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Generating De Novo Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain
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Immunology and Infection
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JoVE Journal Immunology and Infection
Generating De Novo Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain

Generating De Novo Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain

DOI:
10.3791/54697-v

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08:48 min

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October 25, 2016

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, , , , , , , , ,
1Department of Immunology,University of Toronto, 2Princess Margaret Cancer Centre,University Health Network

Chapters

  • 00:05Title
  • 00:35T Cell Receptor (TCR) Hemichain-encoded Retroviral Construct Preparation
  • 01:41Stable Packaging Cell Line Generation
  • 03:08Human T Cell Activation and Transduction
  • 05:04Novel Antigen-specific TCR Clone Reconstitution
  • 06:25Results: Centric TCR Hemichains Pair with Different Counter-chains to Form Antigen-specific TCRs with Varying Affinity
  • 07:59Conclusion

Summary

Automatic Translation

Automatic Translation

Herein we describe a novel method to generate antigen-specific T cell receptors (TCRs) by pairing the TCRα or TCRβ of an existing TCR, possessing the antigen-specificity of interest, with complementary hemichain of the peripheral T cell receptor repertoire. The de novo generated TCRs retain antigen-specificity with varying affinity.

Tags

De Novo T Cell ReceptorTCR Hemi-chainRetroviral TransductionAntigen-specific T CellsGene EngineeringImmunotherapyPCR CloningBacterial TransformationPlasmid Purification293GPG Cell TransfectionPG-13 Packaging Cell LinePBMC Activation

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