NOD.Cg-B2m^tm1Unc Tg(B2M)55Hpl Tg(HLA-B*0702)#Chmb/TdilJ

Stock number: 018907
Product name: NOD.mβ2mnull.hβ2m.HLA-B7
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-B7 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 human HLA-B*0702 MHC class I heavy chain. These mice may be useful for identifying pancreatic β cell antigens/epitopes in the context of the HLA-B*0702 human MHC class I molecule.

Detailed description

NOD.mβ2m^null.hβ2m.HLA-B7 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 human HLA-B*0702 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-B*0702 transgene. Splenocytes exhibit cell-surface expression of the human β2m and HLA-B7 molecules, as well as endogenous NOD MHC class I molecules H-2K^d and H-2D^b. NOD.mβ2m^null.hβ2m.HLA-B7 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-B7 mice have greatly diminished diabetes susceptibility (~5% diabetic by 35 weeks). Treatment with cyclophosphamide induces diabetes. The peptide binding motif described for HLA-B*0702 has a proline (P) at position 2 and leucine (L) at the C terminus (SYFPEITHI database).

Note that when NOD.mβ2m^null.hβ2m.HLA-B7 females are maintained as homozygous for mβ2m^null mutation, female breeding success and viability are diminished.

Development

NOD.mβ2m^null.hβ2m.HLA-B7 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 at 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 human major histocompatibility complex (MHC) class I heavy chain (α chain) is encoded by the human leukocyte antigen B gene (HLA-B). In humans, the HLA-B7 serotype defined by the HLA-B*0702 allele is expressed in 15-20% of the population. Dr. John W. Chamberlain (Yale University School of Medicine) used human HLA-B7 genomic clone pJY150 to isolate a 6.0 kbp EcoRI-BamHI fragment (fragment a) encoding the human HLA-B*0702 heavy chain sequence (α1, α2 α3, transmembrane, and cytoplasmic domains from HLA-B*0702) with 0.66 kbp of 5' flanking DNA and 2.0 kbp of 3' flanking DNA. The transgene was microinjected into the male pronucleus of fertilized one-cell embryos (either (B6SJL)F1/J, (B6SJL)F2/J or (B6D2)F1/J). Founders from several lines were then bred to C57BL/6J mice. Transgenic mice (specific founder line not documented) were sent to Dr. Chella S. David (Mayo Clinic) where they were backcrossed with C57BL/6 inbred mice for several generations. Mice were then sent to Dr. Pere Santamaria (University of Calgary) who backcrossed them to NOD inbred mice for 6-8 generations. Next, the mice were sent to Dr. Teresa P. DiLorenzo (Albert Einstein College of Medicine) who backcrossed them with NOD inbred mice for several more generations (~N10 total). The mice were homozygous for linkage markers delineating all known insulin dependent diabetes susceptibility (Idd) loci of NOD origin. These NOD.HLA-B*0702 transgenic mice were used for breeding as below.

To generate the NOD.mβ2m^null.hβ2m.HLA-B7 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-B*0702 transgenic mice. Mice homozygous for mβ2m^null mutation, hemizygous for the hβ2m transgene, and hemizygous for the HLA-B*0702 transgene were sent to The Jackson Laboratory Repository as Stock No. 018907. Upon arrival, males were used to cryopreserve sperm. To establish the living NOD.mβ2m^null.hβ2m.HLA-B7 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
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
Marker symbol: B2m
Marker name: beta-2 microglobulin
Chromosome: 2
Strain of origin: 129P2/OlaHsd
Molecular note: Insertion of a neomycin-resistance gene into the second exon.

Allele

Symbol: Tg(HLA-B*0702)#Chmb
Name: transgene insertion, Jeffrey S Chamberlain
MGI accession ID: MGI:5440508
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(HLA-B*0702)#Chmb
Marker name: transgene insertion, Jeffrey S Chamberlain
Chromosome: UN
Strain of origin: Not Specified
Expressed genes: HLA-B,major histocompatibility complex, class I, B,human
Molecular note: The human major histocompatibility complex (MHC) class I heavy chain (alpha chain) is encoded by the human leukocyte antigen B gene (HLA-B). In humans, the HLA-B7 serotype defined by the HLA-B*0702 allele is expressed in 15-20% of the population. The transgene used human HLA-B7 genomic clone pJY150 to isolate a 6.0 kbp EcoRI-BamHI fragment (fragment a) encoding the human HLA-B*0702 heavy chain sequence (alpha1, alpha2, alpha3 , transmembrane, and cytoplasmic domains from HLA-B*0702) with 0.66 kbp of 5' flanking DNA and 2.0 kbp of 3' flanking DNA.
General note: The transgene was microinjected into the male pronucleus of fertilized one-cell embryos (either (B6SJL)F1/J, (B6SJL)F2/J or (B6D2)F1/J).