Interferon-gamma modulates cAMP-induced mucin exocytosis without affecting mucin gene expression in a human colonic goblet cell line.
The regulation of intestinal mucin secretion by cytokines, soluble factors released by mucosal activated immune cells, is so far unknown. The aim of the present study was (1) to investigate the regulatory effects of interferon-gamma on baseline and stimulated mucin secretion elicited by an increase in intracellular cAMP, either a short-term increase (induced by vasoactive intestinal peptide or by forskolin) or a long-term increase (cholera toxin-induced), and (2) to attempt to delineate the site of action of interferon-gamma. The in vitro model used was the human colonic goblet cell line Cl.16E, which has already been shown to respond to physiological secretagogues in terms of mucin secretion. We examined the effects of interferon-gamma 1) on mucin exocytosis, measured as release of [3H]glucosamine-labeled macromolecules trapped at the stacking/running gel interface of polyacrylamide gels, and 2) on mucin biosynthesis, examined at the RNA level using a cDNA probe directed to the MUC2 mucin gene. We demonstrated that, while interferon-gamma did not alter baseline Cl.16E mucin secretion and MUC2 gene expression, it strongly inhibited the protein kinase A-dependent secretory response to VIP, forskolin, or cholera toxin. However, interferon-gamma had no effect on the protein kinase A-dependent MUC2 over-expression induced by cholera toxin. We thus concluded that the target for interferon-gamma inhibition of cAMP-stimulated Cl.16E mucin secretion is distal to protein kinase A and might be a component of the exocytotic machinery. Together, our results establish interferon-gamma as a pharmacologically powerful tool to specifically inhibit stimulated secretory processes without affecting baseline secretion.
Jarry
A
A
INSERM U 239, Faculté Bichat, Paris, France.
Merlin
D
D
Velcich
A
A
Hopfer
U
U
Augenlicht
L H
LH
Laboisse
C L
CL
D015152:Blotting, Northern; D002470:Cell Survival; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D003106:Colon / Q000166:cytology / Q000187:drug effects*; D000242:Cyclic AMP / Q000494:pharmacology*; D004594:Electrophysiology; D005089:Exocytosis / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D009077:Mucins / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000032:analysis; D013936:Thymidine / Q000378:metabolism; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
D009077:Mucins; D005576:Colforsin; D014660:Vasoactive Intestinal Peptide; D012313:RNA; D007371:Interferon-gamma; D002772:Cholera Toxin; D000242:Cyclic AMP; D013936:Thymidine