A murine model of IgE-mediated cow’s milk hypersensitivity,☆☆,

https://doi.org/10.1016/S0091-6749(99)70492-6Get rights and content

Abstract

Background: Cow’s milk allergy (CMA) is one of the leading causes of food allergy in children. Understanding the mechanisms involved in the development of CMA has been hampered by the lack of suitable animal models. Objective: We sought to develop a mouse model of IgE-mediated cow’s milk hypersensitivity (CMH) that mimics the clinical features of immediate CMA in humans. Methods: Three-week-old C3H/HeJ mice were sensitized by intragastric administration of cow’s milk (CM) plus cholera toxin and boosted 5 times at weekly intervals. Results: CM-specific IgE antibody levels were significantly increased at 3 weeks and peaked at 6 weeks after the initial feeding. Intragastric challenge with CM at week 6 elicited systemic anaphylaxis accompanied by vascular leakage, significantly increased plasma histamine, and increased intestinal permeability to casein. Histologic examination of intestinal tissue revealed marked vascular congestion, edema, and sloughing of enterocytes. The role of IgE in mediating CMH was confirmed by abrogation of passive cutaneous anaphylaxis reactions by heat inactivation of immune sera. Development of IgE-mediated CMH in this model is likely to be TH2 cell mediated because in vitro stimulation of spleen cells from mice allergic to CM induced significant increases in the levels ofIL-4 and IL-5, but not IFN-γ. Conclusion: This model should provide a useful tool for evaluating the immunopathogenic mechanisms involved in CMA and for exploring new therapeutic approaches. (J Allergy Clin Immunol 1999;103:206-14.)

Section snippets

METHODS

Female C3H/HeJ mice, 3 weeks of age (immediately after weaning), were purchased from the Jackson Laboratory (Bar Harbor, Me) and maintained on regular mouse chow under specific pathogen-free conditions. Guidelines for the care and use of the animals were followed.25

Homogenized CM (GAF Seelig Inc) was used. CT was purchased from List Biological Laboratories, Inc (Campbell, Calif). Concanavalin (Con A) and albumin, human-dinitrophenyl (DNP-albumin) were purchased from Sigma (St Louis, Mo).

CM-specific IgE responses after intragastric CM sensitization

To investigate the kinetics of IgE production in the development of CMH, serum CM-specific IgE was monitored weekly by ELISA. Mice sensitized with the medium dose (1 mg/g) of CM plus CT developed significant (P < .01) increases in antigen-specific IgE by 3 weeks, which peaked at 6 weeks after the initial sensitization (Fig 1).

. Serum levels of CM-specific IgE. Sera from different groups of mice (n = 5) as indicated were obtained weekly after CM and CT sensitization. CM-specific IgE levels in

DISCUSSION

In this study we identified certain conditions required to effectively generate a mouse model of CMA by oral sensitization and challenge. This model exhibited the characteristics of type 1 hypersensitivity reactions. Symptoms of anaphylactic reactions were apparent 15 to 30 minutes after oral challenge. The symptoms involved several organ systems, including the skin and respiratory and gastrointestinal tracts, with the most severe reactions being fatal. Increased vascular leakage and intestinal

Acknowledgements

We thank Ludmilla Bardina, MS, for technical assistance, and Scott Sicherer, MD, for his assistance in the preparation of this manuscript.

References (58)

  • SH Sicherer et al.

    The role of food allergy in childhood asthma

    Immunol Allergy Clin North Am

    (1998)
  • D Robinson et al.

    Activation of CD4+ T cells, increased TH2-type cytokine mRNA expression, and eosinophil recruitment in bronchoalveolar lavage after allergen inhalation challenge in patients with atopic asthma

    J Allergy Clin Immunol

    (1993)
  • A. Host

    Cow’s milk allergy

    J R Soc Med

    (1997)
  • HA. Sampson

    Food allergy

    JAMA

    (1997)
  • A. Host

    Cow’s milk protein allergy and intolerance in infancy. Some clinical, epidemiological and immunological aspects

    Pediatr Allergy Immunol

    (1994)
  • A Host et al.

    The natural history of cow’s milk protein allergy/intolerance

    Eur J Clin Nutr

    (1995)
  • OM Poulsen et al.

    Murine passive cutaneous anaphylaxis test (PCA) for the ‘all or none’ determination of allergenicity of bovine whey proteins and peptides

    Clin Allergy

    (1987)
  • SH Gavett et al.

    Interleukin 12 inhibits antigen-induced airway hyperresponsiveness, inflammation, and Th2 cytokine expression in mice

    J Exp Med

    (1995)
  • XM Li et al.

    Mucosal IFN-γ gene transfer inhibits pulmonary allergic responses in mice

    J Immunol

    (1996)
  • XM Li et al.

    Induction of pulmonary allergic responses by antigen-specific Th2 cells

    J Immunol

    (1998)
  • OM Poulsen et al.

    Comparison of intestinal anaphylactic reactions in sensitized mice challenged with untreated bovine milk and homogenized bovine milk

    Allergy

    (1990)
  • OM Poulsen et al.

    Effect of homogenization and pasteurization on the allergenicity of bovine milk analysed by a murine anaphylactic shock model

    Clin Allergy

    (1987)
  • K Ito et al.

    Murine model of IgE production with a predominant Th2-response by feeding protein antigen without adjuvants

    Eur J Immunol

    (1997)
  • H Kiyono et al.

    Lack of oral tolerance in C3H/HeJ mice

    J Exp Med

    (1982)
  • DG. Hanson

    Ontogeny of orally induced tolerance to soluble proteins in mice. I. Priming and tolerance in newborns

    J Immunol

    (1981)
  • S Strobel et al.

    Immune responses to fed protein antigens in mice. 3. Systemic tolerance or priming is related to age at which antigen is first encountered

    Pediatr Res

    (1984)
  • S. Strobel

    Neonatal oral tolerance

    Ann N Y Acad Sci

    (1996)
  • AG Lamont et al.

    Priming of systemic and local delayed-type hypersensitivity responses by feeding low doses of ovalbumin to mice

    Immunology

    (1989)
  • NF Pierce et al.

    Determinants of the localization, magnitude, and duration of a specific mucosal IgA plasma cell response in enterically immunized rats

    J Immunol

    (1982)
  • Cited by (334)

    • Laboratory skills for immunologists: utility and limitations with emphasis on allergy research

      2022, Allergic and Immunologic Diseases: A Practical Guide to the Evaluation, Diagnosis and Management of Allergic and Immunologic Diseases
    View all citing articles on Scopus

    Supported by AI 24439 NIAID, NIH.

    ☆☆

    Reprint requests: Xiu-Min Li, MD, Pediatric Allergy and Immunology, The Mount Sinai School of Medicine, Box 1198, One Gustave L. Levy Place, New York, NY 10029-6574.

    0091-6749/99 $8.00 + 0  1/1/96137

    View full text