Document Detail

Live colonocytes in newborn stool: surrogates for evaluation of gut physiology and disease pathogenesis.
MedLine Citation:
PMID:  21544008     Owner:  NLM     Status:  MEDLINE    
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.
Dinesh S Chandel; Gheorghe T Braileanu; June-Home J Chen; Hegang H Chen; Pinaki Panigrahi
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Pediatric research     Volume:  70     ISSN:  1530-0447     ISO Abbreviation:  Pediatr. Res.     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-07-07     Completed Date:  2011-11-15     Revised Date:  2014-09-17    
Medline Journal Info:
Nlm Unique ID:  0100714     Medline TA:  Pediatr Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  153-8     Citation Subset:  IM    
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MeSH Terms
Antigens, CD45 / metabolism
Biological Markers / metabolism*
Colon / cytology*
Epithelial Cells / cytology,  metabolism*
Feces / cytology*
Flow Cytometry
Gastrointestinal Tract / metabolism,  physiology*,  physiopathology*
Immunoglobulin A / metabolism
Immunoglobulin G / metabolism
Infant, Newborn
Keratin-19 / metabolism
Membrane Proteins / metabolism
Phosphoproteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Ribosomal Proteins / metabolism
Toll-Like Receptor 2 / metabolism
Toll-Like Receptor 4 / metabolism
Zonula Occludens-1 Protein
Grant Support
Reg. No./Substance:
0/Biological Markers; 0/Immunoglobulin A; 0/Immunoglobulin G; 0/Keratin-19; 0/Membrane Proteins; 0/Phosphoproteins; 0/Ribosomal Proteins; 0/TJP1 protein, human; 0/Toll-Like Receptor 2; 0/Toll-Like Receptor 4; 0/Zonula Occludens-1 Protein; EC, CD45

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