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Related Experiment Video

Updated: Jun 20, 2026

Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
06:13

Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens

Published on: November 23, 2010

ELISPOT and functional T cell analyses using HLA mono-specific target cells.

Claire Horlock1, Bryony Stott, Julian Dyson

  • 1Department of Immunology, Imperial College, London W12 0NN, United Kingdom.

Journal of Immunological Methods
|September 5, 2009
PubMed
Summary
This summary is machine-generated.

Developing HLA mono-specific cells simplifies T cell assays for cancer immunotherapy and infectious disease research. These cells provide highly specific T cell activity measurements with minimal background, improving functional analyses across all HLA allotypes.

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The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

Published on: December 14, 2016

Area of Science:

  • Immunology
  • Molecular Biology
  • Biotechnology

Background:

  • T cell assays are crucial for cancer immunotherapy, vaccine development, and infectious disease research.
  • Reliable T cell activity measurement is challenging due to HLA allele variations and APC characteristics.
  • Standard methods often struggle with non-HLA-A2 specificities and high background noise.

Purpose of the Study:

  • To develop a simplified and highly specific method for T cell functional analysis across diverse HLA allotypes.
  • To overcome limitations of existing T cell assays, particularly for non-HLA-A2 restricted T cells.
  • To establish a versatile platform for reliable T cell epitope identification and characterization.

Main Methods:

  • Generation of HLA mono-specific cells by coating T2 cells with recombinant HLA peptide complexes.
  • Utilizing ELISPOT, intracellular cytokine staining (ICS), and FACS-based killing assays.
  • Testing T cell responses against HLA-A2/peptide, HLA-A24/peptide, and HLA-DR15/peptide complexes.

Main Results:

  • HLA mono-specific cells produced minimal background immune activation, comparable to standard methods for HLA-A2/peptide specific T cells.
  • Successful T cell assays were demonstrated for non-HLA-A2 specificities, including HLA-A24 and HLA-DR15.
  • ELISPOT, ICS, and killing assays consistently showed high specificity and low background activity.

Conclusions:

  • HLA mono-specific cells offer a simple, reliable, and versatile system for improved T cell functional analyses.
  • This approach enhances T cell assays across all HLA allotypes, advancing immunotherapy and research.
  • The method provides a significant improvement for generating dependable functional T cell data.