的一种，可能与金黄色葡萄球菌超抗原激活瘢痕疙瘩信号通路有关。“,”Objective:To study the correlation between superantigen of Staphylococcus aureus and ear keloid.Methods:From June 2017 to March 2018, 10 patients with ear keloid were treated in the Department of Dermatology, affiliated Hospital of Nantong University. The following experiments were carried out. (1) Culture and identification of ear skin secretion before operation. (2) Anti staphylococcal enterotoxin A + B + TSST-1 polyclonal antibody was used to detect the presence of common staphylococcal superantigen in ear keloid by Western blot. (3) T cell Vβ receptor (TCR V β) antibody was used to detect the difference of TCR V β expression between superantigen positive group and superantigen negative group by Western blot. (4) The difference between superantigen positive group and superantigen negative group was observed by histopathology. (5) The monoclonal antibodies of enterotoxin SEA、SEB and TSST-1 were detected by ELISA in the superantigen positive group.Results:(1) Secretions were cultured under the following conditions: aseptic growth, a small amount of miscellaneous bacteria growth, dominant bacteria growth, and the dominant bacteria were selected and identified as Staphylococcus aureus after re-culture. (2) Staphylococcal enterotoxin A + B + TSST-1 polyclonal antibody detected 4 cases of ear keloid protein positive expression, protein expression was 0.47 ± 0.01, divided into superantigen positive group, 6 cases of ear keloid protein negative expression, protein expression was 0.25 ± 0.004, divided into superantigen negative group, normal skin reference group protein negative expression, protein expression was 0.26 ± 0.01, The protein expression in superantigen positive group was significantly higher than that in superantigen negative group and normal skin(P < 0.001, t = 15.22, 8.63, respectively). (3) The expression of TCR Vβ protein in superantigen positive group was 0.38 ± 0.02, which was higher than that in superantigen negative group (0.16 ± 0.01, P < 0.001, t = 8.62). (4) Under Masson staining, a large number of collagen fibers proliferated in the dermis of the superantigen positive group and the superantigen negative group, with no obvious abnormality; under HE staining, collagen proliferation and a small number of inflammatory cells were found in the superantigen negative group; in addition to collagen proliferation, a large number of inflammatory cells infiltrated around the blood vessels in the superantigen positive group. (5) In the 4 cases of superantigen positive keloid, 2 cases were SEA positive, 3 cases were SEB positive, one case was SEA and SEB positive, and TSST-1 was not detected.Conclusion:The superantigen secreted by Staphylococcus aureus is one of the causes of ear keloid,and it is speculated that the superantigen of Staphylococcus aureus may activate the signal pathway of keloid.