Abstract
Oral tolerance, the antigen-specific immunologic unresponsiveness after antigen (Ag) feeding, is of physiologic importance in preventing antibody (Ab) responses to dietary proteins. This is important in the young, especially at weaning when numerous dietary Ag are encountered for the first time. Two related enterotoxins responsible for much diarrhea in the infant, cholera toxin (CT) and Escherichia coli heat-labile enterotoxin (LT), have been shown to abrogate oral tolerance to an unrelated Ag fed simultaneously. The main objective of this study was to determine whether the gut flora can play a role in the CT- or LT-mediated abrogation of oral tolerance to the dietary protein ovalbumin (OVA), on a short-term and long-term basis. Conventional and germ-free mice were fed once or twice with toxin plus OVA. After two intraperitoneal immunizations with OVA, anti-OVA IgG and IgE Ab levels were measured. Because IgG and IgE Ab responses were detected, both CT and LT abrogated oral tolerance to OVA in conventional and germ-free mice. As time progressed (observations over 3 mo), whereas the specific IgG Ab response in the germ-free mice remained similar to that of the bicarbonate-fed controls, a hyporesponsive state was observed in conventional mice. The results showed that, although the gut flora did not prevent the CT- and LT-mediated abrogation of oral tolerance, it did shorten the effect and allow oral tolerance to be recovered.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- Ag:
-
antigen
- Ab:
-
antibody
- CT:
-
cholera toxin
- LT:
-
Escherichia coli heat-labile enterotoxin
- OVA:
-
ovalbumin
- Bic:
-
bicarbonate solution
- i.p.:
-
intraperitoneal
References
Challacombe SJ, Tomasi TB 1980 Systemic tolerance and secretory immunity after oral immunization. J Exp Med 152: 1459–1472.
Mowat AM 1987 The regulation of immune responses to dietary protein antigens. Immunol Today 8: 93–98.
Friedman A, Al-Sabbagh A, Santos LMB, Fishman-Lobell J, Polanski M, Prabhu Das M, Khoury SJ, Weiner HL 1994 Oral tolerance: a biologically relevant pathway to generate peripheral tolerance against external and self antigens. Chem Immunol 58: 259–290.
Husby S, Jensenius JC, Svehag SE 1985 Passage of undegraded dietary antigen into the blood of healthy adults. Quantification, estimation of size distribution and relation of uptake to levels of specific antibodies. Scand J Immunol 22: 83–92.
Jarrett EEE 1984 Immunoregulation of IgE responses: the role of the gut in perspective. Ann Allergy 53: 550–556.
Metcalfe DD 1991 Food allergy. Curr Opin Immunol 3: 881–886.
Crowe SE, Perdue MH 1992 Gastrointestinal food hypersensitivity: basic mechanisms of pathophysiology. Gastroenterology 103: 1075–1095.
Strobel S 1991 Mechanisms of gastrointestinal immunoregulation and food induced injury to the gut. Eur J Clin Nutr 45: 1–9.
Simon GL, Gorbach SL 1984 Intestinal flora in health and disease. Gastroenterology 86: 174–193.
Moreau CM, Coste M 1993 Immune responses to dietary protein antigens. World Rev Nutr Diet 74: 22–57.
Hudault S, Bridonneau C, Ducluzeau R, Raibaud P 1991 Effect of bovine or human milk on the establishment of Bifidobacterium bifidum and some other bacteria from the human neonatal microflora in gnotobiotic mice. Microb Ecol Hum Health Dis 4: 1–8.
Van der Waaij D, Van der Waaij BD 1990 The colonization resistance of the digestive tract in different animal species and in man; a comparative study. Epidemiol Infect 105: 237–243.
Grady GF, Keusch GT 1971 Pathogenesis of bacterial diarrheas. N Engl J Med 285: 831–841.
Sack RB 1980 Enterotoxigenic Escherichia coli: identification and characterization. J Infect Dis 142: 279–286.
Penny ME, Scotland SM, Smith HR, McConnell MM, Knutton SK, Sack RB 1992 Virulence properties of Enterobacteriaceae isolated from the small intestine of children with diarrhea. Pediatr Infect Dis 11: 623–630.
Elson CO, Ealding W 1984 Cholera toxin feeding did not induce oral tolerance in mice and abrogated oral tolerance to an unrelated protein antigen. J Immunol 133: 2892–2897.
Clements JD, Hartzog NM, Lyon FL 1988 Adjuvant activity of Escherichia coli heat-labile enterotoxin and effect on the induction of oral tolerance in mice to unrelated protein antigens. Vaccine 6: 269–277.
Snider DP, Marshall JS, Perdue MH, Liang H 1994 Production of IgE antibody and allergic sensitization of intestinal and peripheral tissues after oral immunization with protein Ag and cholera toxin. J Immunol 153: 647–657.
Moreau MC, Corthier G 1988 Effect of the gastrointestinal microflora on induction and maintenance of oral tolerance to ovalbumin in C3H/HeJ mice. Infect Immun 56: 2766–2768.
Strobel S, Ferguson A 1987 Persistence of oral tolerance in mice fed ovalbumin is different for humoral and cell-mediated immune responses. Immunology 60: 317–318.
Saklayen MG, Pesce AJ, Pollak VE, Michael JG 1983 Induction of oral tolerance in mice unresponsive to bacterial lipopolysaccharide. Infect Immun 41: 1383–1385.
Pierre P, Denis O, Bazin H, Mbella EM, Vaerman JP 1992 Modulation of oral tolerance to ovalbumin by cholera toxin and its B subunit. Eur J Immunol 22: 3179–3182.
Spangler BD 1992 Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin. Microbiol Rev 56: 622–647.
Verma M, Majumdar S, Ganguly NK, Walia BNS 1994 Effect of Escherichia coli enterotoxins on macromolecular absorption. Gut 35: 1613–1616.
Abrams GD 1977 Microbial effects on mucosal structure and function. Am J Clin Nutr 30: 1880–1886.
Heyman M, Dumontier AM, Desjeux JF 1986 Intestinal barrier to intact horseradish peroxidase in experimental secretory diarrhoea. J Pediatr Gastroenterol Nutr 5: 463–466.
Strobel S 1989 Mechanisms of immunologically mediated injury to the gut. Allergologie 5: 163–169.
Boudraa G, Touhami M, Pochart P, Soltana R, Mary JY, Desjeux JF 1990 Effect of feeding yogurt versus milk in children with persistent diarrhea. J Pediatr Gastroenterol Nutr 11: 509–512.
Kvale D, Brandtzaeg P 1995 Constitutive and cytokine induced expression of HLA molecules, secretory component, and intercellular adhesion molecule-1 is modulated by butyrate in the colonic epithelial cell line HT-29. Gut 36: 737–742.
Elson CO, Ealding W 1984 Generalized systemic and mucosal immunity in mice after mucosal stimulation with cholera toxin. J Immunol 132: 2736–2741.
Lycke N, Holmgren J 1986 Strong adjuvant properties of cholera toxin on gut mucosal immune responses to orally presented antigens. Immunology 59: 301–308.
Acknowledgements
The authors thank Catherine Dubuquoy for her technical assistance, and Geneviève Milon and Michel Coste for their helpful discussions and critical reading of the manuscript.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gaboriau-Routhiau, V., Moreau, MC. Gut Flora Allows Recovery of Oral Tolerance to Ovalbumin in Mice after Transient Breakdown Mediated by Cholera Toxin or Escherichia coli Heat-Labile Enterotoxin. Pediatr Res 39, 625–629 (1996). https://doi.org/10.1203/00006450-199604000-00011
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-199604000-00011


