Vowels B R, Yang S, Leyden J J
Department of Dermatology, School of Medicine, University of Pennsylvania, Philadelphia 19104-6142, USA.
Infect Immun. 1995 Aug;63(8):3158-65. doi: 10.1128/iai.63.8.3158-3165.1995.
Although many cytokines have been implicated in the development and persistence of inflammatory immune responses, it is unknown if any of these are important in inflammatory acne. This study investigated the production of the proinflammatory cytokines interleukin-8 (IL-8), IL-1 beta, and tumor necrosis factor alpha (TNF-alpha) by human monocytic cell lines, ThP-1 and U937, and by freshly isolated peripheral blood mononuclear cells from acne patients. Both Propionibacterium acnes and supernatants obtained from 72-h P. acnes cultures could induce significant concentrations of IL-1 beta, TNF-alpha, and IL-8 by both cell lines and by peripheral blood mononuclear cells as determined by enzyme-linked immunosorbent assay. There was no significant difference between acne and non-acne subjects. Endotoxin quantification and addition of polymyxin B to assays indicated no lipopolysaccharide (LPS) contamination. P. acnes supernatant was fractionated into components with molecular weights of < 3,000, < 10,000, and < 30,000 and assayed for the ability to induce IL-8 and TNF production in ThP-1 cells. Nearly 90% of the original activity was found in the < 30,000-molecular-weight fraction, 50% was in the < 10,000-molecular-weight fraction, and only 15% remained in the < 3,000-molecular-weight fraction. The effluent from the < 3,000-molecular-weight fraction contained about 70% activity, indicating that the inducing factor was not retained in the membrane. Incubation of P. acnes supernatant with various concentrations of mutanolysin or lysozyme resulted in a loss of 60% of the original activity. The addition of jimson lectin, which binds peptidoglycan, resulted in a loss of 70% of the activity in a dose-response manner, whereas peanut lectin had little or no effect on the activity. Heating of the P. acnes supernatant to 65 degrees C also had no effect on the activity. Blocking of CD14, a receptor for both LPS and peptidoglycan, reduced cytokine production by > 50%, suggesting that the soluble stimulating factor may be a secreted form of peptidoglycan-polysaccharide.
尽管许多细胞因子与炎症性免疫反应的发生和持续存在有关，但尚不清楚其中是否有任何一种在炎性痤疮中起重要作用。本研究调查了人单核细胞系ThP-1和U937以及痤疮患者新鲜分离的外周血单核细胞中促炎细胞因子白细胞介素-8（IL-8）、IL-1β和肿瘤坏死因子α（TNF-α）的产生情况。痤疮丙酸杆菌和从痤疮丙酸杆菌72小时培养物中获得的上清液均可诱导两种细胞系以及外周血单核细胞产生显著浓度的IL-1β、TNF-α和IL-8，这通过酶联免疫吸附测定法得以确定。痤疮患者和非痤疮患者之间没有显著差异。内毒素定量以及在测定中添加多粘菌素B表明不存在脂多糖（LPS）污染。将痤疮丙酸杆菌上清液分级分离成分子量<3000、<10000和<30000的组分，并检测其诱导ThP-1细胞中IL-8和TNF产生的能力。几乎90%的原始活性存在于分子量<30000的组分中，50%存在于分子量<10000的组分中，而只有15%保留在分子量<3000的组分中。分子量<3000的组分的流出物含有约70%的活性，这表明诱导因子没有保留在膜中。将痤疮丙酸杆菌上清液与不同浓度的变溶菌素或溶菌酶孵育导致原始活性丧失60%。添加结合肽聚糖的曼陀罗凝集素导致活性以剂量反应方式丧失70%，而花生凝集素对活性几乎没有影响。将痤疮丙酸杆菌上清液加热至65℃对活性也没有影响。阻断CD14（LPS和肽聚糖的受体）可使细胞因子产生减少>50%，这表明可溶性刺激因子可能是肽聚糖-多糖的分泌形式。
