In vitro regulation of FcεRIα expression on human basophils by IgE antibody

Donald Macglashan, Jane McKenzie-White, Kristine Chichester, Bruce S. Bochner, Frances M. Davis, John Thomas Schroeder, Lawrence M. Lichtenstein

Research output: Contribution to journalArticle

Abstract

In vivo studies suggested the possibility of an IgE-dependent regulation of high-affinity (FcεRI) IgE receptor expression on basophils. The current studies extend these observations to in vitro cultures of human basophils. Incubation of basophils for 3 to 4 weeks resulted in a slow dissociation of IgE antibody, during which time FcεRI expression decreased, as measured by flow cytometry using the anti-FcεRIα monoclonal antibody, 22E7, or by measuring FcεRIα mass by Western blotting of whole-cell lysates. Culture of basophils with IgE resulted in upregulation of FcεRIα expression by both flow cytometry end Western blotting of whole-cell lysates. Upregulation followed a linear time course during 2 weeks of culture. The relative increase in FcεRIα density depended on the starting density; with starting densities of FcεRIα of 10,000 to 170,000 per basophil, the upregulation varied 20- to 1.1-fold, respectively. Upregulation occurred in high-purity basophils, was not influenced by IgG at concentrations up to 1 mg/mL, and was inhibited by dimeric IgE. Heat-inactivated IgE was less effective and the monoclonal antibody CGP51901 that prevents IgE binding to FcεRIα blocked the ability of IgE to induce upregulation. The dose-response curve for IgE- induced upregulation had an effective concentration50 of 230 ng/mL. Although the receptor through which IgE induces this upregulation is not yet known, several characteristics suggest that the upregulation is mediated by IgE interacting through FcεRIα itself.

Original languageEnglish (US)
Pages (from-to)1633-1643
Number of pages11
JournalBlood
Volume91
Issue number5
StatePublished - Mar 1 1998

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Basophils
Immunoglobulin E
Up-Regulation
Antibodies
Flow cytometry
Flow Cytometry
Western Blotting
Monoclonal Antibodies
In Vitro Techniques
Immunologic Techniques
IgE Receptors
Immunoglobulin G
Hot Temperature

ASJC Scopus subject areas

  • Hematology

Cite this

Macglashan, D., McKenzie-White, J., Chichester, K., Bochner, B. S., Davis, F. M., Schroeder, J. T., & Lichtenstein, L. M. (1998). In vitro regulation of FcεRIα expression on human basophils by IgE antibody. Blood, 91(5), 1633-1643.

In vitro regulation of FcεRIα expression on human basophils by IgE antibody. / Macglashan, Donald; McKenzie-White, Jane; Chichester, Kristine; Bochner, Bruce S.; Davis, Frances M.; Schroeder, John Thomas; Lichtenstein, Lawrence M.

In: Blood, Vol. 91, No. 5, 01.03.1998, p. 1633-1643.

Research output: Contribution to journalArticle

Macglashan, D, McKenzie-White, J, Chichester, K, Bochner, BS, Davis, FM, Schroeder, JT & Lichtenstein, LM 1998, 'In vitro regulation of FcεRIα expression on human basophils by IgE antibody', Blood, vol. 91, no. 5, pp. 1633-1643.
Macglashan D, McKenzie-White J, Chichester K, Bochner BS, Davis FM, Schroeder JT et al. In vitro regulation of FcεRIα expression on human basophils by IgE antibody. Blood. 1998 Mar 1;91(5):1633-1643.
Macglashan, Donald ; McKenzie-White, Jane ; Chichester, Kristine ; Bochner, Bruce S. ; Davis, Frances M. ; Schroeder, John Thomas ; Lichtenstein, Lawrence M. / In vitro regulation of FcεRIα expression on human basophils by IgE antibody. In: Blood. 1998 ; Vol. 91, No. 5. pp. 1633-1643.
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