Development of an Inflammation-Triggered In Vitro “Leaky Gut” Model Using Caco-2/HT29-MTX-E12 Combined with Macrophage-like THP-1 Cells or Primary Human-Derived Macrophages

Abstract

The “leaky gut” syndrome describes a damaged (leaky) intestinal mucosa and is considered a serious contributor to numerous chronic diseases. Chronic inflammatory bowel diseases (IBD) are particularly associated with the “leaky gut” syndrome, but also allergies, autoimmune diseases or neurological disorders. We developed a complex in vitro inflammation-triggered triple-culture model using 21-day-differentiated human intestinal Caco-2 epithelial cells and HT29-MTX-E12 mucus-producing goblet cells (90:10 ratio) in close contact with differentiated human macrophage-like THP-1 cells or primary monocyte-derived macrophages from human peripheral blood. Upon an inflammatory stimulus, the characteristics of a “leaky gut” became evident: a significant loss of intestinal cell integrity in terms of decreased transepithelial/transendothelial electrical resistance (TEER), as well as a loss of tight junction proteins. The cell permeability for FITC-dextran 4 kDa was then increased, and key pro-inflammatory cytokines, including TNF-alpha and IL-6, were substantially released. Whereas in the M1 macrophage-like THP-1 co-culture model, we could not detect the release of IL-23, which plays a crucial regulatory role in IBD, this cytokine was clearly detected when using primary human M1 macrophages instead. In conclusion, we provide an advanced human in vitro model that could be useful for screening and evaluating therapeutic drugs for IBD treatment, including potential IL-23 inhibitors.

Enhanced Cellular Uptake and Transport of Bovine Lactoferrin Using Pectin- and Chitosan-Modified Solid Lipid Nanoparticles

“…A more advanced in vitro intestinal permeability model involving a triple-cell-culture system was developed. This system incorporates enterocytes, mucus-secreting HT29-MTX cells, and M cells [ 67 , 68 ]. M cells exhibit a notable capacity for transcytosis and have the ability to transport a diverse array of materials, encompassing nanoparticles as well [ 69 , 70 ].…”

Understanding the Immunomodulatory Effects of Bovine Colostrum: Insights into IL-6/IL-10 Axis-Mediated Inflammatory Control

“…Therefore, we employed a well-established Caco-2/THP-1 intestinal model to investigate the innate immune responses to bovine colostrum [ 42 ]. To induce inflammatory processes, we utilized phorbol 12-myristate 13-acetate (PMA), a non-specific mitogen that acts through protein kinase C (PKC), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and mito-gen-activated protein kinase (MAPK) signaling pathways, ultimately resulting in the production of pro-inflammatory cytokines across various cell lines, including Caco-2/THP-1 co-culture [ 30 , 43 , 44 ].…”

Understanding the Immunomodulatory Effects of Bovine Colostrum: Insights into IL-6/IL-10 Axis-Mediated Inflammatory Control

“…Therefore, we employed a well-established Caco-2/THP-1 intestinal model to investigate the innate immune responses to bovine colostrum [40]. To induce inflammatory processes, we utilized phorbol 12-myristate 13-acetate (PMA), a non-specific mitogen that acts through protein kinase C (PKC), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and mito-genactivated protein kinase (MAPK) signalling pathways, ultimately resulting in the production of pro-inflammatory cytokines across various cell lines, including Caco-2/THP-1 co-culture [28,41,42].…”