|
KMID : 0923620130130060264
|
|
Immune Network
2013 Volume.13 No. 6 p.264 ~ p.274
|
|
|
Detection of Foreign Antigen-specific CD4＋Foxp3＋ Regulatory T Cells by MHC Class II Tetramer and Intracellular CD154 Staining
|
|
Choi Jin-Young
Eo Seong-Kug
|
|
|
Abstract
|
|
|
|
The unrestricted population of CD4＋Foxp3＋ regulatory T (Treg) cells, which have been known to control the expression of autoimmune diseases and protective immunity to in-flammatory reactions, has led to greater appreciation of functional plasticity. Detecting and/or isolating Ag-specific CD4＋Foxp3＋ Tregs at the single cell level are required to study their function and plasticity. In this study, we estab-lished and compared both MHC class II tetramer and intra-cellular CD154 staining, in order to detect CD4＋Foxp3＋ Treg specific for foreign Ag in acute and chronic infections with lymphocytic choriomeningitis virus (LCMV). Our results revealed that MHC class II tetramer staining showed a lower detection rate of LCMV GP66-77-specific CD4＋ T cells be-cause most of MHC class II tetramers were unbound and un-stable when combined staining was performed with intra-cellular cytokines. In contrast, intracellular CD154 staining was revealed to be easier and simple for detecting LCMV GP66-77-specific CD4＋ T cells, compared to MHC class II tet-ramer staining. Subsequently, we employed intracellular CD154 staining to detect LCMV GP66-77-specific CD4＋Foxp3＋ Tregs using Foxp3GFP knock-in mouse, and found that LCMV GP66-77-specific CD4＋Foxp3＋ Tregs and polyclonal CD4＋Foxp3＋ Tregs showed differential expansion in mice in-fected with LCMV Arms or Cl13 at acute (8 and 13 days pi) and chronic phases (35 days pi). Therefore, our results pro-vide insight into the valuable use of intracellular CD154 staining to detect and characterize foreign Ag-specific CD4＋Foxp3＋ Treg in various models.
|
|
|
KEYWORD
|
|
|
CD4＋Foxp3＋ regulatory T cells, MHC class II tetramer, Intracellular CD154 staining, Exhausted T cell
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
   
|