[Application of HLA-A*0201/WT1 pentamer combined with intracellular IFNgamma+ staining in detecting circulating WT1 specific T cells in leukemia].
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.
Wei
Li
L
Third Department of Internal Medicine, General Hospital of Tibet Military Area, Lhasa 850003, Tibet Autonomous Region, China.
Sun
Xue-Dong
XD
Zuo
Hong-Li
HL
Liu
Tie-Qiang
TQ
Guo
Mei
M
Liu
Guang-Xian
GX
Sun
Qi-Yun
QY
Qiao
Jian-Hui
JH
Wang
Dan-Hong
DH
Yu
Chang-Lin
CL
Hu
Kai-Xun
KX
Dong
Zheng
Z
Ai
Hui-Sheng
HS
D000293:Adolescent; D000328:Adult; D002648:Child; D005260:Female; D005434:Flow Cytometry; D015234:HLA-A Antigens / Q000032:analysis*; D015789:HLA-A2 Antigen; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis*; D007938:Leukemia / Q000097:blood / Q000235:genetics / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D013194:Staining and Labeling; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D025721:WT1 Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D055815:Young Adult
D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D025721:WT1 Proteins; D007371:Interferon-gamma