Flow cytometric assay of phosphotyrosine levels in Bcr-Abl-positive chronic myelogenous leukemias: a potential prognostic marker |
| |
Authors: | Xuemei Sun Jianyong Li Junhao Chen Dong Li Leilei Chen Wei Xu Yonggong Yang Yujie Wu Pengjun Jiang Wei Xie |
| |
Institution: | (1) The Key Laboratory of Developmental Genes and Human Disease, Southeast University Medical School, Nanjing, People’s Republic of China;(2) Department of Hematology, Jiangsu Provincial Hospital of Traditional Chinese Medicine, affiliated with Nanjing University of Traditional Chinese Medicine, Nanjing, People’s Republic of China;(3) Department of Hematology, First Affiliated Hospital of Nanjing Medical University/Jiangsu Province Hospital, Nanjing, Jiangsu Province, 210029, People’s Republic of China;(4) Department of Hematology, Drum Tower Hospital, affiliated with Medical School, Nanjing University, Nanjing, People’s Republic of China; |
| |
Abstract: | Assay of phosphotyrosine levels using flow cytometry has been used to identify patients with chronic myelogenous leukemia
positive for the Bcr-Abl fusion gene. We hypothesized that clinical monitoring could identify treatment response through reductions in intragranulocyte
phosphotyrosine. Initially, we studied cell lines FDC-P1 and HL60 (Bcr-Abl-negative) and FDrv210 and K562 (Bcr-Abl-positive) with our technique. A fluorescein isothiocyanate-conjugated monoclonal antibody was used along with fluorescence-conjugated
microspheres for reference (ratio of sample fluorescence: bead fluorescence = relative fluorescence unit RFU]). Samples from
20 controls and 32 patients undergoing treatment were analyzed using the same method. Bcr-Abl status for each patient was confirmed using fluorescent in situ hybridization or polymerase chain reaction gene amplification
(PCR). Testing of cell lines consistently produced expected results. Patient values were found to be significantly higher
than control values (P < 0.001) and values for patients with advanced disease were significantly higher than for patients with chronic-phase disease
(P < 0.05). Results of clinical monitoring were consistent with results from PCR. Two patients who received allogeneic stem
cell transplantation had molecular remission confirmed by PCR and had a marked decrease in RFU value (from 62 to five and
from 131 to 23). No such fluorescence change was observed in patients who achieved clinical remission. Flow cytometric analysis
of phosphotyrosine levels is a reliable and convenient adjuvant technique for diagnosis of Bcr-Abl-positive leukemias and shows promise for serial evaluation of patients undergoing treatment.
This work was supported by a grant for society development from Jiangsu Science and Technology Bureau (Fund number BS2003603)
and a grant for key medical experts from Jiangsu Public Health Bureau (Fund number RC2007002, recipient Xuemei Sun). |
| |
Keywords: | Phosphorylated tyrosine Bcr-Abl Chronic myelogenous leukemia Flow cytometry Diagnosis Disease status |
本文献已被 PubMed SpringerLink 等数据库收录! |
|