Abstract:
:Although a variety of assays have been used to examine T cell responses in vitro, standardized ex vivo detection of antigen-specific CD4+ T cells from human circulatory PBMCs remains constrained by low-dimensional characterization outputs and the need for polyclonal, mitogen-induced expansion methods to generate detectable response signals. To overcome these limitations, we developed a novel methodology utilizing antigen-pulsed autologous human dendritic target cells in a rapid and sensitive assay to accurately enumerate antigen-specific CD4+ T cell precursor frequency by multiparametric flow cytometry. With this approach, we demonstrate the ability to reproducibly quantitate poly-functional T cell responses following both primary and recall antigenic stimulation. Furthermore, this approach enables more comprehensive phenotypic profiling of circulating antigen-specific CD4+ T cells, providing valuable insights into the pre-existing polarization of antigen-specific T cells in humans. Combined, this approach permits sensitive and detailed ex vivo detection of antigen-specific CD4+ T cells delivering an important tool for advancing vaccine, immune-oncology and other therapeutic studies.
journal_name
Cytotechnologyjournal_title
Cytotechnologyauthors
Carrio R,Zhang G,Drake DR 3rd,Schanen BCdoi
10.1007/s10616-018-0222-8subject
Has Abstractpub_date
2018-10-01 00:00:00pages
1325-1335issue
5eissn
0920-9069issn
1573-0778pii
10.1007/s10616-018-0222-8journal_volume
70pub_type
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