Flow Cytometry in Neoplastic Hematology: Morphologic-Immunophenotypic-Genetic Correlation

Cover
Half Title
Title Page
Copyright Page
Dedication
Contents
Preface
Abbreviations
1. Introduction to Flow Cytometry
	The major roles of flow cytometry
	Flow cytometry in hematological emergencies
	10-color flow cytometry analysis
	Automated flow cytometry data analysis
	Flow cytometry parameters
	Limitations of flow cytometry
	Classification of hematopoietic neoplasms
2. Flow Cytometry Analysis of Blood and Bone Marrow
	Bone marrow: Normal structure and hematopoiesis
	Antigen expression during hematopoietic maturation
	Flow cytometry of blood and bone marrow: Gating strategies
	Blood: Normal flow cytometry pattern
	Bone marrow: Normal flow cytometry pattern
3. Identification of Clonal B-Cell Population
	Polyclonal versus monoclonal B-cells
	Monoclonal B-cell lymphocytosis
	B-cells versus plasma cells
	Mature versus immature B-cells
4. Identification of Abnormal T-Cell Population
	Introduction
	Aberrant T-cell phenotype by flow cytometry
	Lack of CD45 or very dim CD45 expression
	Presence of additional markers
	Peripheral (mature) disorders versus T-ALL
	T-cell markers expressed by non-T-cell processes
5. Identification of Myeloblasts
	Introduction
	Identification of myeloblasts by flow cytometry
	Flow cytometric pattern in major AML subtypes
	Leukemia-associated immunophenotype (LAIP)
	Differential diagnosis of AML based on flow cytometry pattern
6. Identification of Lymphoblasts
	Identification of B-lymphoblasts
	Identification of T-lymphoblasts
7. Immunophenotypic Markers in Flow Cytometry
	BCL2 (B-cell lymphoma/leukemia 2)
	CD1a (cluster of differentiation 1a)
	CD2
	CD3
	CD4 and CD8
	CD5
	CD7
	CD10
	CD11b
	CD11c
	CD13 and CD33
	CD14
	CD15
	CD16
	CD19
	CD20
	CD22
	CD23
	CD24
	CD25
	CD26
	CD27
	CD30
	CD34
	CD36
	CD38
	CD43
	CD45
	CD49d
	CD52
	CD56
	CD57
	CD64
	CD65
	CD71 (transferrin receptor)
	CD79
	CD81
	CD103
	CD117 (c-Kit)
	CD123
	CD133
	CD138
	CD157
	CD200
	CD235a/GPHA (glycophorin A)
	FLAER
	HLA-DR (human leukocyte antigen-DR isotype)
	LILRB1 (leukocyte immunoglobulin-like receptor B1)
	TCR beta F1 (T-cell receptor beta F1)
	TCR gamma/delta
	TdT (terminal deoxynucleotidyl transferase)
8. Morphologic-Flow Cytometric Correlation in Blood
	Normal blood
	Pancytopenia/cytopenia
	Neutrophilia
	Eosinophilia
	Lymphocytosis
	Erythrocytosis/polycythemia
	Basophilia
	Thrombocytosis
	Circulating blasts
	Monocytosis
	Leukoerythroblastosis
	Circulating plasma cells
9. Morphologic-Flow Cytometric Correlation in Bone Marrow
	Benign bone marrow
	Increased blasts
	Diffuse small cell infiltrate
	Diffuse intermediate/large cell infiltrate
	Diffuse blastoid (“high-grade”) infiltrate
	Increased number of scattered large cells (other than megakaryocytes)
	Paratrabecular lymphoid infiltrate
	Intrasinusoidal infiltrate
	Lymphoid aggregates (nodular lymphoid infiltrate)
	Pleomorphic lymphoid infiltrate
	Megakaryocytosis
	Eosinophilia
	Erythroid hyperplasia with dyserythropoiesis
	Marrow infiltrate with fibrosis
	Plasmacytosis
	Metastatic tumors and other infiltrative processes
10. Morphologic-Flow Cytometric Correlation in Lymph Nodes
	Reactive follicular hyperplasia
	Diffuse pattern with mostly small cells
	Diffuse pattern with medium-sized and/or large cells
	Mixed (pleomorphic) infiltrate
	Blastoid and/or “high-grade” infiltrate
	Nodular pattern
	Paracortical (interfollicular; T-zone) pattern
	Intrasinusoidal pattern
	Anaplastic infiltrate
	Histiocyte-rich infiltrate
11. Molecular-Flow Cytometric Correlation
	Introduction – Overview of genetic testing
	Molecular-flow cytometric correlation
12. Phenotypic Classification of Mature B-Cell Neoplasms
	Introduction
	Classification of B-cell neoplasms based on CD5 and CD10 expression
	Additional phenotypic markers in B-cell neoplasms
	Classification of B-cell neoplasms based on forward scatter (grading)
	B-cell neoplasms with plasmacytic differentiation
13. Mature B-Cell Lymphoproliferations
	Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL)
	B-cell prolymphocytic leukemia (B-PLL)/splenic B-cell lymphoma/leukemia with prominent nucleoli (SBLPN)
	Nodal marginal zone lymphoma (MZL)
	Gastric MZL (MALT lymphoma)
	Splenic marginal zone lymphoma (SMZL)
	Marginal zone lymphoma with blood and/or BM involvement
	Splenic B-cell lymphoma/leukemia with prominent nucleoli/hairy cell leukemia variant (SBLPN/HCL-v)
	Hairy cell leukemia (HCL)
	Splenic diffuse red pulp small B-cell lymphoma (SDRPL)
	Lymphoplasmacytic lymphoma (LPL)
	Follicular lymphoma
	Mantle cell lymphoma (MCL)
	Diffuse large B-cell lymphoma (DLBCL)
	Specific variants of DLBCL
	Burkitt lymphoma (BL)
	High-grade B-cell lymphomas
	Plasmablastic lymphoma (PBL)
	Primary effusion lymphoma (PEL)
14. Plasma Cell Neoplasms
	Plasma cell myeloma (PCM)
	Introduction
	Morphology
	Immunohistochemistry
	Flow cytometry
	Measurable (minimal) residual disease (MRD)
	Differential diagnosis of PCM
	Plasmacytoma
	Monoclonal gammopathy of undetermined significance
	Plasma cell leukemia
15. Phenotypic Classification of Mature T/NK-Cell Lymphoproliferations
	Introduction
	Nodal T-cell lymphomas with CD10 expression (T-cell lymphomas with T follicular helper phenotype)
	T-cell lymphoproliferations with CD30 expression
	T-cell lymphoproliferations with CD25 expression
	T-cell lymphoproliferations with CD56 expression
	Additional phenotypic markers in T-cell lymphoproliferations
16. Mature T/NK-Cell Lymphoproliferative Disorders
	T-cell prolymphocytic leukemia (T-PLL)
	Adult T-cell lymphoma/leukemia (ATLL)
	T-large granular lymphocyte leukemia (T-LGLL)
	Peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS)
	Nodal TFH cell lymphoma, angioimmunoblastic-type (AITL)
	Anaplastic large cell lymphoma (ALCL)
	Aggressive NK-cell leukemia (ANKL)
	Extranodal NK/T-cell lymphoma, nasal type (ENKTL)
	Enteropathy-associated T-cell lymphoma (EATL)
	Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL)
	Indolent T-cell lymphoproliferative disorder of the gastrointestinal tract (indolent T-LPD)
	Mycosis fungoides (MF)
	Sézary’s syndrome (SS)
	Hepatosplenic T-cell lymphoma (HSTL)
17. B-Cell Acute Lymphoblastic Leukemia/Lymphoma
	Introduction
	Morphology
	Immunophenotyping
	Genetic features
	Measurable (minimal) residual disease (MRD) in B-ALL
	Differential diagnosis of B-ALL
18. T-Cell Acute Lymphoblastic Leukemia/Lymphoma
	Introduction
	Morphology
	Flow cytometry
	Differential diagnosis of T-ALL
	Early T-cell precursor T-ALL (ETP-ALL)
19. Myelodysplastic Neoplasms
	Introduction
	Minimal diagnostic criteria
	Morphology
	Immunohistochemistry
	Flow cytometry
	Flow cytometry scoring systems in MDS
	Genetic features
	Differential diagnosis of MDS
20. Mixed Myelodysplastic-Myeloproliferative Neoplasms
	Chronic myelomonocytic leukemia
	Juvenile myelomonocytic leukemia
	MDS/MPN with SF3B1 mutation and thrombocytosis
	MDS/MPN with neutrophilia (MDS/MPN-n)
21. Myeloproliferative Neoplasms
	Introduction to myeloproliferative neoplasms
	Chronic myeloid leukemia
	Polycythemia vera
	Essential thrombocythemia
	Primary myelofibrosis
	Chronic neutrophilic leukemia
	Chronic eosinophilic leukemia
22. Acute Myeloid Leukemia, Defined by Differentiation
	Acute myeloid leukemia with minimal differentiation
	AML without maturation
	AML with maturation
	Acute myelomonocytic leukemia (AMML)
	Acute monoblastic (monocytic) leukemia
	Acute erythroid leukemia (AEL)
	Acute megakaryoblastic leukemia
	Acute panmyelosis with myelofibrosis
	Acute basophilic leukemia
	Extramedullary myeloid tumor (EMT)
	AML with myelodysplasia-related changes (AML-MRC)
	Therapy-related myeloid neoplasms
23. Acute Myeloid Leukemia With Defining Genetic Abnormalities
	Acute promyelocytic leukemia (APL)
	Acute myeloid leukemia with t(8;21) [RUNX1-RUNX1T1]
	AML with CBFB-MYH11
	AML with MLLT3-MLL
	AML with RPN1-EVI1
	AML with mutated NPM1
	AML with mutated CEBPA
	AML with DEK-NUP214
24. Measurable (Minimal) Residual Disease in AML
	Introduction
	Methods of MRD detection
	Leukemia-associated immunophenotype (LAIP)
	Flow cytometry analysis of MRD
25. Acute Leukemias of Ambiguous Lineage
	Acute undifferentiated leukemia (AUL)
	Mixed phenotype acute leukemia
26. Other Neoplasms and PNH
	Blastic plasmacytoid dendritic cell neoplasm (BPDCN)
	Follicular dendritic cell neoplasms
	Histiocytic sarcoma
	Langerhans cell histiocytosis
	Langerhans cell sarcoma
	Metastatic tumors to the bone marrow
	Mastocytosis
	Simultaneous bi-lineage hematologic malignancies
	Paroxysmal nocturnal hemoglobinuria (PNH)
Index