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.
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.
Institute of Physiology, University of Maribor, Faculty of Medicine, Maribor, Slovenia.
Martin
M
Trapecar
Department of laboratory diagnostics, University Clinical Center Maribor, Maribor, Slovenia.
Ales
A
Goropevsek
University of Maribor, Faculty of Medicine, Maribor, Slovenia.
Mario
M
Gorenjak
Institute of Physiology, University of Maribor, Faculty of Medicine, Maribor, Slovenia.
Lidija
L
Gradisnik
Institute of Physiology, University of Maribor, Faculty of Medicine, Maribor, Slovenia.
Marjan
M
Slak Rupnik
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D018920:Coculture Techniques; D020895:Enterocytes / Q000276:immunology*; D006801:Humans; D007108:Immune Tolerance; D056747:Immunomodulation; D007421:Intestine, Small / Q000166:cytology / Q000276:immunology / Q000382:microbiology; D007778:Lactobacillus / Q000276:immunology / Q000502:physiology*; D008264:Macrophages / Q000276:immunology*; D064307:Microbiota / Q000276:immunology; D021381:Protein Transport; D050794:STAT1 Transcription Factor / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000378:metabolism*
C498143:RELA protein, human; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D051996:Transcription Factor RelA