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痤疮患者和正常健康受试者对痤疮丙酸杆菌粗细胞裂解物的抗体反应及促炎细胞因子的诱导

Antibody response to crude cell lysate of propionibacterium acnes and induction of pro-inflammatory cytokines in patients with acne and normal healthy subjects.

作者信息

Basal E, Jain A, Kaushal G P

机构信息

Department of Microbiology, KGMU, Lucknow, INDIA.

出版信息

J Microbiol. 2004 Jun;42(2):117-25.

Abstract

Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to > 1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, ranging from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92% (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-alpha) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypeptide reactant was eluted from the gel and was found to cause dose dependent stimulation of the productions of IL-8 and TNF-alpha. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

摘要

痤疮丙酸杆菌(P. acnes)在寻常痤疮的发病机制中起着重要作用,寻常痤疮是一种主要见于青少年年龄组的毛囊皮脂腺疾病。在本研究中,使用斑点酶联免疫吸附测定(dot-ELISA)和蛋白质印迹分析检测了痤疮患者(n = 50)和无病对照者(n = 25)中抗痤疮丙酸杆菌(MTCC1951)抗体的存在情况。还分析了从痤疮患者和健康受试者获取的人外周血单核细胞(PBMCs)诱导促炎细胞因子的痤疮丙酸杆菌的能力。皮肤拭子培养呈阳性的患者(n = 26)与痤疮丙酸杆菌阴性患者(n = 24)和正常对照者(n = 25)相比,病情更严重且抗体滴度更高(1:4000至>1:16000)。通过蛋白质印迹分析对患者血清进行分析,识别出痤疮丙酸杆菌的多种抗原成分,范围从29 kDa至205 kDa。主要的反应性成分是一种约96 kDa的多肽,在92%(26例中的24例)患者血清中被识别。此外,通过细胞因子酶联免疫吸附测定分析发现,从72小时培养物中获得的痤疮丙酸杆菌培养上清液、粗细胞裂解物和热灭活的痤疮丙酸杆菌全细胞,在健康受试者和患者中均能诱导PBMCs产生大量白细胞介素-8(IL-8)和肿瘤坏死因子α(TNF-α)。患者体内细胞因子水平显著高于健康受试者。从凝胶上洗脱下来的一种主要的96 kDa多肽反应物被发现可引起IL-8和TNF-α产生的剂量依赖性刺激。因此,上述结果表明体液免疫反应和促炎反应在痤疮发病机制中均起主要作用。

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