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Lactobacillus rhamnosus Isolated from “amabere amaruranu” Cultured Milk Modulated the Production of Inflammatory Cytokines in Caco‐2 Cells
Author(s) -
Ngeny Beverly,
Onyango E M
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1022.1
Subject(s) - lactobacillus rhamnosus , microbiology and biotechnology , probiotic , caco 2 , immune system , interleukin 8 , tumor necrosis factor alpha , cytokine , lipopolysaccharide , biology , lactobacillus , lysis , chemistry , cell , bacteria , immunology , biochemistry , genetics
Probiotic microorganisms have been shown to confer health benefits on the host when administered in adequate amounts. In the gastrointestinal tract, they promote host immunity, regulate immune signaling pathways and produce anti‐pathogenic factors. The immune effects resulting from interactions between intestinal epithelial cells and commensal bacteria have not yet been fully elucidated. This study examined the effects of a Lactobacillus rhamnosus isolate from “amabere amaruranu” a Kenyan traditional cultured milk, on the production of pro‐inflammatory cytokines (Interleukin 8 and Tumor Necrosis Factor‐α) and anti‐inflammatory cytokines (Interleukin 10 and Transforming Growth Factor‐β) in enterocytes. The L. rhamnosus isolate was cultured in de‐Mann Rogosa Sharpe media overnight, the broth was collected and filtered to obtain filtered spent broth (FSB). Cell pellets resuspended in PBS were sonicated and filtered to obtain cytoplasmic fraction (CF). Differentiated Caco‐2 cell monolayer in transwells, after 21 days post‐confluence were co‐cultured with live L. rhamnosus (MRS6AN) isolate or apically treated with PBS control or CF or FSB. After 6–36 hrs of incubation, cells were collected and lysed and cell supernatants were collected. Enzyme Linked Immunosorbent Assay was used to determine the cytokine content in the cell lysate, apical and basolateral supernatants. Effects of the various treatments (PBS Control, MRS6AN, CF and FSB) on Caco‐2 cell lysate levels of TGF‐β were 14.5, 191.6, 153.3 and 128.8 ng/ml, respectively; of IL10 were 7.1, 44, 33.6 and 31 pg/ml, respectively; of IL8 were 200, 1014, 1489.1 and 1659.6 pg/ml, respectively, and of TNF‐α were 4.1, 27.6, 33.1 and 30.1 pg/ml, respectively. There were very low levels of TGFβ, IL10, IL8 and TNFα cytokines in both the apical and basolateral supernatants. In conclusion, L. rhamnosus isolate and its cytoplasmic fraction or spent media seemed to cause a higher production of anti‐inflammatory TGF‐β and pro‐inflammatory IL8 cytokines by differentiated Caco‐2 cells. Support or Funding Information East Tennessee State University