PMID- 34303196
OWN - NLM
STAT- MEDLINE
DCOM- 20211117
LR  - 20211117
IS  - 1090-2104 (Electronic)
IS  - 0006-291X (Linking)
VI  - 571
DP  - 2021 Sep 24
TI  - Simultaneous monitoring assay for T-cell receptor stimulation-dependent 
      activation of CD4 and CD8 T cells using inducible markers on the cell surface.
PG  - 53-59
LID - S0006-291X(21)01074-3 [pii]
LID - 10.1016/j.bbrc.2021.07.037 [doi]
AB  - Isolation of antigen (Ag)-specific T cells is an important step in the 
      investigation of T-cell immunity. Activation-induced markers (AIMs), such as 
      CD154/tumor necrosis factor (TNF)/CD107A/CD134/CD137 enable the sorting of 
      Ag-specific T cells without using human leukocyte antigen (HLA)-multimers. 
      However, optimal conditions suitable for simultaneous detection of both 
      Ag-specific CD4 and CD8 T cells have not been investigated. Here, conditions were 
      optimized to simultaneously detect the maximum number of activated CD4 and CD8 T 
      cells in a TCR-dependent manner. First, the frequency of total pools of 
      AIM-positive cells induced by superantigen, staphylococcal enterotoxin B (SEB), 
      stimulation in various culture conditions was monitored and compared 
      side-by-side. The total amount of AIM-positive CD4 T cells, but not CD8 T cells, 
      was significantly abrogated by addition of brefeldin A. TNF-alpha converting 
      enzyme inhibitor treatment effectively increased the TNF-positive population, 
      without affecting other markers' positivity. AIM-positive CD4 T cells and CD8 T 
      cells were detected at least 3 h after stimulation. Furthermore, examination of 
      the multiple combination of each marker revealed that minimum contribution of 
      CD134 on the total pool of AIM-positive cells at this setting, suggesting the 
      essential and non-essential AIMs to maximize the detected number of AIM-positive 
      cells. Taken together, this optimized method will be a useful tool for the 
      simultaneous monitoring the T-cell receptor stimulation-dependent activation of 
      CD4 and CD8 T cells using inducible markers on the cell surface including 
      Ag-specific T cells.
CI  - Copyright (c) 2021 Elsevier Inc. All rights reserved.
FAU - Takahama, Shokichi
AU  - Takahama S
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan. Electronic address: stakahama@nibiohn.go.jp.
FAU - Nogimori, Takuto
AU  - Nogimori T
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan.
FAU - Higashiguchi, Masaya
AU  - Higashiguchi M
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan; Department of Gastroenterological Surgery, 
      Graduate School of Medicine, Osaka University, Osaka, Japan.
FAU - Murakami, Hirotomo
AU  - Murakami H
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan; Department of Gastroenterological Surgery, 
      Graduate School of Medicine, Osaka University, Osaka, Japan.
FAU - Akita, Hirofumi
AU  - Akita H
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan; Department of Gastroenterological Surgery, 
      Graduate School of Medicine, Osaka University, Osaka, Japan.
FAU - Yamamoto, Takuya
AU  - Yamamoto T
AD  - Laboratory of Immunosenescence, National Institutes of Biomedical Innovation, 
      Health and Nutrition, Osaka, Japan. Electronic address: yamamotot2@nibiohn.go.jp.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210721
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (Biomarkers)
RN  - 0 (Receptors, Antigen, T-Cell)
SB  - IM
MH  - Biomarkers/metabolism
MH  - CD4-Positive T-Lymphocytes/*metabolism
MH  - CD8-Positive T-Lymphocytes/*metabolism
MH  - Healthy Volunteers
MH  - Humans
MH  - Receptors, Antigen, T-Cell/*metabolism
MH  - Surface Properties
OTO - NOTNLM
OT  - Activation-induced markers
OT  - Antigen-specific T cells
OT  - Flow cytometry
OT  - TCR-Dependent activation
EDAT- 2021/07/25 06:00
MHDA- 2021/11/18 06:00
CRDT- 2021/07/24 20:24
PHST- 2021/06/15 00:00 [received]
PHST- 2021/07/10 00:00 [accepted]
PHST- 2021/07/25 06:00 [pubmed]
PHST- 2021/11/18 06:00 [medline]
PHST- 2021/07/24 20:24 [entrez]
AID - S0006-291X(21)01074-3 [pii]
AID - 10.1016/j.bbrc.2021.07.037 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 2021 Sep 24;571:53-59. doi: 
      10.1016/j.bbrc.2021.07.037. Epub 2021 Jul 21.