Application of HLA-A*0201/WT1 pentamer combined with intracellular IFNgamma+ staining in detecting circulating WT1 specific T cells in leukemia.
- Author:
Li WEI
1
;
Xue-Dong SUN
;
Hong-Li ZUO
;
Tie-Qiang LIU
;
Mei GUO
;
Guang-Xian LIU
;
Qi-Yun SUN
;
Jian-Hui QIAO
;
Dan-Hong WANG
;
Chang-Lin YU
;
Kai-Xun HU
;
Zheng DONG
;
Hui-Sheng AI
Author Information
1. Third Department of Internal Medicine, General Hospital of Tibet Military Area, Lhasa 850003, Tibet Autonomous Region, China.
- Publication Type:Journal Article
- MeSH:
Adolescent;
Adult;
Child;
Female;
Flow Cytometry;
HLA-A Antigens;
analysis;
HLA-A2 Antigen;
Humans;
Interferon-gamma;
analysis;
Leukemia;
blood;
genetics;
immunology;
Male;
Middle Aged;
Staining and Labeling;
T-Lymphocytes, Cytotoxic;
immunology;
metabolism;
WT1 Proteins;
genetics;
immunology;
metabolism;
Young Adult
- From:
Journal of Experimental Hematology
2010;18(2):505-509
- CountryChina
- Language:Chinese
-
Abstract:
This study was purposed to investigate the value of combination of pentamer and intracellular IFNgamma staining in the qualitative and quantitative detection of circulating antigen-specific T cells. WT1 expressions in 14 HLA-A*0201+ patients and their matched donors were detected by RT-PCR, and circulating WT1 specific T cells were assayed by HLA-A*0201/WT1 pentamer combined with intracellular IFNgamma+ staining. The results showed that the low level of WT1 expression was found only in 2 cases out of 14 donors, but different levels of WT1 expression could be observed in all leukemic patients. The WT1+CD8+ CTL and WT1+IFNgamma+ cells did not detected in all 14 donors, but WT1+CD8+ CTL cells in 2 patients and WT1+IFNgamma+ cells in 3 patients could be detected before transplantation respectively, there was no significant difference between them, while the WT1+CD8+ CTL cells and WT1+IFNgamma+ cells both could be detected in all 14 patients after transplantation, the positive detection rate after transplantation was obviously higher than that before transplantation. The WT1+CD8+ and WT1+ IFNgamma+ cells could be detected within 30 days after transplantation, but the positive detection rate of WT1+IFNgamma+ cells was higher than that of WT1+CD8+ CTL cells (p=0.014). The median peak value of WT1+CD8+ CTL cells was 0.18% in 14 patients, and the median peak value of WT1+IFNgamma+ cells was 0.83% in 14 patients, the later was significantly higher than former. The median peak time of WT1+CD8+ CTL cells was 75 days after transplantation, while the WT1+IFNgamma+ cells was 105 days after transplantation, there was no significant difference between them. It is concluded that pentamer and intracellular IFNgamma staining may effectively detect circulating WT1 specific T cells in leukemic patients, and the combination of these two methods profit to the exact qualitation and quantitation of circulating antigen-specific T cells.
