Cochrane, S.W.J.; O'Donoghue, D.P.; Baird, A.W.
April 2003
Gut;Apr2003 Supplement 1, Vol. 52, pA8
Academic Journal
M cells are of paramount importance in microbial-intestinal cross talk. There is sparse data on human M cells due to their distribution and the lack of specific markers, however, in vitro co-culture models have been described. The nature of lymphocyte epithelial cell interactions required for M cell transformation as well as the degree of phenotype specific expression in vitro never-the-less remain unclear. We compared co-cultures of polarized Caco-2 cells and Raji B lymphocytes with monocultures of Caco-2 cells in two configurations: one in which there is direct contact between the two cell types (mode 1) and one in which the epithelial cells are exposed to B cell secreted factors only (mode 2). M like cell transformation was assessed by (a) scanning and transmission electron microscopy (EM); (b) apical membrane enzyme activity; (c) translocation of FITC-labelled latex microparticles; and (d) assessing the interaction of the co-cultures with Salmonella typhimurium and Clostridium difficile. EM demonstrated cells with M like morphology in both co-culture models. Down regulation of alkaline phosphatase expression could only be proven in mode 1 (p < 0.005). Microparticle transport was increased in both co-culture configurations compared to caco-2 cell monoculture but was much greater in mode 1 (mode 1 1629+/-453 v 104 +/-11 events, n = 36, p < 0.0001; mode 2 265+/-99 v 121 +/-29 events, n = 34, p = NS). Adherence of S typhimurium to mode 1 was increased compared to caco-2 monoculture > 6-fold (p < 0.005) but that of C. difficile was unchanged. Transformation of absorptive enterocytes to M like cells can be achieved by co-culturing immortalised cell lines of human origin. This transformation is more reproducible if a model using direct contact between epithelial cells and B lymphocytes is employed. Such studies will prove useful in vitro examining the interaction of bacteria which exploit the M cell pathway with polarized epithelia.


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