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

Description

This fourth edition presents an updated and expanded text and illustrations to reflect continued morphologic, immunophenotypic, and especially molecular advances in the field of neoplastic hematology, mostly due to the rapidly expanding application of next-generation sequencing. Those advances not only allow a more reliable diagnosis of the majority of tumors and identification of early changes such as monoclonal B-cell lymphocytosis or clonal hematopoiesis of indeterminate potential (CHIP), but also in many cases identify mutations or phenotypic changes in tumors that can be targeted by mutation-specific or antigen-specific drugs. This edition incorporates the updated WHO classification of hematopoietic tumors and new immunophenotypic and molecular markers to provide a thorough pathologic overview of hematologic neoplasms while focusing on flow cytometric features. Special emphasis has been put on hematological neoplasms with crucial clinical significance such as acute promyelocytic leukemia, other acute leukemias, and difficult areas in flow cytometry. Flow cytometric features in AML, MDS, CMML, CLL and measurable residual disease were significantly expanded. There are many new comparative tables, illustrations, and diagrams of algorithmic approaches. 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