title: In Vitro Investigation of the Immunomodulatory Potential of Probiotic Lactobacillus casei.
abstract: The current study investigated the immunomodulatory potential of ethyl acetate soluble supernatant of Lactobacillus casei (LC-EAS) in vitro. The effect of LC-EAS on nitric oxide release was analyzed in RAW 264.7 cells, wherein, an inhibition in nitric oxide production through suppression of inducible nitric oxide synthase mRNA expression was observed. Evaluation of LC-EAS on LPS-induced peripheral blood mononuclear cells showed a down-regulation in TNF-α and IL-6 genes and an upregulation of IL-10. An inhibition in the protein expression of NF-κB, ERK1/2 and STAT3 phosphorylation confirms the immunomodulatory potential of LC-EAS. The effect of LC-EAS on in vitro intestinal epithelial cells was investigated using HT-29 human colon adenocarcinoma cancer cells. LC-EAS exhibited an inhibition of NF-κB and ERK1/2 phosphorylation, whereas STAT3 phosphorylation was unregulated. To evaluate the downstream target of STAT3 upregulation, expression of the intestinal trefoil factor TFF3 which is a NF-κB regulator and STAT3 downstream target was studied. LC-EAS was observed to elevate TFF3 mRNA expression. Overall the study shows that the anti-inflammatory potential of LC-EAS is through inhibition of NF-κB in different cell types.
authors.affiliation: Tissue Culture and Drug Discovery Lab, Centre for Biotechnology, Anna University, Chennai, 600 025, Tamilnadu, India.
authors.forename: Thirugnanam
authors.initials: T
authors.lastname: Karthikeyan
authors.affiliation: Tissue Culture and Drug Discovery Lab, Centre for Biotechnology, Anna University, Chennai, 600 025, Tamilnadu, India.
authors.forename: Mariappan
authors.initials: M
authors.lastname: Pravin
authors.affiliation: Tissue Culture and Drug Discovery Lab, Centre for Biotechnology, Anna University, Chennai, 600 025, Tamilnadu, India.
authors.forename: Velusamy Shanmuganathan
authors.initials: VS
authors.lastname: Muthusamy
authors.affiliation: Tissue Culture and Drug Discovery Lab, Centre for Biotechnology, Anna University, Chennai, 600 025, Tamilnadu, India.
authors.forename: Rajaganapathy
authors.initials: R
authors.lastname: Bharathi Raja
authors.affiliation: Tissue Culture and Drug Discovery Lab, Centre for Biotechnology, Anna University, Chennai, 600 025, Tamilnadu, India. lakshmibs@annauniv.edu.
authors.forename: Baddireddi Subhadra
authors.initials: BS
authors.lastname: Lakshmi
keywords: HT-29; Immunomodulation; Lactobacillus casei; NF-κB; PBMCs; Probiotics; TFF3