A 78.8 kb deletion disrupting Class II MHC genes was induced in 129S2/SvPas-derived H1 embryonic stem (ES) cells via Cre recombination. A hygro-resistance cassette was inserted at the deletion site. The deletion spans from the second exon of the H2-Ab1 gene the third exon of the H2-Ea gene. The H2-Aa, H2-Eb1 and H2-Eb2 genes are completely deleted.
Location on chromosome (bp):
cytoband B-C
MGI Gene Accession ID
Please also use this link to determine if other mutants have been registered with MGI (Mouse Genome Informatics)
IIdelta, MHC class IIdelta, MHC II KO, MHC II-, MHC II0, MHC IIdelta, MHC-IIdelta, MHCII-, MHCIIdelta
1. Transgene:
human leukocyte antigen class II
Symbol
HLA DRA - DQB2
Genetic alteration of transgene:
A 320kb YAC DNA clone containing an intact region from HLA DRA to DQB2 was transfected into C57BL/6 ES cells. Two clones in which the relevant HLA loci had integrated into the ES genome without detectable deletion or rearrangement and which lacked yeast chromosome 3. A single founder was established and expanded by breeding with C57BL/6 mice bearing a class II knockout background.
2. Transgene:
transgene insertion 2362, Dan R Littman
Symbol
Tg(Cd4-CD4)2362Litt
Promoter
murine Cd4 and human CD4
Transgene Expression Level
equivalent to human CD4+ cells
Genetic alteration of transgene:
A 4.5 kb BglII-EcoRI fragment encompassing a T cell-specific enhancer element located 13 kb upstream of the transcription initiation site in the murine CD4 gene was ligated to a 30 kb human CD4 minigene that includes the promoter and all of the human protein coding region. Line 2362 carries approximately six copies of this transgenic construct and expresses a level of CD4 equivalent to that seen on human peripheral CD4+ T cells. Transgenic mice do not express human CD4 on their peripheral CD8+ T cells, consistent with appropriate cis-acting transcriptional silencing of the human CD4 transgene from the T cell-specific enhancer element.
Genomic DNA encoding the TCR β-chain and cDNA encoding the TCR α-chain were prepared from HH8.2 hybridomas and cloned into the p3A9cβTCR and pES4 plasmids, respectively. Vector sequences were removed by restriction enzyme digestion and constructs were coinjected into fertilized hCD4.DR3-DQ2.MHCIIΔ/Δ oocytes. Oocytes were transferred into surrogate mothers (F1 of CBA × C57BL/6) and pups were screened by PCR with primers for the transgenes. Two TCR Tg founders were identified by PCR and confirmed by flow cytometry and two TCR Tg lines were established by backcrossing to hCD4.DR3-DQ2.MHCIIΔ/Δ Tg mice.
Expression of HLA DR and DQ evident on B cells, dendritic cells and a proportion of macrophages.
These mice Tg for hCD4.DR3-DQ2.MHCIIΔ/Δ express functional HLA-DR3 and DQ2 molecules that mediate thymic selection generating Ag-specific CD4+ T cell immunity. Although about one in five Europeans homozygous for HLA-DQ2 develops celiac disease, homozygous and heterozygous hCD4.DR3-DQ2.MHCIIΔ/Δ Tg mice do not spontaneously develop hypersensitivity to gluten. However, standard immunization protocols do produce HLA-DQ2-restricted CD4+ T cells in hCD4.DR3-DQ2.MHCIIΔ/Δ Tg mice that preferentially recognize the deamidated α-gliadin 17-mer immunodominant in human celiac disease. Nevertheless, we have found that preexisting DQ2-restricted CD4+ T cell immunity to deamidated α-gliadin 17-mer in hCD4.DR3-DQ2.MHCIIΔ/Δ Tg mice is not sufficient for gluten ingestion to cause measurable intestinal pathology.
TCR Tg T cells proliferated following systemic administration of Α-gliadin57–73 Q65E or chymotrypsin-digested gliadin in vivo and secreted Th1-type cytokines implicated in Celiac disease in vitro. But ingestion of gluten or Α-gliadin57–73 Q65E did not trigger intestinal pathology in TCR Tg mice, either spontaneously or induced by immunization strategies that have produced autoimmune disease in other TCR Tg models.
Findings indicate that gluten ingestion in humanized mice expressing functional HLA-DQ2 and possessing a substantial population of CD4+ T cells specific for gluten is not sufficient to cause celiac-like enteropathy.
Original Genetic Background
C57BL/6
Genetic Background Currently Maintained
C57BL/6
Strain identification
How is this strain characterised?
Genotyping
Phenotyping
Fertility and Strain maintenance
Fertility and Strain maintenance
Female
Male
Are homozygous mice viable?
Yes
Yes
Are homozygous mice fertile?
Yes
Yes
Are heterozygotes / hemizygotes* fertile?
Yes
Yes
* Hemizygote as used here refers to males carring a mutation on the X chromosome or mice of either sex
carrying an inserted transgene with no homologous allele on the other chromosome.
Generations Backcrossed
20
Mating Scheme
intercross
Homozygous Matings Required?
No
Development Summary
Cross of Tg(Cd4-CD4)2362Litt x Tg(HLA DR3-DQ2)303 x H2dlAb1-Ea all on C57BL/6 background. Crossed to B6-Ly5.1 mice to homozygosity at Ly5.1 and H2dlAb1-Ea.
Male Fertility Compromised?
No
Average Number of Pups at Birth
6.0
Average Number of Pups Surviving to Weaning
6.0
Breeding Performance
Good
Special Husbandry Requirements
May be raised of gluten-free diet to eliminate the impact of gluten on the development of gluten-specific T cells, although these T cells develop in the presence of dietary gluten
The potential for human MHC class II haplotypes to function efficiently in transgenic mice and should provide valuable tools for developing humanized models of MHC-associated autoimmune diseases.
Applicable Research Areas
Immunology and inflammation
Keywords
MHC
B cell
selection
autoimmunity
celiac disease
Is the strain available in any form to other researchers
