Intro to Leukocyte Neoplasms

Lymphoblast vs. Myeloblast Oncogenes; tumor suppressor genes Chemotherapy, Radiation therapy, supportive therapy, targeted therapy, stem cell transplantation (allogenic, autologous)

Cytochemistry
Acceptable Specimens and Fixatives
Enzymatic: fresh Non enzymatic: smears stable at RT for months Methanol, ethanol, acetone, formaldehyde

Myeloperoxidase
In primary granules of neutro, eos >>> mono MPX oxidizes dye when H2O2 is present black to red brown Substrate : benzidine (carcinogenic), DAB, pphenylenediamine diHCl

Sudan Black B
More sensitive for early myeloid cells Stains lipids; found in primary and secondary granules of neutrophils & lysosomal granules of monocytes More intense staining as myeloblast matures (more primary & secondary granules)

Esterases
Myeloblasts & neutrophils vs. monocytic Non specific (for monyctes): a-naphthyl acetate; a-naphthyl butyrate (less sensitive but more specific) Specific: naphthol AS-D choloroacetate esterase (specificity-staining of only myelocytic cells) + in myeloblasts Hydrolysis of ester; a naphthol compound is released & combines w/ diazonium salt

PAS
ALL, M6, granulocytes Oxidizes glycogen; aldehydes react with colorless Schiff rgt bright red-pink ALL lymphoblasts: coarse block or finely diffuse pattern Erythroblasts: coarse and granular

 Factor VIII Acid Phosphatase (TRAP) LAP

Cytogenetics
Reproductive loss, birth defects, cancer Metaphase chromosomes Cell cycle: G1SG2mitosis Q banding: bands of differing lengths and relative brightness G banding(Giemsa), R banding (reverse Giemsa), C banding (centromere/primary constriction)

Cytogenetics
FISH Cancer cytogenetics
Clonal proliferation (lymphoid vs. myeloid) Clinical course (acute vs. chronic) Philiadelphia seen in: about 20% of adults with ALL 2-5 % of children with ALL 1% with AML Solid Tumors HER2 invasive breast cancer

Molecular Diagnostics
PCR Detecting amplified DNA gel electrophoresis NA hybridization Southern blot Assessment of Minimal Residual Disease

Flow Cytometry; Gating

Myeloid lineage: CD34, CD38, HLA-DR, stem cell receptor CD117 Panmyeloid markers: CD13, CD33 Myeloblastpromyelocyte: loss of CD34 & HLADR; acquires CD15 Myelocyte: CD11b; temporary loss of CD13; gradual decrease in density of CD33 Band: CD16; CD13

 Monocytic Linage
Increase in density of CD13, CD33, CD11b Promonocyte to monocyte: DD15 and CD14 Bright expression of CD64 and HLA-DR

Erythroid Lineage
Transferrin receptor CD71 Glycophorin A (from basophilic normoblast stage)

Megakaryocytic Lineage
CD41, CD61 (GPIIb/IIIa complex) CD31, CD36 / CD42, CD62P, CD63

Lymphoid Lineage
CD34, TdT, HLA-DR B cell markers
Early: CD19, cytoplasmic CD22, cytoplasmic CD79 Later: CD10, CD20 (as CD10 decreases), cytoplasmic u chain

T cell markers
CD34, TdT, CD2, CD7, cytoplasmic CD3; then CD1a, CD5, CD4, CD8; then CD3 on cell surface while CD4 and CD8 is lost NK derived neoplasm: CD56

Myeloproliferative Disorders
CML Polycythemia Vera Essential Thrombocythemia Chronic Idiopathic Myelofibrosis

CML
Translocation of an ABL protooncogene from band q34 of chromosome 9 to the BCR region of band q11 of chromosome 22 Hyperuricemia, uricosuria, secondary gout, lithiasis Peripheral blood: refractory anemia-like

Polycythemia Vera
Panmyelosis, splenomegaly Normal or high LAP Stable phase Spent phase splenomegaly or hypersplenism
pancytopenia

Essential Thrombocythemia
> 600 or 1000 x 10 9/L Erythromelalgia (throbbing and burning pain in the hands and feet

Chronic Idiopathic Myelofibrosis

Myelofibrosis with myeloid metaplasia

MDS
Refractory anemia, smolderng leukemia, obligate leukemia or preleukemia De novo (primary); therapy-related MDS Subtypes Dyserythropoiesis- oval macrocytes, hypochromic, dimorphic, ringed sideroblasts Dysmyelopoiesis - abn nuclear shapes Dysmegakaryopoiesis giant plts

Mature Lymphoid Neoplasms

Lymph Nodes Lymphoma Plasma Cell neoplasms