Value of 4-color and 10-antibody panel of flow cytometry immunophenotyping in the diagnosis of multiple myeloma_West China Medical Journal

Authors：

 TANG Jie ¹ , JIANG Nenggang ² , LIANG Xiaogong ³ , XUE Li ¹ , XUE Bingrong ¹

1. Department of Clinical Laboratory Examination, Mianyang Central Hospital, Mianyang, Sichuan 621000, P. R. China;
2. Department of Laboratory Medicine, West China Hospital, Sichuan university, Chengdu, Sichuan 610041, P. R. China;
3. Department of Hematology, Mianyang Central Hospital, Mianyang, Sichuan 621000, P. R. China;

Corresponding author：

TANG Jie, Email: tjandbetty@aliyun.com

Keywords：

Multiple myeloma; Flow cytometry; Immunophenotyping; Diagnosis

DOI：

10.7507/1002-0179.201512161

Video：

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Objective To investigate the value of a 4-color and 10-antibody flow cytometry immunophenotyping panel using 10 antibodies including CD45, CD38, CD19, CD56, CD20, CD5, CD10, human leukocyte antigen-DR (HLA-DR), κ antibody and λ antibody marked by four kinds of fluorescein including R-phycoerythrin (PE), fluorescein isothiocyanate (FITC), peridinin chlorophy Ⅱ protein (PerCP) and allophycocyanin (APC) in the diagnosis of multiple myeloma (MM). Methods A 4-color and 10-antibody flow cytometry immunophenotyping panel which used CD45^dim/-/CD38^high as gating strategy supplemented by CD19, CD56, CD20, CD10, CD5, HLA-DR, κ antibody and λ antibody was used to test the bone marrow (BM) specimens of 45 MM patients treated between December 2013 and March 2015. Then by morphological examination, we analyzed the quantitative results and characteristics of myeloma cells. Results In all the 45 MM patients, the myeloma cell detection rate was 100% by flow cytometry. The proportion range of myeloma cells in BM was between 1.17% and 72.31%, which showed a good consistency with the results of 7.5%-90.0% detected by morphological examination. The positive expression rates of antigen on myeloma cells were: 100.00% for CD38, 11.11% for CD45, 2.22% for CD19, 73.33% for CD56, 17.78% for CD20, 42.22% for HLA-DR, and 0% for CD10 and CD5. About 64.44% of the MM patients were restricted cytoplasmic λ light chain typing, and 35.56% were restricted cytoplasmic κ light chain typing. There was no obvious phenotype difference among the 3 Durie-Salmon stages of MM (P>0.05). The expression of CD56 was different among different immunoglobulin types of MM, and the types of immunoglobulin with an expression from high to low were non-secretory, IgA, IgG, and light chain (P<0.05). Conclusion The 4-color and 10-antibody flow cytometry immunophenotyping panel using 10 antibodies including CD45, CD38, CD19, CD56, CD20, CD5, CD10, HLA-DR, κ antibody and λ antibody marked by four kinds of fluorescein including PE, FITC, PerCP and APC has a good diagnostic value for MM.

Citation： TANG Jie, JIANG Nenggang, LIANG Xiaogong, XUE Li, XUE Bingrong. Value of 4-color and 10-antibody panel of flow cytometry immunophenotyping in the diagnosis of multiple myeloma. West China Medical Journal, 2017, 32(3): 419-424. doi: 10.7507/1002-0179.201512161 Copy

1.	Rajkumar SV, Dimopoulos MA, Palumbo A,et al. International myeloma working group updated criteria for the diagnosis of multiple myeloma. Lancet Oncol, 2014, 15(12): E538-E548.
2.	张明慧, 孙薏, 张俊琪, 等. 硼替佐米联合地塞米松和沙利度胺治疗老年多发性骨髓瘤疗效观察. 华西医学, 2014, 29(4): 603-606.
3.	Biran N, Ely S, Chari A,et al. Controversies in the assessment of minimal residual disease in multiple myeloma: clinical significance of minimal residual disease negativity using highly sensitive techniques. Curr Hematol Malig Rep. 2014, 9(4): 368-378.
4.	中国医师协会血液科医师分会, 中华医学会血液学分会, 中国多发性骨髓瘤工作组. 中国多发性骨髓瘤诊治指南(2013年修订). 中华内科杂志, 2013, 52(9): 791-795.
5.	Carulli G, Ottaviano V, Cannizzo E,et al. Multiparameter immunophenotyping by flow cytometry as a diagnostic tool in multiple myeloma and monoclonal gammopathy of undetermined significance. Clin Ter, 2012, 163(5): 387-392.
6.	Bataille R, Jégo G, Robillard N,et al. The phenotype of normal, reactive and malignant plasma cells. Identification of “many and multiple myelomas” and of new targets for myeloma therapy. Haematologica, 2010, 91(9): 1234-1240.
7.	Paíno T, Sarasquete ME, Paiva B,et al. Phenotypic, genomic and functional characterization reveals no differences between CD138⁺⁺ and CD138^low subpopulations in multiple myeloma cell lines. PLoS One, 2014, 9(3): e92378.
8.	Caraux A, Klein B, Paiva B,et al. Circulating human B and plasma cells. Age-associated changes in counts and detailed characterization of circulating normal CD138^- and CD138⁺ plasma cells. Haematologica, 2010, 95(6): 1016-1020.
9.	Chillemi A, Zaccarello G, Quarona V,et al. CD38 and bone marrow microenvironment. Front Biosci (Landmark Ed), 2014, 19: 152-162.
10.	Paiva B, Almeida J, Pérez-Andrés M,et al. Utility of flow cytometry immunophenotyping in multiple myeloma and other clonal plasma cell-related disorder. Cytometry B Clin Cytom. 2010, 78(4): 239-252.
11.	Berdeja JG. Lorvotuzumab mertansine: antibody-drug-conjugate for CD56⁺ multiple myeloma. Front Biosci (Landmark Ed). 2014, 1(19): 163-170.
12.	Naseem S, Kaur S, Gupta R,et al. Plasma cell leukemia: case series from a tertiary center with review of literature. Indian J Hematol Blood Transfus, 2012, 28(1): 10-14.
13.	Kumar S, Kimlinger T, Morice W. Immunophenotyping in multiple myeloma and related plasma cell disorders. Best Pract Res Clin Haematol, 2010, 23(3): 433-451.
14.	Chastain EC, Duncavage EJ. Clinical prognostic biomarkers in chronic lymphocytic leukemia and diffuse large B-cell lymphoma. Arch Pathol Lab Med, 2015, 139(5): 602-607.
15.	刘艳荣. 实用流式细胞术. 北京: 北京大学医学出版社, 2010: 348.
16.	Manasanch EE, Salem DA, Yuan CM,et al. Flow cytometric sensitivity and characteristics of plasma cells in patients with multiple myeloma or its precursor disease: influence of biopsy site and anticoagulation method. Leuk Lymphoma, 2015, 56(5): 1416-1424.

1. Rajkumar SV, Dimopoulos MA, Palumbo A,et al. International myeloma working group updated criteria for the diagnosis of multiple myeloma. Lancet Oncol, 2014, 15(12): E538-E548.
2. 张明慧, 孙薏, 张俊琪, 等. 硼替佐米联合地塞米松和沙利度胺治疗老年多发性骨髓瘤疗效观察. 华西医学, 2014, 29(4): 603-606.
3. Biran N, Ely S, Chari A,et al. Controversies in the assessment of minimal residual disease in multiple myeloma: clinical significance of minimal residual disease negativity using highly sensitive techniques. Curr Hematol Malig Rep. 2014, 9(4): 368-378.
4. 中国医师协会血液科医师分会, 中华医学会血液学分会, 中国多发性骨髓瘤工作组. 中国多发性骨髓瘤诊治指南(2013年修订). 中华内科杂志, 2013, 52(9): 791-795.
5. Carulli G, Ottaviano V, Cannizzo E,et al. Multiparameter immunophenotyping by flow cytometry as a diagnostic tool in multiple myeloma and monoclonal gammopathy of undetermined significance. Clin Ter, 2012, 163(5): 387-392.
6. Bataille R, Jégo G, Robillard N,et al. The phenotype of normal, reactive and malignant plasma cells. Identification of “many and multiple myelomas” and of new targets for myeloma therapy. Haematologica, 2010, 91(9): 1234-1240.
7. Paíno T, Sarasquete ME, Paiva B,et al. Phenotypic, genomic and functional characterization reveals no differences between CD138⁺⁺ and CD138^low subpopulations in multiple myeloma cell lines. PLoS One, 2014, 9(3): e92378.
8. Caraux A, Klein B, Paiva B,et al. Circulating human B and plasma cells. Age-associated changes in counts and detailed characterization of circulating normal CD138^- and CD138⁺ plasma cells. Haematologica, 2010, 95(6): 1016-1020.
9. Chillemi A, Zaccarello G, Quarona V,et al. CD38 and bone marrow microenvironment. Front Biosci (Landmark Ed), 2014, 19: 152-162.
10. Paiva B, Almeida J, Pérez-Andrés M,et al. Utility of flow cytometry immunophenotyping in multiple myeloma and other clonal plasma cell-related disorder. Cytometry B Clin Cytom. 2010, 78(4): 239-252.
11. Berdeja JG. Lorvotuzumab mertansine: antibody-drug-conjugate for CD56⁺ multiple myeloma. Front Biosci (Landmark Ed). 2014, 1(19): 163-170.
12. Naseem S, Kaur S, Gupta R,et al. Plasma cell leukemia: case series from a tertiary center with review of literature. Indian J Hematol Blood Transfus, 2012, 28(1): 10-14.
13. Kumar S, Kimlinger T, Morice W. Immunophenotyping in multiple myeloma and related plasma cell disorders. Best Pract Res Clin Haematol, 2010, 23(3): 433-451.
14. Chastain EC, Duncavage EJ. Clinical prognostic biomarkers in chronic lymphocytic leukemia and diffuse large B-cell lymphoma. Arch Pathol Lab Med, 2015, 139(5): 602-607.
15. 刘艳荣. 实用流式细胞术. 北京: 北京大学医学出版社, 2010: 348.
16. Manasanch EE, Salem DA, Yuan CM,et al. Flow cytometric sensitivity and characteristics of plasma cells in patients with multiple myeloma or its precursor disease: influence of biopsy site and anticoagulation method. Leuk Lymphoma, 2015, 56(5): 1416-1424.

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