Primary human colonic myofibroblasts are resistant to Clostridium difficile toxin A-induced, but not toxin B-induced, cell death.
Colonic inflammation in Clostridium difficile infection is mediated by released toxins A and B. We investigated responses to C. difficile toxins A and B by isolated primary human colonic myofibroblasts, which represent a distinct subpopulation of mucosal cells that are normally located below the intestinal epithelium. Following incubation with either purified toxin A or B, there was a change in myofibroblast morphology to stellate cells with processes that were immunoreactive for alpha-smooth muscle actin. Most of the myofibroblasts remained viable, with persistence of stellate morphology, despite exposure to high concentrations (up to 10 μg/ml) of toxin A for 72 h. In contrast, a majority of the toxin B-exposed myofibroblasts lost their processes prior to cell death over 24 to 72 h. At low concentrations, toxin A provided protection against toxin B-induced cell death. Within 4 h, myofibroblasts exposed to either toxin A or toxin B lost expression of the nonglucosylated form of Rac1, and there was also a loss of the active form of RhoA. Despite preexposure to high concentrations of toxin A for 3 h, colonic myofibroblasts were able to recover their morphology and proliferative capacity during prolonged culture in medium. However, toxin B-preexposed myofibroblasts were not able to recover. In conclusion, primary human colonic mucosal myofibroblasts are resistant to toxin A (but not toxin B)-induced cell death. Responses by colonic myofibroblasts may play an important role in mucosal protection, repair, and regeneration in colitis due to C. difficile infection.
Institute of Infection, Immunity and Inflammation, University of Nottingham Nottingham, UK. yash.mahida@nottingham.ac.uk
N
N
Mullan
K R
KR
Hughes
Y R
YR
Mahida
D000368:Aged; D000369:Aged, 80 and over; D001426:Bacterial Proteins / Q000633:toxicity*; D001427:Bacterial Toxins / Q000633:toxicity*; D015153:Blotting, Western; D016923:Cell Death; D002460:Cell Line; D016360:Clostridioides difficile / Q000737:chemistry*; D003106:Colon / Q000382:microbiology / Q000473:pathology; D004768:Enterotoxins / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008855:Microscopy, Electron, Scanning; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D058628:Myofibroblasts / Q000473:pathology*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile
J Immunol Methods. 1991 Jun 3;139(2):271-9
1710634
Infect Immun. 1982 Mar;35(3):1032-40
7068210
Am J Pathol. 1997 Nov;151(5):1469-76
9358773
Am J Physiol. 1999 May;276(5):G1087-93
10329998
J Immunol Methods. 1983 Dec 16;65(1-2):55-63
6606682
Annu Rev Cell Dev Biol. 2002;18:315-44
12142277
Infect Immun. 1995 Apr;63(4):1421-6
7890404
Clin Exp Immunol. 2008 Sep;153(3):439-47
18505424
Cell. 2004 Jan 23;116(2):167-79
14744429
J Immunol. 1996 Jun 1;156(11):4146-53
8666781
Clin Microbiol Rev. 2005 Apr;18(2):247-63
15831824
J Clin Invest. 1995 May;95(5):2004-11
7738167
Am J Physiol. 1999 Jul;277(1):C1-9
10409103
J Biol Chem. 1986 Jan 25;261(3):1316-21
3944088
Nat Rev Mol Cell Biol. 2008 Sep;9(9):690-701
18719708
Am J Physiol. 1989 Aug;257(2 Pt 1):G274-83
2764111
Infect Immun. 1998 Nov;66(11):5462-9
9784558
Infect Immun. 2005 Mar;73(3):1625-34
15731062
Am J Physiol Lung Cell Mol Physiol. 2002 Oct;283(4):L830-8
12225960
Gastroenterology. 2003 Nov;125(5):1341-54
14598250
Cell. 1992 Aug 7;70(3):389-99
1643657
Gastroenterology. 1997 Jun;112(6):1887-94
9178681
J Clin Pathol. 1977 Jan;30(1):1-12
838865
Glycobiology. 2007 Apr;17(4):15R-22R
17237138
J Med Microbiol. 1989 Sep;30(1):69-77
2778794
Cell. 1992 Aug 7;70(3):401-10
1643658
Infect Immun. 1988 Jul;56(7):1708-14
3384474
Gastroenterology. 2002 Apr;122(4):1048-57
11910356
Am J Physiol. 1997 Dec;273(6):G1341-8
9435560
J Clin Invest. 1988 Nov;82(5):1516-24
3141478
J Infect Dis. 2004 Jun 1;189(11):2110-9
15143480
Oncogene. 1997 Oct;15(18):2179-89
9393976
J Infect Dis. 2002 Nov 15;186(10):1438-47
12404159
Am J Pathol. 2003 Apr;162(4):1355-60
12651627
Inflamm Bowel Dis. 2011 Jan;17(1):213-20
20848536
Am J Physiol Cell Physiol. 2002 Jan;282(1):C172-82
11742810
Am J Physiol Gastrointest Liver Physiol. 2002 Jun;282(6):G1035-44
12016129
FEBS Lett. 2006 Jun 12;580(14):3565-9
16730714
Gut. 1996 Mar;38(3):337-47
8675084
Am J Physiol. 1999 Aug;277(2):C183-201
10444394
Infect Immun. 2007 Feb;75(2):801-9
17145947
Infect Immun. 1986 Oct;54(1):70-6
2428753
J Immunol. 2000 Feb 15;164(4):1952-60
10657645
Infect Immun. 1990 Jul;58(7):2329-36
2114363
Am J Physiol. 1999 Aug;277(2):C271-9
10444403
Gut. 2009 Jun;58(6):777-89
19201776