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Evaluating frequency and quality of pathogen-specific T cells.

Nadia Anikeeva1, Dolores Grosso2, Neal Flomenberg2,3

  • 1Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.

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|October 28, 2016
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Summary

We developed a new assay to measure antigen-specific CD8 T cells by tracking intracellular calcium (Ca2+) changes. This method assesses T cell potency and immune response quality, crucial for evaluating therapies and vaccines.

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Area of Science:

  • Immunology
  • Cellular Biology
  • Medical Diagnostics

Background:

  • Enumeration and characterization of antigen-specific T cells are vital for assessing immune response potency.
  • Existing methods may not fully capture the dynamic nature of T cell responses.
  • Understanding T cell function is critical for immune monitoring and therapeutic development.

Purpose of the Study:

  • To introduce a novel assay for determining the frequency and potency of antigen-specific CD8 T cells.
  • To evaluate the kinetics of intracellular calcium (Ca2+) signaling as a measure of T cell response quality.
  • To demonstrate the assay's utility in assessing immune responses post-transplantation.

Main Methods:

  • Utilizes fluorescent microscopy to monitor real-time intracellular Ca2+ changes in individual CD8 T cells.
  • Employs a unique monolayer cell formation allowing T cells to present peptides to each other.
  • Measures the kinetics of Ca2+ signaling to characterize T cell response quality.

Main Results:

  • Successfully determined the frequency and quality of cytomegalovirus-specific CD8 T cells.
  • Demonstrated the assay's effectiveness in samples from healthy donors and patients post-haploidentical stem cell transplantation.
  • The assay provides insights into T cell response kinetics linked to peptide recognition.

Conclusions:

  • The developed assay offers a novel approach to quantify and characterize antigen-specific CD8 T cells.
  • This technique has the potential to provide critical information on immune system status, disease, therapeutic efficacy, and vaccine effectiveness.
  • Real-time Ca2+ signaling kinetics serve as a valuable indicator of T cell response quality.