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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.
Insights
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.
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.
Abstract:
It is generally accepted that enumeration and characterization of antigen-specific T cells provide essential information about potency of the immune response. Here, we report a new technique to determine the frequency and potency of antigen-specific CD8 T cells. The assay measures changes of intracellular Ca2+ in real time by fluorescent microscopy in individual CD8 T cells responding to cognate peptides. The T cells form continuous monolayer, enabling the cells to present the peptides to each other. This approach allows us to evaluate the kinetics of intracellular Ca2+ signalling that characterizes the quality of T cell response. We demonstrate the usefulness of the assay examining the frequency and quality of cytomegalovirus-specific CD8 T cells from healthy donor and patient after haploidentical stem cell transplantation. The new assay has a potential to provide essential information determining the status of the immune system, disease morbidity, potency of therapeutic intervention and vaccine efficacy.
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