STOCK Il10^tm1Flv H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ

Stock number: 037372
Product name: Tg4 Tiger; Tg4 IL10/GFP
Strain synonyms: Tg4 Tiger
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Tg4 Tiger mice express an Il10 allele with a GFP reporter (Tiger) and rearranged Vα 4/Vβ 8.2 T cell receptor chains generated from T cell hybridoma cDNA raised against the acetylated N-terminal peptide of murine myelin basic protein (MBP), Ac1-9, and directed by human CD2 regulatory elements on the H2-A^u background. Tg4 Tiger mice are highly susceptible to induction of experimental autoimmune encephalomyelitis (EAE). This strain may be useful in studies of IL10 and antigen specific immunotherapy in autoimmune disease including CNS inflammation and EAE/multiple sclerosis.  

Detailed description

The Tg4 Tiger transgenic strain carries a rearranged Vα 4/Vβ 8.2 T cell receptor transgene specific for the H-2^u-restricted acetylated N-terminal peptide of murine myelin basic protein (MBP), Ac1-9 ((Ac-ASQKRPSQR). The Ac1-9 epitope has a low affinity for the class II MHC H2-A^u restriction element, potentially allowing autoreactive T cells to escape self-tolerance. The Tg4 thymic T cell repertoire is composed predominantly of CD4+ cells expressing Vβ 8.2. In addition, this strain carries the IL-10 IRES eGFP (Il10^tm1Flv) KI reporter allele. Fluorescence of GFP correlates positively with IL-10 cytokine protein expression. The donating investigator indicates that spontaneous experimental autoimmune encephalomyelitis (EAE) occurs at a frequency of 5.2% in this strain. Mice homozygous for the transgene and homozygous for the Il10 reporter (Tiger) are viable and fertile.

Tolerance induction against the MBP peptide using dose escalation in the Tg4 strain results in anergic IL10-secreting regulatory CD4+ T cells that function to suppress autoimmunity.  Although peptide-treated Tg4 Tiger CD4+ T cells are anergic, they secrete interfeuron gamma (but not IL-2) at equivalent or higher levels than untreated Tg4 mice. This strain may be useful in studies of IL10 and antigen specific immunotherapy in autoimmune disease including CNS inflammation and EAE/multiple sclerosis.  

The Jackson Laboratory distributes the following strains with the Tg4 allele:
Stock No. 037369 B10.Cg-H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ
Stock No. 037370 B10.Cg-Rag1^tm1Bal H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ
Stock No. 037371 B10.Cg-Ptprc^a Pepc^b H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ
Stock No. 037372 B10.Cg-Il10^tm1Flv H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ

Development

Tg4 mice were crossed to B6.129S6-Il10^tm1Flv/J mice (Stock No. 008379) by Dr. David Wraith at the University of Birmingham.

The Tg4 transgenic construct contains rearranged Vα 4/Vβ 8.2 T cell receptor chain leader and coding sequences under the control of human CD2 regulatory elements. RNA splice sites and a polyA signal were introduced by ligating a genomic segment of the human beta-globin gene 3’ to the TCR sequence. T cell receptors were isolated from 1934.4, and H2-A^u restricted myelin basic protein Ac1-9-specific T cell hybridoma derived from an encephalitogenic T cell clone. The linearized construct was microinjected into fertilized (CBA x C57BL/10)F2 oocytes. Founder Tg4 mice were bred to B10.PL-H2^u H2-T18^a/(73NS)SnJ (Stock No. 000458).

The tiger or IL-10 IRES eGFP targeting vector contains an IRES-GFP (internal ribosome entry site-green fluorescent protein) cassette followed by a loxP-flanked neomycin resistance cassette inserted between the stop codon and polyadenylation signal of exon 5 of the Il10 gene.  A floxed neo was removed via cre mediated recombination. Mice were bred to the F2 generation and then backcrossed on B10.Cg-H2^u H2-T18^a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/Wdc for 10 generations by the donating laboratory (see SNP note below). Upon arrival at The Jackson Laboratory, embryos were cryopreserved.

In 2022, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the males sent to The Jackson Laboratory Repository. Three of the 43 markers throughout the genome were segregating with CBA or 129, suggesting an incomplete backcross. This strain is also still segregating at two markers on Chromosome 17, consistent with results seen in the parental strain (Stock No. 000458) which shows segregation with PL/J on Chromosome 17, likely a passenger segment from H2 mutations.

Allele

Symbol: Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio
Name: transgene insertion 4, Dimitris Kioussis
MGI accession ID: MGI:4354587
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio
Marker name: transgene insertion 4, Dimitris Kioussis
Chromosome: UN
Strain of origin: (CBA x C57BL/10)F2
Molecular note: The TCR alpha and beta transgenic constructs derived from a Ac1-9-specific T cell hybridoma were generated by inserting cDNAs containing the leader and coding sequences into an expression cassette containing the human CD2 promoter. RNA splice sites and a poly(A) signal sequence were introduced by ligating a genomic segment of the human beta-globin gene 3' to the TCR sequences. Transgenic mice express the Vbeta8.2 TCR specific for myelin basic protein (MBP) epitope Ac1-9 in the context of I-A^u in T cells. In addition to line 4, line Tg3 was also created.

Allele

Symbol: H2^u
Name: u variant
MGI accession ID: MGI:3834991
Generation method: Not Applicable
Marker symbol: H2
Marker name: histocompatibility-2, MHC
Chromosome: 17
Strain of origin: Not Applicable
General note: The q variant has been observed in the PL/J strain.

Allele

Symbol: H2-T18^a
Name: a variant
MGI accession ID: MGI:4820507
Generation method: Not Applicable
Marker symbol: H2-T18
Marker name: histocompatibility 2, T region locus 18
Chromosome: 17
Strain of origin: Not Applicable
Molecular note: The allele H2-T18^a determines presence of the antigenic specificities TL.1, 2, 3, 5, 6, 7 and occurs in strains A, C57BR, C58, SJL, and others.

Allele

Symbol: Il10^tm1Flv
Name: targeted mutation 1, Richard A Flavell
MGI accession ID: MGI:3695482
Generation method: Targeted
Attributes: Reporter; No functional change
Marker symbol: Il10
Marker name: interleukin 10
Chromosome: 1
Strain of origin: 129S6/SvEvTac
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: GFP expression recapitulates normal Il10 expression patterns (including activated macrophages and dendritic cells, and some lymphocytes) but is preferentially observed in some populations of the intestinal tissue.
Molecular note: A targeting vector containing IRES-GFP was inserted into the locus between the stop codon and the polyA signal in exon 5. A floxed neo was removed via cre mediated recombination.