Boonma Prapaporn, Spinler Jennifer K, Venable Susan F, Versalovic James, Tumwasorn Somying
Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
BMC Microbiol. 2014 Jul 2;14:177. doi: 10.1186/1471-2180-14-177.
Clostridium difficile is the main cause of hospital-acquired diarrhea and colitis known as C. difficile-associated disease (CDAD).With increased severity and failure of treatment in CDAD, new approaches for prevention and treatment, such as the use of probiotics, are needed. Since the pathogenesis of CDAD involves an inflammatory response with a massive influx of neutrophils recruited by interleukin (IL)-8, this study aimed to investigate the probiotic effects of Lactobacillus spp. on the suppression of IL-8 production in response to C. difficile infection.
We screened Lactobacillus conditioned media from 34 infant fecal isolates for the ability to suppress C. difficile-induced IL-8 production from HT-29 cells. Factors produced by two vancomycin-resistant lactobacilli, L. rhamnosus L34 (LR-L34) and L.casei L39 (LC-L39), suppressed the secretion and transcription of IL-8 without inhibiting C. difficile viability or toxin production. Conditioned media from LR-L34 suppressed the activation of phospho-NF-κB with no effect on phospho-c-Jun. However, LC-L39 conditioned media suppressed the activation of both phospho-NF-κB and phospho-c-Jun. Conditioned media from LR-L34 and LC-L39 also decreased the production of C. difficile-induced GM-CSF in HT-29 cells. Immunomodulatory factors present in the conditioned media of both LR-L34 and LC-L39 are heat-stable up to 100°C and > 100 kDa in size.
Our results suggest that L. rhamnosus L34 and L. casei L39 each produce factors capable of modulating inflammation stimulated by C. difficile. These vancomycin-resistant Lactobacillus strains are potential probiotics for treating or preventing CDAD.
艰难梭菌是医院获得性腹泻和结肠炎(即艰难梭菌相关性疾病,CDAD)的主要病因。随着CDAD病情加重及治疗失败情况增多,需要新的预防和治疗方法,如使用益生菌。由于CDAD的发病机制涉及炎症反应,其中白细胞介素(IL)-8招募大量中性粒细胞大量涌入,本研究旨在调查乳酸杆菌属对艰难梭菌感染后IL-8产生的抑制作用。
我们从34株婴儿粪便分离株中筛选乳酸杆菌条件培养基,以检测其抑制艰难梭菌诱导HT-29细胞产生IL-8的能力。两种耐万古霉素的乳酸杆菌,即鼠李糖乳杆菌L34(LR-L34)和干酪乳杆菌L39(LC-L39)产生的因子,可抑制IL-8的分泌和转录,而不抑制艰难梭菌的活力或毒素产生。LR-L34的条件培养基可抑制磷酸化核因子κB的激活,而对磷酸化c-Jun无影响。然而,LC-L39条件培养基可抑制磷酸化核因子κB和磷酸化c-Jun的激活。LR-L34和LC-L39的条件培养基还可降低艰难梭菌诱导HT-29细胞中粒细胞-巨噬细胞集落刺激因子(GM-CSF)的产生。LR-L34和LC-L39条件培养基中存在的免疫调节因子在高达100°C时热稳定,大小>100 kDa。
我们的结果表明,鼠李糖乳杆菌L34和干酪乳杆菌L39各自产生能够调节艰难梭菌刺激的炎症的因子。这些耐万古霉素的乳酸杆菌菌株是治疗或预防CDAD的潜在益生菌。