Mice expressing RHAG and RHD human blood group genes

Dominique Goossens; Nelly da Silva; Sylvain Metral; Ulrich Cortes; Isabelle Callebaut; Julien Picot; Isabelle Mouro-Chanteloup; Jean-Pierre Cartron

doi:10.1371/journal.pone.0080460

Mice expressing RHAG and RHD human blood group genes

PLoS One. 2013 Nov 18;8(11):e80460. doi: 10.1371/journal.pone.0080460. eCollection 2013.

Authors

Dominique Goossens¹, Nelly da Silva, Sylvain Metral, Ulrich Cortes, Isabelle Callebaut, Julien Picot, Isabelle Mouro-Chanteloup, Jean-Pierre Cartron

Affiliation

¹ Institut National de la Transfusion Sanguine, Paris, France ; Inserm UMR_S 665, Paris, France ; Université Paris Diderot, Sorbonne Paris Cité, UMR-S665, Paris, France.

Abstract

Anti-RhD prophylaxis of haemolytic disease of the fetus and newborn (HDFN) is highly effective, but as the suppressive mechanism remains uncertain, a mouse model would be of interest. Here we have generated transgenic mice expressing human RhAG and RhD erythrocyte membrane proteins in the presence and, for human RhAG, in the absence, of mouse Rhag. Human RhAG associates with mouse Rh but not mouse Rhag on red blood cells. In Rhag knockout mice transgenic for human RHAG, the mouse Rh protein is "rescued" (re-expressed), and co-immunoprecipitates with human RhAG, indicating the presence of hetero-complexes which associate mouse and human proteins. RhD antigen was expressed from a human RHD gene on a BAC or from RHD cDNA under control of β-globin regulatory elements. RhD was never observed alone, strongly indicative that its expression absolutely depends on the presence of transgenic human RhAG. This first expression of RhD in mice is an important step in the creation of a mouse model of RhD allo-immunisation and HDFN, in conjunction with the Rh-Rhag knockout mice we have developed previously.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ammonium Compounds / metabolism
Animals
Blood Proteins / chemistry
Blood Proteins / genetics*
Blood Proteins / metabolism
Cation Transport Proteins / genetics
Cation Transport Proteins / metabolism
Cell Line
Erythrocyte Membrane / metabolism
Erythrocytes / metabolism
Gene Expression Regulation
Gene Expression*
Humans
Male
Membrane Glycoproteins / chemistry
Membrane Glycoproteins / genetics*
Membrane Glycoproteins / metabolism
Mice
Mice, Knockout
Mice, Transgenic
Promoter Regions, Genetic
Protein Binding
Protein Multimerization
Rh-Hr Blood-Group System / chemistry
Rh-Hr Blood-Group System / genetics*
Rh-Hr Blood-Group System / metabolism
Transcription, Genetic
beta-Globins / metabolism

Substances

Ammonium Compounds
Blood Proteins
Cation Transport Proteins
Membrane Glycoproteins
RHAG protein, human
Rh-Hr Blood-Group System
Rho(D) antigen
beta-Globins

Grants and funding

N.d.S. and U.C. were supported by post-doctoral funding from Laboratoire Français du Fractionnement et des Biotechnologies, France. This work was supported by grants from the Association Recherche Transfusion (Contracts 25-2001 and 34-2003) (http://www.art-hemobiologie.org/) and the GIS Maladies Rares (support for part of the initial analysis of the Rh/Rhag knockout mouse models at the ICS) (http://fondation-maladiesrares.org/). No individuals employed or contracted by the funders (other than the named authors) played any role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.