Development and characterization of a unique anti-IgE mouse monoclonal antibody cross-reactive between human and canine IgE.

Abstract

BACKGROUND:

The efficacy assessment of human anti-IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross-reactivity of anti-IgE mAbs between species.

OBJECTIVE:

We developed CRE-DR (an anti-dog IgE monoclonal antibody), an anti-IgE mouse mAb that recognizes canine and human IgE, and then examined its IgE specificity and cross-reactivity between three animal and human species.

METHODS:

After mouse immunization with a synthetic peptide derived from canine IgE (282 NTNDWIEGETYYC294 ), we generated a hybridoma producing CRE-DR. The CRE-DR purified from the ascites of hybridoma-inoculated mice was used for ELISA and Western blot analysis to examine reactivity to dog, human, and rodent IgEs as well as recombinant bovine serum albumin (BSA)-conjugated to canine, human, and rodent IgE amino acid peptides corresponding to the immunizing sequence. We then performed enzyme-linked immunosorbent assays (ELISAs) for dog IgE using sera from dogs with atopic dermatitis (AD) after inhibition with canine IgE and IgG. The amino acid sequence recognized by CRE-DR was identified by ELISA using synthetic peptides.

RESULTS:

CRE-DR is a monoclonal mouse IgG1κ specific for dog IgE, and the ELISA values in atopic dog sera were inhibited by dog IgE, but not dog IgG. The binding of CRE-DR to human IgE was relatively maintained, but not to rodent IgEs, which results were confirmed with the BSA-conjugated IgE peptides of the various species. The CRE-DR reactivity was supported by the comparison of amino acid sequence of CRE-DR epitope, DWIEGETYYC, in dog IgE; one, two, and three amino acids were substituted in the human, rat, and mouse IgE epitopes, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE:

CRE-DR is a mAb cross-reactive to dog and human IgEs, which can allow the use of a dog model of allergy to test the efficacy of a CRE-DR-derived anti-IgE therapeutic mAb before human clinical trials.

• Authors: Akiko Kumagai, Takuya Nara, Mizuho Uematsu, Yoko Kakinuma, Takashi Saito, Kenichi Masuda
• Journal: Immunity, inflammation and disease (Immun Inflamm Dis) Vol. 9 Issue 4 Pg. 1740-1748 (12 2021) ISSN: 2050-4527 [Electronic] England
• PMID: 34533288 (Publication Type: Journal Article)
• Copyright: © 2021 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.

Chemical References

• Antibodies, Anti-Idiotypic
• Antibodies, Monoclonal
• anti-IgE antibodies
• Immunoglobulin E

Topics

• Animals
• Antibodies, Anti-Idiotypic
• Antibodies, Monoclonal
• Antibody Specificity
• Cross Reactions
• Dogs
• Humans
• Immunoglobulin E
• Mice
• Rats