Abstract
histology), circularity values increased up to 0.9. C57BL/6 enteroids showed a dose dependent increase in circularity in response to TNF (10-1000ng/ml) or IFN γ (1-100ng/ml) after 24 and 48 hours. Circularity did not however increase in Nfkb2-/enteroids to the same extent following these stimuli. C57BL/6 BMDCs or their CM or LPS alone did not induce an increase in the circularity of C57BL/6, Nfkb2 or Nfkb1-/enteroids. However, when stimulated with 1μg/ml LPS, C57BL/6 BMDCs and their CM induced a significant >2-fold increase in the circularity of C57BL/6 and Nfkb1-/enteroids. Nfkb2 derived enteroids showed no such increase. LPS stimulated BMDCs from Nfkb2-/mice were able to induce a significant increase in C57BL/6 enteroid circularity of a similar magnitude to C57BL/6 BMDCs. Conclusions:Circularity correlates well with cell death and shedding in murine enteroid culture. Deletion of Nfkb2 from intestinal epithelial cells prevents epithelial cell destruction in response to secreted immune cell factors. Therapeutics that specifically target NFKB2 signalling in intestinal epithelial cells may therefore prevent loss of intestinal barrier function and thus ameliorate intestinal inflammation.
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