NOD.Cg-H2-Ab1^b-tm1Doi Tg(HLA-DRA,HLA-DRB1*0301)#Gjh/DvYtomJ

Stock number: 030434
Product name: NOD-DR3 Ab⁰ (NOD-DR3 , Ab⁰/DR3/NOD , DR3⁺ g7^-)
Strain synonyms: NOD-DR3 Ab<0>
Research areas: Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

NOD-DR3 with MHC class II knock-out mice (also called also called Ab⁰/DR3/NOD, DR3+ g7-, NOD-DR3 Ab⁰ or simply "NOD-DR3") are NOD-congenic animals with endogenous mouse MHC class II expression eliminated (via homozygosity of the Ab⁰ knock-out allele [H2-Ab1^b-tm1Doi]) and replaced with human HLA class II molecule expression (via the DR3(DRw17) transgene). These mice are a model for studying experimental autoimmune thyroiditis (EAT), and also facilitate engraftment of human immune cells.

Detailed description

NOD-DR3 with MHC class II knock-out mice (also called also called Ab⁰/DR3/NOD, DR3+ g7-, NOD-DR3 Ab⁰ or simply "NOD-DR3") are NOD-congenic animals with endogenous mouse MHC class II expression eliminated (via homozygosity of the Ab⁰ knock-out allele [H2-Ab1^b-tm1Doi]) and replaced with human HLA class II molecule expression (via the DR3(DRw17) transgene).

The DR3(DRw17) transgene (Tg(HLA-DRA,HLA-DRB1*0301)#Gjh) expresses the human leukocyte antigen MHC class II sequences of the HLA-DR3 genotype (HLA-DRA*0101 and HLA-DRB1*0301) that is associated with increased susceptibility to autoimmune disease (including autoimmune thyroid diseases, Graves' disease and Hashimoto's thyroiditis). The HLA-DRB1*0301 sequence encodes the HLA-DRβ1-Arg74 variant, which is able to present pathogenic human thyroglobulin peptides (hTg) to T cells more efficiently - thereby triggering thyroid autoimmunity. As such, mice homozygous for Ab⁰ knock-out allele and hemizygous for the DR3 transgene (NOD-DR3 Ab⁰ mice) are a model for studying experimental autoimmune thyroiditis (EAT), and also facilitate engraftment of human immune cells. Specifically, following immunization with thyroglobulin peptides from either human (hTg) or mouse (mTg), NOD-DR3 Ab⁰ mice develop thyroiditis and anti-Tg titres.

When mice homozygous for Ab⁰ knock-out allele and hemizygous for the DR3 transgene (NOD-DR3 Ab⁰ mice), the donating investigator reports NOD-DR3 mice are viable and fertile with no gross physical or behavioral abnormalities. NOD-DR3 Ab⁰ mice are resistant to the spontaneous diabetic phenotype normally observed for NOD mice.

 

Development

NOD-DR3 with MHC class II knock-out mice (also called also called Ab⁰/DR3/NOD, DR3+ g7-, NOD-DR3 Ab⁰ or simply "NOD-DR3") are NOD-congenic animals with endogenous mouse MHC class II expression eliminated (via the Ab⁰ knock-out allele) and replaced with human HLA class II molecule expression (via the DR3(DRw17) transgene). The individual transgenes/mutations are described below.

The "MHC class II knock-out" allele (Ab⁰; H2-Ab1^b-tm1Doi) on chromosome 17 was created by Drs. Diane Mathis and Christophe Benoist (while at INSERM, Strasbourg, France) - see allele description for Stock No. 010966. Ab⁰ mice on a C57BL/6;129 genetic background were used for breeding as below.

The human major histocompatibility complex (MHC) class II α chain is encoded by the human leukocyte antigen DRα gene (HLA-DRA), while the human MHC class II β chain is encoded by different polymorphic alleles of the DRβ1 gene (HLA-DRB1). In humans, the HLA-DR3 genotype (HLA-DRA*0101 / HLA-DRB1*0301) confers increased susceptibility to autoimmune disease (including autoimmune thyroid diseases, Graves' disease and Hashimoto's thyroiditis) - because the HLA-DRB1*0301 sequence encodes the HLA-DRβ1-Arg74 variant. The DR3(DRw17) transgene (Tg(HLA-DRA,HLA-DRB1*0301)#Gjh) was created by Dr. Gunter Hammerling (while at German Cancer Research Center, Heidelberg) by co-injection of the two human MHC class II sequences encoding the HLA-DR3 genotype (HLA-DRA*0101 and HLA-DRB1*0301). Following co-injection into (C57BL/6 x DBA/2)F1 embryos, founder mice were obtained. Founder mice were then backcrossed several generations to B10.Q mice (see Stock No. 002024 ; B10.D1-H2^q/SgJ) to create the B10.Q.DR3 mice. Next, they were bred to C57BL/6;129-Ab⁰ animals to create the Ab⁰/DR3 mice. Those animals were then backcrossed several generations to NOD inbred mice (substrain not identified) - this generated the NOD-DR3 with MHC class II knock-out mice (also called also called Ab⁰/DR3/NOD, DR3+ g7-, NOD-DR3 Ab⁰ or simply "NOD-DR3"). In 2011, Dr. Chella S. David (Mayo Clinic) sent some NOD-DR3 Ab⁰ mice with white coat color to Dr. Yaron Tomer (while at Icahn School of Medicine at Mount Sinai). Dr. Tomer maintained the colony by breeding NOD-DR3 Ab⁰ mice together each generation as homozygous for Ab⁰ and hemizygous for the DR3 transgene. In 2017, Dr. Tomer (while at Albert Einstein College of Medicine) sent NOD-DR3 Ab⁰ animals with white coat color to The Jackson Laboratory Repository as Stock No. 030434. Upon arrival, males were used to cryopreserve sperm. To establish our live colony, an aliquot of frozen sperm was used to fertilize NOD/ShiLtJ oocytes (Stock No. 001976).

Of note, it is not known if the Y chromosome has been fixed to the NOD background during backcrossing.

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Two of the 43 markers, on Chromosomes 2 and 17, were segregating. The marker on Chromosome 17 is likely linked to the mutant allele.

Allele

Symbol: Tg(HLA-DRA,HLA-DRB1*0301)#Gjh
Name: transgene insertion, Gunter J Hammerling
MGI accession ID: MGI:4460916
Generation method: Transgenic
Attributes: Inserted expressed sequence
Synonyms: DR3(DRwl7)
Marker symbol: Tg(HLA-DRA,HLA-DRB1*0301)#Gjh
Marker name: transgene insertion, Gunter J Hammerling
Chromosome: UN
Strain of origin: (C57BL/6 X DBA/2)F1 x C57BL/6
Expressed genes: HLA-DRA,major histocompatibility complex, class II, DR alpha,human; HLA-DRB1,major histocompatibility complex, class II, DR beta 1,human
Molecular note: A 6kb fragment from a genomic clone of HLA-DRA and a 24 kb fragment of HLA-DRB1*0301 were coinjected into fertilized eggs. Transgene expression was confirmed by Northern blot analysis.

Allele

Symbol: H2-Ab1^b-tm1Doi
Name: targeted mutation 1, Christophe Benoist and Diane Mathis
MGI accession ID: MGI:1927483
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: A beta⁰; A^bbeta^-; AB0; Abeta-; Class II⁰; H2-Ab1^tm1Dim; H2-Ab1^tm1Doi; H2-Ab1tm1Doi; II⁰; MHC class II-; MHCII^-; MHC-II-k.o.
Marker symbol: H2-Ab1
Marker name: histocompatibility 2, class II antigen A, beta 1
Marker synonyms: A beta; Abeta; Bb; CELIAC1; DQB2; H-2Ab; H2-Ab; HLA-DQB; HLA-DQB1; HLA-DXB; I-A^b; Ia2; Ia-2; IAb; I-Abeta; IDDM1; Rmcs1; RT1.B
Chromosome: 17
Strain of origin: 129S2/SvPas
Molecular note: The second exon was disrupted by the insertion of a neomycin resistance gene. In addition, the ES cell line used was derived from the 129S2/SvPas strain, which carries a deletion in the promoter region of H2-Ea. Consequently, these MHC class II molecule-deficient mice lacked cell surface expression of both class II-A and class II-E MHC proteins.