Hematopathology Pearls Da Zhang
INDEX
Page numbers followed by f refer to figure and t refer to table.
A
Abciximab 64
Aberrant antigen expression 270
Acanthocyte 10, 14f
Acid electrophoresis 43, 44f
Acidified serum test 46
Acquired coagulation disorders 55
Acquired immunodeficiency, disorders with 262
Acyltransferase deficiency 338
Addison's disease 29
Adenomatous polyposis, multiple 308
Adenosine diphosphate 65
Adhesion abnormalities 69
Alagille syndrome 309
ALCL See Anaplastic large cell lymphoma
Alder-Reilly anomaly 70, 72, 73f
Alkaline electrophoresis 43
ALL See Acute lymphocytic leukemia
Alpha satellite DNA 307
Alpha thalassemia 42t
Aluminum hydroxide 342
Alveolar
rhabdomyosarcoma 317
soft part sarcoma 317
AML See Acute myeloid leukemia
AMML See Acute myelomonocytic leukemia
Amyloidosis 233, 236f, 237f
primary 57
Anaplastic large cell 230f
lymphoma 220, 224t, 225227f
Anaplastic malignant lymphocytes 227f
Anemia 23
of chronic disease 25
related syndromes 45
related to hemolysis 33
severe 34
with acquired immunodeficiency syndrome 28
with chronic
alcoholism 30
renal failure 28
with hemoglobinopathies 37
with lead poisoning 28
Angelman syndrome 309
Angioimmunoblastic T-cell lymphoma 217, 221224f
Angiomatoid fibrous histiocytoma 317
Angiomyxoma, aggressive 317
Angiosarcoma 317
Anisocytosis with erythrocytes 13f
Anticoagulant therapy monitoring 63t
Antigen presenting cells 239t
Antiplasmin deficiency 55
Antithrombin 52
deficiency 63
APL See Acute promyelocytic leukemia
Aplastic anemia 30, 32f, 32t, 45, 158
acquired 32
Argatroban 64
Asymptomatic myeloma 232
Ataxia telangiectasia 262
Atrophic gastritis 29
Autoimmune
disorders 77
hemolytic anemia 46, 47t
lymphoproliferative
syndrome 92
system 96t
Autopsy lung section 203f
Autosomal dominant
disorders 307
inheritance 307
Autosomal recessive
disorders 308
inheritance 308
Axillary lymph node 230f
B
Bacillary angiomatosis 83
Barium sulfate 342
Bartonella
henselae 83
quintana 83
species 83
Basal cells and mast cells, comparison of 75t
Basophil 2
Basophilia 75
causes of 75
Basophilic leukemia, acute 145, 146f
Basophils
and mast cells, comparison of 5t
cells 4
B-cell 2
large 250f
lymphoma 99f, 100f, 177, 208
diffuse large 186, 189, 191194f
high-grade 82, 206
intravascular large 202, 203f, 204f
large 196f, 202, 205f, 206, 207f, 208f, 250f
of eye, large 199f
primary diffuse large 197f
primary mediastinal large 204f, 205f
prolymphocytic leukemia 161, 275f, 276f
β-chain mutation 38t
Bernard-Soulier syndrome 59, 60
Beta thalassemia 43f, 43t
Biphenotypic acute leukemia 131
Birbeck granules 239
Bite cell 10, 15f
Blackfan-Diamond anemia 110
Blast cell with nucleoli 145f
Blastic mantle cell lymphoma 159
Blastic plasmacytoid dendritic
cell neoplasm 147, 383
neoplasm 147150f
Bleeding
disorders 333
time 51
Blood cell 1
Bloom syndrome 110
B-lymphoblastic leukemia/lymphoma 152, 153, 158t, 383
B-lymphocytes 6
Bone marrow 79f, 99, 101, 112, 165, 182f, 240f, 271, 287f, 290f, 292f, 306f
aspirate 26f, 47f, 78f, 125f, 135f, 137f, 142f, 143f, 149f, 167f, 236f, 266f
normal 266f
morphology 46
section 79f, 188f
Bone tumors 316
Brain biopsy 197f
Breast
biopsy 86f
fluid accumulation 229f, 230f
tissue 86f
tumors 320
Burkitt's leukemia 158
Burkitt's lymphoma 158, 208, 209f, 262, 346
Burns, severe 21f
Burr cells 11, 17f, 329
C
Cabot's ring 10, 16f, 329
Castleman's disease 81, 87, 9093f, 243f
Cat scratch disease 88f
Cell
clumping 20f
lymphoma, large 258f
migration response, disorders of decreased 69
pattern, small 225f
Chain mutation hemoglobinopathies 38
Charcot-Leyden
crystals 4
protein 4
Chédiak-Higashi syndrome 62, 67, 68, 68f, 70
Chondroma 317
Christmas disease 54
Chromosome microdeletion 308
Clear cell sarcoma of soft parts 317
CLL See Chronic lymphocytic leukemia
Clonal lymphoid disorders 126f
Clopidogrel bisulfate 65
CNS tumors 320
Coagulation factors 48
antibodies to 56
function of 48t
production 48
Coagulation tests 49
Cobalamin 29
Colony-forming units 1
Congenital dyserythropoietic anemia type I, II, and III, comparison of 46t
Cortical region (cortex) 80
Cowden syndrome 307, 320
Cryoglobulinemia 233
types of 233
Cutaneous mastocytosis 124
Cystic fibrosis 308
Cytochemical stain 131, 131t
Cytogenetic abnormalities 161t, 231t
Cytokine receptors 154
Cytoplasmic granules 289f
Cytoskeletal defect 36t
Cytoskeletal protein, abnormal 19f
D
Dabigatran 65
Dacryocytes 23f
Deep venous thrombosis 53
Degranulation abnormalities 67
Delta/beta thalassemia, type of 333
Dendritic cell
neoplasm 175
sarcoma 245f
Dense body 58
Dermal-epidermal junction 216f
Dermatopathic lymphadenopathy 90, 94f
Detecting methylation of DNA 316
Diamond-Blackfan anemia 32
Digeorge syndrome 309
Dilute Russell's viper venom time 51, 56
Dimorphic erythrocyte population 16f
Dimorphic RBC population 11
Disseminated intravascular coagulopathy 56
Distended sinuses 82
DNA sequencing 315
Döhle bodies 72
Donath-Landsteiner antibody 47, 332
Down's syndrome 110, 146, 383
Dyserythropoietic anemia, congenital 46, 47f
Dysfibrinogenemia 63, 337
Dyskeratosis congenita 30, 67
E
EBV infection 76t
Echinocytes 11, 17f
Effusion lymphoma, primary 262
Elliptocyte 11
Embden-Meyerhof pathway 35
Embryo, newborn and normal adult haemoglobin, comparison of 3t
Endometrial stromal
sarcoma 318
tumor 321
Endothelial cells 97
Enteropathy associated t-cell lymphoma 210
Enzyme 336
Eosinophil 2, 3
granules, component of 326
ultrastructure of 4f
Eosinophilia 74
Eosinophilic
cytoplasm 206f, 213f
debris 96f
granuloma 239
leukemia, chronic 121, 123
Eosinophils, granule contents of 4t
Epidermotropism 216f
Epithelioid hemangioendothelioma 318
Epstein syndrome 62
Epstein-Barr virus 202f
Erdheim-Chester disease 246
Erroneous, potential causes of 9t
Erythroblasts 1
Erythrocyte 2, 20f
protoporphyrin, free 9
Erythrocytosis, secondary 117t
Erythroid
cells 47f
hypoplasia 33t
leukemia
acute 141, 144f, 271, 295f, 345
pure 141, 295f
lineage 1
Erythropoietic porphyria, congenital 45
Erythropoietic protoporphyria, congenital 45
Erythropoietin 326
Essential thrombocythemia 120, 122f
Ewing's sarcoma 318
Exon 310
Extranodal Rosai-Dorfman disease 83
Extrarenal rhabdoid tumor 318
Extraskeletal mesenchymal chondrosarcoma 318
Extraskeletal myxoid chondrosarcoma 318
F
Familial adenomatous polyposis 307
Familial breast-ovarian cancer 307
Fanconi anemia 30, 67, 110, 341
Fanconi syndrome 30
FDC See Follicular dendritic cells
Fechtner syndrome 62
Fetal haemoglobin, hereditary persistence of 42
Fibrinolytic system 52, 53t
Fibrin-stabilizing factor deficiency 55
Fine needle aspirate 198f
Florid follicular hyperplasia 330
Flow cytometry 99, 101, 265, 266f, 271
applications of 265
phenotyping 265
results, interpretation of 268
Flower cells 173f
Fluorescence
in situ hybridization 316
resonance energy transfer 53
Follicles, primary 184f
Follicular cell lymphoma 271, 281f
Follicular dendritic cell 80, 87, 91f, 223f, 239
sarcoma 242, 243f, 244f
tumor 242
Follicular hyperplasia 81,
reactive 81, 82, 82t
Follicular lymphoma 82, 82t, 180, 180184f, 185, 346
grading of 185
high-grade 181f
low-grade 181f
Follicular thyroid carcinoma 323
Fragmented cells 43f
Friedreich's ataxia 308
Fusion gene 310
G
Galactosemia 308
Gamna-Gandy bodies 40f
Gastric mucosa 178f
Gastrointestinal cancers 320
Gaucher's disease 341
Gene
chips 316
microarray technology 316
mutations 136
Genetic abnormalities 316
in bone 317t
recurrent 132, 153
Germinal centers, progressive transformation of 83, 269
Giant platelet disorder 62
Glanzmann thrombasthenia 60
Glucose-6-phosphate dehydrogenase deficiency 36
Glutamic acid 39
Glycolytic pathway, deficiencies of 35
Golgi-staining patterns 230f
Granular lymphocyte leukemia, large 166, 170f, 171f
Granulocyte 1, 266f
colony stimulating factor 1
left shift of 3
right shift of 3
Granulomas 200f
Granulomatous inflammation, chronic 261
Gray platelet syndrome 59
Gynecologic tumors 321
H
Hairy cell
leukemia 98, 101, 101t, 165, 167169f, 271, 282f, 283f, 346
variant 101
variant 101t
Hand-Schüller-Christian disease 239
Hashimoto thyroiditis 179f
Hassall's corpuscles 81
HBH disease 338
Heavy chain
deposition disease 233
disease 349
Heinz body 11
Hematogone 158, 158t
maturation 267t
Hematology testing 1, 6
Hematopoiesis cell 1
maturation 268
sequence of 1
Hematopoietic neoplasms 98, 323
with thrombocytosis 121
Hemoglobin 18f, 38
alkaline 44f
C 39
crystal 11, 17f, 40f
disease 40f
concentration 7
D 39
E 39
H disease 42f
synthesis 38t
Hemoglobinopathy, evaluation of 43
Hemolytic anemia in G6PD deficiency 37
Hemolytic disease 18f
Hemophagocytic
lymphohistiocytosis 77, 78f, 79f
syndrome 332
Hemophilia A 54
acquired 57
Hepatoblastoma 321
Hepatocellular carcinoma 321
Hepatosplenic T-cell lymphoma 101, 102104f
Hepcidin 25
Hereditary diffuse gastric carcinoma 307
Hereditary elliptocytosis 35, 35f, 36
Hereditary erythrocyte
disorders 35
membrane defects 33
Hereditary granulocytic disorders 70t
Hereditary hemochromatosis 308
Hereditary nonpolyposis colon cancer syndrome 308
Hereditary spherocytosis 3336, 331
Hereditary thrombotic disorders 62, 63t
Hermansky-Pudlak syndrome 62
Hexagonal phase phosphatidylethanolamine test 57
Hibernoma 318
High reticulocyte count 24
Histiocytic and dendritic cell neoplasms 239, 385
Histiocytic sarcoma 239, 243f
HIV infection 262
HIV-associated lymphadenopathy 88f
Hodgkin's disease 333
nodular lymphocyte predominant 269
Hodgkin's lymphoma 58, 98, 195t, 239, 247, 247t, 248, 249252f, 340, 348, 385
classical 208, 248, 254259f, 260, 260f, 260t
classification of 248
composite 253
mixed cellularity classical 253
nodular sclerosis classical 252
Hodgkin-like pattern 226f, 227f
Homocystinemia 63
Howell-Jolly bodies 11, 18f
Huntington's disease 307
Hyaline-vascular variant 89, 90f
Hypochromasia 42f
Hypochromatic target cells 39f
Hypochromic anemia 24
Hypoparathyroidism 29
Hypoplastic anemia 30, 32t
congenital 32
I
IDC See Interdigitating dendritic cells
Imatinib 124
Immature cells, clusters of 157f
Immune disorders, primary 261
Immune-mediated hemolysis 334
Immunodeficiency 261
disorder 261
congenital 77
Immunoglobulins 234f
Immunohemolytic anemia 21f
Immunohistochemical stains 190t
Immunophenotype
abnormalities 173
of small B-cell lymphomas, comparison of 176t
In situ hybridization for EBV 211f
In situ mantle cell neoplasia 186
Indolent T-lymphoblastic proliferation 159
Infectious mononucleosis 76t, 331, 332
Inherited aplastic anemia 32
Inherited bleeding disorders 51t
Inherited coagulation disorders 53
Inherited pure red blood cell aplasia 32
Inherited thrombocytopenias 59t
Inhibitory cytokines 25
Insensitive restriction enzyme 310
Interdigitating dendritic cell 80, 239
sarcoma 242, 245f
Interdigitating reticulum cells 327
Intestine biopsy, small 213f
Intrachromosomal amplification of chromosome 21 154
Intravascular hemolysis 332
Iron
deficiency 24, 25
anemia 26f
in hypochromic anemias 25t
Irregular cell membranes 167f
Isochromosome 310
J
Juvenile
myelomonocytic leukemia 111, 343
polyposis 307
xanthogranuloma 242
K
Kaolin clotting time 56
Kaposi's sarcoma 87, 89f
Kearns-Sayre syndrome 311
Kidney
biopsy 236f
core biopsy f-238f
Kikuchi disease 92, 96f
Kikuchi lymphadenitis 92
Kikuchi-Fujimoto disease 92
Kimura disease 92, 95f
Kimura lymphadenopathy 92
Kleihauer-Betke
stain 44
test 337
Kostmann syndrome 67, 110
L
Langer-Giedion syndrome 309
Langerhans cell 239, 240242f
histiocytosis 239, 240f242f
Leber's hereditary 311
Lepore disorder 342
Letterer-Siwe disease 239
Leukemia
acute undifferentiated 148
bilinear acute 131
development of 340
of ambiguous lineage, acute 131 147, 383
phase, acute 115
Leukemic cells 137f, 147f, 165f, 167f
with polylobated nuclei 173f
Leukemic monoblasts and monocytes 143f
Leukemic promyelocytes 135f
Leukocyte alkaline phosphatase 67, 115, 343
Leukocytosis 351
Leukoerythroblastic reaction 343
Li-Fraumeni syndrome 307
Littoral cells 97
Liver
disease 55
mass biopsy 194f, 244f
sinusoids of 103f
Low-molecular-weight heparin 63
Lung 178f
biopsy 225f
tumors 321
Lupus anticoagulant 57, 57t
antibody 56
Lymph node 87, 88f, 89f, 155f, 210, 227f, 230f, 250f, 271, 277f, 278f, 359
biopsy 8789f, 91f, 92f, 96f, 155157f, 164f, 180f, 181f, 187f, 189f, 193f, 196f, 207f, 209f, 221224f, 226f, 227f, 243245f, 249252f, 254260f
capsule of 221f
function, normal 80
hyperplasia 81
normal 80, 164f, 245f
structure, normal 80
with granuloma 82
with necrosis 82
Lymphoblastic
leukemia, acute 131t, 151, 349
leukemia/lymphoma
molecular abnormalities of acute 152t
prognosis of acute 153t
lymphoma, acute 151, 349
Lymphocyte 331
nuclear remnant 162f
predominant cells 247, 248, 252t
rich classical Hodgkin's lymphoma 253
small round 164f
Lymphocyte-depleted classical Hodgkin's lymphoma 253
Lymphocytic
infiltration, diffuse small 178f
leukemia
acute 131
chronic 100, 159, 160t, 161t, 162f, 163f, 271, 272274f
lymphoma, small 100, 159, 160t, 161t, 164f, 271, 272274f
Lymphocytopenia 77
causes of 77
Lymphocytosis 329
benign 76
causes of 76
reactive 158
Lymphoepithelial lesions 179f
Lymphoid
follicles 183f
enlarged 88f
hyperplasia 80
atypical 261
infiltration 200f
neoplasms with eosinophilia 128, 381
Lymphoma 101, 172, 179f
cells 206f, 207f, 211f, 213f
large 199f
malignant 261
of mucosa-associated lymphoid tissue 179f
stages of 175t
Lymphomatoid granulomatosis 195, 200202f
Lymphoplasmacytic lymphoma 98, 175
Lymphoproliferative disorder 98, 261
chronic 271, 303f, 304f, 347
M
Macrocytic anemia 24
Malignant cells, large 198f
Mantle cell lymphoma 98, 186, 187189f, 277280f
immunophenotype of 346
Massive lymphadenopathy 83, 84f
Mast cell 4
aggregates of 126f
circulating 125f
Mastocytosis 124
WHO classification of 124
Mature B-cell
leukemia 159, 175
neoplasms 383
Mature lymphocytes, small 162f
Mature T- and B-cell leukemias 151
Mature T-cell 210, 384
leukemia 166
May-Hegglin anomaly 59, 62, 70, 72f, 340
MCV, RDW diseases, comparison of 7t
Mean corpuscular hemoglobin 7
concentration 7
Mean corpuscular volume 7
Mediastinal biopsy 204f, 205f
Mediastinal mass 298f
Medulla of thymus 82t
Medullary region (medulla) 80
Medullary thyroid carcinoma 323
Megakaryoblastic leukemia 345
acute 145, 271, 296f
Megakaryocyte lineage 2
Megakaryocytic leukemia, acute 145f
Megaloblastic anemia 13, 18f, 29, 31f, 329
causes of 29
Metastasizing leiomyoma, benign 317
Metastatic small cell tumor 159
Methylation of DNA 310
Microbicidal activity, disorders of 69
Microcytic anemia 23, 24
Microcytosis 42f
Microsatellite instability assays 316
Miller-Dieker syndrome 309
Minimal residual disease 270, 271, 305f
detection of 131
Missense mutation 310
Mitochondrial inheritance 311
Mixed-phenotype acute leukemia 147
Molecular basis of diseases 316
Molecular pathology 307
methods in 314
Monoblastic leukemia, acute 141, 142f, 143f, 271, 293f, 294f
Monoclonal b-cell lymphocytosis 160
Monoclonal immunoglobulin 231t
Monocytes 1, 5
Monocytic blasts 141f
Monocytic leukemia, acute 141, 142f, 143f
Monocytoid B-cells 100f
Monocytosis 75
Monomorphic epitheliotropic intestinal T-cell lymphoma 212
Morphology, abnormal 70t
Mucoepidermoid tumor 322
Mucosa-associated lymphoid tissue 177
Multicentric Castleman's disease 207f, 208f, 262
Multilineage dysplasia 106
Multiple myeloma 231t, 305f, 350
Mycosis fungoides 212, 216f, 217f
Myelodysplasia-related changes 137, 382
Myelodysplastic
neoplasms 108, 381
syndrome 45, 105, 107, 108f, 112, 344, 382
in children 110
prognosis in 108t
Myelodysplastic/myeloproliferative neoplasms, WHO classification of 108
Myelofibrosis, primary 119, 119f, 120f
Myelogenous leukemia, chronic 67t, 112, 113115f
Myeloid
acute 131t
and lymphoid neoplasms 129
blasts 141f
leukemia 141
acute 45, 130, 137f, 138f, 139f, 158, 271, 284f, 285287f, 344, 382
atypical chronic 110
diagnosis of acute 130
lineage 2
maturation 267t
neoplasms 129
therapy-related 139
with eosinophilia 128, 381
progenitor, common 1
proliferations 146, 383
sarcoma 146, 383
Myelokathexis 67, 72, 74f
Myeloma light chain cast nephropathy 238f
Myelomonocytic leukemia
acute 140, 140f, 141f, 271, 291f, 292f
chronic 109
Myeloproliferative
disorders 98
neoplasm 108, 112, 129, 381
Myoclonic epilepsy 311
Myoepithelial tumor of soft tissue 319
Myxoid inflammatory fibroblastic sarcoma 319
N
Nasal mass biopsy 211f, 234f, 235f
Natural inhibitors of coagulation 52, 52t
Natural killer cell 6, 166, 268, 347
leukemia 169t
aggressive 171
neoplasm 210, 384
Neoplastic cells 149f, 204f
Neoplastic mast cells 306f
Neoplastic monocytosis, causes of 76
Neoplastic T-cells 104f
clusters of 221f
Nephrotic syndrome 58, 339
Neurofibroma 319
Neurofibromatosis type 1 110
Neutropenia 66
causes of 67
diagnostic criteria of 67t
Neutrophils 2, 67, 96, 326
disorders 66
functional defects of 67
granule contents of 3t
morphology, abnormal 70
Neutrophilia 66, 118f
diagnostic criteria of 66t
reactive 67t
Neutrophilic leukemia, chronic 116
Next-generation sequencing 316
NK-cell See Natural killer cell
NLPHD See Nodular lymphocyte predominant Hodgkin disease
Nodal marginal zone lymphoma 176
Nodular fasciitis 319
Non-Hodgkin lymphoma 175, 239
Non-neoplastic white blood cells, disorders of 66
Normocytic anemia 23
Normocytic red blood cells 27f
Nuclear stain 157f
Nuclear staining pattern 189f
Nucleic acid, sources of 314
Numerous mast cells 125f
Numerous target cells 40f
O
Occasional teardrop cells 118f
Orthochromic cell 2
Osteosclerotic myeloma 233
Ovoid tumor cells 244f
P
Panmyelosis with myelofibrosis, acute 146
Pappenheimer bodies 11, 19f
Paracortical hyperplasia 82
Paracortical region (paracortex) 80
Parotid gland 179f
Paroxysmal nocturnal hemoglobinuria 45
Pas stain 221f
PDGFRA, abnormalities of 381
Pelger-Huët anomaly 70, 71f
Peptide chains 38
Peripheral blood 5, 99, 101, 112, 271, 276f, 283f, 300f, 304f
cell counts, anticoagulant agents used for 6
of thalassemia major 13
Peripheral nerve sheath tumor, malignant 319
Peripheral neuroectodermal tumor 318
Peripheral T-cell lymphoma 215, 218f, 219f
Peutz-Jeghers syndrome 308
Phenylketonuria 308
Plasma cell 81, 91f, 205f, 234f, 305f, 306f, 327
disorders 231t
leukemia 234f
myeloma 232
international staging system for 231t
variants of 232
neoplasms 175, 228
classification of 228
population of 235f
variant 89, 93f
Plasmablastic lymphoma 202, 205f, 206f, 262
Plasmacytoid dendritic cell 147
Plasmacytoma 234f, 235f
Platelet 58
aggregation study 60t
disorders 48
additional 60
function analyser 51
glycoprotein adhesion receptors, abnormalities of 60
storage pool disorder 62
Pleomorphic
adenoma 322
cells, large 192f, 206f
lymphoid infiltrate 200f, 221f
tumor cells 192f
Pluripotential stem cell 1
POEMS syndrome 233
Poikilocytosis 10, 42f
Polychromasia 42f
Polyclonal B-cell lymphocytosis 100
Polycythemia vera 116, 117t, 118f, 345
WHO diagnostic criteria for 117
Polymerase chain reaction 315
Popcorn cells 248, 252f
Porphyrias 45
Post-polycythemic myelofibrosis 118f
WHO diagnostic criteria for 117
Post-transplant lymphoproliferative disorder 262, 264f, 347, 386
classification of 263
Pradaxa 65
Prader-Willi syndrome 309
Prazaxa 65
Precursor cells 1
Precursor lymphoid neoplasms, WHO classification of 151
Predominantly refractory anemia 105
Prolymphocytic leukemia 165f, 166f
Promyelocytic leukemia, acute 133, 134f, 135f, 271, 288f, 289f, 290f
Protamine sulfate 63
Protein
C 52
deficiency 63
in eosinophils, major basic 327
S 52
deficiency 63
truncation test 316
Prothrombin
mutation 63
time assay 49
Pseudo-Chediak-Higashi granules 68
PTGC See Progressive transformation of germinal centers
PTLD See Post-transplant lymphoproliferative disorder
Pyruvate kinase deficiency 36
R
Reactive monocytosis, causes of 75
Red blood cell 7, 13f, 14f, 23, 35f, 89f, 118f, 356
anisopoikilocytosis 118f
aplasia 33t
acquired pure 33
pure 32
defect of 15f
disorders of 10
morphology 10t
Red pulp 97
Reed-Sternberg cell 247, 252t, 253, 254f256f, 259f, 348
Renal tumors 322
Restriction enzyme 312
Restriction fragment length polymorphism 312
Reticulin fiber stain 120f
Reticulocyte count 8
corrected 8
Ribosomal RNA 312
Robertsonian translocation 312
Rosai-Dorfman disease 83, 84f86f
Rouleaux 20f
Round cell liposarcoma 319
Russell viper venom 56
S
Salivary gland tumors 322
Schistocyte 12, 20f
Sebastian syndrome 62
Secretory type ductal carcinoma 320
Serous atrophy 28f, 29f
Serum
ferritin 8
iron 8
Sézary disease 350
Sézary syndrome 173, 271, 299f, 300f
Shwachman-Bodian-Diamond syndrome 30
Shwachman-Diamond syndrome 30, 67, 110
Sickle cell 12, 21, 39f
disease 38, 39f, 40f
Sideroblastic anemia 25, 27f
Single blast population 268t
Single lineage dysplasia 105, 106
Sinus 81
histiocytosis 83, 84f86f
vascular transformation of 83
Sjögren syndrome 179f
Skin
biopsy 86f, 147f, 148f
punch biopsy 216f, 217f
SLL See Small lymphocytic lymphoma
Smith-Magenis syndrome 309
Smudge cells 162f
SMZL See Splenic marginal zone lymphoma
Soft tissue 316, 317
tumors 317t
Solitary fibrous tumor 319
Soluble serum transferrin receptor 9
Southern blot 314
Specific granule deficiency 70
Spherocyte 12, 21f
Spindle cell 190
morphology 193f, 194f
proliferation 89f
in sinus 88f
rhabdomyosarcoma 319
Spleen 40f, 97, 99, 102f, 165, 168f, 169f
function of 330
normal 97
red pulp 102f
section 102f
structure, normal 97
Splenectomy 40f
indications for 97
Splenic B-cell marginal zone lymphoma 99
Splenic lymphomas 98
Splenic marginal zone lymphoma 100
Splenic reactive follicular hyperplasia, causes of 98
Sporadic colon cancer 320
Stain reticulocytes 327
Stomach 178f
Stomatocyte 12, 22f
Stroma-rich variant 89
Subcutaneous
biopsy 215f
panniculitis-like T-cell lymphoma 212, 214f, 215f
tissue 86f, 214f, 215f
Sucrose lysis test 46
Sugar water test 46
Supravital stain 44
Symptomatic plasma cell myeloma 232
Syndromes with
microdeletion 309
microduplication 309
Systemic mastocytosis 124, 125f, 126f, 271, 306f
T
T- and B-cell populations 269
T lymphoblastic lymphoma/leukemia, acute 297f, 298f
Target cell 12, 22f, 42f
T-cell 2
acute lymphoblastic lymphoma 155f157f
diffuse infiltration of 222f
large granular
leukemia 271, 301f, 302f
lymphocyte leukemia 170, 172t
leukemia 169t
adult 172, 173f
lymphoma 213f
lymphoproliferative disorders 215
maturation 267t
phenotypes, comparison of 82t
prolymphocytic leukemia 169
T-cell/histiocyte rich large B-cell lymphoma, comparison of 195t
Teardrop cell 12, 23f
Tenosynovial giant cell tumor, malignant 319
Testicular germ cell tumors 322
Thalassemia 25, 41
Thrombocytopenia 58, 62
acquired 58
causes 58
Thrombocytosis 120
causes of 121
Thromboelastometry 49
Thrombosis, causes of 339
Thymoma 327
Thymus 80
Thyroid 179f
gland 179f
tumors 322
Tissue plasminogen activator 53
T-lymphoblastic leukemia/lymphoma 154, 383
T-lymphocytes 5
Tonsil 183f, 184f
Total iron-binding capacity 8
Transfused cells 21f
Tumor cell 157f, 187f, 188f
pleomorphism 245f
small 217f
Tyrosine kinases 154
U
Unfractionated heparin 63
Uniparental
disomy 313
heterodisomy 313
isodisomy 313
Urine hemosiderin test 46
Uterus mass biopsy 194f
V
Vitamin
B12 29
deficiency 23, 29
E deficiency 341
K
deficiency 55
participates 55
K dependent 339
coagulation factors 48, 49
Vitreoretinal lymphoma, primary 195
Vitreous fluid 198f
von Willebrand
disease 49, 59, 59t
factor 58
W
Warfarin 64
Warthin's tumor 157f
in parotid gland 157f
William's syndrome 309
Wilms’ tumor 309
Wiskott-Aldrich syndrome 59, 62, 261, 339
X
X-linked lymphoproliferative syndrome 262
×
Chapter Notes

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Hematopoiesis and Hematology TestingCHAPTER 1

 
HEMATOPOIESIS AND BLOOD CELLS
Hematopoietic stem cells are self-renewing cells that differentiate and become committed to different cell lineage.
Pluripotent stem cells and early progenitor cells can be cultured on culture media. In culture media, progenitor cells are defined as colony-forming units (CFUs). CFUs are earliest detectable progenitor cells that give rise to granulocytes, erythroblasts, monocytes and megakaryocytes, which together are termed CFUGEMM or CFUCMP (common myeloid progenitor). The more mature and specialized precursor cells are termed CFUGM (granulocytes and monocytes), CFUE (erythroblasts), and CFUEO (eosinophil). The burst-forming units (BFUE) of the erythroid lineage are early progenitor cells committed to erythrocyte differentiation and are the early ancestors of the CFUE. The BFUE has a limited capacity of proliferation and gives rise only to erythrocyte colonies. BFUE is insensitive to erythropoietin, and its progeny must go through as many as 12 divisions before they become mature erythrocytes.
The proliferation of stem cells and progenitor cells is under the control of cytokines. IL-3 and GM-CSF are nonlineage specific cytokines, which act on pluripotent and early progenitor cells; they are required for self-renewal and differentiation throughout the hematopoietic process. In contrast, cytokines such as granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), IL-5, thrombopoietin and erythropoietin all act on more mature and specific hematopoietic lineage cells. Erythropoietin, a glycosylated peptide with its gene located on the chromosome 7, is predominantly derived from the kidney and with a small amount from the liver. It stimulates erythropoiesis at the stage of CFUE. Thrombopoietin, a polypeptide with its gene located on chromosome 3, is mainly formed in the liver and small portion in the kidney. It stimulates the formation of megakaryocytes and the release of platelets. It also works with erythropoietin to stimulate erythroid progenitor cells. The elimination half-life of thrombopoietin is approximately 30 hours. It is the longest half-life of the hematopoietic growth factors.
 
The Sequence of Hematopoietic Cell Maturation
  • Erythroid lineage: Pluripotential stem cell → myeloid stem cell → BFUE → CFUE → pronormoblast → basophilic normoblast → polychromatic 2normoblast → orthochromic normoblast → polychromatic erythrocyte (reticulocyte) → erythrocyte.
    The last stage of erythroid lineage, which is still capable of division, is the orthochromic cell stage.
  • Megakaryocyte lineage: Pluripotential stem cell → myeloid stem cell → CFUGEMM → CFUMEG → megakaryoblast → megakaryocyte → platelets.
  • Myeloid lineage: Pluripotential stem cell → myeloid stem cell → CFUGEMM → CFUGM → myeloblast → promyelocyte → myelocyte → metamyelocyte → band → polymorphonuclear neutrophil.
    Monocyte maturation follows the following sequence: from CFUGM monoblast → promonocyte → monocyte then moves to circulation and tissue.
    The last stage of myeloid lineage, which is still capable of division, is the myelocyte stage. The earliest detectable, specific myeloid antigen is CD33.
  • Eosinophil: Pluripotential stem cell → myeloid stem cell → CFUGEMM → CFUEO → CFUGM → myeloblast → promyelocyte → eosinophilic myelocyte → eosinophilic metamyelocyte → eosinophilic band → eosinophil.
  • Basophil: Pluripotential stem cell → myeloid stem cell → CFUGEMM → CFUBASO → myeloblast → promyelocyte → basophilic myelocyte → basophilic metamyelocyte → basophilic band → basophil → mast cell.
  • B-cell: Pluripotential stem cell → lymphoid stem cell → pre-B → B-lymphoblast → B-prolymphocyte → B-lymphocyte → plasma cell.
    Cytoplasmic CD22, CD10, CD19, TdT and HLA-DR are present on the very early B-cells.
  • T-cell: Pluripotential stem cell → lymphoid stem cell → pre-T → T-lymphoblast → T-prolymphocyte → T-lymphocyte.
    CD7 is the earliest antigen present on the T-cell surface, and cCD3 (cytoplasmic CD3) is the earliest T-cell lineage specific antigen in the cytoplasm.
 
Erythrocytes
The main function of red blood cell is to carry oxygen to tissues and return carbon dioxide to the lung. Hemoglobin molecules present in the RBC contain four peptide chains. The lifespan of a normal RBC is 120 days.
When the oxygen is released, the hemoglobin β-chains are pulled apart so that 2,3 DPG can move in and lower the affinity of hemoglobin for oxygen, resulting in improved delivery of oxygen to the tissue.
Hemoglobin affinity for oxygen is increased by HbF, increased pH and decreased the level of 2,3 DPG. As the affinity for oxygen increases, the disassociation curve will shift to the left.
Hemoglobin affinity is decreased by HbS, decreased pH, increased the level of 2,3 DPG. As the affinity for oxygen decreases, the disassociation curve will shift to the right.
 
Neutrophils
Neutrophil production and differentiation in the bone marrow take 6–10 days. Large numbers of band and segmented neutrophils are stored in the bone marrow 3as a reserve pool (10–15 times of the peripheral neutrophils). After being released from the bone marrow, neutrophils typically spend 6–12 hours in the peripheral blood circulation before migrating into tissues. Neutrophils then survive about 2–4 days in the tissue before being destroyed.
Table 1.1   Comparison of embryo, newborn and normal adult hemoglobin
Hemoglobin
Peptide chain
Site of erythropoiesis
Embryo
Gower1
Portland
Gower2
 
(2ζ2ε)
(2ζ2γ)
(2α2ε)
Yolk sac
ζ is equivalent to the α-chain
€ is equivalent to the β-chain
Newborn
HbA
HbA2
HbF
 
(2α2β)
(2α2δ)
(2α2γ)
Bone marrow and spleen
 
 
 
Normal adult
HbA
HbA2
HbF
 
(2α2β)
(2α2δ)
(2α2γ)
Bone marrow
 
 
 
Table 1.2   Granule contents of neutrophils
Primary (azurophilic) granules
Specific granules
Myeloperoxidase
Serine protease
Acid hydrolase
Lysozyme (muramidase, etc.)
Lysozyme (muramidase, etc.)
Collagenase
Gelatinase
 
 
The Left and Right Shift of Granulocytes
The degree of nuclear lobulation of polymorphonuclear neutrophils (PMNs) is an indication of the cell's maturity. A predominance of hypolobulated cells is called a left shift. Conversely, a predominance of cells with four nuclear lobes is called a right shift. For practical purposes, a left shift is usually noted when more than 10–12% bands are seen on the CBC differential count, or when the total PMN count (segmented and band forms) is more than 80%.
Potential causes of left shift: Bacterial infection, toxemia, hemorrhage, and myeloproliferative disorders.
Potential causes of right shift: Liver disease, megaloblastic anemia, iron deficiency anemia, glucocorticoid use, and reaction to stress.
 
Eosinophils
Eosinophils have receptors for IgE, histamine, the Fc portion of immuno- globulin, and complement. They are capable of phagocytosis. Their granules are membrane bound organelles with a “crystalloid” core. Eosinophils are particularly 4important in allergic and parasitic disease. Eosinophils release arylsulfatase and histaminase, which inactive histamine and SRS-A released from mast cells.
Table 1.3   Granule contents of eosinophils
Primary granules
Secondary/special granules
Charcot-Leyden crystal (lysophospholipase)
Eosinophil peroxidase
Eosinophil peroxidase
Major basic protein (forms the crystalline core of the granules)
Eosinophil cationic protein
Eosinophil-derived neurotoxin (as known as eosinophil protein X)
Gelatinase
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Fig. 1.1: Ultrastructure of an eosinophilA. Primary granules contain Charcot-Leyden crystals. B. Rectangular amorphous crystalloid bar in secondary granules contain major basic protein, eosinophil peroxidase, and other secondary/special granules.
A major constituent of the eosinophil is the Charcot-Leyden protein, which is composed of lysophospholipase. Massive infiltration of eosinophils often leads to the disintegration of cells and formation of Charcot-Leyden crystals. Charcot-Leyden crystals are bipyramidal, hexagonal crystals that may be seen in tissue and fluid specimen (Fig. 1.1).
 
Basophils and Mast Cells
They are both derived from the bone marrow; however, their relationship is not entirely clear.
Mature and immature basophils both contain coarse, densely stained blue-black granules with scattered red-purple granules of varying shapes and sizes.
The mast cells are also referred as tissue basophils.5
Table 1.4   Comparison of basophils and mast cells
Basophils
Mast cells
Size
Mitotic potential
Location
Nuclear
Cytoplasm
Cytochemistry
Granule color
10–15 µm
No
Blood or bone marrow
Round
Moderate amount
Chloroacetate esterase (−)
Myeloperoxidase (+)
Blue-black, some purple to red
15–30 µm
Yes
Tissue or bone marrow
Round in earlier stage and segmented in later stage
Abundant granules
Chloroacetate esterase (+)
Myeloperoxidase (−)
Black, bluish-black, or reddish-purple
Granule content Both contain heparin, SRS-A, histamine, tryptase
Mast cells are larger than basophils, contain black, bluish-black, or reddish- purple metachromatic granules. The granules may overlay the cytoplasm and obscure the nuclear borders.
 
Monocytes
Monocytes share a common stem cell origin with myeloid cells. Mature monocytes have gray-blue cytoplasm and indented nuclei. The cytoplasm may contain vacuoles or fine granules. Monocytes spend only a short time in the bone marrow. After circulating in the blood for 12–24 hours, monocytes migrate into the tissue as antigen presenting cells (APC) or macrophages without dividing.
 
T-lymphocytes
Mature T-cells comprise 65–80% of the circulating lymphocytes in peripheral blood. There are two major subsets, CD8+ suppressor (as known as cytotoxic) T-cells (predominantly in bone marrow), and CD4+ helper T-cells (predominantly in peripheral blood).
Maturation of T-cells occurs in the thymus. T-cells that have migrated from the bone marrow begin the thymic maturation process in the subcapsular region of the lobules of the thymus. As they mature, T-cells progress inward from the subcapsular region, to the cortical region, and finally to the medullary region where they are released into the peripheral blood. As the T-cells mature, cytoplasmic CD3, TdT, and CD7 are expressed first, followed by CD2, and then CD5. When the maturing T-cells have reached the cortex region, they express CD1a and co-express CD4/CD8. When they have reached full maturity in the medullary region, the T-cells express surface CD3, and either CD4 or CD8. Rearrangement of the T-cell receptor (TCR) occurs in a specific sequence following the order of δ, γ, β, and α. The delta (δ) gene (14q11) is rearranged first, followed by rearrangement of the gamma (γ) gene (7p14). This leads to expression of γ/δ T-cells (a minority of the circulating T-cell population), or in the case of the majority of T-cells, the T-cells progress on to rearrange the beta (β) gene (7q34), followed by deletion of the delta gene, and finally, rearrangement of the alpha (α) gene (14q11) to produce α/β T-cells (95% of the circulating T-cell population).6
 
B-lymphocytes
B-cells comprise 5–15% of the circulating lymphocyte population in the peripheral blood. B-cells express CD10, CD19, CD20, CD22, CD79a, PAX-5, and HLA-DR. Plasma cells are mature B-cells that secrete immunoglobulin.
B-cell maturation occurs in the bone marrow and follows a sequential B-cell gene rearrangement process. The first step in the production of a functional immunoglobulin involves recombination at the heavy chain locus (IgH) on chromosome 14q32 by joining a diversity (D) segment with a joining (J) segment to form a D-J fusion, followed by joining a IgH variable (V) region to the D-J fusion to form VDJ fusion. Following rearrangement of IgH, the next rearrangement is the light chain. The kappa light chain (located on chromosome 2p12) rearrangement occurs first, if unsuccessful, then lambda light chain (located on chromosome 22q11) rearrangement occurs.
 
Natural Killer Cells (NK Cells)
Natural killer cells are a minor population of cells, which do not carry either T- or B-cell markers. Large granular lymphocytes seen in peripheral blood smears comprise a major proportion of NK cells. Interferon-γ and IL-2 stimulate NK cell proliferation.
 
HEMATOLOGY TESTING
 
Anticoagulant Agents Used for Peripheral Blood Cell Counts
  • EDTA is the anticoagulant of choice. EDTA can cause platelet clumping and/or satellitism in automated blood counters which is referred to as EDTA-pseudothrombocytopenia. EDTA is also the preferred anticoagulant agent for the extraction of DNA from plasma.
  • Citrate causes dilution of the specimen, which requires mathematical correction.
  • Heparin may cause WBC and platelet clumping; and therefore is unsatisfactory as an anticoagulant.
 
Terminology and Important Calculations
The hematology analyzer is a laboratory instrument that generates a histogram showing size distribution on the X-axis and relative number of particles on the Y-axis. RBC and MCV are directly measured. MCHC and MCH are calculated values.
Spurious analyzer readings may occur. The causes of spuriously increased WBC count include cryoglobulin, Heparin, monoclonal protein, nucleated red cells, platelet clumping, and unlysed red cells. The causes of spuriously decreased WBC count include clotting, smudge cells, and fragmented WBCs.
  • Absolute WBC count: Total WBC × % (neutrophil, monocyte or lymphocytes)
  • Red blood cell (RBC): The normal reference range for males is 4.6–6.0 × 106/µL, and for females is 4.1–5.4 × 106/µL. The presence of cryoglobulin, a WBC count >50,000/µL, or large platelets may lead to spuriously increased 7RBC count. Microcytic red blood cells (schistocytes, iron deficiency anemia, thalassemia) clotting, or agglutination may lead to spuriously decreased RBC counts.
  • Mean corpuscular volume (MCV): The normal reference range is 80–100 fL. It is a measurement of the red blood cell volume or size. In cases of red blood cell clumping (warm or cold agglutinins), or osmotic abnormalities (hyperglycemia, hypernatremia), the MCV may spuriously elevate.
  • Red blood cell distribution width (RDW): The normal reference range is 11.5–14.5%. It measured anisocytosis (difference in cell size). An increase RDW may indicate a mixed cell population.
    RDW is calculated as following:
Table 1.5   Comparison of MCV, RDW and their corresponds diseases
MCV (fL)
Normal RDW
High RDW
<70
Thalassemia
Anemia of chronic disease
Iron deficiency HbH
Normal
Anemia of chronic disease
Hereditary spherocytosis
Bleeding
Early or partially treated iron or vitamin D deficiency
Sickle cell disease
>100
Aplastic anemia
Myelodysplastic syndrome
B12 or folic acid deficiency
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  • Hemoglobin concentration (Hb): The normal reference range for males is 14–18 g/dL, and for females is 12–16 g/dL. A commonly used method is to convert hemoglobin to cyanhemoglobin, and then measure the absorbance at a wavelength of 540 nm.
    Hgb is measured spectrophotometrically; therefore, causes of increased sample turbidity such as paraprotein, lipids, abnormal hemoglobins, or nucleated cells can lead to erroneous results.
  • Hematocrit (Hct): The normal reference range for males is 40–50%, and for females is 37–47%. It is the percentage of blood volume occupied by RBCs. Errors can occur in centrifugation due to plasma trapping, over dilution by anticoagulant, or by prolonged tourniquet application. The hematocrit is calculated as following:
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  • Mean corpuscular hemoglobin (MCH): The normal reference range is 27–31 pg. It is the average amount of hemoglobin per cell.
    The MCH is calculated as follows:
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    Mean corpuscular hemoglobin concentration (MCHC): The normal reference range is 32–36 g/dL. MCHC is increased in spherocytosis and decreased in microcytic or hypochromic anemia. Since this is a calculated value, an erroneous value for hemoglobin may affect the MCHC value.
    MCHC is calculated as follows:8
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  • Reticulocyte count used as a general indicator of bone marrow erythropoiesis and release. A normal or decreased reticulocyte count associated with moderate or marked anemia is strong evidence that the bone marrow is not responding appropriately. An increased reticulocyte count indicates a rapid erythroid cells turnover, which may be associated with blood loss, or acute/chronic hemolysis. The lifespan of reticulocytes is approximately 3–4 days in the bone marrow and 24 hours in peripheral blood. Reticulocytes cannot be visualized with the regular Giemsa stain; therefore special stains such as new methylene blue and brilliant cresol blue are used to precipitate the residual RNA material in these cells to visualize reticulocytes. For automated or flow cytometry analysis, reticulocytes can be stained with auramine O and thiazole orange.
    The corrected reticulocytes count (CRC) is calculated as follows:
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    The reticulocyte production index (RPI) is calculated as following:
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    Correction factor
    Hct
    Correction factor
    40–45
    1.0
    35–39
    1.5
    25–34
    2.0
    15–24
    2.5
    <15
    3.0
    The normal range of RPI is 1 to 2. In an anemic patient, RPI < 1 indicates a decreased production of reticulocytes and red blood cells. RPI > 2 indicates an increased production of reticulocytes to compensate the loss of red blood cells (destruction or bleeding).
  • Serum iron measures iron bounded to transferrin (ferric form). There is diurnal variation (highest level in the morning); therefore, serum iron levels should be drawn in the morning.
  • Total iron-binding capacity (TIBC) measures the concentration of transferrin. It indicates the iron concentration needed to saturate all transferrin-binding sites.
    The normal percentage of transferrin saturation (PTS) is 30% and is calculated as follows:
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  • Serum ferritin: Ferritin is a storage complex of apoferritin and iron. It is a relatively sensitive and reliable indicator for iron-deficiency anemia. It is elevated in iron overload (sideroblastic anemia, hemochromatosis), Gaucher's disease, and inflammatory diseases (ferritin is also an acute phase 9reactant). If iron deficiency and inflammatory disease co-exist, the serum ferritin level may be normal.
    Table 1.6   Potential causes of erroneous results of automated hematology analyzer
    Falsely increase
    Falsely decrease
    WBC
    Cryoglobulin
    Extremely elevated protein
    Nucleated RBC
    Unlysed RBC
    Malaria parasites in RBC
    Platelet clumping
    WBC aggregates
    Fragmented WBC
    RBC
    Cryoglobulin
    Giant platelets
    High WBC count
    Autoagglutination
    Microcytic RBC
    Platelet
    Cryoglobulin
    Microcytic RBC
    Fragmented WBC
    Giant platelets
    Platelet clumping
    Platelet satellitosis
    Hemoglobin
    Cryoglobulin
    High WBC count
    Severe lipidemia
    Heparin
    MCV
    Autoagglutination
    Hyperglycemia
    High WBC count
    Cryoglobulin
    Giant platelets
  • Soluble serum transferrin receptor (STFR): The transferrin receptor acts as an iron-transporting molecule, it present on the most cell surface. The expression of transferrin receptors is dependent on the concentration of iron in the cellular cytoplasm. STFR is increased in iron deficiency anemia and hemolytic anemia.
  • Free erythrocyte protoporphyrin (FEP): FEP is useful in distinguishing iron deficiency anemia and thalassemia minor. FEP is elevated, when there is a failure of iron incorporation to heme (iron deficiency, sideroblastic anemia, anemia of chronic disease, lead poisoning). Thalassemia is associated with abnormal hemoglobin synthesis, but not abnormal heme synthesis, so the FEP level is normal.
  • High performance liquid chromatography (HPLC): HPLC measures HbA2 (increased in most β-thalassemia) and globin chain ratio.
  • Interfering substances of automated hematology analyzer (Table 1.6): Check for hemolysis and clotting before using the specimen.