NOD.Cg-(D9rs4135590-D9rs13480186)H2^k2Tg(ILK3mHEL)3Ccg Tg(TcrHEL3A9)1Mmd/SlsgJ

Stock number: 026243
Strain synonyms: 3A9 TCR: iHEL NOD.H2^k-Chr9S
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research

Description

3A9 TCR:iHEL NOD.H2^k2-Chr9S mice may be useful when identifying new candidate genes in the Idd2 locus for the development of therapeutic avenues aimed at modulating DN T cell number for the prevention of autoimmune diseases.

Detailed description

3A9 TCR:iHEL NOD.H2^k2-Chr9S transgenic mice contain the TcrHEL3A9 transgene, encoding the alpha and beta chains of a T cell receptor (TCR) specific for hen egg lysozyme (HEL). In addition, they also carry the iHEL transgene expressing HEL under the direction of the rat insulin promoter. Together, these two transgenes promote insulitis.

The proportion of 3A9 CD4^-CD8^- double-negative(DN) T cells has been found to be significantly higher in the lymph nodes of TcrHEL3A9 Tg mice on a B10.BR background compared to the NOD.H2^k2 background. It has also been observed that 3A9 TCR:iHEL NOD.H2^k2 mice are highly susceptible to autoimmune diabetes, whereas the 3A9 TCR:iHEL B10.BR mice are resistant. To identify modifiers influencing the proportion of 3A9 DN T cells observed, genome-wide linkage analysis was performed using 3A9 TCR F2 (B10.BR x NOD.H2^k2) mice. A locus was identified exhibiting the highest logarithm of odds (LOD) score on chromosome 9 which coincides with the Idd2 (insulin dependent diabetes susceptibility 2) locus. NOD.H2^k2 congenic mice were generated by backcrossing for 6 generations. At the sixth generation, a recombination event occurred between the D9Mit129 and D9Mit328 markers. One of the mice presented with a B10.BR allele only at distal D9Mit129 marker. This mouse, with the shorter B10.BR segment, was the founder of this 3A9 TCR:iHEL NOD.H2^k2-Chr9S congenic strain. The other mice, bearing B10.BR alleles at both D9Mit129 and D9Mit328 markers, were founders for the 3A9 TCR:iHEL NOD.H2^k2-Chr9L congenic strain (see Stock No. 026624).

3A9 TCR:iHEL NOD.H2^k2-Chr9S mice exhibit a 2 fold increase of 3A9 DN T cell number in both the spleen and the lymph nodes relative to that of 3A9 TCR:iHEL NOD.H2^k2 mice. 3A9 DN T cells make up 2% in spleen and 6% in lymph nodes of 3A9 TCR:iHEL NOD.H2^k2-Chr9S mice, only partially restoring 3A9 DN T cells relative to 3A9 TCR B10.BR mice, which present approximately 9% 3A9 DN T cells in spleen and 30% in lymph nodes. However, 3A9 TCR:iHEL NOD.H2^k2-Chr9S exhibit a lower proportion of 3A9 DN T cells relative to the 3A9 TCR:iHEL NOD.H2k-Chr9L congenic mice (Stock No. 026624). The presence of a self-ligand in this transgenic mouse model thus revealed that at least one gene within the Chr9S interval defines the proportion of 3A9 DN T cells in the lymph nodes.

Development

NOD.H2^k2 mice were outcrossed to B10.BR and backcrossed to NOD.H2^k2 mice carrying the TcrHEL3A9 transgene, encoding the alpha and beta chains of a T cell receptor (TCR) specific for hen egg lysozyme (HEL) (Stock No. 002597) for six generations. TcrHEL3A9 mice were maintained on a NOD.H2^k2 background in which the H2^k2 allele was derived from the B10.BR mice (Stock No. 000465).

At the fifth generation mice were bred to iHEL transgenic mice expressing HEL under the control of the rat insulin promoter. This transgenic line was developed in C57BL/6J oocytes and was backcrossed onto the NOD.H2^k2 background for at least 18 generations.

At the sixth generation, a recombination event occurred between the D9Mit129 and D9Mit328 markers, where one of the mice presented with a B10.BR allele only at D9Mit129 marker. This mouse, with the shorter B10.BR segment, was the founder of this 3A9 TCR:iHEL NOD.H2^k2-Chr9S congenic strain. The other mice, bearing B10.BR alleles at both D9Mit129 and D9Mit328 markers, were founders for the 3A9 TCR:iHEL NOD.H2^k2-Chr9L congenic strain (see Stock No. 026624).

Allele

Symbol: Tg(TcrHEL3A9)1Mmd
Name: transgene insertion 1, Mark M Davis
MGI accession ID: MGI:2385683
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(TcrHEL3A9)1Mmd
Marker name: transgene insertion 1, Mark M Davis
Chromosome: UN
Strain of origin: C57BL/6J
Expressed genes: Tcra,T cell receptor alpha chain,mouse, laboratory; Tcrb,T cell receptor beta chain,mouse, laboratory
Molecular note: Transgenic mice produced from this construct carry transgenes encoding the T cell receptor alpha and beta (Tcra and Tcrb, respectively) chains from the hen egg lysozyme (HEL)-specific T cell hybridoma 3A9. Two vectors were constructed and subsequently co-injected into C57BL/6J eggs. The first vector contained the Tcra variable and joining regions (V and J, respectively) and a 3.5kb segment 3' to the Tcra constant region. The second vector contained the Tcrb variable, diversity, and joining regions (V, D, and J, respectively) and a 7.5kb segment 3' to the Tcrb constant region, which included the Tcrb enhancer region. Transgenic mice expressed the alpha and beta chains of a T cell receptor specific for hen egg lysozyme. Two female founders were generated.

Allele

Symbol: Tg(ILK3mHEL)3Ccg
Name: transgene insertion 3, Christopher C Goodnow
MGI accession ID: MGI:2385454
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(ILK3mHEL)3Ccg
Marker name: transgene insertion 3, Christopher C Goodnow
Chromosome: UN
Strain of origin: C57BL/6JSfd
Expressed genes: HEL,HEL,chicken; H2-K,histocompatibility 2, K region,mouse, laboratory
Molecular note: A genomic fragment containing the coding exons of the chicken lysozyme gene (also known as hen egg lysozyme, HEL) was linked to a 600bp fragment of the rat insulin promoter and to the mouse H2-K^b transmembrane sequence (which included the extracellularspacer sequence, transmembrane segment, and cytoplasmic tail). The rat insulin promoter was used to direct expression of the inserted sequence specifically to pancreatic islet beta cells. Islets from mice carrying the transgene showed expression of chicken lysozyme by Western blot; spleen, thyroid, and thymus did not.

Allele

Symbol: H2^k2
Name: k2 variant
MGI accession ID: MGI:4936842
Generation method: Not Applicable
Marker symbol: H2
Marker name: histocompatibility-2, MHC
Chromosome: 17
Strain of origin: Not Specified
Molecular note: The H2^k2 haplotype differs from the prototype H2^k in having the a, rather than the canonical b, alleles of the Class Ib genes T18 (formerly Tla) and Qa-1. The H2^k2 haplotype is shared by the inbred strains C57BR/cdJ and C58/J and the H2 congenic strain B10.BR-H2^k2 H2-T18^a/SgSnJ (formerly B10.BR-H2^k/SgSnJ).

Allele

MGI accession ID: MGI:96404
Marker symbol: Idd2
Marker name: insulin dependent diabetes susceptibility 2
Chromosome: 9