Monitoring the initiation and kinetics of human dendritic cell-induced polarization of autologous naive CD4+ T cells

A crucial step in generating de novo immune responses is the polarization of naive cognate CD4+ T cells by pathogen-triggered dendritic cells (DC). In the human setting, standardized DC-dependent systems are lacking to study molecular events during the initiation of a naive CD4+ T cell response. We...

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Bibliographic Details
Published in:	PloS one 2014-08, Vol.9 (8), p.e103725-e103725
Main Authors:	Oth, Tammy, Schnijderberg, Melanie C A, Senden-Gijsbers, Birgit L M G, Germeraad, Wilfred T V, Bos, Gerard M J, Vanderlocht, Joris
Format:	Article
Language:	eng
Subjects:	Antigen-presenting cells Antigens Biology and Life Sciences Blood CD4 antigen CD4-Positive T-Lymphocytes - cytology CD4-Positive T-Lymphocytes - metabolism Cell Differentiation - physiology Cell Polarity - physiology Cells, Cultured Cytokines Dendritic cells Dendritic Cells - cytology Dendritic Cells - metabolism Developmental biology Flow Cytometry Hematology Humans Immune response Immunology Induced polarization Internal medicine Kinetics Ligands Lymphocytes Lymphocytes T Medicine Molecular modelling Monocytes - cytology Oncology Pathogens Pharmacology Polarization Polymerase Chain Reaction Reagents Research and Analysis Methods Signal transduction T cell receptors T-cell receptor Transcription factors
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Summary:	A crucial step in generating de novo immune responses is the polarization of naive cognate CD4+ T cells by pathogen-triggered dendritic cells (DC). In the human setting, standardized DC-dependent systems are lacking to study molecular events during the initiation of a naive CD4+ T cell response. We developed a TCR-restricted assay to compare different pathogen-triggered human DC for their capacities to instruct functional differentiation of autologous, naive CD4+ T cells. We demonstrated that this methodology can be applied to compare differently matured DC in terms of kinetics, direction, and magnitude of the naive CD4+ T cell response. Furthermore, we showed the applicability of this assay to study the T cell polarizing capacity of low-frequency blood-derived DC populations directly isolated ex vivo. This methodology for addressing APC-dependent instruction of naive CD4+ T cells in a human autologous setting will provide researchers with a valuable tool to gain more insight into molecular mechanisms occurring in the early phase of T cell polarization. In addition, it may also allow the study of pharmacological agents on DC-dependent T cell polarization in the human system.
ISSN:	1932-6203 1932-6203