NOD.Cg-Prkdc^scid H2-Ab1^tm1Doi Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/SzJ

Stock number: 026561
Product name: NSG-HLA-DQ8
Research areas: Cancer Research, Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-HLA-DQ8 mice lack murine MHC class II and express mutant human leukocyte antigen (HLA)-DQ8 on the immunodeficient NOD scid gamma (NSG) background. This strain may be useful in studies utilizing an autoantigen-specific vaccination approach for the prevention of islet autoimmunity in individuals susceptible to type I diabetes.

Detailed description

NSG-HLA-DQ8 mice are NOD.scid.Il2Rγc^null ("NSG") animals also harboring the H2-Ab1 knock-out allele (Ab^o) and the human leukocyte antigen DQ8A transgene (HLA-DQ8A).


NSG-HLA-DQ8 females homozygous for all three mutations and homozygous for the HLA-DQ8A transgene, and males homozygous for scid, homozygous for Ab^o, homozygous for HLA-DQ8A and hemizygous for the X-linked Il2Rγc^null may be collectively referred to as homozygous NSG-HLA-DQ8 mice.


Homozygous NSG-HLA-DQ8 express HLA-DQ8 and lack murine MHC class II, on an NSG background (see Stock No. 005557 for NSG mouse description). Homozygous NSG-HLA-DQ8 mice are viable and fertile. Human haematopoietic stem cell-engraftment in NSG-HLA-DQ8 mice results in human insulin-specific Foxp3⁺ Treg-induction upon subimmunogenic vaccination with strong agonistic insulin mimetopes in vivo. These Tregs are stable and show increased expression of Treg signature genes while suppressing effector T cells.

Development

NSG-HLA-DQ8 mice (Stock No. 026561) are NOD.scid.Il2Rγc^null ("NSG") animals with the H2-Ab1 knock-out allele (Ab^o; see below) and the HLA-DQ8A transgene. NSG-HLA-DQ8 mice harbor several mutations, as described below.

NSG mice are non-obese diabetic animals (NOD/ShiLtJ) harboring the spontaneous severe combined immunodeficiency mutation (Prkdc^scid) on chromosome 16 and the Il2Rγc^null mutation on the X chromosome (Il2rg^tm1Wjl created by Dr. Warren J. Leonard [NIH]). NSG mice are described and available from The Jackson Laboratory Repository as Stock No. 005557.

The H2-Ab1 knock-out allele in Stock No. 026561 was originally reported to be H2-Ab1^tm1Gru (created by Dr. Michael J. Grusby; while at Harvard) - harboring a forward-oriented neomycin resistance cassette that disrupts H2-Ab1 exon 2 (and made in ES cells with a deletion in the promoter region of H2-Ea). In 2018, quality control efforts at The Jackson Laboratory were completed that revealed Stock No. 026561 instead has the very similar H2-Ab1^tm1Doi knock-out allele (created by Drs. Christophe Benoist and Diane Mathis; while at INSERM) - harboring a reverse-oriented neomycin resistance cassette that disrupts H2-Ab1 exon 2 (and made in ES cells with a deletion in the promoter region of H2-Ea). The Ab^o knock-out allele is on chromosome 17.

The HLA-DQ8A transgene (Tg(HLA-DQA1,HLA-DQB1)1Dv) was created by Dr. Chella David (Mayo Clinic). A 30kb DNA fragment containing the DQA*0301 gene from cosmid H11A and a 38kb DNA fragment containing the DQB*0302 gene from cosmid X10A were microinjected into (CBA/J x B10.M)F2 embryos. To establish germline transmission, founders were mated to the B10.M strain. Next, HLA-DQ8A transgenic mice were mated to NSG mice (Stock No. 005557) for at least ten generations. Subsequently, they were intercrossed with NSG mice lacking mouse MHC class II (NSG-Ab^o; Stock No. 021885). The resulting NSG-Ab^o HLA-DQ8A mice, called NSG-HLA-DQ8, are available as Stock No. 026561.

Allele

Symbol: Tg(HLA-DQA1,HLA-DQB1)1Dv
Name: transgene insertion 1, Chella David
MGI accession ID: MGI:3586633
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(HLA-DQA1,HLA-DQB1)1Dv
Marker name: transgene insertion 1, Chella David
Chromosome: UN
Strain of origin: (CBA/J x B10.M-H2^f)F2
Expressed genes: HLA-DQA1,major histocompatibility complex, class II, DQ alpha 1,human; HLA-DQB1,major histocompatibility complex, class II, DQ beta 1,human
Molecular note: A 30-kb human genomic DNA fragment containing the entire HLA-DQA1*0301 gene and a promoter-truncated HLA-DQB1*0302 gene from cosmid H11A and a 38-kb DNA fragment containing the entire HLA-DQB1*0302 gene from cosmid X10A were coinjected.
General note: Flow cytometric analysis of peripheral blood leukocytes (PBL) using a monoclonal antibody to HLA-DQ demonstrated that transgenic mice express the transgenes. 25-40% of the PBLs of mixed-background transgenic mice homozygous for the targeted MHC class II mutation H2-Ab1^tm1Dim express the transgenes. Lymph nodes of such mice contain three-fold the levels of CD4⁺ CD44⁺ T-cells as do those of nontransgenic MHC class II-deficient mice, levels similar to those of class II-sufficient mice. Immunization of transgenic H2-Ab1^tm1Dim homozygous mice with bovine collagen II results in a strong IgG antibody response to the immunogen, and 66% of these mice develop severe, persistent arthritis. (J:88296)

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Chromosome: X
Strain of origin: 129S4/SvJae
Site of expression: Primarily lymphoid cells.
Molecular note: A neomycin resistance cassette replaced part of exon 3 and all of exons 4 - 8 of the gene, resulting in the loss of most of the extracellular domain and all of the transmembrane and cytoplasmic domains of the protein.

Allele

Symbol: H2-Ab1^tm1Doi
Name: targeted mutation 1, Christophe Benoist and Diane Mathis
MGI accession ID: MGI:1927483
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: H2-Ab1
Marker name: histocompatibility 2, class II antigen A, beta 1
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.