多参数流式细胞术检测多发性骨髓瘤细胞免疫表型及微小残留病灶的临床价值
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摘要
目的探讨多参数流式细胞术在多发性骨髓瘤(multiple myeloma,MM)患者细胞免疫表型及其微小残留病灶(MRD)检测中的应用价值。方法选取我院2014年1月至2016年7月期间收治的多发性骨髓瘤患者65例作为本研究观察组,并选取同期入我院健康体检正常者40例作为本研究对照组。所有受试对象均进行多参数流式细胞术检测,所有患者均接受临床治疗,比较观察组治疗前与对照组免疫表型的差异,观察MM患者治疗后的MRD阴性与阳性情况并分析多参数流式细胞术检测免疫表型对MRD诊断的敏感度、特异度及ROC曲线分析;对患者随访,比较MM治疗后MRD阳性与阴性组患者无症状进展期PFS。结果观察组受试对象CD19阳性表达率明显低于对照组,CD56、CD117以及CD138阳性表达率均明显高于对照组,比较差异均有统计学意义(P<0. 01);两组患者CD38、CD13、CD33、CD20以及CD34阳性表达率之间比较无统计学意义(P>0. 05)。CD38 (+)/CD138 (+)为MM阳性诊断标准可知其敏感度为89. 23%(58/65),CD38 (+)/CD138 (+)与CD19 (-)联合检测作为新的标准显示其诊断敏感度为86. 15%(56/65),特异度为76. 92%(50/65),准确度为76. 92%[(56+50)/(56+9+50+15)],ROC曲线分析显示其曲线下面积为0. 823 (0. 747,0. 899)>0. 5。经治疗后,65例MM患者中MRD阴性患者16例,阳性患者49例。分别以CD38 (+)/CD138 (+)/CD19 (-)、CD38 (+)/CD138 (+)/CD19 (-)/CD56 (+)以及CD38 (+)/CD138 (+)/CD19 (-)/CD117 (+)为诊断标准对治疗后MM患者进行MRD诊断,其中CD38 (+)/CD138 (+)/CD19 (-)诊断的敏感度(93. 75%)最高,ROC曲线下面积最大[0. 795 (0. 681,0. 909)],CD38 (+)/CD138 (+)/CD19 (-)/CD117 (+)的特异度最高(95. 92%),比较差异均有统计学意义(P<0. 01)。MRD阳性患者的平均PFS为(l5. 47±4. 29)月,MRD阴性患者的平均PFS为(24. 93±5. 46)月,MRD阴性患者PFS明显长于MRD阳性患者,比较差异有统计学意义(t=11. 337,P=0. 000)。结论多参数流式细胞术检测在多发性骨髓瘤患者细胞免疫表型及其微小残留病灶检测中均具有较高应用价值,值得在临床推广应用。
Objective To investigate the value of multi-parameter flow cytometry in the detection of cellular immunophenotype and minimal residual lesion in patients with multiple myeloma( MM). Methods 65 patients with MM treated in our hospital from January 2014 to July 2016 were selected as the observation group and 40 patients with normal physical examinations in our hospital were selected as the control group. All the subjects were subjected to the test of multi-parameter flow cytometry. All patients received clinical treatment. The difference of immunophenotype between the observation group before the treatment and the control group was compared. The negative and positive status of MRD after treatment in patients with MM were observed and analyzed. The sensitivity,specificity and ROC curve of MRP were analyzed by flow cytometry. The patients were followed up for 3 years. Comparison was made of asymptomatic PFS in patients with positive and negative MRD after MM therapy. Results The positive expression rate of CD19,CD56,CD117 and CD138 in the observation group were significantly higher than that in the control group( P<0. 01). There was no statistically significant difference in the expression of CD38,CD13,CD33,CD20,CD9 and CD34( P>0. 05). With CD38( +)/CD138( +) as the new criteria for MM-positive diagnosis,the sensitivity was 89. 23%( 58/65),while with combination of CD38( +)/CD138( +) and CD19(-) as the new criteria,its sensitivity was 86. 15%( 56/65),the specificity was 76. 92%( 50/65)and the accuracy was 76. 92% [( 56 +50)/( 56 +9 +50 +15) ]. The ROC curve showed that the area under the curve was 0. 823( 0. 747,0. 899)( >0. 5). After treatment,16 of 65 patients with MM were MRD negative and 49 were positive. With CD38( +)/CD138( +)/CD19(-),CD38( +)/CD138( +)/CD19(-)/CD56( +) as criteria for MRD diagnosis in MM patients,the sensitivity of CD38( +)/CD138( +)/CD19(-) was the highest( 93. 75%),and the area under ROC curve was the largest[0. 795( 0. 681,0. 909) ], and the specificity of CD38( +)/CD138( +)/CD19(-)/CD117( +) was the highest( 95. 92%),with statistically significant difference( P<0. 01). The mean PFS of MRD-positive patients was( 15. 47±4. 29) months,and that of MRD-negative patients was( 24. 93±5. 46) months. The PFS of MRD-negative patients was significantly longer than that of MRD-positive patients,with statistically significant difference( t = 11. 337,P <0. 001). Conclusion Multi-parameter flow cytometry has high application value in detecting cellular immunophenotypes and minimal residual lesions in patients with multiple myeloma,which is worth popularizing in clinical practice.
Objective To investigate the value of multi-parameter flow cytometry in the detection of cellular immunophenotype and minimal residual lesion in patients with multiple myeloma( MM). Methods 65 patients with MM treated in our hospital from January 2014 to July 2016 were selected as the observation group and 40 patients with normal physical examinations in our hospital were selected as the control group. All the subjects were subjected to the test of multi-parameter flow cytometry. All patients received clinical treatment. The difference of immunophenotype between the observation group before the treatment and the control group was compared. The negative and positive status of MRD after treatment in patients with MM were observed and analyzed. The sensitivity,specificity and ROC curve of MRP were analyzed by flow cytometry. The patients were followed up for 3 years. Comparison was made of asymptomatic PFS in patients with positive and negative MRD after MM therapy. Results The positive expression rate of CD19,CD56,CD117 and CD138 in the observation group were significantly higher than that in the control group( P<0. 01). There was no statistically significant difference in the expression of CD38,CD13,CD33,CD20,CD9 and CD34( P>0. 05). With CD38( +)/CD138( +) as the new criteria for MM-positive diagnosis,the sensitivity was 89. 23%( 58/65),while with combination of CD38( +)/CD138( +) and CD19(-) as the new criteria,its sensitivity was 86. 15%( 56/65),the specificity was 76. 92%( 50/65)and the accuracy was 76. 92% [( 56 +50)/( 56 +9 +50 +15) ]. The ROC curve showed that the area under the curve was 0. 823( 0. 747,0. 899)( >0. 5). After treatment,16 of 65 patients with MM were MRD negative and 49 were positive. With CD38( +)/CD138( +)/CD19(-),CD38( +)/CD138( +)/CD19(-)/CD56( +) as criteria for MRD diagnosis in MM patients,the sensitivity of CD38( +)/CD138( +)/CD19(-) was the highest( 93. 75%),and the area under ROC curve was the largest[0. 795( 0. 681,0. 909) ], and the specificity of CD38( +)/CD138( +)/CD19(-)/CD117( +) was the highest( 95. 92%),with statistically significant difference( P<0. 01). The mean PFS of MRD-positive patients was( 15. 47±4. 29) months,and that of MRD-negative patients was( 24. 93±5. 46) months. The PFS of MRD-negative patients was significantly longer than that of MRD-positive patients,with statistically significant difference( t = 11. 337,P <0. 001). Conclusion Multi-parameter flow cytometry has high application value in detecting cellular immunophenotypes and minimal residual lesions in patients with multiple myeloma,which is worth popularizing in clinical practice.
引文
[1]李翰卿,翟勇平.流式细胞术检测多发性骨髓瘤免疫表型和残留病灶的研究进展[J].中国实验血液学杂志,2015,23(1):241-245.
[2]唐洁,梁效功,薛丽,等.多参数流式细胞术在多发性骨髓瘤早期诊断中的应用价值[J].中国卫生检验杂志,2016,26(2):236-239.
[3] Paiva B,Merino J,San Miguel JF. Utility of flow cytometry studies in the management of patients with multiple myeloma[J]. Current Opinion in Oncology,2016,28(6):511-517.
[4]俞晴,施菊妹,陶怡.微小残留病灶在多发性骨髓瘤中的临床意义及其检测方法[J].中国实验血液学杂志,2017,25(3):961-964.
[5] Marchesi F,Masi S,Summa V,et al. Flow cytometry characterization in central nervous system and pleural effusion multiple myeloma infiltration:an Italian national cancer institute experience[J]. British Journal of Haematology,2016,172(6):980.
[6]罗文丰,邹兴立,杨竹.流式细胞术检测多发性骨髓瘤中骨髓细胞表面免疫表型的价值[J].中国肿瘤临床与康复,2017,12(8):410-412.
[7] Maga S,Brosseau C,Descamps G,et al. A simple flow cytometry-based barcode for routine authentication of multiple myeloma and mantle cell lymphoma cell lines[J]. Cytometry Part A,2015,87(4):285-288.
[8]张之南,沈悌.血液病诊断及疗效标准[M].第3版.北京:科学出版社,2007:110.
[9] Papanikolaou X,Alapat D,Rosenthal A,et al. The flow cytometry-defined light chain cytoplasmic immunoglobulin index and an associated 12-gene expression signature are independent prognostic factors in multiple myeloma[J]. Leukemia,2015,29(8):1713-1720.
[10]张宏,范银银,戴晓莉,等. 3种设门方法在多发性骨髓瘤免疫表型分析中的应用比较[J].临床检验杂志,2015,33(3):181-184.
[11] Paiva B,Cedena MT,Puig N,et al. Minimal residual disease monitoring and immune profiling using second generation flow cytometry in elderly multiple myeloma[J]. Blood,2016,127(25):3165.
[12] Paiva B,Chandia M,Puig N,et al. The prognostic value of multiparameter flow cytometry minimal residual disease assessment in relapsed multiple myeloma[J]. Haematologica,2015,100(2):53-55.
[13]张翠萍,汪鹏,李庆,等.发性骨髓瘤免疫表型特征及其临床意义[J].中国临床保健杂志,2016,19(3):228-231.
[14] Flores-Montero J,Sanoja-Flores L,Paiva B,et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma[J]. Leukemia,2017,2(14):131-132.
[15]芦晗,王志银,李强. 3种实验技术在多发性骨髓瘤检测中的应用[J].检验医学与临床,2015,12(9):1278-1280.
[16] Gonsalves WI,Rajkumar SV,Dispenzieri A,et al. Quantification of circulating clonal plasma cells via multiparametric flow cytometry identifies patients with smoldering multiple myeloma at high risk of progression[J]. Leukemia, 2016, 31(1):130.
[17]常子维,朱华峰,冯苗娟,等. 160例多发性骨髓瘤患者免疫表型特征及其临床意义[J].现代生物医学进展,2016,16(26):5179-5183.
[18] Iii HRB,Dasari S,Dispenzieri A,et al. Clonotypic Light Chain Peptides Identified for Monitoring Minimal Residual Disease in Multiple Myeloma Without Bone Marrow Aspirates[J].Clinical Chemistry,2016,62(1):243-251.
[19] Pandiella A,Carvajal-Vergara X,Tabera S,et al. Imatinib mesylate(STI571)inhibits multiple myeloma cell proliferation and potentiates the effect of common antimyeloma agents[J].British Journal of Haematology,2015,123(5):858-868.
[20] Sonneveld P,Broijl A. Treatment of relapsed and refractory multiple myeloma[J]. Haematologica,2016,101(4):396-397.
[2]唐洁,梁效功,薛丽,等.多参数流式细胞术在多发性骨髓瘤早期诊断中的应用价值[J].中国卫生检验杂志,2016,26(2):236-239.
[3] Paiva B,Merino J,San Miguel JF. Utility of flow cytometry studies in the management of patients with multiple myeloma[J]. Current Opinion in Oncology,2016,28(6):511-517.
[4]俞晴,施菊妹,陶怡.微小残留病灶在多发性骨髓瘤中的临床意义及其检测方法[J].中国实验血液学杂志,2017,25(3):961-964.
[5] Marchesi F,Masi S,Summa V,et al. Flow cytometry characterization in central nervous system and pleural effusion multiple myeloma infiltration:an Italian national cancer institute experience[J]. British Journal of Haematology,2016,172(6):980.
[6]罗文丰,邹兴立,杨竹.流式细胞术检测多发性骨髓瘤中骨髓细胞表面免疫表型的价值[J].中国肿瘤临床与康复,2017,12(8):410-412.
[7] Maga S,Brosseau C,Descamps G,et al. A simple flow cytometry-based barcode for routine authentication of multiple myeloma and mantle cell lymphoma cell lines[J]. Cytometry Part A,2015,87(4):285-288.
[8]张之南,沈悌.血液病诊断及疗效标准[M].第3版.北京:科学出版社,2007:110.
[9] Papanikolaou X,Alapat D,Rosenthal A,et al. The flow cytometry-defined light chain cytoplasmic immunoglobulin index and an associated 12-gene expression signature are independent prognostic factors in multiple myeloma[J]. Leukemia,2015,29(8):1713-1720.
[10]张宏,范银银,戴晓莉,等. 3种设门方法在多发性骨髓瘤免疫表型分析中的应用比较[J].临床检验杂志,2015,33(3):181-184.
[11] Paiva B,Cedena MT,Puig N,et al. Minimal residual disease monitoring and immune profiling using second generation flow cytometry in elderly multiple myeloma[J]. Blood,2016,127(25):3165.
[12] Paiva B,Chandia M,Puig N,et al. The prognostic value of multiparameter flow cytometry minimal residual disease assessment in relapsed multiple myeloma[J]. Haematologica,2015,100(2):53-55.
[13]张翠萍,汪鹏,李庆,等.发性骨髓瘤免疫表型特征及其临床意义[J].中国临床保健杂志,2016,19(3):228-231.
[14] Flores-Montero J,Sanoja-Flores L,Paiva B,et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma[J]. Leukemia,2017,2(14):131-132.
[15]芦晗,王志银,李强. 3种实验技术在多发性骨髓瘤检测中的应用[J].检验医学与临床,2015,12(9):1278-1280.
[16] Gonsalves WI,Rajkumar SV,Dispenzieri A,et al. Quantification of circulating clonal plasma cells via multiparametric flow cytometry identifies patients with smoldering multiple myeloma at high risk of progression[J]. Leukemia, 2016, 31(1):130.
[17]常子维,朱华峰,冯苗娟,等. 160例多发性骨髓瘤患者免疫表型特征及其临床意义[J].现代生物医学进展,2016,16(26):5179-5183.
[18] Iii HRB,Dasari S,Dispenzieri A,et al. Clonotypic Light Chain Peptides Identified for Monitoring Minimal Residual Disease in Multiple Myeloma Without Bone Marrow Aspirates[J].Clinical Chemistry,2016,62(1):243-251.
[19] Pandiella A,Carvajal-Vergara X,Tabera S,et al. Imatinib mesylate(STI571)inhibits multiple myeloma cell proliferation and potentiates the effect of common antimyeloma agents[J].British Journal of Haematology,2015,123(5):858-868.
[20] Sonneveld P,Broijl A. Treatment of relapsed and refractory multiple myeloma[J]. Haematologica,2016,101(4):396-397.