02778nas a2200325   4500000000100000000000100001008004100002260001500043653001300058653001500071100001700086700001600103700002000119700001900139700002200158700001700180700003000197700001900227700001900246700001600265700002100281700002100302700002200323245012400345856005500469300000900524490000700533520189800540022001402438       2024                            d  c2024-03-2910abacteria10amicrobiota1 aJianbo Zhang1 aYu-Ja Huang1 aMartin Trapecar1 aCharles Wright1 aKirsten Schneider1 aJohn Kemmitt1 aVictor Hernandez-Gordillo1 aJun Young Yoon1 aMathilde Poyet1 aEric J. Alm1 aDavid T. Breault1 aDavid L. Trumper1 aLinda G. Griffith00aAn immune-competent human gut microphysiological system enables inflammation-modulation by Faecalibacterium prausnitzii  uhttps://www.nature.com/articles/s41522-024-00501-z  a1-110 v103 aCrosstalk of microbes with human gut epithelia and immune cells is crucial for gut health. However, there is no existing system for a long-term co-culture of human innate immune cells with epithelium and oxygen-intolerant commensal microbes, hindering the understanding of microbe-immune interactions in a controlled manner. Here, we established a gut epithelium-microbe-immune (GuMI) microphysiological system to maintain the long-term continuous co-culture of Faecalibacterium prausnitzii/Faecalibacterium duncaniae with colonic epithelium, antigen-presenting cells (APCs, herein dendritic cells and macrophages), and CD4+ naive T cells circulating underneath the colonic epithelium. In GuMI-APC condition, multiplex cytokine assays suggested that APCs contribute to the elevated level of cytokines and chemokines secreted into both apical and basolateral compartments compared to GuMI condition that lacks APC. In GuMI-APC with F. prausnitzii (GuMI-APC-FP), F. prausnitzii increased the transcription of pro-inflammatory genes such as toll-like receptor 1 (TLR1) and interferon alpha 1 (IFNA1) in the colonic epithelium, without a significant effect on cytokine secretion, compared to the GuMI-APC without bacteria (GuMI-APC-NB). In contrast, in the presence of CD4+ naive T cells (GuMI-APCT-FP), TLR1, IFNA1, and IDO1 transcription levels decreased with a simultaneous increase in F. prausnitzii-induced secretion of pro-inflammatory cytokines (e.g., IL8) compared to GuMI-APC-FP that lacks T cells. These results highlight the contribution of individual innate immune cells in regulating the immune response triggered by the gut commensal F. prausnitzii. The integration of defined populations of immune cells in the gut microphysiological system demonstrated the usefulness of GuMI physiomimetic platform to study microbe-epithelial-immune interactions in healthy and disease conditions.  a2055-5008