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Quantification of CD4 + T Cell Alloreactivity and Its Control by Regulatory T Cells Using Time‐Lapse Microscopy and Immune Synapse Detection
Author(s) -
Juvet S. C.,
Sanderson S.,
Hester J.,
Wood K. J.,
Bushell A.
Publication year - 2016
Publication title -
american journal of transplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.89
H-Index - 188
eISSN - 1600-6143
pISSN - 1600-6135
DOI - 10.1111/ajt.13607
Subject(s) - cd28 , t cell , immunological synapse , flow cytometry , immune system , immunology , transplantation , major histocompatibility complex , microbiology and biotechnology , context (archaeology) , immunosuppression , biology , medicine , t cell receptor , paleontology
Assays designed to select transplant recipients for immunosuppression withdrawal have met with limited success, perhaps because they measure events downstream of T cell–alloantigen interactions. Using in vitro time‐lapse microscopy in a mouse transplant model, we investigated whether transplant outcome would result in changes in the proportion of CD 4 + T cells forming prolonged interactions with donor dendritic cells. By blocking CD 4– MHC class II and CD 28–B7 interactions, we defined immunologically relevant interactions as those ≥500 s. Using this threshold, T cell–dendritic cell (T‐ DC ) interactions were examined in rejection, tolerance and T cell control mediated by regulatory T cells. The frequency of T‐ DC contacts ≥500 s increased with T cells from mice during acute rejection and decreased with T cells from mice rendered unresponsive to alloantigen. Regulatory T cells reduced prolonged T‐ DC contacts. Importantly, this effect was replicated with human polyclonally expanded naturally occurring regulatory T cells, which we have previously shown can control rejection of human tissues in humanized mouse models. Finally, in a proof‐of‐concept translational context, we were able to visualize differential allogeneic immune synapse formation in polyclonal CD 4 + T cells using high‐throughput imaging flow cytometry.