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- Sprache: Englisch
Flow Cytometry of Hematological Malignancies
Flow cytometric analysis is often integral to the swift and accurate diagnosis of leukemias and lymphomas of the blood, bone marrow, and lymph nodes. However, in the fast-moving and expanding field of clinical hematology, in can be challenging to remain up to speed with the latest biological research and technological innovations. Flow Cytometry of Hematological Malignancies has been designed to provide all those working in hematological oncology with a practical, cutting-edge handbook, featuring clear and fully illustrated guidance on all aspects of cytometry’s role in diagnosis and analysis. This essential second edition includes:
- Explorations of more than 70 antigens
- Full-color illustrations throughout
- New descriptions of recently discovered markers
- WHO classifications of hematological neoplastic diseases
- Helpful tips for result interpretation and analysis
Featuring all this and more, Flow Cytometry of Hematological Malignancies, Second Edition, is an invaluable resource for both trainee and experienced hematologists, hematopathologists, oncologists, and pathologists, as well as medical students and diagnostic lab technicians.
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Table of Contents
Cover
Dedication Page
Title Page
Copyright Page
Foreword to the Second Edition
Foreword to the First Edition
Foreword to the First Edition
Preface to the Second Edition
Preface to the First Edition
Abbreviations
1 Antigens
CD1 Antigens
CD2 Antigen
CD3 Antigen
CD4 Antigen
CD5 Antigen
CD7 Antigen
CD8 Antigen
CD10 Antigen
CD11b Antigen
CD11c Antigen
CD13 Antigen
CD14 Antigen
CD15 Antigen
CD16 Antigen
CD19 Antigen
CD20 Antigen
CD22 Antigen
CD23 Antigen
CD24 Antigen
CD25 Antigen
CD26 Antigen
CD27 Antigen
CD28 Antigen
CD30 Antigen
CD33 Antigen
CD34 Antigen
CD38 Antigen
CD43 Antigen
CD45 Antigen
CD45 Isoforms
CD49 Antigens
CD56 Antigen
CD57 Antigen
CD61 Antigen
CD62L Antigen
CD64 Antigen
CD65 Antigen
CD66c Antigen
CD71 Antigen
CD79 Antigen
CD81 Antigen
CD103 Antigen
CD117 Antigen
CD123 Antigen
CD138 Antigen
CD200 Antigen
CD305 Antigen
CD307 (IRTA) Family
CD371 Antigen
BCL‐2 Protein
Chemokines and Chemokine Receptors
CRLF2 Antigen
Cytotoxic Proteins
HLA‐DR Antigen
Immunoglobulins
KIRs, CD158 Isoforms
Myeloperoxidase
NG2 Antigen
PCA‐1 Antigen
ROR1 Antigen
SLAM Molecules and SLAM‐Associated Protein
SOX11
T‐Cell Receptor (TCR)
Terminal Deoxy‐Nucleotidyl Transferase
Toll‐Like Receptors
VS38 Antigen
ZAP‐70 Protein
2 Diseases
Myeloproliferative Neoplasms
Mastocytosis
Myelodysplastic/Myeloproliferative Neoplasms
Myelodysplastic Syndromes
Myeloid Neoplasms with Germline Predisposition
Acute Myeloid Leukemias
Blastic Plasmacytoid Dendritic Cell Neoplasm (BPDCN/PDCL)
Acute Leukemias of Ambiguous Lineage
Neoplastic Diseases of B and T Lymphatic Precursors
Neoplastic Diseases of Mature B Cells
Neoplastic Diseases of Mature T and NK Cells
Hodgkin Lymphomas
Neoplasms of Histiocytic and Dendritic Cells
3 Appendix
Acute Leukemias Not Recognized by the 2017 WHO Classification
Composite Lymphomas
Hypereosinophilic Syndrome (HES), Lymphocyte Variant
Indolent T Lymphoblastic Proliferation (iT‐LBP)
Polyclonal Lymphocytoses of B Lymphocytes
Small‐Round (Blue) Cell Tumors (SR(B)CT)
References
Index
End User License Agreement
List of Tables
Chapter 1
Table 1.1 Differential diagnosis of CD5(−) CD10(−) B‐CLPDs
Table 1.2 Functional subsets of CD4(+) lymphocytes
Table 1.3 Human memory T‐cell differentiation
Chapter 2
Table 2.1 EGIL scoring system for lineage assignment
Table 2.2 EGIL classification of lymphoblastic leukemias
Table 2.3 Correlations between genotype and phenotype in B‐ALL/LBL
List of Illustrations
Chapter 1
Figure 1.1 Peripheral blood from a subject affected by T‐ALL. The blasts (re...
Figure 1.2 The histogram produced by the cytometric analysis of CD2 is bimod...
Figure 1.3 Pattern of expression of T‐specific CD3 antigen on peripheral T l...
Figure 1.4 Different patterns of expression of T‐specific CD3 antigen on per...
Figure 1.5 If lymphocytes are stained with an anti CD3ε antibody (MoAb SK7 i...
Figure 1.6 Without the steric hindrance caused by the anti CD3ε antibody, Mo...
Figure 1.7 When conjugated with FITC, MoAb T3 (A, B, F) does not recognize γ...
Figure 1.8 Comparison between WT31 and T3‐FITC monoclonal antibodies. Neithe...
Figure 1.9 Combined membrane and cytoplasmic CD3 staining in a case of T‐ALL...
Figure 1.10 A putative case of M/NK‐AL whose blasts (red) display SSC values...
Figure 1.11 Aberrant CD3 expression on neoplastic CD4(+) cells (red) in four...
Figure 1.12 Differential CD4 expression on T lymphocytes in a subject affect...
Figure 1.13 CD4 antigen is expressed by monocytes (red) at a lower intensity...
Figure 1.14 γδ(+) T lymphocytes (red) tend to express CD5 at a lower intensi...
Figure 1.15 Aberrantly low CD5 expression on neoplastic CD3(+) cells (red) i...
Figure 1.16 Analysis of peripheral lymphocytes stained for CD3, CD8α, and CD...
Figure 1.17 CD8(dim+) lymphocytes (red) display CD3(−) (A), CD5(−) (B), CD16...
Figure 1.18 Analysis of peripheral blood lymphocytes from a subject affected...
Figure 1.19 Multicolor analysis of normal CD10(+) haematogones stained for C...
Figure 1.20 Multicolor analysis of peripheral granulocytes in a subject affe...
Figure 1.21 Multicolor analysis of a case of CD10(+) T‐ALL (C) whose blasts ...
Figure 1.22 Analysis of peripheral blood lymphocytes from a subject affected...
Figure 1.23 CD11b analysis carried out on peripheral blood of three normal d...
Figure 1.24 Bivariate analysis carried out for CD11b and CD16 on the normall...
Figure 1.25 The staining of a sample of normal peripheral blood with an anti...
Figure 1.26 The staining of a sample of normal peripheral blood with an anti...
Figure 1.27 Bivariate analysis carried out for CD13 and CD16 on the normally...
Figure 1.28 Analysis of neoplastic cells (red) in peripheral blood (C) and b...
Figure 1.29 Aberrant CD13 expression on neoplastic cells (red) in three case...
Figure 1.30 The staining of a sample of normal peripheral blood with an anti...
Figure 1.31 Multicolor analysis of a case of CD15(+) B‐ALL, whose blasts (re...
Figure 1.32 Multicolor analysis of peripheral granulocytes in a subject affe...
Figure 1.33 Anti CD16 MoAb B73.1 binds to neutrophils of some donors (B), bu...
Figure 1.34 Analysis of neoplastic cells (red) in peripheral blood from thre...
Figure 1.35 Analysis of peripheral blood myeloid blasts (red) from a subject...
Figure 1.36 The analysis carried out on a sample of peripheral blood of a su...
Figure 1.37 Distribution of CD20 antigen expression on neoplastic cells (red...
Figure 1.38 Multicolor analysis of peripheral basophils (red) in a subject a...
Figure 1.39 Distribution of CD22 antigen expression on neoplastic cells (red...
Figure 1.40 CD23 expression on the neoplastic B lymphocytes (red) of two cas...
Figure 1.41 Distribution of CD24 antigen expression on the CD19(+) neoplasti...
Figure 1.42 Multicolor analysis of peripheral granulocytes in a subject affe...
Figure 1.43 Multicolor analysis of a case of CD10(+) (D), CD19(+) (B, C), CD...
Figure 1.44 Multicolor analysis of a case of CD2(−) (E), mCD3(−) (E), cyCD3(...
Figure 1.45 Analysis of a sample of normal bone marrow. A bivariate cytometr...
Figure 1.46 Pattern of CD45 expression on blasts (red) in six cases of acute...
Figure 1.47 Pattern of CD45 expression on blasts (red) in three cases of acu...
Figure 1.48 Pattern of CD45 expression on neoplastic cells (red) in six case...
Figure 1.49 Pattern of CD45 expression on neoplastic cells (red) in six case...
Figure 1.50 Aberrantly high CD45 expression on neoplastic CD3(−) CD4(+) cell...
Figure 1.51 CD45RA expression (B) on the CD3(+) CD5(−) cells (red, A) of a c...
Figure 1.52 CD56 expression (B) on the CD45(−) cells (A) of a rhabdomyosarco...
Figure 1.53 Pattern of CD64 expression on the granulocytes from a subject af...
Figure 1.54 Expression of CD66c (C) recognized by KOR‐SA3544 MoAb on the bla...
Figure 1.55 Distribution of CD79b antigen expression on neoplastic cells (re...
Figure 1.56 Expression of CD103 on the blasts (red) of two CD103(+) cases of...
Figure 1.57 Expression of CD103 on the cells (red) of a leukemized case of E...
Figure 1.58 Typical expression of HLA‐DR on T‐lymphocytes during acute activ...
Figure 1.59 Different pattern of expression of immunoglobulin light chains o...
Figure 1.60 Pattern of expression of CD158a and CD158b isoforms in a case of...
Figure 1.61 T‐cell repertoire assay on the neoplastic cells (red) in a case ...
Figure 1.62 Analysis of ZAP‐70 expression in a case of B‐CLL. ZAP‐70 was ana...
Chapter 2
Figure 2.1 Phenotypic analysis of polymorphonuclear granulocytes (red) in a ...
Figure 2.2 Phenotypic analysis of eosinophils in a subject affected by CEL. ...
Figure 2.3 Phenotypic analysis of mast cells (red) in the BM of a subject af...
Figure 2.4 Distribution of CD14 and CD16 expression in two population of per...
Figure 2.5 Analysis of a sample of peripheral blood in a case of CMML. The n...
Figure 2.6 Analysis of a sample of peripheral blood in a case of CMML. Most ...
Figure 2.7 Combined analysis of SSC and CD45 in a normal sample of periphera...
Figure 2.8 Combined analysis of CD11b and CD16 in the normal maturing BM mye...
Figure 2.9 Combined analysis of CD13 and CD16 in the normal maturing BM myel...
Figure 2.10 Analysis of a sample of BM (A–E) and peripheral blood (F–I) from...
Figure 2.11 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.12 Negative control analysis of a sample of BM from a subject affec...
Figure 2.13 Analysis of a sample of BM from a subject affected by APL with t...
Figure 2.14 Analysis of a sample of BM from a subject affected by APL “varia...
Figure 2.15 Analysis of a sample of BM from a subject affected by AML with m...
Figure 2.16 Analysis of a sample of BM from a subject affected by AML with m...
Figure 2.17 Analysis of samples of BM (A–D) or peripheral blood (E–H) from f...
Figure 2.18 Analysis of a sample of BM from a subject affected by AMoL (AML‐...
Figure 2.19 Analysis of a sample of BM from a subject affected by AMoL (AML‐...
Figure 2.20 Analysis of a sample of BM from a subject affected by AMKL (AML‐...
Figure 2.21 Analysis of a sample of BM from a subject affected by ABL. The n...
Figure 2.22 Analysis of a sample of BM from a subject affected by BPDCN, als...
Figure 2.23 Analysis of a sample of BM from a subject affected by BPDCN, als...
Figure 2.24 From top to bottom, differential behavior of the antigens CD10, ...
Figure 2.25 Analysis of a sample of BM from a subject affected by B‐ALL with...
Figure 2.26 Analysis of a sample of BM from a subject affected by B‐ALL with...
Figure 2.27 Analysis of a sample of BM from a subject affected by T‐ALL char...
Figure 2.28 Analysis of a sample of BM from a subject affected by T‐ALL char...
Figure 2.29 Analysis of a sample of BM from a subject affected by ETP‐ALL. T...
Figure 2.30 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.31 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.32 Analysis of a sample of BM from a subject affected by LPL. The n...
Figure 2.33 Analysis of a sample of BM from a subject affected by LPL. The n...
Figure 2.34 Analysis of a sample of BM from a subject affected by HCL. The n...
Figure 2.35 Comparison of the phenotypical features of the neoplastic lympho...
Figure 2.36 Analysis of a sample of BM from a subject affected by leukemized...
Figure 2.37 Analysis of a sample of BM from a subject affected by leukemized...
Figure 2.38 Analysis of a sample of BM from a subject affected by FCL. The n...
Figure 2.39 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.40 Analysis of a sample of BM (A–D) and of a sample of peripheral b...
Figure 2.41 Analysis of a sample of BM from a subject presenting reactive pe...
Figure 2.42 Analysis of a sample of BM from a subject affected by MGUS. The ...
Figure 2.43 Analysis of a sample of BM from a subject affected by MM. The ne...
Figure 2.44 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.45 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.46 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.47 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.48 Analysis of a sample of BM from a subject affected by ATLL. The ...
Figure 2.49 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.50 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.51 Analysis of a sample of peripheral blood from a subject affected...
Figure 2.52 Analysis of a sample of lymph node from a subject affected by AI...
Figure 2.53 Analysis of a sample of peripheral blood from a subject affected...
Chapter 3
Figure 3.1 Analysis of a sample of peripheral blood from a subject affected ...
Figure 3.2 Analysis of a sample of peripheral blood from a subject affected ...
Figure 3.3 Analysis of a sample of peripheral blood from a subject affected ...
Figure 3.4 Analysis of a sample of peripheral blood from a subject affected ...
Guide
Cover Page
Dedication
Flow Cytometry of Hematological Malignancies
Copyright
Foreword to the Second Edition
Foreword to the First Edition
Foreword to the First Edition
Preface to the Second Edition
Preface to the First Edition
Abbreviations
Table of Contents
Begin Reading
References
Index
WILEY END USER LICENSE AGREEMENT
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To my wife, Angela, and to my sons Stefano, Paola, and Marzia
Flow Cytometry of Hematological Malignancies
Second Edition
CLAUDIO ORTOLANI MD
Adjunct Professor of Laboratory of Diagnostic Cytometry
Urbino University
Urbino, Italy;
Consultant Clinical Pathologist (retired)
Ospedale dell’Angelo
Venice, Italy
This edition first published 2021© 2021 John Wiley & Sons Ltd
Edition HistoryBlackwell Publishing Ltd (1e, 2011)
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by law. Advice on how to obtain permission to reuse material from this title is available at http://www.wiley.com/go/permissions.
The right of Claudio Ortolani be identified as the author of this work has been asserted in accordance with law.
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Library of Congress Cataloging‐in‐Publication Data
Names: Ortolani, C. (Claudio), author.Title: Flow cytometry of hematological malignancies / Claudio Ortolani.Description: Second edition. | Hoboken, NJ : Wiley‐Blackwell, 2021. |Includes bibliographical references and index. | Description based onprint version record and CIP data provided by publisher; resource notviewed.Identifiers: LCCN 2020028333 (print) | LCCN 2020028334 (ebook) | ISBN9781119611301 (epub) | ISBN 9781119611271 (Adobe PDF) | ISBN 9781119611257(cloth) Subjects: | MESH: Hematologic Neoplasms–diagnosis | Flow Cytometry–methodsClassification: LCC RC280.H47 (ebook) | LCC RC280.H47 (print) | NLM WH 525 |DDC 616.99/418–dc23LC record available at https://lccn.loc.gov/2020028333LC record available at https://lccn.loc.gov/2020028334
Cover Design: WileyCover Image: courtesy of Claudio Ortolani
Foreword to the Second Edition
It has been about a decade since Dr. Claudio Ortolani gave us the first edition of Flow Cytometry in Hematologic Malignancies, and in that time flow cytometry has solidified its place as a routine diagnostic test in the diagnosis and classification of leukemias and lymphomas. Over the decade, dozens of new markers have been discovered, many of which are now a standard part of the flow cytometry arsenal. It is thus timely that Dr. Ortolani has chosen to update his work with this new second edition. As before, this work is extensively researched and prodigiously referenced, with more than 1300 new references not included in the first edition, bringing the total to more than 4500 relevant citations. These references not only cover the new markers, but also include new information about common markers in use for years, and descriptions of the markers covered by the first edition have been expanded to discuss these new advances.
In compiling this book, Dr. Ortolani has maintained the unique structure of the first edition, with the first part of the book oriented around individual antigens, and the second around diseases, supplementing the comprehensive text with characteristic images to illustrate the key points raised. The first section includes detailed information about normal tissue distribution of antigens, and also includes a discussion of some key pitfalls in the interpretation of cytometric findings of many antigens, before discussing pathologic conditions in which the antigens may be found. The disease section has been extensively revised and now follows the 2016 version of the WHO classification, while also including some newer entities not mentioned in the WHO monograph. In keeping with the changes in classification, this book now includes a much more extensive discussion of phenotypic properties of the different types of T‐cell lymphomas and histiocytic neoplasms than was available previously.
This book will be a valuable resource for anyone interested in flow cytometry and hematologic malignancies. Trainees can easily find detailed phenotypic information about any disease they are learning about, while even an expert, struggling with an unusual case with a phenotype that appears contradictory to an expected diagnosis, can quickly learn whether others have found similar patterns in that diagnosis; moreover, because the citations are so extensive, it is possible simply by considering the numbers of references to a phenotype to assess the weight of evidence for a particular interpretation. Those of us in the field are grateful that Dr. Ortolani took the time to update his work and produce a reference that will be useful for many years to come.
Michael J. Borowitz, MD, PhD
Professor of Pathology and Oncology
Director of Hematopathology and Flow Cytometry
Johns Hopkins Medical Institutions
Baltimore, MD, USA
Foreword to the First Edition
Flow cytometry is a crucial tool in the diagnosis of hematolymphoid neoplasms, determining prognosis and monitoring response to therapy. Clinical flow cytometric immunophenotyping, however, is a complex field requiring extensive expertise in normal and abnormal patterns before clinical tests can be appropriately interpreted. Those new to the field are left with the conundrum of how best to achieve this expertise. At particular disadvantage is the resident or clinical fellow seeking to interpret flow cytometric data on a specific patient. The typical flow cytometry reference text is written in an encyclopedic format with extensive narrative that is not conducive to looking up the meaning of unusual test results. Furthermore, the general flow cytometry textbook, although a useful reference, cannot completely cover all the aspects needed to interpret clinical flow cytometry data. Therefore, Flow Cytometry of Hematological Malignancies fills a much needed role in hematopathology and hematology/oncology. The presentation is oriented toward the diagnostic laboratory in the academic center as well as in the general hospital.
Flow Cytometry of Hematological Malignancies is organized in a novel manner that makes it especially useful for the medical student and residents/fellows still in training, while still providing a valuable resource for hematopathologists, hematologists/oncologists and experts in the field of clinical flow cytometry. It lists antigens typically studied in clinical flow cytometry laboratories, from CD1 to CD138, followed by a discussion of general as well as flow cytometric features and hematolymphoid neoplasms expressing each antigen. Thus, when interpreting a clinical flow cytometry report, one can easily research an unusual antigen expressed by a leukemia or lymphoma. This pattern of organization makes more sense than only presenting lists of neoplastic processes and the expected flow cytometric findings. One has to first know the diagnosis on a particular patient before such a reference can be useful. Flow Cytometry of Hematological Malignancies also provides the usual description of typical flow cytometric immunophenotypical findings in the various hematolymphoid neoplasms. This is useful as a reference for panel design as well as diagnosis.
Flow Cytometry of Hematological Malignancies is being published at a time when the field is expanding rapidly and flow cytometry is assuming an even greater role in management of patients with hematolymphoid neoplasia. Dr Ortolani, an outstanding flow cytometrist, possesses extensive expertise in the clinical arena. For over 30 years he was employed in the Clinical Pathology Department of the Venice General Hospital, running one of the first diagnostic flow cytometry units in Italy. His main clinical activity was the diagnosis of hematological neoplasms, with a particular interest in lymphoproliferative diseases. Dr Ortolani has also been very active in teaching flow cytometry in many national and international courses. He has written what I believe to be an outstanding textbook covering the essential aspects of clinical flow cytometry. Dr Ortolani is to be commended for this brilliant contribution that is sure to become a well‐used textbook in clinical centers around the world.
Maryalice Stetler‐Stevenson, PhD, MD
Chief, Flow Cytometry Laboratory
National Cancer Institute, National Institutes of Health
Bethesda, MD, USA
Foreword to the First Edition
The European Society for Clinical Cell Analysis (ESCCA) is proud to present this volume by Dr Claudio Ortolani. Flow Cytometry of Hematological Malignancies is a benchtop companion for all who are involved in the complex process of characterization and diagnosis of leukemias and lymphomas by immunophenotypical techniques and flow cytometry.
This volume is a useful quick reference text for the matching of CD antigens with malignant hematological diseases, as defined by the WHO 2008 classification, taking into account antibody clones, features and behavior, with particular emphasis on variant forms and unexpected presentations.
After several decades of clinical and laboratory practice in this field, Dr Claudio Ortolani has meticulously prepared this book under the auspices of the ESCCA. It represents a major achievement for the dissemination of knowledge in one of the most important specialties within clinical cell analysis, as the book aims to improve and standardize the diagnostic process of malignant blood diseases. As a result, communication between clinicians and laboratory operators should benefit!
Bruno Brando
ESCCA President
Director, Hematology Laboratory and Transfusion Center,
Legnano Hospital, Milan, Italy
Preface to the Second Edition
Ten years have passed since the first edition of this book, 10 fruitful years full of new acquisitions in the field of Hematology. Many doubts have been clarified, new questions have been raised, and – above all – new therapeutic targets have been reached. Although molecular biology keeps assuming ever greater importance, especially in the field of acute leukemias and myeloproliferative neoplasms; nevertheless, flow cytometry still remains one of the fundamental pillars of hematopathology and becomes even more and more important in particular areas such as the evaluation of the minimal residual disease and the study of the pharmacodynamic characteristics of experimental therapies with monoclonal antibodies.
In these 10 years, technology has also made substantial progress, and today we have much more efficient tools than before, but also much more complex ones, and consequently the pre‐analytical and analytical components have become considerably complicated. Even if an attempt to account for these problems has been made, in‐depth treatment of most of them is beyond the scope of this book; they will be dealt with – hopefully – in the next edition, together with the many other topics neglected this time.
Claudio Ortolani MD
Adjunct Professor of Laboratory of Diagnostic Cytometry
Urbino University
Urbino, Italy
Consultant Clinical Pathologist (retired)
Ospedale dell’Angelo
Venice, Italy
Preface to the First Edition
The cytometric analysis of hematological malignancies is one of the most difficult applications of flow cytometry, requiring both a good knowledge of hematopathology and good control of the technique. Moreover, the effort of operators is made harder by the continuous evolution of the technology and by the continuous progress in the comprehension of the nature of the diseases.
This book was compiled from a series of notes originally intended for people practically involved in the field of diagnostic flow cytometry, and it is an example of what the author would have liked to consult at the beginning of his own career. The goal of this book is to offer the reader a quick and updated source of information on the phenotype of the hematological malignancies recognized by the last WHO classification, with the major exception of Hodgkin lymphoma which because of its peculiar nature is still beyond the limits of flow cytometry, even if things promise to change in the next few years.
The author may have unwittingly sown a number of mistakes and imprecisions, and he will be grateful to all colleagues who report these to him. He also realizes that this book could not have been written without the help of many friends and colleagues. Being unable to cite all of them, the author wants to particularly thank his friend Bruno Brando, current President of the European Society for Clinical Cell Analysis, for the continuous moral and practical support he has given over the years.
Claudio Ortolani MD
Former Director of the Flow Cytometry Unit
Clinical Pathology Department
Venice General Hospital
Abbreviations
3‐FAL
3‐fucosyl‐
N
‐acetyllactosamine
ABC
antibody‐binding capacity
ABC
age/autoimmune‐associated B cell
ABC‐like
activated B‐cell like
a‐B‐CLL
Atypical B‐cell chronic lymphatic leukemia
ABL
acute basophilic leukemia
acPGP
N‐acetyl Proline–Glycine–Proline
aCML
atypical chronic myeloid leukemia
ADA
adenosine deaminase
ADP
adenosine diphosphate
AITL
angioimmunoblastic T‐cell lymphoma
aka
also known as
ALAL
acute leukemia of ambiguous lineage
ALALnos
acute leukemia of ambiguous lineage not otherwise specified
ALCL
anaplastic large cell lymphoma
ALDH
aldehyde dehydrogenase
ALK
anaplastic lymphoma kinase
ALL
acute lymphoblastic leukemia
ALPS
autoimmune lymphoproliferative syndrome
AMCL
acute mast cell leukemia
AMKL
acute megakaryoblastic leukemia
AML
acute myeloid leukemia
AMLL
acute mixed lineage leukemia
AML‐MRC
AML with myelodysplasia‐related changes
AMM
agnogenic myeloid metaplasia
AMoL
acute monoblastic and monocytic leukemia
ANKL
aggressive NK‐cell leukemia
ANXA‐1
Annexin‐1
APC
antigen‐presenting cell
APL
acute promyelocytic leukemia
ASM
aggressive systemic mastocytosis
ATLL
adult T‐cell leukemia/lymphoma
atMSC
adipose‐tissue‐derived mesenchymal stromal cells
ATRA
all‐trans retinoic acid
AUL
acute undifferentiated leukemia
AZT
azidothymidine (zidovudine)
BALF
bronchoalveolar lavage fluid
B‐ALL
B‐cell acute lymphoblastic leukemia
BCL
B‐cell lymphoma (gene)
B‐CLL
B‐cell chronic lymphocytic leukemia
B‐CLL/LPL
B‐CLL in lymphoplasmacytoid transformation
B‐CLL/PL
B‐CLL in prolymphocytoid transformation
B‐CLPD
B‐cell chronic lymphoproliferative disease
BCR
B‐cell receptor
BFU‐E
burst‐forming units/erythroid
biaALCL
breast implant–associated anaplastic large cell lymphoma
BIM
bcl‐2‐interacting mediator of cell death
BL
Burkitt lymphoma
B‐LBL
B‐cell lymphoblastic lymphoma
BM
bone marrow
B‐NHL
B‐cell non‐Hodgkin lymphoma
BPDCN
blastic plasmacytoid dendritic cell neoplasm
B‐PLL
B‐cell prolymphocytic leukemia
B‐SLL
B‐cell small lymphocytic lymphoma
BVMCL
blastic variant mantle‐cell lymphoma
C9RP
complement 9 related protein
CA I
carbonic anhydrase isoenzyme I
CBCL
cutaneous B‐cell lymphoma
CBL‐MZ
clonal B‐cell lymphocytosis of marginal zone origin
CCND1
cyclin D1
CEA
carcinoembryonic antigen
CEL
chronic eosinophilic leukemia
CEL nos
chronic eosinophilic leukemia not otherwise specified
CFU
colony‐forming unit
CFU‐EO
colony‐forming unit/eosinophil
CFU‐G
colony‐forming unit/granulocyte
CFU‐GEMM
colony‐forming unit/granulocyte, erythrocyte, monocyte, megakaryocyte
CFU‐GM
colony‐forming unit granulocyte‐macrophage
CFU‐M
colony‐forming unit/macrophage
CGH
comparative genomic hybridization
CHL
classic Hodgkin lymphoma
CINCA
chronic infantile neurological cutaneous articular syndrome
CLA
cutaneous lymphocyte antigen
CLPD
chronic lymphoproliferative disease
CLPD‐NK
chronic lymphoproliferative disorders of NK cells
CM
cutaneous mastocytosis
CMCL
chronic mast cell leukemia
CML
chronic myeloid leukemia
CML‐BC
CML in blastic crisis
CMML
chronic myelomonocytic leukemia
CMPD
chronic myeloproliferative disease
CMPN
chronic myeloproliferative neoplasm
CMV
cytomegalovirus
CNKL
chronic NK cell lymphocytosis
CNL
chronic neutrophilic leukemia
CNS
central nervous system
CPLD
chronic lymphoproliferative disease
CRLF2
cytokine receptor‐like factor 2
CRS
chemokine receptor status
CSF
cerebrospinal fluid
CSR
class switch recombination
CTCL
cutaneous T‐cell lymphoma
CY5.5
cyanine 5.5
CY7
cyanine 7
cyμ
cytoplasmic μ heavy chain
DFS
disease‐free survival
DH‐JH
diversity and joining segments of the Ig heavy chain gene
DHL
double‐hit B‐cell lymphoma
DLBCL
diffuse large B‐cell lymphoma
DLBCLnos
diffuse large B‐cell lymphoma not otherwise specified
DLBCL‐SS
sanctuary site DLBCL
DPP IV
dipeptidyl peptidase‐IV
DRESS
DRug‐induced Eosinophilia with Systemic Symptoms
EATCL
enteropathy‐associated T‐cell lymphoma
EBER
Epstein–Barr virus‐encoded RNA
EBV
Epstein–Barr virus
ECD
Erdheim–Chester disease
EFS
event‐free survival
EGIL
European Group for the Immunological Characterization of Leukemias
EMA
epithelial membrane antigen
EML
extramedullary myeloid leukemia
EMZL
extranodal marginal zone lymphoma
ENKTL
extranodal NK/T‐cell lymphoma
EPC
endothelial progenitor cells
ET
essential thrombocythemia
ETP
early T‐cell precursor
FAB
French–American–British
FCCL
follicular cell cutaneous lymphoma
FCL
follicular lymphoma
FCM
flow cytometry
FcRH
Fc receptor homologues
FCRL
Fc receptor–like proteins
FDC
follicular dendritic cell
FDCS
follicular dendritic cell sarcoma
FISH
fluorescence in situ hybridization
FITC
fluorescein isothiocyanate
FL
follicular lymphoma
FLT3
fms‐related tyrosine kinase 3 (CD135)
FLT3‐ID
FLT3 internal tandem duplication
FOXP
FOX (forkhead box) protein
FSC
forward scatter
FTCL
follicular T‐cell lymphoma
FTH
follicular helper T cells
FUT
fucosyl‐transferase
GCB‐like
germinal center B‐cell like
GCC
germinal center cell
G‐CSF
granulocyte‐colony stimulating factor
GEP
gene expression profile
GLPD
germinotropic lymphoproliferative disorder
GM‐CSF
granulocyte‐macrophage colony‐stimulating factor
HABP4
hyaluronan binding protein 4
HAL
hybrid acute leukemia
HBME‐1
human bone marrow endothelium marker‐1
HBLD
hairy B‐cell lymphoproliferative disorder
HCD
heavy chain disease
HCL
hairy cell leukemia
HCL‐J
hairy cell leukemia, Japanese variant
HCL‐v
hairy cell leukemia, variant
HCV
hepatitis C virus
HE
hematoxylin–eosin
HES
hypereosinophilic syndrome
HEV
high endothelial venules
HGAL protein
human germinal center–associated lymphoma protein
HGBL
high‐grade B‐cell lymphoma
HGL
high‐grade lymphoma
HLH
hemophagocytic lymphohistiocytosis
HN
hematodermic neoplasm
HPC
hemopoietic precursor cell
HRS
Hodgkin and Reed–Sternberg
HS
histiocytic sarcoma
HSTCL
hepatosplenic T‐cell lymphoma
HUMARA
human androgen receptor
ICOS
inducible T‐cell COStimulator (CD278)
IDCS
interdigitating dendritic cell sarcoma
IDCT
indeterminate dendritic cell tumor
IEL
intraepithelial intestinal lymphocyte
Ig
immunoglobulin
i‐GIT‐T‐LPD
indolent T‐cell lymphoproliferative disorder of the gastrointestinal tract
IHC
immunohistochemistry
IL
interleukin
iNK
immature NK cells
IPSID
immune proliferative small intestinal disease
IPTCLB
intralymphatic proliferation of T‐cell lymphoid blasts
IRF4
interferon regulatory factor 4 protein, aka mum1
IRTA
immune receptor translocation‐associated proteins
ISM
indolent systemic mastocytosis
ITAM
immunoreceptor tyrosine‐based activation motif
ITCL
intestinal T‐cell lymphoma
ITIM
immunoreceptor tyrosine‐based inhibition motif
iT‐LBP
indolent T‐lymphoblastic proliferation
IT‐LPD
indolent T‐cell lymphoproliferative disorder
ITSM
immunoreceptor tyrosine‐based switch motif
IVL
intravascular lymphoma
IVLBCL
intravascular large B‐cell lymphoma
IWCLL
International Workshop on Chronic Lymphocytic Leukemia
JAM
junctional adhesion molecule
JMML
juvenile myelomonocytic leukemia
KIR
killer cell immunoglobulin‐like receptor
LAIP
leukemia‐associated immune phenotype
LANA
latency‐associated nuclear antigen
LBC
lymphoid blastic crisis
LBCL
large B‐cell lymphoma
LBP
lipopolysaccharide‐binding protein
LCA
leukocyte common antigen
LCH
Langerhans cell histiocytosis
LDBCL
large diffuse B‐cell lymphoma
LDCHL
lymphocyte‐depleted classic Hodgkin lymphoma
L/DCS
Langerhans/dendritic cell sarcoma
LDH
lactate dehydrogenase
LEF1
lymphoid enhancer‐binding factor 1
L&H cells
lymphocytic & histiocytic Reed–Sternberg variant cells
LHL
lymphoepithelioid lymphoma
LinAg
lineage antigen
LIR
leukocyte Ig‐like receptors
LMO2
LIM domain only 2 (rhombotin‐like 1)
LP cells
lymphocyte predominant cells
LPL
lymphoplasmacytic lymphoma
LPS
lipopolysaccharide
LRCHL
lymphocyte‐rich classic Hodgkin lymphoma
LRP
lung resistance protein
LSC
leukemic stem cell
LyAg
lymphoid antigen
LYG
lymphomatoid granulomatosis
LyGa
lymphomatoid gastropathy
LyP
lymphomatoid papulosis
MAIT
mucosal‐associated invariant T cell
MALD1
monoclonal asymptomatic lymphocytosis, cyclin D1–positive
MALT
mucosa‐associated lymphoid tissue
MALToma
lymphoma of the mucosa‐associated lymphoid tissue
MAMP
microbe‐associated molecular patterns
MATK
megakaryocyte‐associated tyrosine kinase
MBC
myeloid blastic crisis
MBCL
monocytoid B‐cell lymphoma
MBL
monoclonal B‐cell lymphocytosis
MCC
Merkel cell carcinoma
MCCHL
mixed cellularity classic Hodgkin lymphoma
MCD
multicentric Castleman disease
MCL
mantle‐cell lymphoma
MCL‐BV
mantle‐cell lymphoma – blastic variant
MDC
myeloid dendritic cell
MDCL
myeloid dendritic cell leukemia
MDS
myelodysplastic syndrome
MDSC
myeloid‐derived suppressor cells
MDS‐U
myelodysplastic syndrome, unclassified
MEITL
monomorphic epitheliotropic intestinal T‐cell lymphoma
MEP
megakaryocyte/erythroid progenitor
MESF
molecules of equivalent soluble fluorochrome
MF
mycosis fungoides
MFI
mean fluorescence intensity
MGUS
monoclonal gammopathy of undefined significance
MHC
major histocompatibility complex
MLBCL
mediastinal large B‐cell lymphoma
MLC
mixed leukocyte culture
MLP
multiple lymphomatous polyposis
MM
multiple myeloma
MML
myelomastocytic leukemia
MMoL
myelomonocytic leukemia
MNDA
myeloid cell nuclear differentiation antigen
M/NK‐AL
acute leukemia of myeloid/NK precursors
MoAb
monoclonal antibody
MoAg
monocytic antigen
MPAL
mixed phenotype acute leukemia
MPDMN
mature PDC proliferations associated with myeloid neoplasms
MPN
myeloproliferative neoplasm
MPO
myeloperoxidase
MRD
minimal residual disease
MS
myeloid sarcoma
mTOR
mammalian target of rapamycin
MUM1
multiple myeloma 1 protein, aka IRF4
MVL
microvillous lymphoma
MyAg
myeloid antigen
MZL
marginal zone lymphoma
NBS
Nijmegen breakage syndrome
NCA
non‐specific cross‐reacting antigen
NCAM
neural cell adhesion molecule
NEC
nucleated erythroid cells
NGC
next‐generation cytometry
NK
natural killer
NKCE
NK‐cell enteropathy
NKP
NK‐cell precursors
NKR
NK‐cell receptors
NLPHL
nodular lymphocyte predominant Hodgkin lymphoma
NMZL
nodal marginal zone lymphoma
NPM
nucleophosmin
NRBC
nucleated red blood cell
NSCHL
nodular sclerosis classic Hodgkin lymphoma
OS
overall survival
PAL
pyothorax‐associated lymphoma
PB
peripheral blood
PBL
plasmablastic lymphoma
PCATCL
primary cutaneous acral T‐cell lymphoma
PCAETL
primary cutaneous aggressive epidermotropic cytotoxic T‐cell lymphoma
pcALCL
primary cutaneous anaplastic large cell lymphoma
PCBCL‐LT
primary cutaneous diffuse large B‐cell lymphoma “leg type”
PC‐DLBCL
primary cutaneous diffuse large B‐cell lymphoma
PC‐FCL
primary cutaneous follicle center lymphoma
PC‐ENKTL
primary cutaneous extranodal NK/T‐cell lymphoma
PCGD‐TCL
primary cutaneous TCRγδ(+) T‐cell lymphoma
PCH
pseudo Chediak–Higashi
PCL
plasma cell leukemia
PCMZL
primary cutaneous marginal zone lymphoma
PCNSL
primary CNS lymphoma
PcP
peridinin‐chlorophyll‐protein
PCSM‐TCL
primary cutaneous lymphoma of medium/small CD4(+) T lymphocytes
PD‐1
programmed death‐1
PDC
plasmacytoid dendritic cell
PDCL
plasmacytoid dendritic cell leukemia
PE
phycoerythrin
PEL
primary effusion lymphoma
PEL
pure erythroid leukemia
PFS
progression‐free survival
PHA
phytohemagglutinin
PID
primary immunodeficiency disease
PLEVA
pityriasis lichenoides et varioliformis acuta
PMA
phorbol myristate acetate
PMBC
peripheral mononuclear blood cells
PMBCL
primary mediastinal (thymic) large B‐cell lymphoma
PMF
primary myelofibrosis
PNET
primitive neuroectodermic tumors
PNH
paroxysmal nocturnal hemoglobinuria
PPBL
persistent polyclonal B‐cell lymphocytosis
pPNET
peripheral primitive neuroectodermic tumors
PPO
platelet peroxidase
PRCA
pure red cell aplasia
PTCL
peripheral T‐cell lymphoma
PTCLnos
peripheral T lymphoma not otherwise specified
PTFL
pediatric‐type follicular lymphoma
PTLD
post‐transplant lymphoproliferative disease
PV
polycythemia vera
RA
refractory anemia
RAEB
refractory anemia with excess of blasts
RALD
RAS‐associated autoimmune leukoproliferative disorder
RARS
refractory anemia with ring sideroblasts
RCMD
refractory cytopenia with multilineage dysplasia
RCUD
refractory cytopenia with unilineage dysplasia
RER
rough endoplasmic reticulum
RN
refractory neutropenia
ROR1
receptor‐tyrosine‐kinase‐like orphan receptor 1
RRV
rhesus rhadinovirus
RT
refractory thrombocytopenia
RTE
recent thymic emigrants
sALCL
systemic anaplastic large cell lymphoma
SAP
SLAM‐associated protein
SCF
stem cell factor
SCLC
small cell lung cancer
SDRPL
splenic diffuse red pulp lymphoma
SFTS
severe fever with thrombocytopenia syndrome
SLAM
signaling lymphocytic activation molecules
SLF
steel factor
SLL
small lymphocytic lymphoma
SLVL
splenic lymphoma with villous lymphocytes (obsolete)
SM
systemic mastocytosis
SM‐AHNMD
systemic mastocytosis with an associated clonal hematological non‐mast cell disorder
SMZL
splenic marginal zone lymphoma
SPTCL
subcutaneous panniculitis‐like T‐cell lymphoma
SR(B)CT
small‐round (blue) cell tumor
SRCT
small‐round‐cell tumor
SS
Sézary syndrome
SSC
side scatter
SSEA
stage‐specific embryonic antigen
T‐ALL
T‐cell acute lymphoblastic leukemia
TAM
transient abnormal myelopoiesis
TCL1
T‐cell leukemia/lymphoma 1
T‐CLL
T‐cell chronic lymphocytic leukemia
T‐CLPD
T‐cell chronic lymphoproliferative disease
TCM
T central memory
TCR
T‐cell receptor
TCRAV
T‐cell receptor A variable (region)
TCRBCL
T‐cell‐rich large B‐cell lymphoma
TCRBV
T‐cell receptor B variable (region)
TdT
terminal deoxy‐nucleotidyl transferase
TEM
T effector memory
TFR
therapy‐free remission
THL
true histiocytic lymphoma (obsolete)
THRLBCL
T‐cell/histiocyte‐rich B‐cell lymphoma
TKI
tyrosine kinase inhibitor
TKR
tyrosine kinase receptor
T‐LBL
T‐cell lymphoblastic lymphoma
T‐LGL
T‐cell large granular lymphocytic leukemia
TLR
Toll‐like receptor
TMD
transient myeloproliferative disorder
tMF
mycosis fungoides in transformation
TMPD
transient myeloproliferative disorder
TN
T naïve
TNF
tumor necrosis factor
TNFAIP2
TNF alpha induced protein 2
T‐NHL
T‐cell non‐Hodgkin lymphoma
T‐PLL
T‐cell prolymphocytic leukemia
TPO
thrombopoietin
TPO‐R
thrombopoietin receptor
TRAF
TNF receptor‐associated factor
TSCM
T stem cell memory
TSLP
stromal thymic lymphopoietin
TTE
T terminal effector
TTM
T transitional memory
TTT
time to treatment
TXR
Texas Red
TZL
T zone lymphoma
VDJ
variable, diversity, joining segments of the Ig heavy chain gene
VH
variable segment of the Ig heavy chain gene
VLA
very late activation
vWF
von Willebrand factor
WBC
white blood cells
WD‐EMT
well‐differentiated extramedullary myeloid tumor
WHO
World Health Organization
ZAP‐70
zeta‐chain‐associated protein‐70
1Antigens
Clustered (CD) Antigens
CD1
CD2
CD3
CD4
CD5
CD7
CD8
CD10
CD11b
CD11c
CD13
CD14
CD15
CD16
CD19
CD20
CD22
CD23
CD24
CD25
CD26
CD27
CD28
CD30
CD33
CD34
CD38
CD43
CD45
CD45 Isoforms
CD48, see SLAM molecules
CD49
CD56
CD57
CD61
CD62L
CD64
CD65
CD66c
CD71
CD79
CD81
CD84, see SLAM molecules
CD103
CD117
CD123
CD138
CD150, see SLAM molecules,
CD158 isoforms, see KIRs
CD181–186, CD191–199, see Chemokines and Chemokine Receptors
CD200
CD229, see SLAM molecules
CD244, see SLAM molecules
CD280–290, see Toll‐like Receptors
CD305
CD307 (IRTA) Antigen Family
CD319, see SLAM molecules
CD352–353, see SLAM molecules
CD371
Non clustered (or primarily known with other names) antigens
Bcl‐2 Protein
Chemokines and Chemokine Receptors
CRLF2
Cytotoxic Proteins
HLA‐DR
Immunoglobulins
KIR, CD158 isoforms
Myeloperoxidase (MPO)
NG2
PCA‐1
ROR‐1
SLAM Molecules and SLAM‐associated Protein (SAP)
SOX11
T‐cell Receptor (TCR)
Terminal Deoxy‐nucleotidyl‐transferase (TdT)
Toll‐like Receptors (TLR)
VS38
ZAP‐70
CD1 Antigens
General features
CD1 antigens are a group of five different glycoproteins that weigh 43–49 kD and are encoded by a group of genes situated on the long arm of chromosome 1 [1].
CD1 antigens play a role in the presentation of lipidic and glycolipidic antigens to T and NKT cells [1,2], and can be divided in three groups, namely:
Group I, encompassing CD1a, CD1b, and CD1c
Group II, encompassing CD1d
Group III, encompassing CD1e, which is an intracytoplasmic protein.
CD1 antigens are mainly expressed on cells belonging to T and B lineages and on antigen‐presenting cells (APC).
As for the T lineage, CD1a, CD1b, and CD1c antigens have been demonstrated on the membrane of the cortical or “common” thymocytes [3] and on the membrane of some T‐lymphocyte subsets in cord and neonatal peripheral blood [4]; according to some authors, a low expression of CD1 antigens can be demonstrated in the cytoplasm of T lymphocytes activated in vitro by phytohemagglutinin (PHA) [5]. CD1d has been reported at a high density on common thymocytes, and at a lower density on medullary thymocytes [6].
As for the B lineage, both CD1c and CD1d have been demonstrated on the precursors and on some subsets of mature B lymphocytes. More precisely:
CD1c has been demonstrated on some subsets of B lymphocytes in the peripheral blood [
7
,
8
], in the spleen [
7
,
8
], and in the mantle of the germinal center
[7]
CD1c(+) B lymphocytes account for the majority of B cells in tonsils
[9]
, in cord blood
[4]
, in the peripheral blood of newborns
[4]
, and in the peripheral blood of subjects submitted to autologous or allogeneic bone marrow transplantation during the first year following transplant
[8]
CD1d has been demonstrated on the membrane of bone marrow B precursors
[10]
, on B lymphocytes in peripheral blood [
11
,
12
], and on a subset of B lymphocytes in the mantle of the germinal center
[11]
and in the spleen
[13]
.
As for the APC, CD1a, CD1b, and CD1c antigens have been commonly reported on dendritic cells [6]. Moreover:
CD1a has been demonstrated on Langerhans cells
[14]
, where it is expressed at an intensity of 1600 molecules per cell
[15]
, on some CD11b(+) CD14(+) mononuclear cells reported in the peripheral blood of burnt subjects and interpreted as Langerhans cell precursors migrating from bone marrow to epidermis
[16]
, on monocytes activated with granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) in vitro
[17]
, and on in vitro monocyte‐derived dendritic cells
[18]
CD1b has been demonstrated on monocytes activated with GM‐CSF in vitro
[17]
and on a subset of Langerhans cells
[19]
CD1c has been demonstrated on monocytes activated with GM‐CSF in vitro, on Langerhans cells
[19]
, and on a minor subset of myeloid dendritic cells (MDC) characterized by CD11c(++) CD123(±) phenotype
[20]
CD1d has been demonstrated on “resting” monocytes
[11]
, on dendritic cells of the dermis
[21]
, on dendritic cells in peripheral blood
[6]
, and on in vitro monocyte‐derived dendritic cells
[21]
.
CD1d has also been demonstrated in keratinocytes of psoriatic skin [22], in the cells of human scalp hair follicles [22], in the epithelial cells of the gut and other organs [23], and in adipocytes [24].
CD1a, CD1b, CD1c, and CD1d have been demonstrated in the “foam cells” of the atherosclerotic plaque [25].
Cytometric features
The cytometric demonstration of molecules belonging to the CD1 family should be performed taking the following points into account:
cytometric studies have demonstrated that activated T lymphocytes express CD1c on the membrane only when kept at room temperature, and fail to mount the molecule on the surface when kept at +4 or to +37°C
[26]
the expression of CD1a on the surface of the leukemic blasts can fluctuate spontaneously after a short period of incubation in vitro
[27]
.
The antibodies specific for CD1 antigens do not behave in the same way.
It should be kept in mind that CD1a features four different epitopes, the first of which is recognized by clones D47, Na1/34, and L119, the second by clone L404, and the third by clone L504 [28]; it should be noted that CD1a on the cells of B‐cell chronic lymphocytic leukemia (B‐CLL) can be demonstrated only with clones other than OKT6 or Na1/34 [29].
The clones 7C4 and IOT6b recognize different epitopes of CD1b [30].
Diagnostic features
CD1 in myelodysplastic and chronic myeloproliferativediseases
CD1a can be detected by immunohistochemistry (IHC) in the neoplastic cells of the cutaneous localization of chronic myelomonocytic leukemia (CMML); a case is known displaying S‐100(−), CD1a(+), CD4(+), CD56(+), CD123(−), and Langerin/CD207(−) phenotype, mimicking the phenotype of the indeterminate dendritic cell tumor (IDCT) [31].
CD1a, CD1b, and CD1c are expressed on the membrane of the blast cells in the 20% of cases of chronic myeloid leukemia in blastic crisis (CML‐BC) [27].
The expression of CD1d has been reported on the cells of the juvenile myelomonocytic leukemia (JMML) [32].
CD1 antigens in neoplastic diseases of B‐cell precursors
In a group of 80 patients affected by childhood B‐cell acute lymphoblastic leukemia (B‐ALL), the expression of CD1d has been demonstrated in 15% of the cases [10]. CD1d expression is significantly associated with pre‐B phenotype, rearrangement of the gene KMT2A (formerly known as MLL), and shorter global survival [10].
CD1 antigens in neoplastic diseases of T‐cell precursors
In T‐cell acute lymphoblastic leukemia (T‐ALL) the expression of CD1a antigen has been detected in a percentage of cases ranging from 40% to 50% of cases [27,4565]. CD1a antigen is generally expressed on the cells of T lymphoblastic leukemia/lymphoma (T‐ALL/LBL) related to the stage of cortical or “common” thymocyte [3,27] (Fig. 1.1). According to the European Group for the Immunological Characterization of Leukemias (EGIL) classification of T‐ALL, CD1a antigen is typically present in the T III form but missing in the T I, T II, and T IV forms [34]. If CD13 is negative, the expression of CD1a is related to good survival [35], while the expression of CD1a together with CD10 is associated with the presence of the t(5;14)(q35;q32) translocation [33,4594,4595].
CD1 antigens in acute myeloid leukemias
The expression of CD1a and of CD1d has been repeatedly reported on the surface of the blasts of the acute myeloid leukemias (AML) [27,32,33]. According to some authors, the expression of CD1a and CD1d is restricted to French–American–British (FAB) subtypes characterized by a monocytic component [32]. In a percentage of cases, the expression of CD1a can be accompanied by the presence of other histiocytic markers (S‐100, CD163, Langerin/CD207) and is interpreted as a sign of histiocytic differentiation [36].
Figure 1.1 Peripheral blood from a subject affected by T‐ALL. The blasts (red) express phenotype CD45(dim+) (A), CD1a(+) (B), CD3(+) (heterogeneous) (B), CD4(+) (C), CD8(+) (partial) (D).
Table 1.1 Differential diagnosis of CD5(−) CD10(−) B‐CLPDs
B‐CLPD
CD1d
CD200
HCL
Positive
Positive
LPL
Dimly positive
Positive
MZL
Positive
Negative
B‐CLPD: B chronic lymphoproliferative disease; HCL: hairy cell leukemia; LPL: lymphoplasmacytic lymphoma; MZL: marginal zone lymphoma.
CD1 antigens in neoplastic diseases of mature B cells
It has been reported that B‐cell prolymphocytic leukemia (B‐PLL) cells express CD1c [9,37], that Burkitt lymphoma (BL) cells do not express CD1c [7], and that hairy cell leukemia (HCL) cells express CD1a [38] and CD1c [9]. The cells of B‐CLL have been reported to express CD1a [9,29], CD1c [7], and CD1d; it is noteworthy that CD1a on B‐CLL cells could be demonstrated only with clones other than OKT6 or Na1/34 [29].
CD1d is usually expressed by the cells of the B‐CLPDs, but at an intensity depending on the type of disease. Its intensity in B‐CLL is typically lower than in normal B cells, but it is progressively higher on MCL, HCL‐variant (HCL‐v), lymphoplasmacytic lymphoma (LPL), splenic marginal zone lymphoma (SMZL), and HCL cells [12]. The combined exploration of CD1d and CD200 seems very promising in the differential diagnosis of CD5(−) CD10(−) B‐CLPDs as well, inasmuch as a recent study has shown that HCL is CD1d(+) CD200(+), LPL is CD1d(−/±) CD200(+), and MZL is CD1d(+) CD200(−) [39] (Table 1.1).
CD1d antigen seems of particular interest in B‐CLL workup, because its intensity of expression and the percentage of CD19(+) CD1d(+) cells are bad prognostic predictors [40,41]; according to some authors [42,43], but not to others [40], CD1d intensity seems to correlate with the absence of somatic hypermutations [43].
Multiple myeloma (MM) cells express CD1d in the early stages but tend to reduce its expression with disease progression [44].
CD1 antigens in neoplastic diseases of mature T and natural killer (NK) cells
The expression of CD1a antigens has been reported in rare cases of peripheral T‐cell lymphoma (PTCL) [45] and in an isolated case of adult T‐cell leukemia/lymphoma (ATLL) [46].
Overexpression of CD1d mRNA has been detected in Sezary's cells by RT‐PCR [47], but this report has not yet been confirmed by phenotypic studies.
CD1 antigens in neoplasms of histiocytes and dendritic cells
As for the neoplasms of histiocytic and dendritic cells, CD1a, CD1b, and CD1c have been demonstrated with immunohistochemical techniques in the Langerhans cell histiocytosis (LCH) [19,48,49]. One of the most typical features of the pulmonary involvement in LCH is the occurrence of more than 5% of CD1(+) cells in the bronchoalveolar lavage fluid (BALF) [50]. Cells with CD1a(+), CD207(+), CD11b(±), and CD11c(++) phenotype have been detected in the peripheral blood of patients with active LCH [51]. CD1a expression has been reported in an anecdotal case interpreted as acute leukemia of Langerhans cell precursors based on the presence of Birbeck granules and of the ability of blasts to develop dendritic processes when cultured in vitro [52].
CD1a has been demonstrated with immunohistochemical techniques in the IDCT [53], but neither in the follicular dendritic cell sarcoma (FDCS) nor in the interdigitating dendritic cell sarcoma (IDCS) [49].
Partial expression of CD1a and expression of CD1c have been reported on the cells of an exceptional case of leukemia classified as Langerhans cell/dendritic cell leukemia, which occurred in a patient suffering from myelodysplastic syndrome (MDS). This case displayed CD11c(+), CD123(−), CD141(+), CD303(−), CD304(±/−), and CD207/Langerin(+/−) phenotype [54].
CD1 antigens in other pathological conditions
CD1a has been reported on the cells of the so called “indolent T‐lymphoblastic proliferation” (iT‐LBP) [55], an entity not recognized by the 2017 WHO classification (see also the dedicated paragraph).
CD2 Antigen
General features
CD2 is a 45–58 kD glycoprotein belonging to the superfamily of the immunoglobulins, which is encoded by a gene situated on the short arm of chromosome 1. CD2 is an adhesion molecule, constitutes the ligand of the CD58 molecule [56] and interacts with CD48 and CD59 molecules as well [57].
CD2 is normally expressed on thymocytes, on whose membrane it begins to appear at the prothymocyte level [58], and on mature T lymphocytes [59].
Not all mature T lymphocytes co‐express CD2; in the peripheral blood small subsets of T lymphocytes exist that show CD3(+) CD2(−) phenotype and are characterized by the expression of T‐cell receptor (TCR) either with αβ [60] or γδ chains [61].
The expression of CD2 is not restricted to the T lineage. Indeed, it is well known that CD2 is expressed:
on 70–90% of the NK cells negative for CD3 [
62
,
63
], where it is upregulated by activation
[64]
on a minority of follicular dendritic cells (FDC)
[65]
on a subset of mononuclear peripheral cells interpreted as precursors of MDCs
[66]
on a subset of peripheral monocytes characterized by the co‐expression of Fcε receptor (FcεRI)
[67]
on a subset of plasmacytoid dendritic cells (PDCs)
[68]
on a small subset of B cells in fetal liver
[69]
, in fetal bone marrow
[69]
, in thymus
[70]
, in peripheral blood
[71]
, and in the bone marrow of normal subjects
[71]
.
Cytometric features
The staining of peripheral normal lymphocytes with an anti‐CD2 monoclonal antibody (MoAb) generates a positive histogram with a narrow gaussian‐like peak, clearly separated from the negative component, with a channel peak representing the presence of 24 ± 7 E03 ABC (antibody‐binding capacity) [72].
Bimodal histograms can often be seen, especially when immune system activation is ongoing, because a higher number of CD2 molecules is expressed on activated cells [73] (Fig. 1.2).
In these cases, the CD2(bright+) population tends to show higher values of forward and side scatter than the CD2(dim+) population.
CD2(bright+) cells nearly exclusively express CD45R0, while the CD2(dim+) population display either CD45R0 or CD45RA [74]; the staining of CD2(+) CD45RA(+) cells with an anti‐CD2 MoAb generates a histogram with a channel peak representing the presence of 21 ± 4 E03 ABC while the staining of CD2(+) CD45R0(+) lymphocytes with the same MoAb generates a histogram with a channel peak representing the presence of 55 ± 9 E03 ABC [72].
In accordance with the state of chronic activation caused by HIV infection, the lymphocytes of HIV‐infected subjects seem to express a higher amount of CD2 molecules [75], whereas a reduced expression has been documented on the lymphocytes of elderly subjects [76]. According to some authors, the expression of CD2 on NK cells is inhomogeneous, being more intense in the CD16 dim CD56 bright subset than in the CD16 bright CD56 dim subset [77].
Not all the anti‐CD2 MoAbs behave in the same way; some clones are able to inhibit the E‐rosette formation [78], while others are able to activate T lymphocytes in vitro [79].
Diagnostic features
CD2 in myelodysplastic and chronic myeloproliferativediseases
CD2 has been reported in a third of cases of CMML [80].
CD2 in neoplastic diseases of mast cells
CD2 has been reported together with CD22 and CD25 on the neoplastic mast cells in systemic mastocytosis (SM) and acute mast cell leukemia (AMCL) [81–83]. However, an aberrant CD2 (and CD25) expression can be found on mast cells in the bone marrow of subjects without evidence of neoplasms of mast or myeloid stem cell [84].
Figure 1.2 The histogram produced by the cytometric analysis of CD2 is bimodal (A), because the activated CD25(+) lymphocytes (red) express more CD2 molecules than CD25(−) lymphocytes (blue) (B).
CD2 in neoplastic diseases of B‐cell precursors
CD2 has been reported in 1–4% of the globally considered neoplastic diseases of B‐cell precursors [85–87], where it is associated with a worse prognosis [88].
Nonetheless, the expression of CD2 has been detected in just less than 25% of the cases of B‐ALL with KMT2A‐MLLT3 translocation [4565].
CD2 in neoplastic diseases of T‐cell precursors
According to the EGIL classification of T‐lymphoblastic leukemias, CD2 is typically present in the T II, T III, and T IV forms, but is missing in the most immature form, T I [34,89]. In a pediatric group of more than 100 cases of T‐ALL, CD2 has been detected in 80% of the patients [4565].
CD2 is generally expressed by the cases with TCRαβ, but only by some cases with TCRγδ [90,91]; its presence in childhood cases is correlated with an increased probability of maintaining complete remission [92].
CD2 in AML and BPDCN
Depending on the survey, the expression of CD2 has been reported in 3–34% of the observed cases [93–100]. CD2 has been reported:
on the blasts of pediatric AML‐M2 negative for translocation t(8;21)
[101]
on the promyelocytes of AML‐M3, with predilection for the microgranular variant (AML‐M3v) [
99
,
102
,
103
], and for the presence of the “short” type of the PML‐RARA fusion gene [
100
,
103
]
on both the monocytic and non‐monocytic neoplastic cells of AML‐M4 [
80
,
99
,
104
]
on the blasts of AML‐M5
[80]
on the blasts of de novo AML with inv3(q21q26.2) and monosomy 7 [
105
–
107
].
The presence of CD2 (and also of CD4, CD7, and CD56) on the blasts of AML is correlated with an increased risk of extramedullary disease (granulocytic sarcoma, and cutaneous, gingival, and meningeal involvement) [108], and with a lower incidence of complete remission [109]. The CD2 expression has been reported in cases of AML with morphological anomalies mimicking the picture of Chediak–Higashi disease (pseudo Chediak–Higashi, PCH) [110], and in some cases of BPDCN, also known as plasmacytoid dendritic cell leukemia (PDCL) [111]. The presence of CD2 on AML‐M3 promyelocytes correlates with the occurrence of thrombotic events [112].
In AML‐M4 with inv(16)/t(16;16), the expression of CD2 is variable, and has been reported as weaker in cases with fusion transcript CBFβ‐MYH11 other than type A [113].
CD2 in neoplastic diseases of mature B cells
Sporadic reports exist signaling the presence of CD2 in isolated cases of B lineage non‐Hodgkin lymphoma [114,115]. Since CD2 has been demonstrated on the surface of normal B lymphocytes [71]
