Overview

This transgenic strain carries a rearranged T cell receptor transgene specific for the mouse homologue (pmel^Si or pmel-17) of human premelanosome protein (referred to as PMEL, SILV or gp100), and the T lymphocyte specific Thy1^a (Thy1.1) allele. About 1/3 of transgenic mice develop lymphomas by 5 to 7 months of age. These transgenic mice provide a tumor model system for studies related to immunotherapy, and for studying in vivo T-cell biology

Donating Investigator

Nicholas P. Restifo, National Cancer Institute

Genetic overview

Genetic Background	Generation
	N?+N3F3 (2019-06-12 00:00:00)

Tg(TcraTcrb)8Rest

Allele Type
Transgenic (Inserted expressed sequence)

Thy1^a

Allele Type	Gene Symbol	Gene Name
Not Applicable	Thy1	thymus cell antigen 1, theta

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Research Applications

Immunology, Inflammation and Autoimmunity Research
Research Tools

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Base Price

Starting at:

$255.00 Domestic price for female 4-week

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Details

Detailed Description

This transgenic strain carries a rearranged T cell receptor transgene specific for the mouse homologue (pmel-17) of human SILV (gp100), an enzyme involved in pigment synthesis that is expressed by the majority of malignant melanoma cells including B16 melanoma, as well as by normal melanocytes. In mice from founder line 8 the transgenic integration was determined to be on Chromosome 2, in the 3' UTR of the serine threonine kinase 39 (Stk39) gene. STK39 is involved in the cellular stress response pathway. The strain is also homozygous for the T lymphocyte specific Thy1^a (Thy1.1) allele. CD8+ T cells express a Tcra-V1/Tcrb-V13- transgenic TCR that recognizes an epitope of pmel-17 corresponding to amino acids 25-33 of gp100 presented by H2-D^b MHC class I molecules. Greater than 95% of the CD8+ T cells in transgenic mice expressed the transgenic TCR based on the expression of Vbeta13, amounting to about 20% of all splenocytes. T cells in blood and spleen generally expressed baseline levels of the activation/effector markers CD25, CD44, and CD69, indicating that most of the transgenic cells were in the naive state.

In The Jackson Laboratory Repository colony, approximately 30% of transgenic mice develop lymphomas, found predominantly in the thymus and lymph nodes, at 5 to 7 months of age. Low incidence of earlier onset (by ~4 months) may be observed.

These transgenic mice in conjunction with the poorly immunogenic B16 melanoma, a highly aggressive tumor in C57BL/6 mice, provide a physiologically relevant tumor model system for studies related to immunotherapy. In addition, these transgenic mice are useful for studying in vivo T-cell biology such as TCR-ligand interactions, T-cell activation, thymic selection, cross-presentation of antigens, process of thymic selection and central and peripheral T-cell tolerance and induction.

Development

Transgenic constructs containing rearranged alpha-chain and beta-chain of the T-cell receptor specific for the mouse homologue (pmel-17) of human SILV (gp100) were co-injected into fertilized C57BL/6 mouse eggs. Transgenic integration was on Chromosome 2, in the 3' UTR of the serine threonine kinase 39 (Stk39) gene. Mice from founder line 8 were bred to C57BL/6 mice, and then crossed with B6.PL-Thy1^a/CyJ (Stock No. 000406) mice.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between C57BL substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggest a C57BL/6 genetic background, 3 of 5 markers that determine different C57BL substrains were found to be segregating. This may be due to contributions from the PL/J regions in B6.PL-Thy1^a/CyJ.

Expression Data

Expressed Gene	Tcrb, T cell receptor beta chain, mouse, laboratory
Expressed Gene	Tcra, T cell receptor alpha chain, mouse, laboratory
Site of Expression
Site of Expression	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells.

Control Suggestions

000406 B6.PL-Thy1^a/CyJ

Additional Information

Considerations for Choosing Controls

Selected References

Ji Y; Abrams N; Zhu W; Salinas E; Yu Z; Palmer DC; Jailwala P; Franco Z; Roychoudhuri R; Stahlberg E; Gattinoni L; Restifo NP. 2014. Identification of the Genomic Insertion Site of Pmel-1 TCR alpha and beta Transgenes by Next-Generation Sequencing. PLoS One 9(5):e96650PubMed: 24827921 MGI: J:208902
Overwijk WW; Theoret MR; Finkelstein SE; Surman DR; de Jong LA; Vyth-Dreese FA; Dellemijn TA; Antony PA; Spiess PJ; Palmer DC; Heimann DM; Klebanoff CA; Yu Z; Hwang LN; Feigenbaum L; Kruisbeek AM; Rosenberg SA; Restifo NP. 2003. Tumor regression and autoimmunity after reversal of a functionally tolerant state of self-reactive CD8+ T cells. J Exp Med 198(4):569-80PubMed: 12925674 MGI: J:85058

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Genetics

Tg(TcraTcrb)8Rest

Allele Symbol: Tg(TcraTcrb)8Rest

Allele Name	transgene insertion 8, Nicholas Restifo
Allele Type	Transgenic (Inserted expressed sequence)
Allele Synonym(s)	Pmel-1; pmel-17; Pmel-I
Gene Symbol and Name	Tg(TcraTcrb)8Rest, transgene insertion 8, Nicholas Restifo
Gene Synonym(s)
Promoter	Tcrb, T cell receptor beta chain, mouse, laboratory
Promoter	Tcra, T cell receptor alpha chain, mouse, laboratory
Expressed Gene	Tcrb, T cell receptor beta chain, mouse, laboratory
Expressed Gene	Tcra, T cell receptor alpha chain, mouse, laboratory
Strain of Origin	C57BL/6
Chromosome	2
Molecular Note	These coinjected transgenes encode an H2-D^b-restricted Valpha1 Vbeta13 T-cell receptor that recognizes a peptide comprising amino acids 25-33 of the protein known as pmel-17/gp100, a pigment biosynthetic enzyme that is encoded by the mouse Si (silver) gene and is expressed both in normal melanocytes and in malignant melanoma. The insertion is accompanied by a 157-bp deletion (chr2: 68,208,872?68,209,029 bp - build unknown).
Mutations Made By	Marc Theoret, NIH

Thy1^a

Allele Symbol: Thy1^a

Allele Name	a variant
Allele Type	Not Applicable
Allele Synonym(s)	thetaAKR; theta-AKR; Thy1.1; Thy-1.1; Thy1a
Gene Symbol and Name	Thy1, thymus cell antigen 1, theta
Gene Synonym(s)
Site of Expression	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells.
Strain of Origin	Not Applicable
Chromosome	9
General Note	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells. The allele Thy1^a determines an antigenic specificity, Thy-1.1, found in the AKR and RF strains; the allele Thy1^b determines an antigenic specificity, Thy-1.2, found in the C3HeB/Fe and many other strains (J:5243, J:5012, J:4469). The Thy1 antigen is probably present on all T lymphocytes and absent from all B lymphocytes, and it thus serves as a valuable T-cell marker (J:5243). It is very widely used in experiments designed to determine the distribution and function of T-cells. Thy1 specifies a T-cell surface glycoprotein, T25, with a molecular weight of 25 kDa (J:5707). The protein appears to be anchored in the cell membrane by a lipid that is either phosphotidylinositol or closely related to it (J:12016). Thy1 may function in the cell membrane as a signal transduction molecule (J:8333). The Thy1 locus, or possibly a gene closely linked to it, controls quantitative expression of a protein that is the same size as Thy1 and is expressed on thymus and brain but not on lymph node and spleen cells (J:7900).
Molecular Note	The allele Thy1^a determines an antigenic specificity, Thy-1.1, found in the AKR and RF strains.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Genotype: Thy1^a related

Immunology, Inflammation and Autoimmunity Research
- CD Antigens, Antigen Receptors, and Histocompatibility Markers
Research Tools
- Immunology, Inflammation and Autoimmunity Research
  - T cell specific surface marker
- Genetics Research
  - Tissue/Cell Markers: T cell specific surface marker
  - Tissue/Cell Markers

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Tg(TcraTcrb)8Rest/?
C57BL/6-Tg(TcraTcrb)8Rest

hematopoietic system phenotype

abnormal CD8-positive, alpha-beta cytotoxic T cell morphology
- over 95 % of the CD8 T cells present in circulation express the transgenic TCR that is specific for the H2-D^b restricted epitope of Silv (gp100), a protein that is highly expressed in melanoma cells
- the baseline levels of activation markers for the transgenic T cells indicate that most of cells are in a na¿ve state
- these transgenic T cells constitute about 20% of spleen cells

abnormal cytotoxic T cell physiology
- upon in vitro stimulation with the immunogenic epitope of gp100, CD8 T cells are activated and proliferate extensively

neoplasm

altered tumor susceptibility
- adoptive transfer of transgenic splenocytes into normal mice bearing melanoma tumors also has no effect on tumor growth
- despite the presence of large number of anti-gp100 CD8 T cells, transplanted melanoma tumors that express gp100 have the same growth kinetics as tumors transplanted into control mice
- mice bearing melanoma tumors that receive vaccine, transgenic T cells, and IL-2 have survival times of over 1 year compared to controls that die within 2 months
- there is a modest delay in tumor growth when mice that are recipients of transgenic T cells are also vaccinated with peptides that mimic the gp100 epitope
- when IL-2 is administered with the vaccine, there is a large regression in tumor size in mice that are also recipients of transgenic T cells

immune system phenotype

abnormal CD8-positive, alpha-beta cytotoxic T cell morphology
- over 95 % of the CD8 T cells present in circulation express the transgenic TCR that is specific for the H2-D^b restricted epitope of Silv (gp100), a protein that is highly expressed in melanoma cells
- the baseline levels of activation markers for the transgenic T cells indicate that most of cells are in a na¿ve state
- these transgenic T cells constitute about 20% of spleen cells

abnormal cytotoxic T cell physiology
- upon in vitro stimulation with the immunogenic epitope of gp100, CD8 T cells are activated and proliferate extensively

increased interferon-gamma secretion
- CD 8 T cells produce large amounts of IFN-gamma when cultured in the presence of melanoma cells
- large number of IFN-gamma producing CD8 T cells are found in melanoma tumors that have regressed due to treatment with gp100 vaccine, IL-2, and adoptive transfer of transgenic CD8 T cells

References

Ji Y; Abrams N; Zhu W; Salinas E; Yu Z; Palmer DC; Jailwala P; Franco Z; Roychoudhuri R; Stahlberg E; Gattinoni L; Restifo NP. 2014. Identification of the Genomic Insertion Site of Pmel-1 TCR alpha and beta Transgenes by Next-Generation Sequencing. PLoS One 9(5):e96650PubMed: 24827921 MGI: J:208902
Overwijk WW; Theoret MR; Finkelstein SE; Surman DR; de Jong LA; Vyth-Dreese FA; Dellemijn TA; Antony PA; Spiess PJ; Palmer DC; Heimann DM; Klebanoff CA; Yu Z; Hwang LN; Feigenbaum L; Kruisbeek AM; Rosenberg SA; Restifo NP. 2003. Tumor regression and autoimmunity after reversal of a functionally tolerant state of self-reactive CD8+ T cells. J Exp Med 198(4):569-80PubMed: 12925674 MGI: J:85058

Additional References

Antony PA; Paulos CM; Ahmadzadeh M; Akpinarli A; Palmer DC; Sato N; Kaiser A; Hinrichs CS; Klebanoff CA; Tagaya Y; Restifo NP. 2006. Interleukin-2-dependent mechanisms of tolerance and immunity in vivo. J Immunol 176(9):5255-66PubMed: 16621991 MGI: J:115150
Klebanoff CA; Finkelstein SE; Surman DR; Lichtman MK; Gattinoni L; Theoret MR; Grewal N; Spiess PJ; Antony PA; Palmer DC; Tagaya Y; Rosenberg SA; Waldmann TA; Restifo NP. 2004. IL-15 enhances the in vivo antitumor activity of tumor-reactive CD8+ T cells. Proc Natl Acad Sci U S A 101(7):1969-74PubMed: 14762166 MGI: J:90388

Additional - Thy1^a related

Additional - Tg(TcraTcrb)8Rest related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- End Point Analysis:Thy1 End Point
- QPCR:Tg(TcraTcrb)8Rest
- Standard PCR:Tg(TcraTcrb)8Rest
- Genotyping resources and troubleshooting
Dietary Information
- LabDiet® 5K52 formulation (6% fat)
Breeding Considerations

Transgenic constructs containing the mouse alpha-chain and beta-chain of a T-cell receptor were used to create transgenic animals on a C57BL/6 background. The mice were then crossed to B6.PL-Thy1^a/CyJ (Stock No. 000406). The strain is maintained homozygous for the transgenic insert and homozygous for the Thy1^a allele.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the B6.Cg-Thy1^a/Cy Tg(TcraTcrb)8Rest/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #005023 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX11 (Maximum)

Pricing & Availability

Available Now

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Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Hemizygous or Non carrier for Tg(TcraTcrb)8Rest

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Tg(TcraTcrb)8Rest

Allele Symbol: Tg(TcraTcrb)8Rest

Thy1a

Allele Symbol: Thy1a

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Genotype: Thy1a related

Mammalian Phenotype Terms by Genotype

Genotype: Tg(TcraTcrb)8Rest/?C57BL/6-Tg(TcraTcrb)8Rest

hematopoietic system phenotype

neoplasm

immune system phenotype

References

Additional References

Additional - Thy1a related

Additional - Tg(TcraTcrb)8Rest related

Genotyping Protocols

Dietary Information

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Thy1^a

Allele Symbol: Thy1^a

Genotype: Thy1^a related

Genotype: Tg(TcraTcrb)8Rest/?
C57BL/6-Tg(TcraTcrb)8Rest

Additional - Thy1^a related