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Related Concept Videos

T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...
Diversity of Antigen Receptors01:28

Diversity of Antigen Receptors

Antigen receptors are essential components of the immune system crucial in defending the body against foreign invaders. These receptors are present on the surface of B and T cells, enabling them to recognize antigens and mount an appropriate immune response.
Before encountering any antigen, lymphocytes express these receptors. On B cells, the antigen receptor is a membrane-bound antibody molecule called BCR; on T cells, it is a T cell receptor or TCR. B and T cell receptors are composed of two...

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Related Experiment Video

Updated: Jun 23, 2026

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
11:21

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Published on: September 10, 2017

A fast and robust method to clone and functionally validate T-cell receptors.

Katrin Birkholz1, Christian Hofmann, Stefanie Hoyer

  • 1Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.

Journal of Immunological Methods
|May 12, 2009
PubMed
Summary
This summary is machine-generated.

Researchers developed a universal PCR method to clone T-cell receptor (TCR) chains and a fast, low-cost assay for functional testing. This accelerates immunotherapy and immunological research by enabling rapid TCR identification and validation.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biotechnology

Background:

  • T-cell receptor (TCR) sequencing and functional testing are crucial for immunotherapy and immunological research.
  • Current methods for TCR alpha- and beta-chain determination are limited by the lack of conserved 5' sequences, and functional testing is often laborious and time-consuming.

Purpose of the Study:

  • To develop a universal PCR-based protocol for amplifying TCR alpha- and beta-chain mRNAs without prior knowledge of variable domain subtypes.
  • To establish a rapid, easy, and low-cost method for functional testing of cloned TCRs.

Main Methods:

  • Adaptation of capswitch technology for a universal PCR protocol to amplify TCR alpha- and beta-chain mRNAs.
  • Development of a novel functional assay involving electroporation of Jurkat T cells with TCR-encoding RNA and an NFAT-reporter construct, followed by stimulation and measurement of cell activation.

Main Results:

  • Successful cloning of MelanA/HLA-A2-specific and HIVgag/HLA-A2-specific TCRs using the universal PCR protocol.
  • The novel functional assay results correlated well with cytokine release, functional avidity, proliferative activity, and tumor cell recognition by T cells.

Conclusions:

  • The developed protocols offer a rapid and cost-effective tool for identifying and functionally testing T-cell receptors.
  • These methods have significant potential for application in immunotherapy and broader immunological research.