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

Stock number: 037369
Product name: Tg4
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Tg4 H-2^u mice express 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^u background. Tg4 mice are highly susceptible to induction of experimental autoimmune encephalomyelitis (EAE). This strain may be useful in studies of immunological tolerance to self-antigen, antigen-specific immunotherapy of autoimmune disease including CNS inflammation and EAE/multiple sclerosis.

Detailed description

The Tg4 transgenic strain carries a rearranged Vα 4/Vβ 8.2 T cell receptor transgene specific for the H2-A^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 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.

Experimental autoimmune encephalomyelitis (EAE) induced by priming of Ac1-9 in CFA results in 2 of 5 mice developing EAE. When combined with pertussis toxin, almost all mice develop EAE. The Tg4 TCR transgenic mouse has the advantage of being able to stimulate lymph node or spleen cells in vitro to elicit an immune response without prior immunization. Mice homozygous for the transgene are viable and fertile. This strain may be useful in studies of immunological tolerance to self-antigen, antigen-specific immunotherapy of 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

The 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, an 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 000458) mice 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. Two of the 43 markers throughout the genome, on chromosomes 5 and 12, were segregating with CBA, 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.