Skip to main content
Top
Published in: Surgery Today 8/2020

01-08-2020 | Ulcerative Colitis | Original Article

Resistin-like molecule beta, a colonic epithelial protein, exhibits antimicrobial activity against Staphylococcus aureus including methicillin-resistant strains

Authors: Kazuhiro Watanabe, Kikuji Itoh, Sang-Hee Park, Mitsuo Kaku, Keiko Ishii, Hironobu Sasano, Takeshi Naitoh, Michiaki Unno, Kouhei Fukushima

Published in: Surgery Today | Issue 8/2020

Login to get access

Abstract

Purpose

Resistin-like molecule beta (RELMβ) is a small cysteine-rich protein secreted by colonic epithelial cells. RELMβ mRNA and protein expressions are dramatically induced by bacterial exposure in germ-free mice. We hypothesized that RELMβ has antimicrobial activity.

Methods

The antimicrobial activity of RELMβ was screened by an agar spot test and confirmed by a liquid broth test. The amount of RELMβ in human stools was semi-quantified by Western blot analysis. The induction of RELMβ mRNA and protein expression by bacteria was measured by quantitative RT-PCR using LS174T cells. Electron microscopic immunohistochemistry was performed using polyclonal anti-RELMβ antibody.

Results

RELMβ showed antimicrobial activity against S. aureus and all MRSAs examined in a dose- and pH-dependent fashion. Western blot study showed that the amount of RELMβ in healthy human stools was comparable to that exhibiting antimicrobial activity in vitro. Both RELMβ mRNA and protein expression were induced by heat-inactivated S. aureus, but not by E. coli in LS174T cells. Electron microscopic immunohistochemistry showed that RELMβ bound to the cell surface of S. aureus, followed by destruction of the bacterial cytoplasm.

Conclusions

RELMβ is a colonic antimicrobial protein and its antibacterial activity is species selective. Because RELMβ is abundant in healthy human stool, RELMβ may modulate gut flora.
Literature
2.
go back to reference Zasloff M. Antimicrobial peptides of multicellular organisms. Nature. 2002;415:389–95.CrossRef Zasloff M. Antimicrobial peptides of multicellular organisms. Nature. 2002;415:389–95.CrossRef
3.
go back to reference Mukherjee S, Hooper LV. Antimicrobial defense of the intestine. Immunity. 2015;42:28–39.CrossRef Mukherjee S, Hooper LV. Antimicrobial defense of the intestine. Immunity. 2015;42:28–39.CrossRef
5.
go back to reference Garcia JR, Krause A, Schulz S, Rodríguez-Jiménez FJ, Klüver E, Adermann K, et al. Human beta-defensin 4: a novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. Faseb J. 2001;15:1819–21.CrossRef Garcia JR, Krause A, Schulz S, Rodríguez-Jiménez FJ, Klüver E, Adermann K, et al. Human beta-defensin 4: a novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. Faseb J. 2001;15:1819–21.CrossRef
6.
go back to reference Glaser R, Harder J, Lange H, Bartels J, Christophers E, Schröder JM. Antimicrobial psoriasin (S100A7) protects human skin from Escherichia coli infection. Nat Immunol. 2005;6:57–64.CrossRef Glaser R, Harder J, Lange H, Bartels J, Christophers E, Schröder JM. Antimicrobial psoriasin (S100A7) protects human skin from Escherichia coli infection. Nat Immunol. 2005;6:57–64.CrossRef
7.
go back to reference Schroeder BO, Ehmann D, Precht JC, Castillo PA, Küchler R, Berger J, et al. Paneth cell alpha-defensin 6 (HD-6) is an antimicrobial peptide. Mucosal Immunol. 2015;8:661–71.CrossRef Schroeder BO, Ehmann D, Precht JC, Castillo PA, Küchler R, Berger J, et al. Paneth cell alpha-defensin 6 (HD-6) is an antimicrobial peptide. Mucosal Immunol. 2015;8:661–71.CrossRef
8.
go back to reference Okumura R, Takeda K. Roles of intestinal epithelial cells in the maintenance of gut homeostasis. Exp Mol Med. 2017;49:e338.CrossRef Okumura R, Takeda K. Roles of intestinal epithelial cells in the maintenance of gut homeostasis. Exp Mol Med. 2017;49:e338.CrossRef
9.
go back to reference Hendrikx T, Schnabl B. Antimicrobial proteins: intestinal guards to protect against liver disease. J Gastroenterol. 2018;54(3):209–17.CrossRef Hendrikx T, Schnabl B. Antimicrobial proteins: intestinal guards to protect against liver disease. J Gastroenterol. 2018;54(3):209–17.CrossRef
10.
go back to reference Ciumac D, Gong H, Hu X, Lu JR. Membrane targeting cationic antimicrobial peptides. J Colloid Interface Sci. 2018;537:163–85.CrossRef Ciumac D, Gong H, Hu X, Lu JR. Membrane targeting cationic antimicrobial peptides. J Colloid Interface Sci. 2018;537:163–85.CrossRef
11.
go back to reference Jones DE, Bevins CL. Paneth cells of the human small intestine express an antimicrobial peptide gene. J Biol Chem. 1992;267:23216–25.PubMed Jones DE, Bevins CL. Paneth cells of the human small intestine express an antimicrobial peptide gene. J Biol Chem. 1992;267:23216–25.PubMed
12.
go back to reference O'Neil DA, Porter EM, Elewaut D, Anderson GM, Eckmann L, Ganz T, et al. Expression and regulation of the human beta-defensins hBD-1 and hBD-2 in intestinal epithelium. J Immunol. 1999;163:6718–24.PubMed O'Neil DA, Porter EM, Elewaut D, Anderson GM, Eckmann L, Ganz T, et al. Expression and regulation of the human beta-defensins hBD-1 and hBD-2 in intestinal epithelium. J Immunol. 1999;163:6718–24.PubMed
13.
go back to reference Xhindoli D, Pacor S, Benincasa M, Scocchi M, Gennaro R, Tossi A. The human cathelicidin LL-37—a pore-forming antibacterial peptide and host-cell modulator. Biochim Biophys Acta. 2016;1858:546–66.CrossRef Xhindoli D, Pacor S, Benincasa M, Scocchi M, Gennaro R, Tossi A. The human cathelicidin LL-37—a pore-forming antibacterial peptide and host-cell modulator. Biochim Biophys Acta. 2016;1858:546–66.CrossRef
14.
go back to reference Zhao C, Wang I, Lehrer RI. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Lett. 1996;396:319–22.CrossRef Zhao C, Wang I, Lehrer RI. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Lett. 1996;396:319–22.CrossRef
15.
go back to reference Frohm Nilsson M, Sandstedt B, Sorensen O, Weber G, Borregaard N, Ståhle-Bäckdahl M. The human cationic antimicrobial protein (hCAP18), a peptide antibiotic, is widely expressed in human squamous epithelia and colocalizes with interleukin-6. Infect Immun. 1999;67:2561–6.CrossRef Frohm Nilsson M, Sandstedt B, Sorensen O, Weber G, Borregaard N, Ståhle-Bäckdahl M. The human cationic antimicrobial protein (hCAP18), a peptide antibiotic, is widely expressed in human squamous epithelia and colocalizes with interleukin-6. Infect Immun. 1999;67:2561–6.CrossRef
16.
go back to reference Hase K, Eckmann L, Leopard JD, Varki N, Kagnoff MF. Cell differentiation is a key determinant of cathelicidin LL-37/human cationic antimicrobial protein 18 expression by human colon epithelium. Infect Immun. 2002;70:953–63.CrossRef Hase K, Eckmann L, Leopard JD, Varki N, Kagnoff MF. Cell differentiation is a key determinant of cathelicidin LL-37/human cationic antimicrobial protein 18 expression by human colon epithelium. Infect Immun. 2002;70:953–63.CrossRef
17.
go back to reference Steppan CM, Brown EJ, Wright CM, Bhat S, Banerjee RR, Dai CY, et al. A family of tissue-specific resistin-like molecules. Proc Natl Acad Sci USA. 2001;98:502–6.CrossRef Steppan CM, Brown EJ, Wright CM, Bhat S, Banerjee RR, Dai CY, et al. A family of tissue-specific resistin-like molecules. Proc Natl Acad Sci USA. 2001;98:502–6.CrossRef
18.
go back to reference He W, Wang ML, Jiang HQ, Steppan CM, Shin ME, Thurnheer MC, et al. Bacterial colonization leads to the colonic secretion of RELMbeta/FIZZ2, a novel goblet cell-specific protein. Gastroenterology. 2003;125:1388–97.CrossRef He W, Wang ML, Jiang HQ, Steppan CM, Shin ME, Thurnheer MC, et al. Bacterial colonization leads to the colonic secretion of RELMbeta/FIZZ2, a novel goblet cell-specific protein. Gastroenterology. 2003;125:1388–97.CrossRef
19.
go back to reference Knight PA, Pemberton AD, Robertson KA, Roy DJ, Wright SH, Miller HR, et al. Expression profiling reveals novel innate and inflammatory responses in the jejunal epithelial compartment during infection with Trichinella spiralis. Infect Immun. 2004;72:6076–86.CrossRef Knight PA, Pemberton AD, Robertson KA, Roy DJ, Wright SH, Miller HR, et al. Expression profiling reveals novel innate and inflammatory responses in the jejunal epithelial compartment during infection with Trichinella spiralis. Infect Immun. 2004;72:6076–86.CrossRef
20.
go back to reference Artis D, Wang ML, Keilbaugh SA, He W, Brenes M, Swain GP, et al. RELMbeta/FIZZ2 is a goblet cell-specific immune-effector molecule in the gastrointestinal tract. Proc Natl Acad Sci USA. 2004;101:13596–600.CrossRef Artis D, Wang ML, Keilbaugh SA, He W, Brenes M, Swain GP, et al. RELMbeta/FIZZ2 is a goblet cell-specific immune-effector molecule in the gastrointestinal tract. Proc Natl Acad Sci USA. 2004;101:13596–600.CrossRef
21.
go back to reference Kitayama T, Fukushima K, Naito H, Funayama Y, Shibata C, Hashimoto A, et al. Possible association of resistin-like molecule beta (RELMβ) in pathogenesis of ulcerative colitis (UC). Gastroenterology. 2002;122(suppl):A261. Kitayama T, Fukushima K, Naito H, Funayama Y, Shibata C, Hashimoto A, et al. Possible association of resistin-like molecule beta (RELMβ) in pathogenesis of ulcerative colitis (UC). Gastroenterology. 2002;122(suppl):A261.
22.
go back to reference Watanabe K, Fukushima K, Itoh K, Park SH, Kaku M, Ishii K, et al. Resistin-like molecule beta (RELMβ), a novel secretory peptide from human colonic epithelial cells, exhibits antimicrobial activity against Staphylococcus aureus including methicillin-resistant S. aureus (MRSA). Gastroenterology. 2005;128(suppl):A196. Watanabe K, Fukushima K, Itoh K, Park SH, Kaku M, Ishii K, et al. Resistin-like molecule beta (RELMβ), a novel secretory peptide from human colonic epithelial cells, exhibits antimicrobial activity against Staphylococcus aureus including methicillin-resistant S. aureus (MRSA). Gastroenterology. 2005;128(suppl):A196.
23.
go back to reference Schillinger U, Lucke FK. Antibacterial activity of Lactobacillus sakei isolated from meat. Appl Environ Microbiol. 1989;55:1901–6.CrossRef Schillinger U, Lucke FK. Antibacterial activity of Lactobacillus sakei isolated from meat. Appl Environ Microbiol. 1989;55:1901–6.CrossRef
24.
go back to reference Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: general principles and contemporary practices. Clin Infect Dis. 1998;26:973–80.CrossRef Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: general principles and contemporary practices. Clin Infect Dis. 1998;26:973–80.CrossRef
25.
go back to reference Yano Y, Matsui T, Hirai F, Okado Y, Sato Y, Tsurumi K, et al. Cancer risk in Japanese Crohn’s disease patients: investigation of the standardized incidence ratio. J Gastroenterol Hepatol. 2013;28:1300–5.CrossRef Yano Y, Matsui T, Hirai F, Okado Y, Sato Y, Tsurumi K, et al. Cancer risk in Japanese Crohn’s disease patients: investigation of the standardized incidence ratio. J Gastroenterol Hepatol. 2013;28:1300–5.CrossRef
26.
go back to reference Song MD, Wachi M, Doi M, Ishino F, Matsuhashi M. Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett. 1987;221:167–71.CrossRef Song MD, Wachi M, Doi M, Ishino F, Matsuhashi M. Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett. 1987;221:167–71.CrossRef
27.
go back to reference Fukushima K, Sasaki I, Hasegawa H, Takahashi K, Naito H, Funayama Y, et al. Sodium butyrate-induced liver-type alkaline phosphatase activity in a small intestinal epithelial cell line, IEC6. Dig Dis Sci. 1998;43:1116–23.CrossRef Fukushima K, Sasaki I, Hasegawa H, Takahashi K, Naito H, Funayama Y, et al. Sodium butyrate-induced liver-type alkaline phosphatase activity in a small intestinal epithelial cell line, IEC6. Dig Dis Sci. 1998;43:1116–23.CrossRef
28.
go back to reference Porter EM, van Dam E, Valore EV, Ganz T. Broad-spectrum antimicrobial activity of human intestinal defensin 5. Infect Immun. 1997;65:2396–401.CrossRef Porter EM, van Dam E, Valore EV, Ganz T. Broad-spectrum antimicrobial activity of human intestinal defensin 5. Infect Immun. 1997;65:2396–401.CrossRef
29.
go back to reference Lee IH, Cho Y, Lehrer RI. Effects of pH and salinity on the antimicrobial properties of clavanins. Infect Immun. 1997;65:2898–903.CrossRef Lee IH, Cho Y, Lehrer RI. Effects of pH and salinity on the antimicrobial properties of clavanins. Infect Immun. 1997;65:2898–903.CrossRef
30.
go back to reference Lai R, Takeuchi H, Lomas LO, Jonczy J, Rigden DJ, Rees HH, et al. A new type of antimicrobial protein with multiple histidines from the hard tick, Amblyomma hebraeum. Faseb J. 2004;18:1447–9.CrossRef Lai R, Takeuchi H, Lomas LO, Jonczy J, Rigden DJ, Rees HH, et al. A new type of antimicrobial protein with multiple histidines from the hard tick, Amblyomma hebraeum. Faseb J. 2004;18:1447–9.CrossRef
31.
go back to reference Michel M, Gutmann L. Methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci: therapeutic realities and possibilities. Lancet. 1997;349:1901–6.CrossRef Michel M, Gutmann L. Methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci: therapeutic realities and possibilities. Lancet. 1997;349:1901–6.CrossRef
32.
go back to reference Guarner F, Malagelada JR. Gut flora in health and disease. Lancet. 2003;361:512–9.CrossRef Guarner F, Malagelada JR. Gut flora in health and disease. Lancet. 2003;361:512–9.CrossRef
33.
go back to reference Moore JE, Corcoran D, Dooley JS, Fanning S, Lucey B, Matsuda M, et al. Campylobacter. Vet Res. 2005;36:351–82.CrossRef Moore JE, Corcoran D, Dooley JS, Fanning S, Lucey B, Matsuda M, et al. Campylobacter. Vet Res. 2005;36:351–82.CrossRef
34.
go back to reference Ohkusa T, Okayasu I, Ogihara T, Morita K, Ogawa M, Sato N. Induction of experimental ulcerative colitis by Fusobacterium varium isolated from colonic mucosa of patients with ulcerative colitis. Gut. 2003;52:79–83.CrossRef Ohkusa T, Okayasu I, Ogihara T, Morita K, Ogawa M, Sato N. Induction of experimental ulcerative colitis by Fusobacterium varium isolated from colonic mucosa of patients with ulcerative colitis. Gut. 2003;52:79–83.CrossRef
35.
go back to reference Kodama T, Santo T, Yokoyama T, Takesue Y, Hiyama E, Imamura Y, et al. Postoperative enteritis caused by methicillin-resistant Staphylococcus aureus. Surg Today. 1997;27:816–25.CrossRef Kodama T, Santo T, Yokoyama T, Takesue Y, Hiyama E, Imamura Y, et al. Postoperative enteritis caused by methicillin-resistant Staphylococcus aureus. Surg Today. 1997;27:816–25.CrossRef
36.
go back to reference Evans DF, Pye G, Bramley R, Clark AG, Dyson TJ, Hardcastle JD. Measurement of gastrointestinal pH profiles in normal ambulant human subjects. Gut. 1988;29:1035–41.CrossRef Evans DF, Pye G, Bramley R, Clark AG, Dyson TJ, Hardcastle JD. Measurement of gastrointestinal pH profiles in normal ambulant human subjects. Gut. 1988;29:1035–41.CrossRef
37.
go back to reference Propheter DC, Chara AL, Harris TA, Ruhn KA, Hooper LV. Resistin-like molecule beta is a bactericidal protein that promotes spatial segregation of the microbiota and the colonic epithelium. Proc Natl Acad Sci USA. 2017;114:11027–33.CrossRef Propheter DC, Chara AL, Harris TA, Ruhn KA, Hooper LV. Resistin-like molecule beta is a bactericidal protein that promotes spatial segregation of the microbiota and the colonic epithelium. Proc Natl Acad Sci USA. 2017;114:11027–33.CrossRef
38.
go back to reference Yang L, Weiss TM, Lehrer RI, Huang HW. Crystallization of antimicrobial pores in membranes: magainin and protegrin. Biophys J. 2000;79:2002–9.CrossRef Yang L, Weiss TM, Lehrer RI, Huang HW. Crystallization of antimicrobial pores in membranes: magainin and protegrin. Biophys J. 2000;79:2002–9.CrossRef
39.
go back to reference Friedrich CL, Moyles D, Beveridge TJ, Hancock RE. Antibacterial action of structurally diverse cationic peptides on gram-positive bacteria. Antimicrob Agents Chemother. 2000;44:2086–92.CrossRef Friedrich CL, Moyles D, Beveridge TJ, Hancock RE. Antibacterial action of structurally diverse cationic peptides on gram-positive bacteria. Antimicrob Agents Chemother. 2000;44:2086–92.CrossRef
40.
go back to reference Patel SD, Rajala MW, Rossetti L, Scherer PE, Shapiro L. Disulfide-dependent multimeric assembly of resistin family hormones. Science. 2004;304:1154–8.CrossRef Patel SD, Rajala MW, Rossetti L, Scherer PE, Shapiro L. Disulfide-dependent multimeric assembly of resistin family hormones. Science. 2004;304:1154–8.CrossRef
Metadata
Title
Resistin-like molecule beta, a colonic epithelial protein, exhibits antimicrobial activity against Staphylococcus aureus including methicillin-resistant strains
Authors
Kazuhiro Watanabe
Kikuji Itoh
Sang-Hee Park
Mitsuo Kaku
Keiko Ishii
Hironobu Sasano
Takeshi Naitoh
Michiaki Unno
Kouhei Fukushima
Publication date
01-08-2020
Publisher
Springer Singapore
Published in
Surgery Today / Issue 8/2020
Print ISSN: 0941-1291
Electronic ISSN: 1436-2813
DOI
https://doi.org/10.1007/s00595-020-01974-z

Other articles of this Issue 8/2020

Surgery Today 8/2020 Go to the issue