NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(HLA-A/H2-D/B2M)Dvs Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/J

Stock number: 035855
Product name: DRAGA
Strain synonyms: DRAGA
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

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DRAGA mice are NOD.Rag1KO.IL2RγcKO ("NRG") animals with transgenes encoding the human HLA class I heavy and light chains (HLA-A2.1), as well as the human HLA class II antigen binding domain molecules (defined by the HLA-DR4 genotype [HLA-DRA/HLA-DRB1*0401]) fused to the I-E^d MHC class II molecule. The presence of these transgenes allows enhanced HLA-matched hematopoietic stem cells (HSC) engraftment and subsequent human T cell and B cell development. These DRAGA mice may be useful as an in vivo model for studying human T cell/B cell development and function, vaccine testing, and generation of "fully human" IgM, IgG, IgA or IgE monoclonal antibodies for prophylactic and/or therapeutic use in autoimmune diseases and infectious diseases.

Detailed description

DRAGA mice are NOD.Rag1KO.IL2RγcKO ("NRG"; Stock No. 007799) animals with transgenic human HLA class I heavy and light chains (HLA-A2.1) and chimeric human-mouse class II transgenes encoding the HLA class II antigen binding domain molecules (defined by the HLA-DR4 genotype [HLA-DRA/HLA-DRB1*0401]) fused to the I-E^d MHC class II molecule. As expression of HLA class I molecules is required for reconstitution of of HLA class I restricted human CD8 T cells, the presence of these transgenes allows irradiated DRAGA mice to be engrafted with HLA-DR-matched human hematopoietic stem cells (HSC) and elicit cytotoxic, HLA class I and II restricted CD8 T cells.

DRAGA females homozygous for the Rag1KO mutation, homozygous for the X-linked IL2RγcKO mutation, homozygous for the Tg(HLA-A/H2-D/B2M)1Dvs transgene and hemizygous for the Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito and DRAGA males homozygous for the Rag1KO mutation, hemizygous for the X-linked IL2RγcKO mutation, homozygous for the Tg(HLA-A/H2-D/B2M)1Dvs transgene and hemizygous for the Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito transgene are viable and fertile, with no mature T cells, B cells, or natural killer (NK) cells. Like NRG mice, DRAGA mice have the same irradiation/engraftment advantages compared to NOD.scidKO.IL2RγcKO ("NSG"; Stock No. 005557), NOD.Rag1KO (Stock No. 003729), and NOD.scidKO (Stock No. 001303) mice. DRAGA mice differ from DRAG mice (Stock No. 017914) in that DRAGA mice have transgenic expression of human HLA class I antigen-binding domain molecules (HLA-A2.1) in addition to HLA class II molecules (HLA-DR4).

Specifically, when irradiated DRAGA, NRG-HLA-A2/HHD, or NRG mice are engrafted with HLA-A2 and HLA-DR-matched umbilical cord blood HSC, Majji et al. (Sci Rep, 2015) report that the HLA-DR4-expression in DRAGA mice results in several advantages compared to NRG-HLA-A2/HHD or NRG mice:
i) DRAGA mice show improved human T cell reconstitution rates and develop higher numbers of human CD4 T cells.
ii) Such engraftment results in homing and development of bone-marrow-derived human T cell precursors in thymus, as well as proficient repopulation of peripheral lymphoid organs with mature CD4 cells and CD8 T cells.
iii) The repopulation of mature CD8 T cells in the periphery is noteworthy as DRAG mice do not express HLA class I molecules (the restriction element for thymic differentiation of human CD8 T cells).
iv) Increased repopulation of T regulatory cell compartment (human CD4+FOXP3+ Tregs).
v) While engrafted DRAGA, NRG-HLA-A2/HHD, and NRG mice develop similar human B cell (periphery or spleen) percentages, the human B cells in DRAGA mice are functional and secrete IgG as well as IgM (whereas human B cells in NRG and NRG-HLA-A2/HHD mice secrete only IgM and have impaired class-switching).

Development

DRAGA mice are NOD.Rag1KO.IL2RγcKO ("NRG") animals expressing human HLA-A2 class I heavy and light chains from the HLA-A2/HHD transgene and human class II HLA-DRB1*0401 molecules from the co-injected HLA-DR4-IE transgenes.
To generate the DRAGA animals (Stock No. 035855), in 2021 The Jackson Laboratory bred NOD.B.2m^null.HHD mice (JAXMice Stock No. 006611) to DRAG mice (JAXMice Stock No. 017914), and the B2m^null allele was subsequently removed.

DRAGA mice harbor several mutations/transgenes, as described below.

Creation of NOD.B.2m^null.HHD mice is described for JAXMice Stock No. 006611. Briefly, NOD.B.2m^null.HHD mice are NOD-congenic animals with the B2m^null mutation on chromosome 2, and carrying the HLA-A2/HHD transgene.
The HLA-A2/HHD transgene was generated by Dr. David Serreze (The Jackson Laboratory) to encode 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 transgene was first injected into NOD/ShiLtDvs embryos as described for JAXMice Stock No. 006604.

Creation of DRAG mice is described for JAXMice Stock No. 017914. Briefly, DRAG mice are NOD-congenic animals with the Rag1^null (Rag1KO or Rag1^tm1Mom; neo cassette replacing 1356 bp genomic fragment) mutation on chromosome 2, the IL2rγ^null mutation on the X chromosome (as described above), and carrying the HLA-DRA / HLA-DRB1*0401 co-injected transgenes.
The MHC class II α chain is encoded by HLA-DRA, while the MHC class II β chain is encoded by different polymorphic alleles of HLA-DRB. 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. Dr. Kouichi Ito (while at Hoffmann-La Roche Inc) generated mice possessing two HLA-DR4-IE chimeric MHC class II genes each encoded by its own HLA-DR4-IE transgene, HLA-DRA-IE^dα and HLA-DRBI*0401-IE^dβ. The HLA-DRA-IE^dα transgene has the mouse IE^dα genomic DNA sequences with its exon 2 replaced by the human HLA-DRA exon 2 sequence; the resulting chimeric human-mouse MHC class II gene encodes the IE^dα membrane-proximal domains with the antigen binding domain from HLA-DRA. The HLA-DRBI*0401-IE^dβ transgene has the mouse IE^dβ genomic DNA sequences with its exon 2 replaced by the human HLA-DRBI*0401 exon 2 sequence; the resulting chimeric human-mouse MHC class II gene encodes the IE^dβ membrane-proximal domains with the antigen binding domain from HLA-DRBI*0401. The two transgenes were co-injected into C57BL/6 fertilized eggs, and founder mice were obtained. Mice from founder line 39-2 were then propagated and maintained as described for JAXMice Stock No. 017914.

Allele

Symbol: Tg(HLA-A/H2-D/B2M)1Dvs
Name: transgene insertion 1, David V Serreze
MGI accession ID: MGI:3711057
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: HDD; HHD
Marker symbol: Tg(HLA-A/H2-D/B2M)1Dvs
Marker name: transgene insertion 1, David Serreze
Marker synonyms: HDD; HHD
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: Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito
Name: transgene insertion 39-2, Kouichi Ito
MGI accession ID: MGI:4429540
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: DR4; DR4-H2E; DRA/DRB1*0401-IE Tg; HLA-DR4; HLA-DR4-IE; HLA-DRA-IEalpha/HLA-DRBI*0401-IEbeta Tg; Tg(HLA-DRA,HLA-DRB1)39-2Kito; Tg(HLA-DRA/H2-Ea,HLA-DRB1*0401/H2-Eb)1Kito
Marker symbol: Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito
Marker name: transgene insertion 39-2, Kouichi Ito
Marker synonyms: Tg(HLA-DRA,HLA-DRB1)39-2Kito
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: HLA-DRA,major histocompatibility complex, class II, DR alpha,human; HLA-DRB1,major histocompatibility complex, class II, DR beta 1,human
Molecular note: A chimeric MHC class II complex was created by fusing the human peptide antigen binding domains of class II alpha and beta chains (HLA-DRA and HLA-DRBI*0401) to the remaining domains of murine class II alpha or beta chains (H2-Ea and H2-Eb1 genes that make up mouse I-Ed). The chimeric gene constructs were manipulated by replacing exon 2 of IE^d-alpha or IE^d-beta genomic DNA with gene fragments containing exon 2 of HLA-DRA or HLA-DRB1*0401 respectively. The native murine promoters were left intact. These transgene constructs were co-injected into blastocytes. Flow cytometry analysis indicates that B220+ cells co-express the chimeric MHC class II molecules. Line 39-2 is a representative of several lines generated. The chosen human alleles represent human serotype HLA-DR4.

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: Rag-; Rag^1tm1Mom; RAG-1-; Rag1^-; RAG1^null; Rag-1KO
Marker symbol: Rag1
Marker name: recombination activating gene 1
Marker synonyms: RAG-1; RNF74
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: [KO]gammac; CD132^-; gammac^-; gc^-; Il2rg^tm1Wjll; IL2Rgamma^null
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Marker synonyms: [g]c; Ab2-183; Cd132; CIDX; common cytokine receptor gamma chain; common gamma chain; gamma C receptor; gamma(c); gammaC; gc; IL-2RG; IMD4; P64; SCIDX; SCIDX1
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.