白细胞分化特异性胞浆抗原在急性白血病细胞的表达及意义
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摘要
背景和目的
在急性白血病的诊断中,细胞形态学联合细胞化学将急性白血病分型的准确性达89%,白血病免疫分型的准确性及灵敏度都较高,能够将急性白血病的分型诊断准确率大大提高,其在急性白血病的现代诊断中起着非常重要的作用。急性白血病的免疫分型是基于不同的急性白血病细胞表达有其特定的细胞表面抗原群,应用抗原抗体特异性结合的原理,借助于流式细胞仪,对急性白血病细胞膜表面表达的抗原进行分型,能够对那些形态学不典型,仅靠细胞形态和化学染色难以诊断的病例做出精确诊断,尤其是对急性淋巴细胞白血病(acute lymphoblastic leukemis, ALL)的诊断,不仅能够将ALL与与急性髓系白血病(acute myeloid leukemis, AML)区别开,更能将B细胞ALL与T细胞ALL进行区分,弥补了ALL形态学诊断的不足。但白血病免疫分型检测中发现有些AML细胞不仅表达髓细胞相关抗原,而且还表达淋巴细胞相关抗原,反之亦然。这种情况是伴系抗原表达,还是混合型白血病,较难确诊。因此选择更加特异的标志物以明确诊断非常必要,这对病人的治疗、预后具有重要的作用。我们在常规免疫分型检测的基础上,加做胞浆抗原染色,以探讨白细胞分化特异性胞浆抗原在急性白血病细胞的表达及意义。
方法
选择2010年9月至2011年5月在我院血液科住院治疗的急性白血病患者138例,男86例,女52例,年龄14岁-76岁,中位年龄37.5岁。
所有患者经血常规、生化、骨髓细胞形态学及化学染色、白血病免疫分型及细胞遗传学诊断。
应用流式细胞术三色荧光标记,对细胞膜抗原进行常规检测,对形态学分类难以诊断,出现跨系列表达的加测胞浆内抗原检测。髓系选择胞浆抗原cMPO、急性B淋巴细胞白血病选择胞浆抗原cCD79a、急性T淋巴细胞白血病选择胞浆抗原cCD3。
结果
1.诊断
按FAB分型138例AL中诊断为AML者76例,ALL者49例,不能明确分型者13例。应用流式细胞术进行白血病免疫分型分析,ALL49例包括B-ALL40例和T-ALL9例。13例根据细胞形态无法确诊的病例最终诊断2例AML-MO、5例B-ALL、2例T-ALL和4例混合细胞白血病。
2.跨系抗原表达
急性白血病中出现跨系抗原表达的占41.3%,其中AML、B-ALL和T-ALL中出现跨系抗原表达的比例分别为50%、26.7%、18.1%。AML中跨系抗原表达以CD7(53.8%)和CD19(30.8%)为主。出现跨系抗原表达的B-ALL中CD33阳性率为83.3%,显著高于CD2及CD13(P<0.05)。
3.敏感性和特异性
本组所用抗体对AML的敏感性从高到低依次为CD33、CD13、cMPO、CD117,特异性依次为CD117、cMPO、CD13、CD33,胞浆抗原cMPO的敏感性低于CD33和CD13,但特异性高于CD33和CD13,cMPO和CD117的特异性无显著差别(P=-0.312),但cMPO敏感性高于CD117(P=-0.027)；本组所用抗体对B-ALL的敏感性依次为CD19、cCD79a、CD10、CD20,cCD79a特异性和CD10无显著差别(P=-0.312),和CD20无显著差别(P=0.287),但显著高于CD19(P=0.018)；本组抗体对T-ALL的敏感性从高到低依次为cCD3、CD7、CD2,特异性依次为cCD3、CD2、CD7,cCD3的敏感性高于CD7(P=0.042)和CD2(P=-0.047),特异性和CD2无差别(P=0.426),显著高于CD7(P=0.022)。
4.胞浆抗原cMPO、cCD79a、cCD3的表达
cMPO在AML、B-ALL和T-ALL中阳性率分别为87.5%、9.5%和0,cMPO在AML的表达率显著高于ALL(P<0.05),其中B-ALL与T-ALL之间的差别无显著性(P>0.05)；cCD79a在B-ALL、T-ALL和AML中的阳性率分别为92.7%、9.0%和0。cCD79a在B-ALL的表达率显著高于T-ALL(P<0.05),其中T-ALL与AML之间的差别无显著性(P>0.05)；cCD3在T-ALL中表达率为100%,在AML和B-ALL中均无表达。
结论
1.胞浆抗原cMPO、cCD79a、cCD3分别是AML、B-ALL、T-ALL的特异性标志,系列特异性高于细胞膜表面抗原。
2.胞浆抗原cMPO、cCD79a、cCD3的检测有助于细胞形态不典型、细胞膜出现跨系抗原表达的白血病的准确分型及混合性白血病的正确诊断。
Background and Objective
Cell morphology and cytochemistry can improve the classification accuracy of acute leukemia up to89%in the diagnosis of acute leukemia. Because of the high accuracy and sensitivity, immunophenotyping plays an important role in the diagnosis of leukemia. The immunophenotype of acute leukemia is method to classify acute leukemia, based on the specific antigens expressed on acute leukemia blasts, which bind to specific antibodies conjugated with fluorescent detected by flow cytometer. This technique is used in the diagnosis and classification of acute leukemia cases with atypical morphology which cannot be diagnosed by bone marrow morphology and cytochemistry. In the diagnosis of acute lymphoblast leukemia, immunophenotype can be used to differentiate ALL from AML as well as to differentiate B-All from T-ALL, which is a compliment to the morphological diagnosis of ALL. However, it is found that AML leukemic cells express not only myeloid-associated antigen, but also the lymphocyte-associated antigen and vice versa. It is difficult to distinguish cross-lineage expression from mixed-lineage acute leukemia (MAL). Therefore it is necessary to choose more specific markers to confirm the diagnosis of acute leukemia, which plays an important role in the patient's treatment and prognosis. On the base of the immunophenotype of cellular surface antigens, we performed a cytoplasmic antigen staining in order to investigate the expression of lineage specific cytoplasmic antigens in patients with acute leukemia and its significance.
Methods
One hundred and thirty-eight cases of acute leukemia were treated in the Department of Hematology of the First Affiliated Hospital of Zhengzhou University form September2010to May2011, including86male cases and52female cases. These patients range from14years old to76years old, with the median age at37.5years old.
All patients were diagnosed by CBC, liver function, renal function, bone marrow cell morphology and histochemistry, immunophenotype and cytogenetics.
The membrane antigens were detected in138patients with acute leukemia using three-color fluorescent by flow cytometry. Cytoplasmic antigens were detected in cases which difficulty to diagnose with morphological classification and cases with cross-lineage expression. Cytoplasmic antigens cMPO was selected for myeloid, Cytoplasmic antigens cCD79a was selected for acute B-lymphoblastic leukemia, and Cytoplasmic antigens cCD3was selected for acute T-lymphoblastic leukemia.
Results
1. Dignosis
According to the FAB classification,76out of138cases were diagnosed as AML and49/138cases were diagnosed as ALL, and other13/138cases were not explicitly classified. By immunophenotype, the49/138cases of ALL were subdivided into B-ALL (40cases) and T-ALL (9cases). In the cases morphologically not defined subtypes,2/13cases were diagnosed as AML-MO,5/13cases as B-ALL,2/13cases as T-ALL and4/13cases as mixed-lineage acute leukemia.
2. The expression of cross-lineage antigens
There were41.3%of acute leukemia express Cross-lineage antigen. The cross-lineage antigen expression rates were50%in AML,26.7%in B-ALL and18.1%in T-ALL. Cross-lineage antigen expression in AML is mainly CD7(53.8%) and CD19(30.8%). CD33was expressed in83.3%of B-ALL, significantly higher than that of CD13and CD2(P<0.05).
3. The specificity and sensitivity
The sensitivity of the antibody used in the diagnosis of AML were listed from high to low as CD33, CD33, cMPO, CD117, and the specificity from high to low as CD117, cMPO, CD13, CD33. The sensitivity of cytoplasmic antigens cMPO was lower than that of CD33and CD13, but the specific was higher than that CD33and CD13.There was no significant difference in the specificity between cMPO and CD117(P=0.312), but the sensitive of cMPO was higher than CD117(P=0.027). The sensitivity of the antibody used in the diagnosis of B-ALL were listed from high to low as CD19, cCD79a, CD10, CD20. There was no significant difference with specificity between cCD79a and CD10(P=0.312), CD20(P=0.287), but it was significantly higher than that of CD19(P=0.018). The sensitivity of antibodies used in the diagnosis of T-ALL were listed from high to low as cCD3, CD7, CD2, the specific from high to low as cCD3, CD2and CD7.The sensitive of cCD3was higher than CD7(P=0.042) and CD2(P=0.047), The specificity of cCD3had no difference with CD2(P=0.426), but it was significantly higher than that of CD7(P=0.022).
4. The expression of Cytoplasmic antigen cMPO, cCD79a and cCD3
The positive rates of cMPO were88.6%in AML,9.5%in B-ALL and0in T-ALL. The expression of cMPO in AML was significantly higher than that in ALL (P<0.05). There was no statistically significance in the expression of cMPO between B-ALL and T-ALL (P>0.05). The positive rates of cCD79a were92.7%in B-ALL,9.0%in T-ALL and0in AML. There was no statistically significance in the expression of cCD79a between T-ALL and AML (P>0.05). The positive rate of cCD3in T-ALL was100%. There were no cCD3expression in AML and B-ALL.
Conclusion
1. Cytoplasmic antigen cMPO, cCD79a, cCD3are specific markers of AML, B-ALL, and T-ALL. Their lineage specificity is higher than the cell surface antigen
2. The detection of Cytoplasmic antigen cMPO, cCD79a, cCD3plays an importment role in the accurate diagnosis and classification of acute leukemia, especially in the cases with atypical morphology and cross-lineage antigen expression, such as mixed-lineage acute leukemia.
在急性白血病的诊断中,细胞形态学联合细胞化学将急性白血病分型的准确性达89%,白血病免疫分型的准确性及灵敏度都较高,能够将急性白血病的分型诊断准确率大大提高,其在急性白血病的现代诊断中起着非常重要的作用。急性白血病的免疫分型是基于不同的急性白血病细胞表达有其特定的细胞表面抗原群,应用抗原抗体特异性结合的原理,借助于流式细胞仪,对急性白血病细胞膜表面表达的抗原进行分型,能够对那些形态学不典型,仅靠细胞形态和化学染色难以诊断的病例做出精确诊断,尤其是对急性淋巴细胞白血病(acute lymphoblastic leukemis, ALL)的诊断,不仅能够将ALL与与急性髓系白血病(acute myeloid leukemis, AML)区别开,更能将B细胞ALL与T细胞ALL进行区分,弥补了ALL形态学诊断的不足。但白血病免疫分型检测中发现有些AML细胞不仅表达髓细胞相关抗原,而且还表达淋巴细胞相关抗原,反之亦然。这种情况是伴系抗原表达,还是混合型白血病,较难确诊。因此选择更加特异的标志物以明确诊断非常必要,这对病人的治疗、预后具有重要的作用。我们在常规免疫分型检测的基础上,加做胞浆抗原染色,以探讨白细胞分化特异性胞浆抗原在急性白血病细胞的表达及意义。
方法
选择2010年9月至2011年5月在我院血液科住院治疗的急性白血病患者138例,男86例,女52例,年龄14岁-76岁,中位年龄37.5岁。
所有患者经血常规、生化、骨髓细胞形态学及化学染色、白血病免疫分型及细胞遗传学诊断。
应用流式细胞术三色荧光标记,对细胞膜抗原进行常规检测,对形态学分类难以诊断,出现跨系列表达的加测胞浆内抗原检测。髓系选择胞浆抗原cMPO、急性B淋巴细胞白血病选择胞浆抗原cCD79a、急性T淋巴细胞白血病选择胞浆抗原cCD3。
结果
1.诊断
按FAB分型138例AL中诊断为AML者76例,ALL者49例,不能明确分型者13例。应用流式细胞术进行白血病免疫分型分析,ALL49例包括B-ALL40例和T-ALL9例。13例根据细胞形态无法确诊的病例最终诊断2例AML-MO、5例B-ALL、2例T-ALL和4例混合细胞白血病。
2.跨系抗原表达
急性白血病中出现跨系抗原表达的占41.3%,其中AML、B-ALL和T-ALL中出现跨系抗原表达的比例分别为50%、26.7%、18.1%。AML中跨系抗原表达以CD7(53.8%)和CD19(30.8%)为主。出现跨系抗原表达的B-ALL中CD33阳性率为83.3%,显著高于CD2及CD13(P<0.05)。
3.敏感性和特异性
本组所用抗体对AML的敏感性从高到低依次为CD33、CD13、cMPO、CD117,特异性依次为CD117、cMPO、CD13、CD33,胞浆抗原cMPO的敏感性低于CD33和CD13,但特异性高于CD33和CD13,cMPO和CD117的特异性无显著差别(P=-0.312),但cMPO敏感性高于CD117(P=-0.027)；本组所用抗体对B-ALL的敏感性依次为CD19、cCD79a、CD10、CD20,cCD79a特异性和CD10无显著差别(P=-0.312),和CD20无显著差别(P=0.287),但显著高于CD19(P=0.018)；本组抗体对T-ALL的敏感性从高到低依次为cCD3、CD7、CD2,特异性依次为cCD3、CD2、CD7,cCD3的敏感性高于CD7(P=0.042)和CD2(P=-0.047),特异性和CD2无差别(P=0.426),显著高于CD7(P=0.022)。
4.胞浆抗原cMPO、cCD79a、cCD3的表达
cMPO在AML、B-ALL和T-ALL中阳性率分别为87.5%、9.5%和0,cMPO在AML的表达率显著高于ALL(P<0.05),其中B-ALL与T-ALL之间的差别无显著性(P>0.05)；cCD79a在B-ALL、T-ALL和AML中的阳性率分别为92.7%、9.0%和0。cCD79a在B-ALL的表达率显著高于T-ALL(P<0.05),其中T-ALL与AML之间的差别无显著性(P>0.05)；cCD3在T-ALL中表达率为100%,在AML和B-ALL中均无表达。
结论
1.胞浆抗原cMPO、cCD79a、cCD3分别是AML、B-ALL、T-ALL的特异性标志,系列特异性高于细胞膜表面抗原。
2.胞浆抗原cMPO、cCD79a、cCD3的检测有助于细胞形态不典型、细胞膜出现跨系抗原表达的白血病的准确分型及混合性白血病的正确诊断。
Background and Objective
Cell morphology and cytochemistry can improve the classification accuracy of acute leukemia up to89%in the diagnosis of acute leukemia. Because of the high accuracy and sensitivity, immunophenotyping plays an important role in the diagnosis of leukemia. The immunophenotype of acute leukemia is method to classify acute leukemia, based on the specific antigens expressed on acute leukemia blasts, which bind to specific antibodies conjugated with fluorescent detected by flow cytometer. This technique is used in the diagnosis and classification of acute leukemia cases with atypical morphology which cannot be diagnosed by bone marrow morphology and cytochemistry. In the diagnosis of acute lymphoblast leukemia, immunophenotype can be used to differentiate ALL from AML as well as to differentiate B-All from T-ALL, which is a compliment to the morphological diagnosis of ALL. However, it is found that AML leukemic cells express not only myeloid-associated antigen, but also the lymphocyte-associated antigen and vice versa. It is difficult to distinguish cross-lineage expression from mixed-lineage acute leukemia (MAL). Therefore it is necessary to choose more specific markers to confirm the diagnosis of acute leukemia, which plays an important role in the patient's treatment and prognosis. On the base of the immunophenotype of cellular surface antigens, we performed a cytoplasmic antigen staining in order to investigate the expression of lineage specific cytoplasmic antigens in patients with acute leukemia and its significance.
Methods
One hundred and thirty-eight cases of acute leukemia were treated in the Department of Hematology of the First Affiliated Hospital of Zhengzhou University form September2010to May2011, including86male cases and52female cases. These patients range from14years old to76years old, with the median age at37.5years old.
All patients were diagnosed by CBC, liver function, renal function, bone marrow cell morphology and histochemistry, immunophenotype and cytogenetics.
The membrane antigens were detected in138patients with acute leukemia using three-color fluorescent by flow cytometry. Cytoplasmic antigens were detected in cases which difficulty to diagnose with morphological classification and cases with cross-lineage expression. Cytoplasmic antigens cMPO was selected for myeloid, Cytoplasmic antigens cCD79a was selected for acute B-lymphoblastic leukemia, and Cytoplasmic antigens cCD3was selected for acute T-lymphoblastic leukemia.
Results
1. Dignosis
According to the FAB classification,76out of138cases were diagnosed as AML and49/138cases were diagnosed as ALL, and other13/138cases were not explicitly classified. By immunophenotype, the49/138cases of ALL were subdivided into B-ALL (40cases) and T-ALL (9cases). In the cases morphologically not defined subtypes,2/13cases were diagnosed as AML-MO,5/13cases as B-ALL,2/13cases as T-ALL and4/13cases as mixed-lineage acute leukemia.
2. The expression of cross-lineage antigens
There were41.3%of acute leukemia express Cross-lineage antigen. The cross-lineage antigen expression rates were50%in AML,26.7%in B-ALL and18.1%in T-ALL. Cross-lineage antigen expression in AML is mainly CD7(53.8%) and CD19(30.8%). CD33was expressed in83.3%of B-ALL, significantly higher than that of CD13and CD2(P<0.05).
3. The specificity and sensitivity
The sensitivity of the antibody used in the diagnosis of AML were listed from high to low as CD33, CD33, cMPO, CD117, and the specificity from high to low as CD117, cMPO, CD13, CD33. The sensitivity of cytoplasmic antigens cMPO was lower than that of CD33and CD13, but the specific was higher than that CD33and CD13.There was no significant difference in the specificity between cMPO and CD117(P=0.312), but the sensitive of cMPO was higher than CD117(P=0.027). The sensitivity of the antibody used in the diagnosis of B-ALL were listed from high to low as CD19, cCD79a, CD10, CD20. There was no significant difference with specificity between cCD79a and CD10(P=0.312), CD20(P=0.287), but it was significantly higher than that of CD19(P=0.018). The sensitivity of antibodies used in the diagnosis of T-ALL were listed from high to low as cCD3, CD7, CD2, the specific from high to low as cCD3, CD2and CD7.The sensitive of cCD3was higher than CD7(P=0.042) and CD2(P=0.047), The specificity of cCD3had no difference with CD2(P=0.426), but it was significantly higher than that of CD7(P=0.022).
4. The expression of Cytoplasmic antigen cMPO, cCD79a and cCD3
The positive rates of cMPO were88.6%in AML,9.5%in B-ALL and0in T-ALL. The expression of cMPO in AML was significantly higher than that in ALL (P<0.05). There was no statistically significance in the expression of cMPO between B-ALL and T-ALL (P>0.05). The positive rates of cCD79a were92.7%in B-ALL,9.0%in T-ALL and0in AML. There was no statistically significance in the expression of cCD79a between T-ALL and AML (P>0.05). The positive rate of cCD3in T-ALL was100%. There were no cCD3expression in AML and B-ALL.
Conclusion
1. Cytoplasmic antigen cMPO, cCD79a, cCD3are specific markers of AML, B-ALL, and T-ALL. Their lineage specificity is higher than the cell surface antigen
2. The detection of Cytoplasmic antigen cMPO, cCD79a, cCD3plays an importment role in the accurate diagnosis and classification of acute leukemia, especially in the cases with atypical morphology and cross-lineage antigen expression, such as mixed-lineage acute leukemia.
引文
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[18]Alvares C L, Schenk T, Hulkki S, et al. Tyrosine kinase inhibitor insensitivity of non-cycling CD34+ human acute myeloid leukaemia cells with FMS-like tyrosine kinase 3 mutations[J]. Br J Haematol,2011,154 (4):457-465.
[19]王兴兵,姚军霞,郑金娥,等.115例急性髓细胞白血病多参数流式细胞术免疫表型分析[J].中国实验血液学杂志,2005,13(2)：250-253.
[20]Kishimoto T, KI KutaniH, Von Dem Borne A E G et al. Leucocyte typing Ⅳ [M].NewYork & London:Garland Publishing Inc,1997:929-965.
[21]王清霞,张秋堂,岳保红,等.成人急性髓性白血病免疫学表型分析[J].郑州大学学报(医学版),2008,43(04)：779-781.
[22]江雁,穆红,唐志琴,等.急性白血病系列特异性抗原表达分析[J].国际检验医学杂志2009,30(6)：532-534.
[23]孔令环,韩梅,鲍彦娜,等.流式细胞术对急性白血病免疫分型的临床意义[J].中国实验诊断学,2010,14(10)：1570-1573.
[24]Roberson JR,Onciu M, Pounds S, et al. Prognostic significance of myeloperoxidase expression in childhood acute myeloid leukemia[J]. Pediatr Blood Cancer,2008, 50(3):542-548.
[25]Higa B, Alkan S, Barton K, et al. Precursor B-cell acute lymphoblastic leukaemia with FAB L3 (i.e., Burkitt's leukaemia/lymphoma) morphology and co-expression of monoclonal surface light chains and Tdt:report of a unique case and review of the literature[J]. Pathology,2009,41 (5):495-498.
[26]张颜,何明生.急性淋巴细胞白血病靶向基因治疗的研究进展[J].当代医学,2010,(02)：35-36+47.
[27]Mason DY, Cordell JL, Brown MH, et al. CD79a:a novel marker for B-cell neoplasms in routinely processed tissue samples [J].Blood,1995,86(4):1453-1459.
[28]Melinda S. Gordon,Cindy M. Kanegai,Jeanette R. Doerr et al.Somatic hypermutation of the B cell receptor genes B29 (Igβ, CD79b) and mb1 (Igα, CD79a) PNAS[J] 2003,100(7)4126-4131.
[29]Abdelhaleem M. The expression of cytoplasmic CD79a correlates with surface CD3 expression in childhood T-ALL lymphoblasts.Leuk Res,2008,32 (3):511-512.
[30]Fujisawa S, Tanioka F, Matsuoka T, et al. CD7/CD19 double-positive T-cell acute lymphoblastic leukemia. Int J Hematol,2006,83(4):324-327.
[31]Gujral S, Subramanian P G, Dasgupta A. Diversity amongst various guidelines for immunopheno typing[J]. Cytometry A,2009,75 (7):560-561.
[32]Zhang J J, Zhang N, Zhang X F, et al. Immunophenotyping characteristics of biphenotypic acute leukemia and analysis of curative effect[J]. Zhongguo Shi Yan Xue Ye Xue Za Zhi,2011,19 (2):317-320.
[33]张之南,沈悌.血液病诊断及疗效标准[M](第3版),科学出版社,2007,184-214.
[34]李晓玲.李睿.陈燕,等.急性混合细胞白血病临床特点及免疫学表达的研究[J].中国实验血液学杂志,2007,15(3)636-639.
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[1]Mazur G,Halon A,Wrobel T, et al.Contribution of flow cytometric immunopheno typing and bone marrow trephine biopsy in the detection of lymphoid bone marrow infiltration in non-Hodgkins lymphoma[J].Neoplasma,2004,51(3):159-163
[2]Tamok A,Hambsch J,Chen R,et al.Cytometric bead array to measure six cytokines in twenty-five microliters of serum[J].ClinChem,2003,49(6):1000-1002.
[3]张艺.流式细胞仪构成与工作原理[J].医疗设备信息,2005,20(8)：25-26.
[4]Grogan WM. Guide to flow cytometry methods [M].New York Marcel Dekker Inc,1990
[5]郑海萍,李艳辉.流式细胞仪的原理和应用[J].医疗器械2003,16(3)：286.
[6]Hirons GT, TO T, YOY. New very flow cytometry[J]. Cytometry,1994,1:129-137.
[7]李超,韩金路,王玉刚,等.流式细胞仪的工作原理及应用[J].中国实用医药,2009,4(20)：235-236.
[8]沈二霞.流式细胞仪的原理和临床应用[J].现代医学仪器与应用,2008,20(1)：49-53.
[9]沈关心,周汝麟.现代免疫学实验技术[M].武汉：湖北科学技术出版社,2002.
[10]杨峥,莫武宁,李山,等.505例白血病应用流式细胞术免疫分型与FAB分型的结果分析[J].中国实验诊断学,2010,14(8)：1238-1241.
[11]Jaffe ES,Harris NL,Stein H,el al.Pathology and genetics of tumours of haematopoietic and lymphoid tissues.IARC Press.Lyon,2001:91-92.
[12]Kaleem Z, White G. Diagnostic criteria for minimally differentiated acute myeloid leukemia(AML-MO). Evaluation and aproposal. Am J Clin Pathol,2001,115:876-884
[13]上海中美白血病协作组,微分化急性髓系白血病21例临床分型及诊断标准复习[J],中华血液学杂志,2008.29(12)847-849.
[14]Bene MC, Bernier M, Casasnovas RO,et al. Acute myeloid leukemia MO:haematological immunophenotypic and cytogenetic characteristics and their prognostic significance analysis in 241 patients[J]. Br J Haematol,2001,113 (3),737-745.
[15]赵菲,陈燕.急性髓系白血病M-1的免疫学特征和预后分析[J].中国实验血液学杂志,2007,15(04)：687-691.
[16]易丽萍,郭新红.急性髓细胞性白血病M2患者形态学及免疫学分型与融合基因染色体检查的价值[J].中国组织工程研究与临床康复,2011,15(10)：1777-1782.
[17]佟海伙,王慧涵,张继红,等.192例急性髓系白血病免疫表型、细胞遗传学及临床特征的研究[J].中国实验血液学杂志,2009,17(05)：1174-1178.
[18]程薇,马保根,翟亚萍,等.139例急性髓系白血病免疫分型特点分析[J].肿瘤防治研究,2009,(03)：235-237.
[19]张颜,何明生.急性淋巴细胞白血病靶向基因治疗的研究进展[J].当代医学,2010,(02)：35-36+47.
[20]Gujral S,Subramanian P G,Dasgupta A.Diversity amongst various guidelines for immunopheno typing[J]. Cytometry A,2009,75 (7):560-561.
[21]Zhang J J, Zhang N, Zhang X F, et al. Immunophenotyping characteristics of biphenotypic acute leukemia and analysis of curative effect[J]. Zhongguo Shi Yan Xue Ye Xue Za Zhi, 2011,19 (2):317-320.
[22]袁婷婷,刘艳荣,常艳,等.95例初治急性T淋巴细胞白血病患者免疫表型特点[J].中国实验血液学杂志,2011,19(05)：1134-1140.
[23]宣恒报,李玉峰,曹维克,等.免疫分型在一些特殊类型急性白血病诊断中的应用[J].现代医学,2008,36(05)：312-315.
[24]Wang QR, Shi JY, Shi L, et al. Mutational Detection of Full-length Mixed Lineage Leukemia Gene in Patients with De Novo AML-M4 and M5 [J].Zhongguo Shi Yan Xue Ye Xue Za Zhi.2012,20 (1):12-17.
[25]Kern W,Voskova D,Schoch C, et al.Determination of relapse riskbased on assessment of minimal residual disease during complete remission by muhiparameter flow cytometry in unseleeted patients with acute myeloid leukemia[J].Blood,2004,104(10):3078-3085.
[26]万岁桂,赵弘,孙雪静,等多参数流式细胞术对急性髓系白血病微小残留病变与疾病复发的监测[J].中国实验血液学杂志,2009,17(03)：557-562.
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[5]杨峥,莫武宁,李山,等.505例白血病应用流式细胞术免疫分型与FAB分型的结果分析[J].中国实验诊断学,2010,14(8)：1238-1241.
[6]Wan SG, Zhao H, Sun XJ, et al. Prognosticating relapse risk based on multiparameter flow cytometric assessment of minimal residual disease in patients with acute myeloid leukemia.Journal of Experimental Hematology,2009,17 (3):557-562.
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[8]Jaffe ES, Harris NL, Stein H, el al. Pathology and genetics of tumours of haematopoietic and lymphoid tissues. IARC Press. Lyon,2001:91-92.
[9]Kaleem Z, White G. Diagnostic criteria for minimally differentiated acute myeloid leukemia(AML-MO). Evaluation and aproposal. Am J Clin Pathol, 2001,115:876-884
[10]上海中美白血病协作组,微分化急性髓系白血病21例临床分型及诊断标准复习,中华血液学杂志,2008.29(12)847-849.
[11]赵菲,陈燕.急性髓系白血病M-1的免疫学特征和预后分析[J].中国实验血液学杂志,2007,15(04)：687-691.
[12]易丽萍,郭新红.急性髓细胞性白血病M2患者形态学及免疫学分型与融合基因染色体检查的价值[J].中国组织工程研究与临床康复,2011,15(10)：1777-1782.
[13]Tong H X, Wang H H, Zhang J H, et al. Immunophenotypes, cytogenetics and clinical features of 192 patients with acute myeloid leukemia [J]. Zhongguo Shiyan XueYe Xue Za Zhi,2009,17(5):1174-1178.
[14]汪永红,黄知平,李滔.急性白血病免疫表型及伴髓系/淋系抗原表达的比较[J].实用医学杂志,2010,26(17)：3231-3233.
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[16]S D de Jonge-Peeters, K van der Weide, Kuipers F, et al. Variability in responsiveness to lovastatin of the primitive CD34+ AML subfraction compared to normal CD34+ cells [J]. Ann Hematol,2009,88 (6):573-580.
[17]H J de Jonge, Woolthuis C M, Vos A Z, et al. Gene expression profiling in the leukemic stem cell-enriched CD34+ fraction identifies target genes that predict prognosis in normal karyotype AML[J]. Leukemia,2011,25 (12):1825-1833.
[18]Alvares C L, Schenk T, Hulkki S, et al. Tyrosine kinase inhibitor insensitivity of non-cycling CD34+ human acute myeloid leukaemia cells with FMS-like tyrosine kinase 3 mutations[J]. Br J Haematol,2011,154 (4):457-465.
[19]王兴兵,姚军霞,郑金娥,等.115例急性髓细胞白血病多参数流式细胞术免疫表型分析[J].中国实验血液学杂志,2005,13(2)：250-253.
[20]Kishimoto T, KI KutaniH, Von Dem Borne A E G et al. Leucocyte typing Ⅳ [M].NewYork & London:Garland Publishing Inc,1997:929-965.
[21]王清霞,张秋堂,岳保红,等.成人急性髓性白血病免疫学表型分析[J].郑州大学学报(医学版),2008,43(04)：779-781.
[22]江雁,穆红,唐志琴,等.急性白血病系列特异性抗原表达分析[J].国际检验医学杂志2009,30(6)：532-534.
[23]孔令环,韩梅,鲍彦娜,等.流式细胞术对急性白血病免疫分型的临床意义[J].中国实验诊断学,2010,14(10)：1570-1573.
[24]Roberson JR,Onciu M, Pounds S, et al. Prognostic significance of myeloperoxidase expression in childhood acute myeloid leukemia[J]. Pediatr Blood Cancer,2008, 50(3):542-548.
[25]Higa B, Alkan S, Barton K, et al. Precursor B-cell acute lymphoblastic leukaemia with FAB L3 (i.e., Burkitt's leukaemia/lymphoma) morphology and co-expression of monoclonal surface light chains and Tdt:report of a unique case and review of the literature[J]. Pathology,2009,41 (5):495-498.
[26]张颜,何明生.急性淋巴细胞白血病靶向基因治疗的研究进展[J].当代医学,2010,(02)：35-36+47.
[27]Mason DY, Cordell JL, Brown MH, et al. CD79a:a novel marker for B-cell neoplasms in routinely processed tissue samples [J].Blood,1995,86(4):1453-1459.
[28]Melinda S. Gordon,Cindy M. Kanegai,Jeanette R. Doerr et al.Somatic hypermutation of the B cell receptor genes B29 (Igβ, CD79b) and mb1 (Igα, CD79a) PNAS[J] 2003,100(7)4126-4131.
[29]Abdelhaleem M. The expression of cytoplasmic CD79a correlates with surface CD3 expression in childhood T-ALL lymphoblasts.Leuk Res,2008,32 (3):511-512.
[30]Fujisawa S, Tanioka F, Matsuoka T, et al. CD7/CD19 double-positive T-cell acute lymphoblastic leukemia. Int J Hematol,2006,83(4):324-327.
[31]Gujral S, Subramanian P G, Dasgupta A. Diversity amongst various guidelines for immunopheno typing[J]. Cytometry A,2009,75 (7):560-561.
[32]Zhang J J, Zhang N, Zhang X F, et al. Immunophenotyping characteristics of biphenotypic acute leukemia and analysis of curative effect[J]. Zhongguo Shi Yan Xue Ye Xue Za Zhi,2011,19 (2):317-320.
[33]张之南,沈悌.血液病诊断及疗效标准[M](第3版),科学出版社,2007,184-214.
[34]李晓玲.李睿.陈燕,等.急性混合细胞白血病临床特点及免疫学表达的研究[J].中国实验血液学杂志,2007,15(3)636-639.
[35]Wang QR, Shi JY, Shi L, Chen SJ. Mutational Detection of Full-length Mixed Lineage Leukemia Gene in Patients with De Novo AML-M4 and M5 [J].Zhongguo Shi Yan Xue Ye Xue Za Zhi.2012,20 (1):12-17.
[1]Mazur G,Halon A,Wrobel T, et al.Contribution of flow cytometric immunopheno typing and bone marrow trephine biopsy in the detection of lymphoid bone marrow infiltration in non-Hodgkins lymphoma[J].Neoplasma,2004,51(3):159-163
[2]Tamok A,Hambsch J,Chen R,et al.Cytometric bead array to measure six cytokines in twenty-five microliters of serum[J].ClinChem,2003,49(6):1000-1002.
[3]张艺.流式细胞仪构成与工作原理[J].医疗设备信息,2005,20(8)：25-26.
[4]Grogan WM. Guide to flow cytometry methods [M].New York Marcel Dekker Inc,1990
[5]郑海萍,李艳辉.流式细胞仪的原理和应用[J].医疗器械2003,16(3)：286.
[6]Hirons GT, TO T, YOY. New very flow cytometry[J]. Cytometry,1994,1:129-137.
[7]李超,韩金路,王玉刚,等.流式细胞仪的工作原理及应用[J].中国实用医药,2009,4(20)：235-236.
[8]沈二霞.流式细胞仪的原理和临床应用[J].现代医学仪器与应用,2008,20(1)：49-53.
[9]沈关心,周汝麟.现代免疫学实验技术[M].武汉：湖北科学技术出版社,2002.
[10]杨峥,莫武宁,李山,等.505例白血病应用流式细胞术免疫分型与FAB分型的结果分析[J].中国实验诊断学,2010,14(8)：1238-1241.
[11]Jaffe ES,Harris NL,Stein H,el al.Pathology and genetics of tumours of haematopoietic and lymphoid tissues.IARC Press.Lyon,2001:91-92.
[12]Kaleem Z, White G. Diagnostic criteria for minimally differentiated acute myeloid leukemia(AML-MO). Evaluation and aproposal. Am J Clin Pathol,2001,115:876-884
[13]上海中美白血病协作组,微分化急性髓系白血病21例临床分型及诊断标准复习[J],中华血液学杂志,2008.29(12)847-849.
[14]Bene MC, Bernier M, Casasnovas RO,et al. Acute myeloid leukemia MO:haematological immunophenotypic and cytogenetic characteristics and their prognostic significance analysis in 241 patients[J]. Br J Haematol,2001,113 (3),737-745.
[15]赵菲,陈燕.急性髓系白血病M-1的免疫学特征和预后分析[J].中国实验血液学杂志,2007,15(04)：687-691.
[16]易丽萍,郭新红.急性髓细胞性白血病M2患者形态学及免疫学分型与融合基因染色体检查的价值[J].中国组织工程研究与临床康复,2011,15(10)：1777-1782.
[17]佟海伙,王慧涵,张继红,等.192例急性髓系白血病免疫表型、细胞遗传学及临床特征的研究[J].中国实验血液学杂志,2009,17(05)：1174-1178.
[18]程薇,马保根,翟亚萍,等.139例急性髓系白血病免疫分型特点分析[J].肿瘤防治研究,2009,(03)：235-237.
[19]张颜,何明生.急性淋巴细胞白血病靶向基因治疗的研究进展[J].当代医学,2010,(02)：35-36+47.
[20]Gujral S,Subramanian P G,Dasgupta A.Diversity amongst various guidelines for immunopheno typing[J]. Cytometry A,2009,75 (7):560-561.
[21]Zhang J J, Zhang N, Zhang X F, et al. Immunophenotyping characteristics of biphenotypic acute leukemia and analysis of curative effect[J]. Zhongguo Shi Yan Xue Ye Xue Za Zhi, 2011,19 (2):317-320.
[22]袁婷婷,刘艳荣,常艳,等.95例初治急性T淋巴细胞白血病患者免疫表型特点[J].中国实验血液学杂志,2011,19(05)：1134-1140.
[23]宣恒报,李玉峰,曹维克,等.免疫分型在一些特殊类型急性白血病诊断中的应用[J].现代医学,2008,36(05)：312-315.
[24]Wang QR, Shi JY, Shi L, et al. Mutational Detection of Full-length Mixed Lineage Leukemia Gene in Patients with De Novo AML-M4 and M5 [J].Zhongguo Shi Yan Xue Ye Xue Za Zhi.2012,20 (1):12-17.
[25]Kern W,Voskova D,Schoch C, et al.Determination of relapse riskbased on assessment of minimal residual disease during complete remission by muhiparameter flow cytometry in unseleeted patients with acute myeloid leukemia[J].Blood,2004,104(10):3078-3085.
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