EXPERIMENTAL MODELS
Contributor Information
- Name Thorbald Van Hall
- Institute Leiden University and Leiden University Medical Center; Stichting Oncode Institute
Tool Details
- Tool name: LnB5 TCR Tg mice
- Tool type: Experimental models
- Tool sub-type: Mouse
- Model: Transgenic
- Conditional: No
- Conditional description: The expression of the TCR is driven by the CD2 promoter and not conditional
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Genetic background and cross history: The LnB5 TCR transgenic mouse strain was generated by transgenesis of the TCRa and TCR
genes of the LnB5 T cell clone, specific for the TRH4/Dbcomplex. Cloning and sequencing of the TCR chains identified the a chain as composed by TRAV9N-3*01 and TRAJ15*01 and the chain composed by V13-2*01, D1*01, and J1-4*02. TheTcraandTcrbchains were separately cloned into pCRII-TOPO plasmid vectors using reverse transcription PCR (RT-PCR). Validation of correct TCR cloning was performed by retroviral transduction of the TCR in C57BL/6 splenocytes using pMX vectors. Next, the 2 chains were separately cloned into VA-hCD2 vectors, the inserts of which were subsequently injected in C57BL/6 oocytes to produce transgenic mice. (PMID:26784543) - Zygosity: Unknown
- Strain: C57BL/6
- Description: Tumor cells frequently escape from CD8+ T cell recognition by abrogating MHC-I antigen presentation. Deficiency in processing components, like the transporter associated with antigen processing (TAP), results in strongly decreased surface display of peptide/MHC-I complexes. A class of hidden self-antigens known as T cell epitopes associated with impaired peptide processing (TEIPP), which emerge on tumor cells with such processing defects can be investigated with this model. Using this mouse model, it is possible to investigate the generation of the TEIPP T cell repertoire specifically. These animals harbour rearranged receptors recognizing the TRH4/Db complex (named LnB5). Efficient selection of this TCR has been observed in the thymus, a strong CD8 skewing of the T cells, and high quantities of naive phenotype in the periphery. T cells have been reported to be readily activated by peptide vaccination and have showed to kill target cells efficiently. Author publication: PMID: 26784543
- Research area: Cancer
- Production details: The LnB5 TCR transgenic mouse strain was generated by transgenesis of the TCRι and TCRβ genes of the LnB5 T cell clone, specific for the TRH4/Db complex. Cloning and sequencing of the TCR chains identified the ι chain as composed by TRAV9N-3*01 and TRAJ15*01 and the β chain composed by V13-2*01, D1*01, and J1-4*02. The Tcra and Tcrb chains were separately cloned into pCRII-TOPO plasmid vectors using reverse transcription PCR (RT-PCR). Validation of correct TCR cloning was performed by retroviral transduction of the TCR in C57BL/6 splenocytes using pMX vectors. Next, the 2 chains were separately cloned into VA-hCD2 vectors, the inserts of which were subsequently injected in C57BL/6 oocytes to produce transgenic mice. (PMID:26784543)
- Breeding information: These mice breed normally in heterozygous or homozygous background
- For Research Use Only
Target Details
- Target: LnB5 TCR
Application Details
Handling
- Shipping conditions: Embryo/Spermatoza- Dry Ice
Documentation
References
- • Doorduijn et al. 2016. J Clin Invest. 126(2):784-94. PMID: 26784543.
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