NOD.Cg-Prkdc^scid H2-Ab1^tm1Doi Il2rg^tm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv Tg(INS*)172Dvs/DvsJ

Stock number: 026936
Product name: NSG-Ab^o-DQ8-INS*VNTR , NSG-Abo-DQ8-INS*VNTR
Research areas: Cancer Research, Cardiovascular Research, Dermatology Research, Developmental Biology Research, Endocrine Deficiency Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-Ab⁰-DQ8-INS*VNTR (or NSG-Ab0-DQ8-INS*VNTR) mice are NOD.scid.Il2Rγc^null ("NSG") animals lacking murine MHC class II (Ab⁰), expressing a humanized HLA-DQ8 molecule linked to type 1 diabetes development and a human proinsulin class I variant (INS*VNTR; 26-63 tandem ACAGGGGTGTGGGG repeats) common to individuals with type 1 diabetes. These mice are useful for studying type 1 diabetes, autoimmunity and transplantation.

Detailed description

NSG-Ab⁰-DQ8-INS*VNTR (or NSG-Ab0-DQ8-INS*VNTR) mice are NOD.scid.Il2Rγc^null ("NSG"; Stock No. 005557) animals also lacking murine MHC class II (Ab⁰) that express the HLA-DQ8A transgene (Tg(HLA-DQA1,HLA-DQB1)1Dv) and the INS*VNTR transgene (Tg(INS*)172Dvs).

The following genotype(s) may be collectively referred to as NSG-Ab⁰-DQ8-INS*VNTR mice: females homozygous for scid, Ab⁰, Il2Rγc^null and HLA-DQ8A and hemizygous for the HLA-DQ8A transgene - and males homozygous for scid and Ab⁰, hemizygous for the X-linked Il2Rγc^null, homozygous for HLA-DQ8A and hemizygous for INS*VNTR.

The human insulin gene (INS) is translated into preproinsulin (the precursor of mature insulin), which is processed to proinsulin by removal of the signal peptide and then to mature biologically active insulin by removal of the C-peptide. In the INS promoter region, a variable number tandem repeat (VNTR) region consisting of a 14-15 bp consensus sequence is associated with three classes of alleles: there is a higher frequency of class I alleles with shorter repeat sequences in individuals with Type 1 diabetes, whereas individuals with longer class III alleles are relatively protected from Type 1 diabetes. The VNTR regulates transcription rates of insulin and its precursors. The INS*VNTR transgene derived from founder line 17-2 (172) expresses a human proinsulin class I variant (26-63 tandem ACAGGGGTGTGGGG repeats).

Therefore, the NSG-Ab⁰-DQ8-INS*VNTR mice may be expected to combine the NSG phenotype (Stock No. 005557) and murine MHC class II knockout phenotype, while expressing a humanized HLA-DQ8 molecule linked to Type 1 diabetes development (see Stock No. 026561) - and also expressing a class I variant human proinsulin (INS*VNTR) common to individuals with Type 1 diabetes.


When hemizygous for the INS*VNTR transgene derived from line 17-2 (172), both female and male NSG-Ab⁰-DQ8-INS*VNTR mice show high expression levels of human insulin and human C-peptide in both serum and pancreas.

Stock No. 005557: Although viable and fertile, homozygous NSG mice are immunodeficient: they have no mature T cells or B cells, lack functional natural killer (NK) cells, have reduced numbers of lymphocytes and myeloid dendritic cells, and are deficient in cytokine signaling.
Stock No. 026561: NSG-Ab⁰-DQ8 are viable and fertile. Ab⁰ homozygotes do not express murine major histocompatibility complex (MHC) class II. Human haematopoietic stem cell-engraftment in NSG-Ab⁰-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.

Similar to other immunodeficient strains, maintaining NSG-Ab⁰-DQ8-INS*VNTR mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If NSG-Ab⁰-DQ8-INS*VNTR animals are maintained in low health barrier rooms, the use of medicated water (e.g., sulfatrim/trimethoprim-sulfa or enrofloxacin/Baytril) is suggested to increase overall colony health.

View our Resources for the NSG mouse model, including discussion forum, immunodeficient model comparison, and categorized, up-to-date references.

Development

NSG-Ab⁰-DQ8-INS*VNTR (or NSG-Ab0-DQ8-INS*VNTR) mice are NOD.scid.Il2Rγc^null ("NSG") animals with the H2-Ab1^tm1Doi knockout allele (Ab⁰), the HLA-DQ8A transgene (Tg(HLA-DQA1,HLA-DQB1)1Dv) and the INS*VNTR transgene (Tg(INS*)172Dvs). NSG-Ab⁰-DQ8-INS*VNTR 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 "MHC class II knockout" allele (Ab⁰; H2-Ab1^tm1Doi) on chromosome 17 was created by Drs. Diane Mathis and Christophe Benoist (while at INSERM, Strasbourg, France) - see allele description for Stock Nos. 004606 or 010966. Mice with this Ab⁰ mutation on the NOD genetic background were used to assemble Stock No. 026936.

To create the HLA-DQ8A transgene (Tg(HLA-DQA1,HLA-DQB1)1Dv), a 30 kbp DNA fragment containing the DQA*0301 gene [from cosmid H11A] and a 38 kbp 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. Mice with the HLA-DQ8A transgene on the NOD genetic background were used to assemble Stock No. 026936.

The INS*VNTR transgene (Tg(INS*)172Dvs) was created by the Type 1 Diabetes Repository (Dr. David V. Serreze) at The Jackson Laboratory. A human insulin genomic sequence was obtained with 26-63 tandem ACAGGGGTGTGGGG repeats in its promoter region. The transgene was microinjected into NSG embryos (Stock No. 005557). Mice from founder line 17-2 (172) were established and then bred to NOD-congenic mice harboring the Ab⁰ mutation and the HLA-DQ8A transgene. The resulting NSG-Ab^o-DQ8-INS*VNTR mutant mice were established at The Jackson Laboratory Repository as Stock No. 026936.

The colony is maintained by breeding females homozygous for scid, homozygous for Ab⁰, homozygous for the X-linked Il2Rγc^null, homozygous for HLA-DQ8A and hemizygous or noncarrier for INS*VNTR with males that are homozygous for scid, homozygous for Ab⁰, hemizygous for the X-linked Il2Rγc^null, homozygous for HLA-DQ8A and noncarrier or hemizygous for INS*VNTR.

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: 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.

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: Tg(INS*)172Dvs
Name: transgene insertion 172, David V Serreze
MGI accession ID: MGI:5903249
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(INS*)172Dvs
Marker name: transgene insertion 172, David V Serreze
Chromosome: UN
Strain of origin: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
Expressed genes: INS,insulin,human
Site of expression: Expression of the transgene is found in all primary cilia of the inner ear sensory epithelium.
Molecular note: The INS*VNTR transgene (Tg(INS*)172Dvs) was created by the Type 1 Diabetes Repository (Dr. David V. Serreze) at The Jackson Laboratory. A human insulin genomic sequence was obtained with 26-63 tandem ACAGGGGTGTGGGG repeats in its promoter region.