The anti-inflammatory effects of a high-frequency oligodeoxynucleotide from the genomic DNA of Lactobacillus casei

Abstract

Genomic DNA has been identified as an anti-inflammatory component of Lactobacillus species, the effects of which are mediated through toll-like receptor (TLR) 9. In this study, we identified 14 novel anti-inflammatory oligodeoxynucleotide (ODN) from the genomic DNA of Lactobacillus casei by measuring their effects on the secretion of interleukin (IL)-8 (CXCL8) in the human epithelial colorectal adenocarcinoma cell line Caco-2 cells. The ODN TTTTGCCG strongly decreased IL-8 secretion. In the genomic DNA of Lactobacillus species, the frequency of TTTTGCCG was highest in the genomic DNA of L. casei and similar among strains of L. casei. Decreases in inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressions in macrophage-like differentiated THP-1 cells confirmed the anti-inflammatory effect of TTTTGCCG. Furthermore, oral administration of TTTTGCCG ameliorated dextran sodium sulfate (DSS)-induced murine colitis and DSS-induced increased expression of inflammatory factor mRNAs, such as macrophage inflammatory protein (MIP)-2 (CXCL2), iNOS, and COX-2. The anti-inflammatory effect of TTTTGCCG was mainly regulated by an increase in heat shock protein (Hsp) 70 expression in the epithelium. TLR9 and Hsp90 may primarily mediate the anti-inflammatory effect of TTTTGCCG on Hsp70 signaling.

The Putative Antidepressant Mechanisms of Probiotic Bacteria: Relevant Genes and Proteins

“…infantis ATCC 15697 are capable of binding to Toll-like receptor 9 [ 4 , 101 ]. This effect was similarly observed in oligonucleotides of L. casei ATCC 27092—namely, TTTTGCCG and the total RNA and genomic DNA of phagocytosed L. gasseri OLL2809 cells [ 102 , 103 ].…”

Lactobacillus plantarum induces genomic DNA-dependent and TLR9-mediated elafin secretion from Caco-2 cells

“…On the other hand, we also showed previously that suppression of TLR9 expression by RNAi abolished the decrease in H 2 O 2 -induced IL-8 secretion by lactobacilli genomic DNA and the antiinflammatory ODN. 6,21 These observations indicated that the TLR9 signaling pathway plays a major role in mediating the decrease in IL-8 secretion. However, at present, we cannot explain the mechanism by which activation of the TLR9 signaling pathway results in the observed decrease in IL-8 secretion.…”

“…6 In addition, we identified an oligodeoxynucleotide (ODN), which markedly decreased H 2 O 2 -induced IL-8 secretion, from the genomic DNA of L. casei, and demonstrated that oral administration of this anti-inflammatory ODN to mice ameliorated DSS-induced murine colitis. 21 Therefore, suppression of IL-8 secretion in vitro is an important factor for ameliorating colitis in this in vivo model of IBD. In our preliminary study, we found that heat-killed L. plantarum and its genomic DNA decreased H 2 O 2 -induced IL-8 secretion from Caco-2 cells without inducing a pro-inflammatory response (data not shown).…”

“…TLR9 expression in Caco-2 cells was suppressed by RNAi, as described previously. 21 Briefly, aliquots of 2 × 10 5 Caco-2 cells plated in 24-well plates were transfected with 50 nM TLR9 -siRNA or scramble TLR9-siRNA (Bonac Corp.) as a negative control using siLentFect Lipid Reagent. After incubation for 24 h, the cells were treated with heat-killed L. plantarum (5 × 10 9 CFU/ml) or transfected with L. plantarum genomic DNA (17 µg/mL) for 48 h as described above.…”

Specific vaginal lactobacilli suppress the inflammation induced by lipopolysaccharide stimulation through downregulation of toll-like receptor 4 expression in human embryonic intestinal epithelial cells

“…[ 21 ] reported that signal activation of TLR9 inhibited LPS-mediated TLR4 signaling and expression via regulation of multiple cyclin-dependent kinase pathways. Moreover, it is known that Lactobacillus strains and the abovementioned bacterial oligodeoxynucleotides bind to TLR9 and induce the innate immune response through TLR9 in human intestinal epithelial cells [ 22 , 23 ], and Ju et al . [ 24 ] reported that INT-407 (Henle 407) cells express the majority of the TLRs (with the exception of TLR8) and that CpG-DNA activated signaling in this cell line.…”