NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ

Stock number: 029295
Product name: NSG DR4
Strain synonyms: NSG-DR4
Research areas: Cancer Research, Cardiovascular Research, Developmental Biology Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG DR4 mice are NOD.scid.Il2Rγc^null ("NSG") animals with the DR4β(NT) transgene that encodes a human HLA-DRB1*0401 engineered to have amino acid mutations in the β2 domain for better interaction with murine CD4. The NSG platform is permissive for xenograft/human tumor growth, with the DR4β(NT) transgene allowing irradiated NSG DR4 mice to be engrafted with HLA-DR-matched hematopoietic stem cells (HSC) - resulting in humanized T cell and B cell populations.

Detailed description

NSG DR4 mice are NOD.scid.Il2Rγc^null ("NSG"; Stock No. 005557) animals with the DR4β(NT) transgene that encodes a human HLA-DRB1*0401 modified to have improved interaction with murine CD4. The presence of the DR4β(NT) transgene allows irradiated NSG DR4 mice to be engrafted with HLA-DR-matched hematopoietic stem cells (HSC) - resulting in humanized T cell and B cell populations.

Females that are homozygous for both mutations and homozygous for the DR4β(NT) transgene, and males that are homozygous for scid, hemizygous for the X-linked Il2Rγc^null mutation and homozygous for the DR4β(NT) transgene, may be collectively referred to as homozygous NSG DR4 mice.

Although viable and fertile, homozygous NSG DR4 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. The NSG platform is permissive for xenograft/human tumor growth.
The DR4β(NT) transgene has the mouse H2-Eα promoter directing expression of the HLA class II antigen binding domain molecule (HLA-DRB1*0401) modified to have amino acid mutations in the β2 domain for better interaction with murine CD4. In humans, the HLA-DR4 genotype (HLA-DRA/HLA-DRB1*0401) is associated with the development of autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, type 1 diabetes, and lyme disease-induced arthritis.

Maintenance Note: Similar to other immunodeficient strains, maintaining homozygous NSG DR4 mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If homozygous NSG DR4 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 DR4 mice are NOD.scid.Il2Rγc^null ("NSG") animals with the DR4β(NT) transgene. Each mutation/transgene is 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 at ~101 Mbp 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 DR4β(NT) transgene (also called DR4β(NT)/pDOI-5) was generated by Dr. Chella S. David (Mayo Clinic) as described in Pan et al. 1998 J Immunol 161:2925. This transgene has the mouse H2-Eα promoter (and rabbit β-globin intron) upstream of "DRB1*0401(NT)" - a human HLA-DRB1*0401 cDNA sequence engineered to have amino acid mutations (Q110N*K139T) in the β2 domain for better interaction with murine CD4. The transgene was microinjected into fertilized eggs from (SWR x B10)F1 mice. Mice from the founder line with five transgene copies were backcrossed to B10.RFB3 animals (C57BL/10-congenic Eβ⁰ Eα^p mice lacking endogenous Eβ but expressing Eα intracytoplasmically). Next, Dr. David backcrossed them to NOD inbred mice for five generations. In 2009, the NOD-congenic DR4 Ab⁰ mice were sent to Dr. Leonard D. Shultz (The Jackson Laboratory). They were then bred to NSG mice (Stock No. 005557) for at least one generation, and the Ab⁰ mutation was replaced with the endogenous H2-Ab1 allele. The resulting NSG DR4 animals were bred together for several generations, and then made available as Stock No. 029295.

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(H2-Ea-HLA-DRB1*0401*)1Dv
Name: transgene insertion 1, Chella David
MGI accession ID: MGI:6117040
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(H2-Ea-HLA-DRB1*0401*)1Dv
Marker name: transgene insertion 1, Chella David
Chromosome: UN
Strain of origin: (SWR x C57BL/10)F1
Expressed genes: HLA-DRB1,major histocompatibility complex, class II, DR beta 1,human
Molecular note: This transgene has the mouse H2-Ealpha promoter (and rabbit beta-globin intron) upstream of a human HLA-DRB1*0401 cDNA sequence engineered to have amino acid mutations (Q110N and K139T) in the beta2 domain to provide an improved interaction with murine CD4.