Abstract
Paneth cell dysfunction has been suggested in necrotizing enterocolitis (NEC). The aim of this study was to i) study Paneth cell presence, protein expression, and developmental changes in preterm infants with NEC and ii) determine Paneth cell products and antimicrobial capacity in ileostomy outflow fluid. Intestinal tissue from NEC patients (n = 55), preterm control infants (n = 22), and term controls (n = 7) was obtained during surgical resection and at stoma closure after recovery. Paneth cell abundance and protein expression were analyzed by immunohistochemistry. RNA levels of Paneth cell proteins were determined by real-time quantitative RT-PCR. In ileostomy outflow fluid, Paneth cell products were quantified, and antimicrobial activity was measured in vitro. In acute NEC, Paneth cell abundance in small intestinal tissue was not significantly different from preterm controls. After recovery from NEC, Paneth cell hyperplasia was observed in the small intestine concomitant with elevated human alpha-defensin 5 mRNA levels. In the colon, metaplastic Paneth cells were observed. Ileostomy fluid contained Paneth cell proteins and inhibited bacterial growth. In conjunction, these data suggest an important role of Paneth cells and their products in various phases of NEC.
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Abbreviations
- A-NEC:
-
acute phase of necrotizing enterocolitis
- CFU:
-
colony forming units
- HD:
-
human defensins
- LGG:
-
Lactobacillus rhamnosus GG
- NEC:
-
necrotizing enterocolitis
- NEC-R:
-
stoma reanastomosis after necrotizing enterocolitis
- post-NEC stricture:
-
stricture formation after acute phase of necrotizing enterocolitis
- preterm CO:
-
preterm control patients
- preterm CO-R:
-
stoma reanastomosis in preterm control patient
- qRT-PCR:
-
real-time quantitative PCR
- sPLA2:
-
secretory phospholipase-A2
- term CO:
-
term control patient
References
Paneth J 1888 [About the secreting cells in the small intestinal epithelium]. Arch Mikroskop Anat 31: 113–191
Sandow MJ, Whitehead R 1979 The Paneth cell. Gut 20: 420–431
Ganz T 2003 Defensins: antimicrobial peptides of innate immunity. Nat Rev Immunol 3: 710–720
Salzman NH, Underwood MA, Bevins CL 2007 Paneth cells, defensins, and the commensal microbiota: a hypothesis on intimate interplay at the intestinal mucosa. Semin Immunol 19: 70–83
Ayabe T, Satchell DP, Wilson CL, Parks WC, Selsted ME, Ouellette AJ 2000 Secretion of microbicidal alpha-defensins by intestinal Paneth cells in response to bacteria. Nat Immunol 1: 113–118
Salzman NH, Ghosh D, Huttner KM, Paterson Y, Bevins CL 2003 Protection against enteric salmonellosis in transgenic mice expressing a human intestinal defensin. Nature 422: 522–526
Fernandez MI, Regnault B, Mulet C, Tanguy M, Jay P, Sansonetti PJ, Pedron T 2008 Maturation of paneth cells induces the refractory state of newborn mice to Shigella infection. J Immunol 180: 4924–4930
Wilson CL, Ouellette AJ, Satchell DP, Ayabe T, Lopez-Boado YS, Stratman JL, Hultgren SJ, Matrisian LM, Parks WC 1999 Regulation of intestinal alpha-defensin activation by the metalloproteinase matrilysin in innate host defense. Science 286: 113–117
Salzman NH, Hung K, Haribhai D, Chu H, Karlsson-Sjoberg J, Amir E, Teggatz P, Barman M, Hayward M, Eastwood D, Stoel M, Zhou Y, Sodergren E, Weinstock GM, Bevins CL, Williams CB, Bos NA 2010 Enteric defensins are essential regulators of intestinal microbial ecology. Nat Immunol 11: 76–83
Lin PW, Simon PO Jr Gewirtz AT, Neish AS, Ouellette AJ, Madara JL, Lencer WI 2004 Paneth cell cryptdins act in vitro as apical paracrine regulators of the innate inflammatory response. J Biol Chem 279: 19902–19907
Lencer WI, Cheung G, Strohmeier GR, Currie MG, Ouellette AJ, Selsted ME, Madara JL 1997 Induction of epithelial chloride secretion by channel-forming cryptdins 2 and 3. Proc Natl Acad Sci U S A 94: 8585–8589
Erlandsen SL, Parsons JA, Taylor TD 1974 Ultrastructural immunocytochemical localization of lysozyme in the Paneth cells of man. J Histochem Cytochem 22: 401–413
Klockars M, Reitamo S 1975 Tissue distribution of lysozyme in man. J Histochem Cytochem 23: 932–940
Kiyohara H, Egami H, Shibata Y, Murata K, Ohshima S, Ogawa M 1992 Light microscopic immunohistochemical analysis of the distribution of group II phospholipase A2 in human digestive organs. J Histochem Cytochem 40: 1659–1664
Jones DE, Bevins CL 1992 Paneth cells of the human small intestine express an antimicrobial peptide gene. J Biol Chem 267: 23216–23225
Jones DE, Bevins CL 1993 Defensin-6 mRNA in human Paneth cells: implications for antimicrobial peptides in host defense of the human bowel. FEBS Lett 315: 187–192
Klockars M, Reitamo S, Adinolfi M 1977 Ontogeny of human lysozyme. Distribution in fetal tissues. Biol Neonate 32: 243–249
Mallow EB, Harris A, Salzman N, Russell JP, DeBerardinis RJ, Ruchelli E, Bevins CL 1996 Human enteric defensins. Gene structure and developmental expression. J Biol Chem 271: 4038–4045
Coutinho HB, da Mota HC, Coutinho VB, Robalinho TI, Furtado AF, Walker E, King G, Mahida YR, Sewell HF, Wakelin D 1998 Absence of lysozyme (muramidase) in the intestinal Paneth cells of newborn infants with necrotising enterocolitis. J Clin Pathol 51: 512–514
Sharma R, Tepas JJ III Hudak ML, Wludyka PS, Mollitt DL, Garrison RD, Bradshaw JA, Sharma M 2005 Portal venous gas and surgical outcome of neonatal necrotizing enterocolitis. J Pediatr Surg 40: 371–376
Blakely ML, Lally KP, McDonald S, Brown RL, Barnhart DC, Ricketts RR, Thompson WR, Scherer LR, Klein MD, Letton RW, Chwals WJ, Touloukian RJ, Kurkchubasche AG, Skinner MA, Moss RL, Hilfiker ML 2005 Postoperative outcomes of extremely low birth-weight infants with necrotizing enterocolitis or isolated intestinal perforation: a prospective cohort study by the NICHD Neonatal Research Network. Ann Surg 241: 984–989; discussion 989–994.
Lin PW, Stoll BJ 2006 Necrotising enterocolitis. Lancet 368: 1271–1283
Lucas A, Cole TJ 1990 Breast milk and neonatal necrotising enterocolitis. Lancet 336: 1519–1523
Claud EC, Walker WA 2001 Hypothesis: inappropriate colonization of the premature intestine can cause neonatal necrotizing enterocolitis. FASEB J 15: 1398–1403
Bell MJ, Ternberg JL, Feigin RD, Keating JP, Marshall R, Barton L, Brotherton T 1978 Neonatal necrotizing enterocolitis. Therapeutic decisions based upon clinical staging. Ann Surg 187: 1–7
Renes IB, Verburg M, Bulsing NP, Ferdinandusse S, Buller HA, Dekker J, Einerhand AW 2002 Protection of the Peyer's patch-associated crypt and villus epithelium against methotrexate-induced damage is based on its distinct regulation of proliferation. J Pathol 198: 60–68
Schaart MW, Yamanouchi T, van Nispen DJ, Raatgeep RH, van Goudoever JB, de Krijger RR, Tibboel D, Einerhand AW, Renes IB 2006 Does small intestinal atresia affect epithelial protein expression in human newborns?. J Pediatr Gastroenterol Nutr 43: 576–583
Meijerink J, Mandigers C, van de Locht L, Tonnissen E, Goodsaid F, Raemaekers J 2001 A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR. J Mol Diagn 3: 55–61
Schaart MW, Schierbeek H, de Bruijn AC, Tibboel D, van Goudoever JB, Renes IB 2006 A novel method to determine small intestinal barrier function in human neonates in vivo. Gut 55: 1366–1367
Tytgat KM, Klomp LW, Bovelander FJ, Opdam FJ, Van der Wurff A, Einerhand AW, Buller HA, Strous GJ, Dekker J 1995 Preparation of anti-mucin polypeptide antisera to study mucin biosynthesis. Anal Biochem 226: 331–341
Holman RC, Stoll BJ, Clarke MJ, Glass RI 1997 The epidemiology of necrotizing enterocolitis infant mortality in the United States. Am J Public Health 87: 2026–2031
Maayan-Metzger A, Itzchak A, Mazkereth R, Kuint J 2004 Necrotizing enterocolitis in full-term infants: case-control study and review of the literature. J Perinatol 24: 494–499
Sankaran K, Puckett B, Lee DS, Seshia M, Boulton J, Qiu Z, Lee SK 2004 Variations in incidence of necrotizing enterocolitis in Canadian neonatal intensive care units. J Pediatr Gastroenterol Nutr 39: 366–372
Salzman NH, Polin RA, Harris MC, Ruchelli E, Hebra A, Zirin-Butler S, Jawad A, Martin Porter E, Bevins CL 1998 Enteric defensin expression in necrotizing enterocolitis. Pediatr Res 44: 20–26
Tanaka M, Saito H, Kusumi T, Fukuda S, Shimoyama T, Sasaki Y, Suto K, Munakata A, Kudo H 2001 Spatial distribution and histogenesis of colorectal Paneth cell metaplasia in idiopathic inflammatory bowel disease. J Gastroenterol Hepatol 16: 1353–1359
Helmrath MA, Fong JJ, Dekaney CM, Henning SJ 2007 Rapid expansion of intestinal secretory lineages following a massive small bowel resection in mice. Am J Physiol Gastrointest Liver Physiol 292: G215–G222
Eisenhauer PB, Harwig SS, Lehrer RI 1992 Cryptdins: antimicrobial defensins of the murine small intestine. Infect Immun 60: 3556–3565
Ouellette AJ, Hsieh MM, Nosek MT, Cano-Gauci DF, Huttner KM, Buick RN, Selsted ME 1994 Mouse Paneth cell defensins: primary structures and antibacterial activities of numerous cryptdin isoforms. Infect Immun 62: 5040–5047
Porter EM, van Dam E, Valore EV, Ganz T 1997 Broad-spectrum antimicrobial activity of human intestinal defensin 5. Infect Immun 65: 2396–2401
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We thank J. Bouma for excellent technical assistance.
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Puiman, P., Burger-van Paassen, N., Schaart, M. et al. Paneth Cell Hyperplasia and Metaplasia in Necrotizing Enterocolitis. Pediatr Res 69, 217–223 (2011). https://doi.org/10.1203/PDR.0b013e3182092a9a
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DOI: https://doi.org/10.1203/PDR.0b013e3182092a9a
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