Live colonocytes in newborn stool: Surrogates for evaluation of gut physiology and disease pathogenesis

Dinesh S. Chandel, Gheorghe T. Braileanu, June Home J Chen, Hegang H. Chen, Pinaki Panigrahi

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Studies of gastrointestinal pathophysiology are not feasible by biopsies in human neonates. We examined the utility of live colonocytes in stool in studying cellular markers during early neonatal life. Expression of IgA, IgG, cluster of differentiation-45 cells (CD45), and toll-like receptors-2 and 4 (TLR2 and TLR4) were analyzed by flow cytometry. Colonocyte RNA extracts were used in quantitative real-time PCR (qRT-PCR) to examine the expression of cytokeratin-19, ribosomal protein-24, and tight-junction (Tj) protein zonula occludens-1 (ZO-1). Colonocyte yield varied between 5 × 104 to 2 × 106 cells/g of stool. Meconium samples yielded a highly enriched population of viable cells. Although low, all samples showed CD45-positive cells during the initial weeks of life. Starting as early as d 2, IgA expression was observed in 69% of the cells. Low to moderate expression of IgG was observed with a linear increase as the infants grew. There was an almost total lack of TLR2 staining; however, <55% of the colonocytes showed TLR4 expression. Although high levels of IgA in gut cells may serve as a natural protectant during neonatal period, increased TLR4 may provide a niche for lipopolysaccharide (LPS)-mediated epithelial damage. Use of stool colonocytes can be a valuable noninvasive approach for studying gut pathophysiology in the neonatal period.

Original languageEnglish (US)
Pages (from-to)153-158
Number of pages6
JournalPediatric Research
Volume70
Issue number2
DOIs
StatePublished - Aug 2011

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

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