NOD.Cg-B2m^tm1Unc Tg(B2M)55Hpl Tg(HLA-A*1101/H2-K^b)#Sette/TdilJ

Stock number: 027849
Product name: NOD.mβ2m^null.hβ2m.HLA-A*11:01/K
Strain synonyms: NOD.mbeta2m.hbeta2m.HLA-A11
Research areas: Diabetes and Obesity Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Metabolism Research, Research Tools

Description

NOD.mβ2m^null.hβ2m.HLA-A11 mice are an NOD/ShiLtJ congenic strain expressing a chimeric MHC class I molecule composed of the human β2-microglobulin (in place of the endogenous mouse β2m) and the human/mouse HLA-A*1101 MHC class I heavy chain. These mice may be useful for identifying pancreatic β cell antigens/epitopes in the context of the HLA-A*1101 human MHC class I molecule.

Detailed description

NOD.mβ2m^null.hβ2m. HLA-A11 mice are an NOD/ShiLtJ congenic strain expressing a human MHC class I molecule composed of the human β2-microglobulin (in place of the endogenous mouse β2m) and the chimeric HLA-A11 MHC class I heavy chain.

The phenotype here describes mice homozygous for the mβ2m^null mutation, hemizygous for the hβ2m transgene, and hemizygous for the HLA-A11 transgene. Splenocytes exhibit cell-surface expression of the human β2m and HLA-A11 molecules, as well as endogenous NOD MHC class I molecules H-2K^d and H-2D^b. NOD.mβ2m^null.hβ2m. HLA-A11 mice are viable and fertile, although both traits are diminished when homozygous for the mβ2m^null mutation. Compared to NOD mice, NOD.mβ2m^null.hβ2m. HLA-A11 mice have greatly diminished diabetes susceptibility (~10% diabetic by about 16 weeks of age). Treatment with cyclophosphamide induces diabetes.

Note that when NOD.mβ2m^null.hβ2m.HLA-A11 females are maintained as homozygous for mβ2m^null mutation, female breeding success and viability are diminished.

Development

NOD.mβ2m^null.hβ2m.HLA-A11 mice harbor several mutations/transgenes, as described below.

The mβ2m^null mutation (B2m^tm1Unc) on chromosome 2 was created by Drs. Beverly Koller and Oliver Smithies (University of North Carolina, Chapel Hill) as described for Stock No. 002070. Mutant mice were sent to The Jackson Laboratory Repository and then bred to NOD/ShiLtJ inbred mice (Stock No. 001976) for at least 11 generations to create the NOD.mβ2m^null (Stock No. 002309). These mice were used for breeding as below

The hβ2m transgene (Tg(B2M)55Hpl) was created by Dr. Hidde Ploegh (University of Amsterdam, The Netherlands). A 4.3 kbp sequence containing the human β2m gene was isolated from an HLA-B27 genomic clone (HLA-B27 subtype). The transgene was microinjected into the pronuclei of (CBA x C57BL/LiA)F2 fertilized eggs, and mice from founder line 55 were propagated. Mice harboring 2-4 copies of the transgene on a B10.D2 congenic background were sent to Dr. David V. Serreze at The Jackson Laboratory in 1996. There, mice were bred to NOD.mβ2m^null mutant mice for at least nine generations. The resulting NOD.mβ2m^null.hβ2m mice (Stock No. 003355) were homozygous for linkage markers delineating all known insulin dependent diabetes susceptibility (Idd) loci of NOD origin, and were used for breeding as below.

The chimeric HLA-A*11:01/K transgene, Tg(HLA-A*1101)#Sette, was generated by Dr. Alessandro Sette (La Jolla Institute for Allergy and Immunology), contains the human leader α1 and α2 domains from HLA-A11 fused to the mouse H-2k^b α3 transmembrane and cytoplasmic domains. This transgenic insert was microinjected into fertilized BALB/c X C57BL/6 eggs. The founder line (number not specified) expressing the highest class I levels were backcrossed to C57BL/6 mice for 4 generations before the line was made homozygous. Transgenic mice (specific founder line not documented) were sent to Dr. Chella S. David (Mayo Clinic). Mice were then sent to Dr. Pere Santamaria (University of Calgary) who backcrossed them with NOD inbred mice for 8 generations. Dr. Teresa P. DiLorenzo (Albert Einstein College of Medicine) obtained mice from Dr. Santamaria and backcrossed the mice to NOD (obtained from JAX) for an additional 3 generations. The mice were homozygous for linkage markers delineating all known insulin dependent diabetes susceptibility (Idd) loci of NOD origin.

To generate the NOD.mβ2m^null.hβ2m.HLA-A*11:01/K animals, Dr. Teresa P. DiLorenzo (Albert Einstein College of Medicine) obtained NOD.mβ2m^null.hβ2m mice from Dr. David V. Serreze (The Jackson Laboratory) and bred them to NOD. HLA-A*11:01/K transgenic mice. Mice homozygous for mβ2m^null mutation, hemizygous for the hβ2m transgene, and carrying the HLA-A*11:01/K transgene were sent to The Jackson Laboratory Repository as Stock No. 027849. Upon arrival, males were used to cryopreserve sperm. To establish the living NOD.mβ2m^null.hβ2m.HLA-A11 mouse colony, an aliquot of the frozen sperm was used to fertilize NOD/ShiLtJ oocytes (Stock No. 001976).

Allele

Symbol: Tg(B2M)55Hpl
Name: transgene insertion 55, Hidde Ploegh
MGI accession ID: MGI:2159101
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: (B2M)55HplTG
Marker symbol: Tg(B2M)55Hpl
Marker name: transgene insertion 55, Hidde Ploegh
Chromosome: UN
Strain of origin: (CBA x C57BL/LiA)F2
Expressed genes: B2M,beta-2-microglobulin,human
Molecular note: A transgene containing the human B2M gene was shown to be expressed in spleen cells of mice carrying the transgene.

Allele

Symbol: B2m^tm1Unc
Name: targeted mutation 1, University of North Carolina
MGI accession ID: MGI:1857133
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: b2m^null; beta2M-; beta2m⁰; beta2m^null; beta2m^o; beta2m^tm1Unc; beta2MKO; beta2-m-KO; I⁰; MHC-I^-
Marker symbol: B2m
Marker name: beta-2 microglobulin
Marker synonyms: beta 2 microglobulin; beta2-m; IMD43; Ly-m11
Chromosome: 2
Strain of origin: 129P2/OlaHsd
Molecular note: Insertion of a neomycin-resistance gene into the second exon.

Allele

Symbol: Tg(HLA-A*1101/H2-K^b)#Sette
Name: transgene insertion, Alessandro Sette
MGI accession ID: MGI:3851009
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: HLA-A11; HLA-A11/K^b
Marker symbol: Tg(HLA-A*1101/H2-K^b)#Sette
Marker name: transgene insertion, Alessandro Sette
Chromosome: UN
Strain of origin: BALB/c or C57BL/6
Expressed genes: HLA-A/H2-K,major histocompatibility complex, class I, A, human and histocompatibility 2, K region fusion gene,
Molecular note: The model was created by microinjecting a chimeric transgene combining a fragment of the genomic clone of the human HLA-A*1101 gene that included leader sequence, alpha1 and alpha2 domains ligated to a fragment of the murine H2-K^b gene containing the alpha3, transmembrane and cytoplasmic domains.