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4146 Reproducible Diagnosis of Chronic Lymphocytic Leukemia (CLL) By Flow Cytometry: An European Research Initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation Project

CLL: Biology and Pathophysiology, excluding Therapy
Program: Oral and Poster Abstracts
Session: 641. CLL: Biology and Pathophysiology, excluding Therapy: Poster III
Monday, December 7, 2015, 6:00 PM-8:00 PM
Hall A, Level 2 (Orange County Convention Center)

Andy C Rawstron1, Karl-Anton Kreuzer2*, Asha Soosapilla3*, Martin Spacek4*, Peter Gambell5*, Neil McIver-brown6*, Katherina Psarra7*, Maria Arroz8*, Raffaella Milani9*, Javier de la Serna10*, M. Teresa Cedena10*, Ozren Jaksic11, Josep F. Nomdedeu12*, Carol Moreno13*, Gian Matteo Rigolin14*, Antonio Cuneo14*, Preben Johansen15*, Hans Erik Johnsen16, Richard Rosenquist17, Carsten Utoft Niemann18, Wolfgang Kern, MD19, David A Westerman20, Marek Trneny21, Stephen P. Mulligan22, Peter Hillmen23, David G Oscier24, Michael Hallek, MD25, Paolo Ghia, MD, PhD26 and Emili Montserrat27

1Haematological Malignancy Diagnostic Service, St. James's n, Leeds, United Kingdom
2Department I of Internal Medicine, University of Cologne, Cologne, Germany
3Laverty Pathology, Macquarie Park, Australia
4University Hospital, Prague, Czech Republic
5Peter MacCallum Cancer Centre, Melbourne, Australia
6Royal Bournemouth Hospital, Bournemouth, United Kingdom
7Evangelismos Hospital, Athens, Greece
8Hospital S. Francisco Xavier, Lisbon, Portugal
9Immunohematology and Transfusion Medicine Unit, San Raffaele Scientific Institute, Milan, Italy
10Hospital Universitario 12 de Octubre, madrid, Spain
11Dubrava University Hospital, Zagreb, Croatia
12Department of Hematology Lab, On behalf of CETLAM Cooperative Group, Barcelona, Spain
13Hematologia, Hospital De La Santa Creu I Sant Pau, Barcelona, Spain
14Hematology Section, Azienda Ospedaliero-Universitaria S. Anna, University of Ferrara, Ferrara, Italy
15Aalborg University Hospital, Aalborg, Denmark
16Clinical Cancer Research Center, Aalborg University Hospital, Aalborg, Denmark
17Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
18Department of Hematology, Rigshospitalet, Copenhagen, Denmark
19MLL Munich Leukemia Laboratory, Munich, Germany
20Department of Haematology, Peter MacCallum Cancer Centre, Melbourne, Australia
21First Dep. of Medicine, Charles University General Hospital, Prague, Czech Republic
22Department of Haematology, Royal North Shore Hospital, St Leonards, Australia
23St. James's Institute of Oncology, Leeds, United Kingdom
24Department of Molecular Pathology, Royal Bournemouth Hospital, Bournemouth, United Kingdom
25Department of Internal Medicine, University of Cologne, Cologne, Germany
26Università Vita-Salute San Raffaele and IRCCS Istituto Scientifico San Raffaele, Milano, Italy
27Department of Hematology, Hospital Clinic, Barcelona, Spain

BACKGROUND: The WHO and iwCLL diagnostic criteria for CLL rely on morphology and immunophenotype based on the co-expression of CD19/CD5/CD23 on B-cells with weak CD20 and monoclonal sIg expression. These diagnostic criteria are likely to persist in the near future because there is no specific diagnostic molecular abnormality for CLL. The current criteria have some limitations affecting reproducibility, particularly flexibility in marker expression with many centres using a scoring system that permits absence of CD5 or CD23. Potentially informative new markers have been identified but there is no consensus yet on which should be routinely assessed.

AIM: To identify reproducible criteria and to achieve a consensus on markers recommended for the diagnosis of CLL

METHODS: ERIC/ESCCA members were invited to classify 35 flow-cytometry markers as being required or recommended for the diagnosis of CLL. Consensus was considered to be achieved if >75% of participants agreed on the marker classification. A diagnostic panel was identified by the steering committee and characteristics of component markers that could be reproducibly validated within an individual laboratory were identified. The proposed panel was assessed in 13 different centres.

RESULTS: Responses were received from 154 members (100 laboratory staff, 14 clinicians and 36 from both laboratory and clinic) with a diagnostic workload >20 cases per week in 23/154 (15%), 5-20 in 82/154 (53%) and <5 cases per week in 49/154 (32%). The consensus minimum diagnostic panel should include : CD19, CD5, CD20, CD23, Kappa and Lambda. Participants recommended the following markers: CD38, CD45, CD79b, CD10, CD22, CD43, CD200, and FMC7. A minimum and recommended panel with reproducible criteria for component reagents were determined and the criteria were applied to 10,876 cases diagnosed with a B-LPD, of which 8120 were CD5+ B-LPD. Out of 5947 sent as a primary referrals for diagnosis, 4493 (75.6%) met the proposed diagnostic criteria for CLL, 821 (13.8%) did not and had a clear alternative diagnosis (e.g. MCL) and 633 (10.6%) would not be readily classified by flow cytometry if the proposed criteria were applied. Out of 2173 cases previously diagnosed as CLL at another centre, 2028 (93.3%) met the proposed diagnostic criteria, 19 (0.9%)  had a clear alternative diagnosis while 126 (5.8%) did not meet the flow-cytometry criteria.

CONCLUSIONS: We present flow-cytometry criteria for the diagnosis of CLL largely consistent with current practice. In addition, reproducible definitions of the required expression pattern and performance characteristics of reagents are provided. Prospective evaluation of the proposed criteria as well as a parallel project to facilitate definitive diagnosis of CD5+ B-LPD cases that do not meet the proposed criteria are underway. 

Table: required and recommended markers for use in the diagnosis of CLL with reagent specification based on expression patterns in normal peripheral blood.

Weak expression = median fluorescence intensity at least 20% lower than median for normal peripheral blood B-cells, reference range determined within each laboratory, based on ICSH/ISLH/CLIA guidelines for reproducibility

*consensus, not specifically validated

 

Disclosures: Rawstron: Abbvie: Honoraria ; Pharmacyclics: Research Funding ; Celgene: Honoraria ; Roche: Honoraria ; BD Biosciences: Patents & Royalties ; Gilead: Honoraria , Research Funding . Cuneo: Roche: Speakers Bureau ; Gilead: Speakers Bureau ; Jannsen: Speakers Bureau ; Celgene: Speakers Bureau ; Novartis: Speakers Bureau . Kern: MLL Munich Leukemia Laboratory: Employment , Equity Ownership . Trneny: Celgene: Consultancy , Honoraria , Other: Travel, accommodations, expenses , Research Funding . Mulligan: Celgene: Consultancy , Honoraria ; Janssen: Consultancy , Honoraria , Speakers Bureau ; Roche: Consultancy , Honoraria , Research Funding , Speakers Bureau ; Sanofi Aventis: Research Funding . Hillmen: Celgene: Research Funding ; Gilead: Consultancy , Honoraria , Research Funding ; AbbVie: Consultancy , Honoraria , Research Funding ; Janssen: Consultancy , Honoraria , Research Funding ; GSK: Consultancy , Honoraria , Research Funding ; Novartis: Honoraria , Research Funding ; Roche: Consultancy , Honoraria , Research Funding . Hallek: Celgene: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Mundipharma: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Roche: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Gilead: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; AbbVie: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Boehringher Ingelheim: Honoraria , Other: Speakers Bureau and/or Advisory Boards ; GSK, Genentech: Membership on an entity’s Board of Directors or advisory committees , Research Funding ; Janssen: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding ; Pharmacyclics: Honoraria , Other: Speakers Bureau and/or Advisory Boards , Research Funding . Ghia: AbbVie: Consultancy ; Janssen: Consultancy ; Roche: Consultancy , Research Funding ; Adaptive: Consultancy ; Gilead: Consultancy , Research Funding , Speakers Bureau ; GSK: Research Funding ; Acerta Pharma BV: Research Funding ; Pharmacyclics: Consultancy .

*signifies non-member of ASH