PY20抗体检测慢性髓系白血病细胞内酪氨酸磷酸水平的临床应用

Detection of Phosphotyrosine in Chronic Myeloid Leukemia Cells with PY20 Antibody and Its Clinical Applications

本研究探讨PY20抗酪氨酸磷酸单克隆抗体检测在慢性髓系白血病(CML)诊断中的特异性及其临床应用的可能性。采用抗PY20和CD45抗体标记,通过流式细胞术检测28例初诊CML患者治疗前后PY20阳性率,比较其与bcr-abl融合基因及Ph染色体结果的符合率。另外,随访监测7例异基因造血干细胞移植后CML患者的PY20阳性率、bcr-abl融合基因及Ph染色体的变化。结果表明:①28例初诊CML患者PY20在慢性期、加速期、急变期表达率分别为(40.31±1.22)%、(77.28±1.14)%,(78.12±1.32)%。CML慢性期PY20表达率低于加速期或急变期(p<0.05),加速期和急变期之间无显著差异(p>0.05)。②28例初诊CML患者经治疗后获得CR、PR、NR患者的PY20阳性率分别为(15.56±1.51)%、(38.73±2.31)%、(60.43%±2.04)%。CR患者PY20阳性率明显低于PR及NR的患者(p<0.05)。PR患者低于NR患者(p<0.05)。③初诊患者PY20与RT-PCRbcr-abl检测的阳性符合率为92.31%,阴性符合率为95.45%;PY20与Ph染色体检测的阳性符合率为88.46%,阴性符合率为95.46%。④7例患者移植后Ph染色体均为阴性,移植后bcr-abl融合基因持续阴性5例,持续阳性2例。此7例PY20细胞表达均为阴性。结论:①流式细胞术检测CML细胞酪氨酸磷酸化水平阳性率有较好的特异性及敏感性,可应用于CML的早期、快速辅助诊断。②PY20阳性率对CML分期、病情监测及疗效判断有一定的临床指导意义。③流式细胞术、RT-PCR及染色体核型分析能相互补充提高CML的诊断率。

The objective of this study was to investigate the specificity of detecting the phosphotyrosine level with anti-phosphotyrosine monoclonal antibody PY20 for diagnosis and prognosis of patients with chronic myeloid leukemia （CML） and the possibility of its clinical application. The positive rate of PY20 in 28 newly diagnosed CML patients was detected by flow cytometry using anti-PY20 antibody, the bcr-abl fusion gene was detected by nested RT-PCR, the Ph chromosome was measured by R-banding cytogenetic analysis, and the coincidence of PY20 positive rate with results of bcr-abl fusion gene and Ph chromosome detection was compared. In addition, the positive rate of PY20, the changes of bcr-abl fusion gene and Ph chromosome were determined in follow up 7 CML patients after allo-hematopoietic stem cell transplantation. The results indicated that the positive rates of PY20 in 28 newly diagnosted CML patients in groups of chronic phase（CP）, accelerated phase（AP）, and blast phase（BP） were （40.31% ± 1.22）%, （77.28 ± 1.14）% and （78.12± 1.32） % respectively. The positive rate of PY20 in CP was lower than that in AP and BP （p 〈 0. 05）. There was no difference in positive rate of PY20 between AP and BP（p 〉 0.05 ）. PY20 expression level of leukocytes from peripheral blood and bone marrow showed no difference （p 〈0.05）. The positive rates of PY20 in patients with CR, PR and NR were （15.56% ± 1.51）% ,（38.73% v2.31）% and （60.43% ±2.04）% respectively. The positive and negative coincidence between PY20 and RT-PCR was 92, 31% and 95.45% respectively. The positive and negative coincidence between PY20 and Ph Chromosome in newly diagnosed patients was 88.46% and 95.46% respectively. Ph chromosome and PY20 were all negative in 7 CML patients after allo-HSCT. Bcr-abl fusion gene was negative persistently in 5 patients, but in the other 2 patients, the fusion gene was persistently positive. In conclusion, the detection of the level of phosphotyrosine in CML ceils has high sensitivity and specificity. The results of PY20 cell positive rate combined with detection of bcr/abl fusion gene and Ph chromosome might be useful in diagnosis as a good index of monitoring.