Zhu Hong-Hu, Liu Yan-Rong, Qin Ya-Zhen, Chang Yan, Li Jin-Lan, Ruan Guo-Rui, Jiang Bin, Chen Shan-Shan, Lu Dao-Pei
Institute of Hematology, People's Hospital, Peking University, Beijing 100044, China.
Zhonghua Xue Ye Xue Za Zhi. 2006 Jul;27(7):441-4.
To explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications.
Bcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI).
Bcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01).
PY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.
探讨抗磷酸酪氨酸单克隆抗体PY20在bcr-abl+细胞中的特异性及其可能的临床应用。
用PY20对bcr-abl细胞系（K562、MEG-01）和bcr-abl-细胞系（Jurkat、MCF-7）进行染色。用伊马替尼处理K562和MEG-01细胞前后，通过流式细胞术检测其磷酸酪氨酸蛋白。检测49例慢性髓性白血病（CML）、Ph+急性淋巴细胞白血病（Ph(+) -ALL）、Ph- ALL、急性髓性白血病（AML-M1、M2、M3、M5，FAB分类）、慢性淋巴细胞白血病（CLL）患者及3名正常供者骨髓细胞中的磷酸酪氨酸蛋白。磷酸酪氨酸蛋白阳性细胞占总细胞数超过5%者为阳性病例。通过中位荧光强度（MFI）测定酪氨酸磷酸化水平。
10例CML患者和8例ALL患者的bcr-abl细胞系及骨髓细胞均为PY20阳性，而18例白血病患者和3名正常供者的bcr-abl-细胞系及骨髓细胞均为PY20阴性。伊马替尼阻断后，K562细胞和MEG-01细胞的MFI显著降低。10例新诊断CML患者和9例伊马替尼敏感CML患者骨髓细胞的阳性细胞百分比分别为（54.20±19.82）%和（14.84±6.17）%（P<0.05），而2例伊马替尼耐药患者分别为64.3%和57.2%。伊马替尼耐药患者与伊马替尼敏感患者之间的MFI有显著差异（99.42±4.87对46.41±4.67，P<0.01）。
PY20单克隆抗体对bcr-abl+细胞具有高度特异性。它可能有助于快速检测bcr-abl+细胞及CML患者对伊马替尼的敏感性。
