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Development and characterization of anti-Gal B cell receptor transgenic Gal-/- mice.

Xu, H; Sharma, A; Lei, Y; Okabe, J; Wan, H; Chong, AS-F; Logan, JS; (2002) Development and characterization of anti-Gal B cell receptor transgenic Gal-/- mice. Transplantation , 73 (10) pp. 1549-1557.

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BACKGROUND: The successful clinical application of pig-to-primate xenotransplantation is currently limited by the development of an acute vascular rejection, which is thought to involve an induced humoral immune response to the galactose alpha1,3 galactose (alpha-Gal) antigen. Successful xenotransplantation may require the development of novel methods for removal or neutralization of anti-Gal antibodies and anti-Gal-producing B cells. The large diversity of the B-cell repertoire makes it difficult, however, to isolate and study anti-Gal B-cell development. METHODS: We have established a transgenic mouse model for investigating anti-Gal B cells by introducing a transgene encoding both heavy and light chains for an anti-Gal IgM antibody into an alpha-galactosyltransferase-deficient (Gal-/-) background. We have characterized the frequency, phenotype, and function of transgenic anti-Gal B cells by multiparameter flow cytometric analysis and ELISA. RESULTS: ELISA analysis of serum from animals with the transgene in an alpha-galactosyltransferase-deficient background (Tg Gal-/-), from transgenic animals with a heterozygous alpha-galactosyltransferase background (Tg Gal-/+), and from nontransgenic alpha-galactosyltransferase-deficient littermates (Gal-/-) demonstrated elevated expression of anti-Gal antibodies in Tg Gal-/- mice compared with nontransgenic Gal-/- animals and a lack of transgene expression in the Tg Gal-/+ mice. Anti-Gal antibody expression in Tg Gal-/- mice could be increased by immunization with an ovalbumin-Gal glycoconjugate in vivo and through stimulation with lipopolysaccharide in vitro. Multiparameter flow cytometric analysis indicates that 50% to 80% of splenic and peritoneal B cells expressed the transgene and excluded endogenous immunoglobulin gene rearrangements. The majority of these B cells expressed anti-Gal receptors on the surface, as identified by staining with a fluorescein isothiocyanate-bovine serum albumin-Gal glycoconjugate. FACS analysis of the Tg Gal-/- B cells identified them as a population of CD21highCD23lowIgMhigh marginal zone B cells in the spleen and CD5-CD23low B1 cells in the peritoneal cavity. CONCLUSIONS: These observations suggest that this model can be used to study the regulation of anti-Gal B cells and can establish a reliable source of functional anti-Gal B cells, which could be used to test the effectiveness of alpha-Gal-specific immunosuppressive reagents.

Type: Article
Title: Development and characterization of anti-Gal B cell receptor transgenic Gal-/- mice.
Location: United States
Keywords: Animals, Antigens, CD, B-Lymphocytes, Bone Marrow Cells, Disaccharides, Immunoglobulin D, Immunoglobulin M, Leukosialin, Mice, Mice, Knockout, Mice, Transgenic, Models, Animal, Receptors, Antigen, B-Cell, Sialoglycoproteins, Spleen, Transplantation, Heterologous
URI: http://discovery.ucl.ac.uk/id/eprint/90877
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