Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response in mice

Abstract

We constructed two megabase-sized YACs containing large contiguous fragments of the human heavy and kappa (kappa) light chain immunoglobulin (Ig) loci in nearly germline configuration, including approximately 66 VH and 32 V kappa genes. We introduced these YACs into Ig-inactivated mice and observed human antibody production which closely resembled that seen in humans in all respects, including gene rearrangement, assembly, and repertoire. Diverse Ig gene usage together with somatic hypermutation enables the mice to generate high affinity fully human antibodies to multiple antigens, including human proteins. Our results underscore the importance of the large Ig fragments with multiple V genes for restoration of a normal humoral immune response. These mice are likely to be a valuable tool for the generation of therapeutic antibodies.

Heterosubtypic Immunity to Influenza A Virus in Mice Lacking IgA, All Ig, NKT Cells, or γδ T Cells

“…Instead of MT mice that have been reported to produce low levels of Ig (29), we chose to use DI mice that are unable to produce mature B cells and Ig due to the knockout of both the heavy and -chain loci (37)(38)(39). Our previous study in DI mice had used B/AA virus.…”

Immunoglobulin gene usage and immunohistochemical characteristics of human monoclonal antibodies to the mitochondrial autoantigens of primary biliary cirrhosis induced in the XenoMouse

“…Thirdly, the 4 clones that recognize both PDC-E2 and OGDE-E2, and 4G6 reacting with OGDC-E2, all use V 1-17, likely as a result of antigen-driven selection of the V in these antibodies. 6,7 the Ig repertoire of the AMA HmAbs suggest that these antibodies are the result of antigen-driven selection. We should re-emphasize that the Ig gene restriction described herein has not been noted in multiple other studies of the XenoMouse 7 (Gallo M, unpublished data, 1998).…”

“…The inguinal and popliteal lymph nodes and spleen were removed from immunized XenoMouse animals; the fusion of lymphocytes and splenocytes and selection of hybridomas were performed as described. 6 AMA-positive hybridomas were identified by ELISA using native proteins and recombinant mitochondrial proteins as antigens, and hybridomas purified from culture media of antibody-producing hybridomas using protein-A or protein-L affinity chromatography kit (Pierce, Rockford, IL).…”

“…To address this issue, we have taken advantage of a unique transgenic animal, XenoMouse (Abgenix Inc., Fremont, CA), that contains the human heavy (H) and light chain loci cloned on yeast artificial chromosomes. 5,6 The yeast artificial chromosomes in XenoMouse include approximately 66 V H (ϳ80%) and 32 V (ϳ75%) gene regions, rendering these mice capable of producing high-affinity, fully functional IgM and IgG2 antibodies, which closely resemble those seen in humans, to multiple antigens. 7,8 Moreover, the Ig gene usage toward immunizing antigens has been shown to be quite diverse.…”

Transgenèse animale et humanisation des anticorps

“…Ce qui améliore l'étendue du répertoire et permet la reconstitution de compartiments lymphocytaires B de taille plus proche de la normale [7,8]. Ces lignées ont été produites tant par transgenèse classique que par fusion de cellules souches embryonnaires (ES) avec des protoplastes pour le transfert de chromosomes de levure porteurs de larges transgènes humains, ou encore par fusion de cellules ES avec des microcellules dérivées de fibroblastes humains et apportant un minichromosome humain.…”