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Related Concept Videos

T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

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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...
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In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues
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Identification and Characterization of Antigen-Specific CD8+ T Cells Using Surface-Trapped TNF-α and Single-Cell

Shaheed Abdulhaqq1, Abigail B Ventura1, Jason S Reed1

  • 1Vaccine and Gene Therapy Institute, Oregon Health and Science University, Beaverton, OR.

Journal of Immunology (Baltimore, Md. : 1950)
|November 23, 2021
PubMed
Summary

This study introduces a novel system for rapidly identifying and characterizing antigen-specific CD8+ T cells using ex vivo stimulation and TNF-α sorting. This method enables efficient T cell receptor sequencing and characterization for disease research and therapeutic development.

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Area of Science:

  • Immunology
  • Cellular immunology
  • T cell biology

Background:

  • CD8+ T cells are crucial for antiviral and antitumor immunity.
  • Studying antigen-specific CD8+ T cells and their MHC restriction is vital for disease research and therapeutic development.
  • Current methods for isolating and characterizing these cells are often cumbersome and limited by sample availability.

Purpose of the Study:

  • To develop a novel, rapid system for the identification and characterization of antigen-specific CD8+ T cells.
  • To enable efficient T cell receptor (TCR) sequencing and functional analysis, especially from limited primary cell samples.
  • To create a versatile platform for epitope mapping and MHC restriction studies.

Main Methods:

  • Ex vivo stimulation of cells with specific antigens.
  • Live cell sorting based on surface-trapped Tumor Necrosis Factor-alpha (TNF-α).
  • Single-cell sequencing for paired TCR alpha and beta chain data, followed by TCR transduction into donor T cells.

Main Results:

  • Successful characterization of CMV-specific CD8+ T cells and an immunodominant Mamu-A1*002:01-restricted epitope in rhesus macaques.
  • Mapping of a novel HLA-A*68:02-restricted HIV Gag epitope from an HIV-infected donor.
  • Demonstration of the system's utility in generating unlimited experimental reagents from identified TCRs.

Conclusions:

  • The developed system offers a rapid and efficient strategy for identifying novel antigens and characterizing antigen-specific CD8+ T cells.
  • This approach has broad applications in infectious disease research, immunotherapeutics, and precision medicine.
  • The method overcomes limitations of primary cell availability and experimental complexity in T cell studies.