Lactobacillus rhamnosus L34 and Lactobacillus casei L39 suppress Clostridium difficile-induced IL-8 production by colonic epithelial cells

Abstract

BackgroundClostridium 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.ResultsWe 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.ConclusionsOur 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.

Next-Generation Probiotics Targeting Clostridium difficile through Precursor-Directed Antimicrobial Biosynthesis

“…A recent U.S.-based surveillance study of the drug susceptibilities of diarrhea-associated C. difficile isolates showed the MIC 90 of vancomycin, metronidazole, and fidaxomicin to be 4, 2, and 0.5 g/ml, respectively (27). L. casei LC-39 and L. rhamnosus LR-34, known to produce factors that modulate the inflammation stimulated by C. difficile in vitro (28), demonstrated substantial resistance to vancomycin and metronidazole, with an MIC value of each drug Ͼ256 g/ml, a concentration 64-or 128-fold greater than the vancomycin or metronidazole MIC 90 for C. difficile, respectively. However, these strains were susceptible to fidaxomicin at 2 g/ml, which is a concentration only 4-fold greater than the fidaxomicin MIC 90 for C. difficile and much lower than the estimated colonic concentrations of fidaxomicin in patients (29).…”

Enterococcus faeciumHDRsEf1 Protects the Intestinal Epithelium and Attenuates ETEC-Induced IL-8 Secretion in Enterocytes

“…Our data are in agreement with recent reports [15, 53] that commensal bacteria or probiotics can downregulate IL-8 released by IECs to fight against the enteropathogens and reduce proinflammatory factors. The supernatants of Lactobacillus rhamnosus L34 and L. casei L39 can inhibit Clostridium difficile -induced IL-8 production in IECs [54]. Some reports had elaborated that probiotics and their components could modulate inflammatory responsiveness and TLR-related gene expression [55, 56], such that L. amylovorus and its supernatant inhibit TLR4 inflammatory signalling triggered by ETEC, and TLR2 is required for the suppression of TLR4 signalling [27].…”

Mechanisms and therapeutic effectiveness of lactobacilli

“…245 246 Boonma and coworkers investigated the probiotic effect of L. rhamnosus L34 and L. casei L39, two vancomycin-resistant lactobacilli, on the suppression of IL-8 production in response to C. difficile infection. 247 While L. casei L39 suppressed the activation of phosphonuclear factor κ-light-chain-enhancer of activated B cells and phospho-c-Jun in HT-29 cells, L. rhamnosus L34 and L. casei L39 decreased the production of C. difficile-induced granulocyte-macrophage colony-stimulating factor. Moreover, L. acidophilus GP1B cell extract decreased transcriptional levels of luxS, tcdA, tcdB and txeR genes of C. difficile, thus reducing virulence in vitro.…”