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Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
Published on: April 19, 2017
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Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry.
Stephen C Juvet1, Sajad Moshkelgosha2, Sharon Sanderson3
1Division of Respirology, Departments of Medicine and Immunology, Toronto Lung Transplant Program, Multiorgan Transplant Program, Toronto General Research Institute, University of Toronto and University Health Network; stephen.juvet@uhn.ca.
Journal of Visualized Experiments : Jove
|April 28, 2017
Summary
Measuring T-cell reactivity to donor antigens using imaging flow cytometry can predict transplant outcomes. This novel method shows promise for guiding immunosuppression reduction and assessing tolerance therapies.
Area of Science:
- Immunology
- Transplantation Biology
- Cellular Immunology
Background:
- Assessing immune responses to donor antigens is vital for managing immunosuppression in transplant recipients.
- Existing methods like mixed leukocyte reaction (MLR) and delayed-type hypersensitivity (DTH) assays have limitations in predictive power and practicality.
- Imaging flow cytometry offers a powerful combination of quantitative analysis and imaging for detailed cellular interaction studies.
Purpose of the Study:
- To evaluate the utility of imaging flow cytometry in quantifying T-cell interactions with antigen-presenting cells (APCs) for predicting transplant outcomes.
- To assess the frequency of immune synapse formation between recipient T cells and donor APCs as a biomarker for allograft rejection and tolerance.
- To investigate the impact of regulatory T cells on T-APC interactions in the context of transplant tolerance.
Main Methods:
- Utilized a mouse heart transplant model.
- Employed imaging flow cytometry to quantify the proportion of recipient T cells forming mature immune synapses with donor APCs.
- Analyzed T-cell-APC contact events in scenarios of acute rejection, tolerance, and regulatory T-cell mediated survival.
- Tested the effect of human regulatory T cells on T-APC interactions in vitro.
Main Results:
- The frequency of in vitro T-APC immune synapses strongly predicted allograft outcomes across rejection and tolerance states.
- Increased T-APC contacts correlated with acute rejection, while decreased contacts were observed in unresponsive recipients.
- Regulatory T cells significantly reduced prolonged T-APC contacts, an effect also observed with human regulatory T cells.
- The findings were validated in a mouse heart transplant model.
Conclusions:
- Imaging flow cytometry provides a predictive measure of T-cell alloreactivity and allograft outcome.
- This technique can characterize the alloreactive T-cell compartment and potentially guide immunosuppression minimization strategies.
- Further development may lead to clinical assays for patient selection and monitoring of tolerogenic therapies.
