A Co-Culture Model of the Developing Small Intestine Offers New Insight in the Early Immunomodulation of Enterocytes and Macrophages by Lactobacillus spp. through STAT1 and NF-kB p65 Translocation

Abstract

The early establishment of a complete microbiome has been shown to play an integral part in the development and maintenance of an intact intestine and its immune system, although much remains unknown about the specific mechanisms of immune modulation in newborns. In our study we show in a co-culture model of the undeveloped small intestine that members of Lactobacillus spp. influence STAT1 and NF-kB p65 nuclear translocation in both intestinal epithelial cells as well as underlying macrophages. Moreover, by using imaging flow cytometry we were able to monitor each individual cell and create a framework of the percentage of cells in which translocation occurred in challenged versus control cell populations. We also observed a significant difference in baseline translocation in intestinal cells when cultured alone versus those in a co-culture model, underpinning the importance of 3D models over monolayer set-ups in epithelial in vitro research. In conclusion, our work offers new insights into the potential routes by which the commensal microbiome primes the early immune system to fight pathogens, and shows how strain-specific these mechanisms really are.

SIRT2 Plays Significant Roles in Lipopolysaccharides-Induced Neuroinflammation and Brain Injury in Mice

“…Moreover, we found that AGK2 can block the translocation of NFκB stimulated by LPS from the cytosol to the nucleus. Due to the critical roles of nuclear translocation of NFκB in inflammation [42][43][44][45][46], our study has suggested that SIRT2 could mediate neuroinflammation by modulating nuclear translocation of NFκB. These observations, together with our previous finding that SIRT2 silencing can lead to decreased LPS-induced microglial activation-a major event in neuroinflammation [16], have collectively suggested that SIRT2 activity may be required for LPS-induced neuroinflammation.…”

Oral Administration of Probiotics Inhibits Absorption of the Heavy Metal Cadmium by Protecting the Intestinal Barrier

“…L. plantarum strains have been proven to be one of the predominant transient lactobacilli in the jejunum and ileum (54). Besides their Cd sequestration and antioxidative stress activities in the small intestine, L. plantarum strains may interact with dendritic cells in the small intestine lamina propria to influence T cell responses and influence STAT1 and NF-B p65 translocation in the epithelial cells and macrophages of small intestine (55,56). Additionally, a previous human study showed that L. plantarum strains can enhance barrier function by rearrangement of the tight-junction protein conformation in response to Toll-like receptor 2 (TLR2) signaling in the small intestine (52).…”

Flow cytometry as the new ‘gold standard’ for detection of free tumour cells in abdominal lavage fluid in gastric cancer patients: A comparative study of molecular and conventional methods

“…21 The infocus single cells were identified by gating on 7-AAD-positive events with high nuclear aspect ratios (minor to major axis ratio, as a measure of circularity) and high nuclear contrast (as measured by the Gradient Max feature). The cells were first stained with the anti-CD45 APC-Cy7 antibody (BD Biosciences, San Jose, CA, USA).…”