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. 1995 Oct;37(4):536–543. doi: 10.1136/gut.37.4.536

Interleukin 8 secretion by colonic crypt cells in vitro: response to injury suppressed by butyrate and enhanced in inflammatory bowel disease.

P Gibson 1, O Rosella 1
PMCID: PMC1382907  PMID: 7489942

Abstract

Epithelia from several sites exhibit inducible secretion of interleukin 8 (IL-8). This study aimed to characterise secretion of IL-8 by colonic epithelial cells in vitro. Colonic crypt cells were isolated enzymatically from resected colon and the IL-8 content of culture supernates was measured by ELISA. The rate of secretion of IL-8 accelerated and levels of IL-8 transcripts increased appreciably during culture. Exposure to tumour necrosis factor alpha (TNF alpha) failed to increase secretion further. Secretion was not induced by the enzymatic digestion or by serum used in the culture medium but was significantly inhibited by butyrate, by a mean of 23%. Control experiments indicated that colonic crypt cells were the likely source. The secretion of IL-8 over 24 hours by cells from uninflamed mucosa of patients with ulcerative colitis or Crohn's disease was more than twofold that from normal cells, while that from cancer bearing colons was normal. TNF alpha (10 mM) significantly suppressed IL-8 secretion only in the ulcerative colitis group and the change was different to those in the normal (p = 0.007) and Crohn's disease groups (p = 0.012). Cells from inflamed areas secreted more IL-8 than those from autologous uninflamed areas (p = 0.009) but responses to modulating factors were no different. The induction of IL-8 secretion by colonic crypt cells in vitro is probably a response to injury associated with isolation and culture. It is suppressed by butyrate and increased in inflammatory bowel disease independently of the presence of mucosal inflammation. Whether epithelial derived IL-8 plays a part in the pathogenesis of inflammatory bowel disease is not yet clear.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barker J. N., Jones M. L., Mitra R. S., Crockett-Torabe E., Fantone J. C., Kunkel S. L., Warren J. S., Dixit V. M., Nickoloff B. J. Modulation of keratinocyte-derived interleukin-8 which is chemotactic for neutrophils and T lymphocytes. Am J Pathol. 1991 Oct;139(4):869–876. [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Choi A. M., Jacoby D. B. Influenza virus A infection induces interleukin-8 gene expression in human airway epithelial cells. FEBS Lett. 1992 Sep 14;309(3):327–329. doi: 10.1016/0014-5793(92)80799-m. [DOI] [PubMed] [Google Scholar]
  5. Dixit V. M., Green S., Sarma V., Holzman L. B., Wolf F. W., O'Rourke K., Ward P. A., Prochownik E. V., Marks R. M. Tumor necrosis factor-alpha induction of novel gene products in human endothelial cells including a macrophage-specific chemotaxin. J Biol Chem. 1990 Feb 15;265(5):2973–2978. [PubMed] [Google Scholar]
  6. Eckmann L., Jung H. C., Schürer-Maly C., Panja A., Morzycka-Wroblewska E., Kagnoff M. F. Differential cytokine expression by human intestinal epithelial cell lines: regulated expression of interleukin 8. Gastroenterology. 1993 Dec;105(6):1689–1697. doi: 10.1016/0016-5085(93)91064-o. [DOI] [PubMed] [Google Scholar]
  7. Gibson P. R., Moeller I., Kagelari O., Folino M., Young G. P. Contrasting effects of butyrate on the expression of phenotypic markers of differentiation in neoplastic and non-neoplastic colonic epithelial cells in vitro. J Gastroenterol Hepatol. 1992 Mar-Apr;7(2):165–172. doi: 10.1111/j.1440-1746.1992.tb00956.x. [DOI] [PubMed] [Google Scholar]
  8. Gibson P. R., Rosella O., Rosella G., Young G. P. Butyrate is a potent inhibitor of urokinase secretion by normal colonic epithelium in vitro. Gastroenterology. 1994 Aug;107(2):410–419. doi: 10.1016/0016-5085(94)90166-x. [DOI] [PubMed] [Google Scholar]
  9. Gibson P. R., van de Pol E., Maxwell L. E., Gabriel A., Doe W. F. Isolation of colonic crypts that maintain structural and metabolic viability in vitro. Gastroenterology. 1989 Feb;96(2 Pt 1):283–291. doi: 10.1016/0016-5085(89)91549-7. [DOI] [PubMed] [Google Scholar]
  10. Gibson P., Folino M., McIntyre A., Rosella O., Finch C., Young G. Dietary modulation of colonic mucosal urokinase activity in rats. J Gastroenterol Hepatol. 1995 May-Jun;10(3):324–330. doi: 10.1111/j.1440-1746.1995.tb01101.x. [DOI] [PubMed] [Google Scholar]
  11. Gibson P., Rosella O., Young G. Serum free medium increases expression of markers of differentiation in human colonic crypt cells. Gut. 1994 Jun;35(6):791–797. doi: 10.1136/gut.35.6.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glotzer D. J., Glick M. E., Goldman H. Proctitis and colitis following diversion of the fecal stream. Gastroenterology. 1981 Mar;80(3):438–441. [PubMed] [Google Scholar]
  13. Izzo R. S., Witkon K., Chen A. I., Hadjiyane C., Weinstein M. I., Pellecchia C. Interleukin-8 and neutrophil markers in colonic mucosa from patients with ulcerative colitis. Am J Gastroenterol. 1992 Oct;87(10):1447–1452. [PubMed] [Google Scholar]
  14. Izzo R. S., Witkon K., Chen A. I., Hadjiyane C., Weinstein M. I., Pellecchia C. Neutrophil-activating peptide (interleukin-8) in colonic mucosa from patients with Crohn's disease. Scand J Gastroenterol. 1993 Apr;28(4):296–300. doi: 10.3109/00365529309090244. [DOI] [PubMed] [Google Scholar]
  15. Janne P., Carpentier Y., Willems G. Colonic mucosal atrophy induced by a liquid elemental diet in rats. Am J Dig Dis. 1977 Sep;22(9):808–812. doi: 10.1007/BF01694512. [DOI] [PubMed] [Google Scholar]
  16. Kripke S. A., Fox A. D., Berman J. M., Settle R. G., Rombeau J. L. Stimulation of intestinal mucosal growth with intracolonic infusion of short-chain fatty acids. JPEN J Parenter Enteral Nutr. 1989 Mar-Apr;13(2):109–116. doi: 10.1177/0148607189013002109. [DOI] [PubMed] [Google Scholar]
  17. Labarca C., Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  18. Oakes J. E., Monteiro C. A., Cubitt C. L., Lausch R. N. Induction of interleukin-8 gene expression is associated with herpes simplex virus infection of human corneal keratocytes but not human corneal epithelial cells. J Virol. 1993 Aug;67(8):4777–4784. doi: 10.1128/jvi.67.8.4777-4784.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roediger W. E. Role of anaerobic bacteria in the metabolic welfare of the colonic mucosa in man. Gut. 1980 Sep;21(9):793–798. doi: 10.1136/gut.21.9.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Roediger W. E. The colonic epithelium in ulcerative colitis: an energy-deficiency disease? Lancet. 1980 Oct 4;2(8197):712–715. doi: 10.1016/s0140-6736(80)91934-0. [DOI] [PubMed] [Google Scholar]
  21. Sakata T. Depression of intestinal epithelial cell production rate by hindgut bypass in rats. Scand J Gastroenterol. 1988 Dec;23(10):1200–1202. doi: 10.3109/00365528809090191. [DOI] [PubMed] [Google Scholar]
  22. Sakata T. Stimulatory effect of short-chain fatty acids on epithelial cell proliferation in the rat intestine: a possible explanation for trophic effects of fermentable fibre, gut microbes and luminal trophic factors. Br J Nutr. 1987 Jul;58(1):95–103. doi: 10.1079/bjn19870073. [DOI] [PubMed] [Google Scholar]
  23. Scheppach W., Sommer H., Kirchner T., Paganelli G. M., Bartram P., Christl S., Richter F., Dusel G., Kasper H. Effect of butyrate enemas on the colonic mucosa in distal ulcerative colitis. Gastroenterology. 1992 Jul;103(1):51–56. doi: 10.1016/0016-5085(92)91094-k. [DOI] [PubMed] [Google Scholar]
  24. Schmouder R. L., Strieter R. M., Wiggins R. C., Chensue S. W., Kunkel S. L. In vitro and in vivo interleukin-8 production in human renal cortical epithelia. Kidney Int. 1992 Jan;41(1):191–198. doi: 10.1038/ki.1992.26. [DOI] [PubMed] [Google Scholar]
  25. Whitehead R. H., Jones J. K., Gabriel A., Lukies R. E. A new colon carcinoma cell line (LIM1863) that grows as organoids with spontaneous differentiation into crypt-like structures in vitro. Cancer Res. 1987 May 15;47(10):2683–2689. [PubMed] [Google Scholar]
  26. de Bruin P. A., Crama-Bohbouth G., Verspaget H. W., Verheijen J. H., Dooijewaard G., Weterman I. T., Lamers C. B. Plasminogen activators in the intestine of patients with inflammatory bowel disease. Thromb Haemost. 1988 Oct 31;60(2):262–266. [PubMed] [Google Scholar]

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