Lymphoma Diagnostic Work-up from a Lab Perspective

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Lymphoma Diagnostic Work-up from a Lab Perspective Mohamed E. Salama M.D. Mohamed.salama@path.utah.edu

Lymphoproliferative Disorders Malignant lymphoma 1. Non-Hodgkin lymphoma (NHL) 2. Hodgkin (disease) lymphoma 3. Multiple myeloma

Basic concepts Lymphomas are solid tumors of the hematopoietic system. Neoplasms of lymphoid origin, typically causing lymphadenopathy leukemia vs. lymphoma Leukemias as systemically distributed neoplasms of white cells

Very important concept lymphomas and leukemias are clonal expansions of cells at certain developmental stages

Mantle zone Follicle / germinal center (B cell) (B-cell) Paracortex (T-cell) Medullary cord (B-cell) Lymphatic sinus NORMAL LYMPH NODE

B-cell development stem cell lymphoid precursor mature naive B-cell germinal center B-cell memory B-cell progenitor-b pre-b immature B-cell plasma cell

B-cell development stem cell lymphoid precursor CLL MCL mature naive B-cell germinal center B-cell memory B-cell MZL CLL progenitor-b MM LBL, ALL pre-b immature B-cell DLBCL, FL, BL, HL plasma cell

A practical way to think of lymphoma Category Survival of untreated patients Curability To treat or not to treat Non- Hodgkin lymphoma Indolent Years Generally not curable Aggressive Months Curable in some Generally defer Rx if asymptomatic Treat Very aggressive Weeks Curable in some Treat Hodgkin lymphoma All types Variable months to years Curable in most Treat

Non-Hodgkin Lymphomas How do we diagnose and classify these types of lymphoproliferative disorders? Architectural pattern Cytologic (cellular) morphologic appearance Immunophenotypic (antigenic) characteristics Molecular / genetic characteristics

Diagnosis requires an adequate biopsy Diagnosis should be biopsy-proven before treatment is initiated Need enough tissue to assess cells and architecture open bx vs core needle bx vs FNA

Lymph Node Protocol Permanent sections Morphologic evaluation Flowcytometry Immunostains Cytogenetics Molecular

Detection enzyme CD20 AntiCD20 antibody

Flow-cytometry

Flow-cytometry Flow-cytometry

Flow-cytometry

Immunostains

Non-Hodgkin Lymphomas Neoplasm of the immune system B-cells, T-cells, histiocytes Usually begin in the lymph nodes, but may arise in other lymphoid tissues such as spleen, bone marrow, or extranodal sites

Clinical Findings Enlarged, painless lymphadenopathy B-symptoms-fever, weight loss Impingement or obstruction of other structures

Subtypes of Non-Hodgkin Lymphoma Frequency % I Diffuse Large B cell Lymphoma 422 31 L Follicular Lymphoma 306 22 L Chronic Lymphocytic Leukemia 88 6 I Mantle Cell Lymphoma 83 6 L Marginal Zone B-cell Lymphoma,MALT-type 72 5 L Marginal Zone B-cell Lymphoma,Nodal 20 1 L Lymphoplasmacytic Lymphoma 15 1 H Burkitt Lymphoma 10 <1 H Burkitt-like Lymphoma 29 2 H Lymphoblastic Lymphoma T/B 26 2 I Peripheral T cell Lymphoma 76 6 I Anaplastic Large T-/Null cell Lymphoma 33 2

Most common types of lymphoma 1. Non-Hodgkin lymphoma (NHL) - SLL/CLL - Follicular lymphoma - Diffuse large B cell lymphoma - Burkitt s lymhoma 2. Hodgkin lymphoma (HL)

Non-Hodgkin lymphoma Incidence Diffuse large B-cell lymphoma Follicular lymphoma Other NHL

General Features Low Grade Lymphomas Adult population affected (median age, 50-70 years) Rare in children High stage disease (III/IV) is most common Indolent course with relatively long survival Generally incurable Transformation to higher grade NHL may occur

Small Lymphocytic Lymphoma Low grade B-cell malignancy Similar to chronic lymphocytic leukemia (CLL) Frequency - ~ 4% of NHL Older age group (median, 60.5 years) Bone marrow involvement: Common Indolent course

Flow cytometry

Follicular Lymphomas Frequency -~40% of NHL (most common) Older age group (median, 55 years) Often asymptomatic Bone marrow involvement: Common Indolent Course Chromosomal translocation, t(14;18)

Follicular Lymphomas Several chemotherapy options if symptomatic Median survival: years Transformation to more aggressive B-cell lymphoma

Follicular Lymphoma

Reactive Follicular Hyperplasia

Architectural Features Distinguishing Reactive Follicular Hyperplasia and Follicular NHL Reactive Follicular Follicular NHL Hyperplasia Nodal Architecture Preserved Effaced Germinal Center Size & Shape Marked variation Slight to moderate variation Capsular infiltration None or minimal Invasion with extension into pericapsular fat Density of follicles Morphology of follicles Low, with intervening lymphoid tissue Sharply defined, mantle zone High, with back to back follicles Ill defined, no mantle zone

Intermediate Grade/Aggressive Mantle cell lymphoma t(11;14) translocation results in over- expression of cyclin D1 protein Diffuse large cell lymphoma

Diffuse Large Cell 60-70% derived from B-cells Often stage I or II at diagnosis More likely to have extranodal sites Peripheral blood involvement is rare

Diffuse large B-cell lymphoma

Diffuse Large B-cell Lymphoma CD20 H & E

MIB-1

High Grade (small non-cleaved) Burkitt lymphoma Endemic in Africa Seen in children and related to Epstein-Barr virus Usually extranodal AIDS associated lymphoma

Clinical Findings Enlarged painless lymphadenopathy B-symptoms, fever, sweats, weight loss Impingement or obstruction of adjacent structures (mass effect) Extranodal presentation (30% of cases) GI tract, spleen, salivary gland

Burkitt lymphoma involving jaw

Burkitt lymphoma - Starry-sky pattern

Burkitt lymphoma tingible-body macrophages

High grade Burkitt lymphoma - Endemic in Africa - Seen in children and related to Epstein-Barr virus - B-cell phenotype - t(8:14) MYC/IgH - Usually extranodal -High mitotic rate (starry-sky) Lymphoblastic lymphoma

Sum Indolent Lymphomas Very slow growing, over years. Follicular lymphoma, grades I/II is prototype. If can t cure, goal is to control disease/symptoms. Decision of WHEN to treat is important. Aggressive Lymphomas Rapidly growing, over days, months. Diffuse large B cell lymphoma is prototype. Cure is possible. About 50% with multi-agent chemotherapy.