• Document: Ten-Color 15 Antibody Flow Cytometry Panel for Immunophenotyping of Lymphocyte Population
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Ten-Color 15 Antibody Flow Cytometry Panel for Immunophenotyping of Lymphocyte Population Amr Rajab BSc, MLT, QCYM (ASCP) Flow cytometry Technical Specialist LifeLabs | 100 International Blvd. | Toronto, ON M9W 6J6 T 416-675-4530 Ext. 42096 | F 416-213-4206 | C 416-617-5910 E Amr.Rajab@LifeLabs.com Disclosure Have no conflict of interest to disclose for this presentation Topics • Development & validation of the of lymphoproliferative disorder screening tube (LPD-ST) • Technology Transfer Using Harmonemia Concept • Pattern recognition and empty spaces analysis GOAL To developed a LPD-ST with the aim to provide comprehensive immunophenotyping of lymphocyte subsets with minimal need for additional testing Why the LPD-ST? 10-color lymphoproliferative disorder (LPD) panel University Health Network, Toronto, Canada Mahdi T, Rajab A, Padmore R, Porwit A. Characteristics of lymphoproliferative disorders with more than one aberrant cell population as detected by 10-color flow cytometry. Cytometry Part B, Clinical cytometry. 2016. Why the LPD-ST? • ~ 5000 samples (PB, BMA, Lymph node cell suspensions and cytology) per year analyzed using LPD panel • 80% of LPD cases we diagnosed in 2014 in blood are CLL/MBL. MYELOID/ LYMPHOID SCREENING TUBE 10 COLORS, 14 ANTIBODIES Rajab A, Porwit A. Screening bone marrow samples for abnormal lymphoid populations and myelodysplasia- related features with one 10-color 14-antibody screening tube. Cytometry Part B, Clinical cytometry. 2015;88(4):253-60. Similar approach has been applied by other groups in 4- color, 6-color, 8-color and 10-color settings • Quijano S, …etc. Spanish Group for the Study of CNS Disease in NHL.Identification of leptomeningeal disease in aggressive B-cell non-Hodgkin's lymphoma: improved sensitivity of flow cytometry. J Clin Oncol. 2009 Mar 20;27(9):1462-9. • Preijers FW…etc. B. OMIP-010: a new 10-color monoclonal antibody panel for polychromatic immunophenotyping of small hematopoietic cell samples. Cytometry A. 2012 Jun;81(6):453-5. • Costa ES, …etc. A new automated flow cytometry data analysis approach for the diagnostic screening of neoplastic B-cell disorders in peripheral blood samples with absolute lymphocytosis. Leukemia. 2006 Jul;20(7):1221-30 • van Dongen JJ,…etc. EuroFlow Consortium (EU-FP6, LSHB-CT-2006-018708). EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia. 2012 Sep;26(9):1908-75. • Hedley BD, Keeney M, Popma J, Chin-Yee I. Novel lymphocyte screening tube using dried monoclonal antibody reagents. Cytometry B Clin Cytom. 2015 May 4 Materials and Methods Development of the LPD screening tube (LPD-ST) Ab FITC CD4 + Kappa PE CD8 + Lambda LPD Screening Tube ver. I ECD CD3 + CD14 PC5.5 CD5 PC7 CD20 + CD56 APC CD10 APC-A700 CD19 APC-A750 CD200 PB CD57 +CD23 KO CD45 Materials and Methods Development of the LPD screening tube (LPD-ST) Pilot study: • Blood (n=10), BMA (n=3) and tissue samples (n=2). • 3 normal specimens, one reactive blood sample with increased γδ T- cells, 5 CLL, 1 CD5-/CD10- B-cell LPD, 1 large B-cell lymphoma, 1 Follicular lymphoma, 1 hairy cell leukemia (HCL), 1 T-cell Large granular lymphocytosis (LGL) and 1 angioimmunoblastic T-cell lymphoma (AITL). Materials and Methods Development of the LPD screening tube (LPD-ST) Results: • Final diagnosis could be rendered in 10 cases (67%) including 3 normal cases, 5 CLL, 1 large B-cell and 1 follicular lymphoma. • 5 cases (33%) required further investigation including a T-cell LGL, a CD5-/CD10- B-cell LPD, AITL, HCL and one case with increased γδ T-cells. ICCS 2015, Abstract #88, Poster Session 2 on Monday, October 12, 2015 Ten-Color 15 Antibody Flow Cytometry Panel for Immunophenotyping of Lymphocyte population Amr Rajab, Graeme Quest, Jenni

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