NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(HLA-A/H2-D/B2M)1Dvs/SzJ

Stock number: 014570
Product name: NSG-HLA-A2/HHD
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-HLA-A2/HHD mutant mice are immunodeficient and express human HLA class I heavy and light chains. This strain may be useful as a human hematopoietic engraftment host that supports the maturation of human T cells with transplantation of human hematopoietic stem cells.

To order this model with CD34+ or PBMC engraftment, please visit our Humanized Mice page.

Detailed description

Mice that are homozygous for the Prkdc^scid and Il2rg^tm1Wjl alleles (males are hemizygous for the Il2rg^tm1Wjl allele) and harbor Tg(HLA-A2/H2-D/B2m)1Dvs/SzJ are immunodeficient and express human HLA class I heavy and light chains. HLA-A2 and B2M proteins are detected on the surface of NSG-HLA-A2/HHD splenocytes by flow cytometry analysis. Transplantation of purified human hematopoietic stem cells into newborn NSG-HLA-A2/HHD mice establishes a humanized immune microenvironment allowing functional maturation of human hematopoietic cells. Epstein-Barr virus (EBV) infection results in EBV-associated B cell proliferation and HLA-restricted EBV antigen-specific effector memory CTLs.

In 2018, protein expression was verified by flow cytometry in male and female CD45+, TER119- splenocytes using an anti-HLA-A2 antibody (Clone B7-2.1). A second monoclonal antibody, CR11-351, also detected expression but was less effective at discriminating HLA+ and HLA- cells.

Development

To generate this spontaneous/targeted mutation/transgenic strain NOD/ShiLtDvs-Tg(HLA-A/H2-D/B2M)1Dvs/J (Stock No. 006604) mice were crossed to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (Stock No. 005557) mice. Mice were subsequently bred to homozygosity for all alleles, with male mice hemizygous for the X-linked Il2rg^tm1Wjl allele. The Donating Investigator maintained the strain as homozygotes for all the alleles: Prkdc^scid Il2rg^tm1Wjl and Tg(HLA-A2/H2-D/B2M)1Dvs (males will be hemizygous for the X-linked Il2rg^tm1Wjl targeted mutation). The mice were crossed to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (Stock No. 005557) for 9 generations.

Allele

Symbol: Tg(HLA-A/H2-D/B2M)1Dvs
Name: transgene insertion 1, David Serreze
MGI accession ID: MGI:3711057
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(HLA-A/H2-D/B2M)1Dvs
Marker name: transgene insertion 1, David Serreze
Chromosome: UN
Strain of origin: NOD/ShiLtDvs
Expressed genes: B2M,beta-2-microglobulin,human; HLA-A,major histocompatibility complex, class I, A,human
Molecular note: This transgene encodes a human B2-microglubulin (B2M) covalently linked to the MHC class 1, alpha1 and alpha2 binding domains of the human HLA-A2.1 gene, and the alpha3, cytoplasmic and transmembrane domains of the murine H2-D^b. The construct was injected into fertilized NOD/ShiLtDvs oocytes. The construct was originally described in J:41077 and used to create a different transgenic line.
General note: The HDD construct was created and used to generate a differnet transgenic mouse line in J:41077.

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.