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Category Archives: Lymphoma
Methods to establish monoclonality
Methods to establish monoclonality
• Immunohistochemistry
• Flow cytometry
• Molecular techniques (eg. PCR)
Posted in Lymphoma
Tagged B cell lymphoma, B cell lymphproliferative lesion, Flow cytometry, Immunohistochemsitry, Molecular techniques, Monoclonal lesion, Monoclonality
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Waldenstrom Macroglobulinemia
Waldenstrom Macroglobulinemia
Lymphoplasmacytic Lymphoma
Also known as Waldenstrom Macroglobulinemia
Monoclonal IgM serum paraprotein
Hyperviscosity symptoms
Morphology
Mixture of cells
Small lymphocytes
Plasmacytoid lymphocytes
Plasma cells
Dutcher bodies
Immunophenotype
CD19, CD20, CD22, CD79a
CD38
Surface IgM
Molecular Genetics
Heavy chain of immunoglobulin rearranged
Posted in Lymphoma
Tagged Lymphoma, lymphoplasmacytic lymphoma, Macroglobulinemia, Waldenstrom lymphoma, Waldenstrom Macroglobulinemia
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Hairy Cell Leukemia
Hairy Cell Leukemia
Males
Middle age
Pancytopenia
Splenomegaly often
Heptaomegaly
NO lymphadenopathy commonly
Spleen
Bloody lakes of red pulp infiltration by neoplastic cells
Peripheral Blood
Medium sized cells
Circumferential hairy projections
Bone Marrow Aspirate
Dry Tap – as neoplastic cells are difficult to aspirate due to reticulin
Immunophenotype
CD19, CD20, CD22, CD79a
CD11c, CD25, CD103, Cyclin D1
Molecular Genetics
Heavy Immunoglobulin chain rearrangements
Small Lymphocytic Lymphoma
Small Lymphocytic Lymphoma
Chronic lymphocytic leukemia Small Lymphocytic Lymphoma
Patients usually asymptomatic.
Many different presentations including fatigue, absolute lymphocytosis, hepatosplenomegaly, anemia, lymphadenopathy.
Morphology
Small lymphocytes
Occasional mixed large lymphocytes termed prolymphocytes
Proliferation centers composed of paraimmunoblasts and prolymphocytes
Peripheral Blood
Small cells with no nucleoli and clumped chromatin
Immunophenotype
CD19, CD20, CD22, CD23, CD79a
CD5
Molecular Genetics
Trisomy 12
Deletions
Somatic hypermutation
Lymphoma Follicular
Follicular Lymphoma
Non-Hodgkin’s Lymphoma
Most common non-hodgkin’s lymphoma
45% of adult lymphomas
Patients middle age
Widespread peripheral and central lymphadenopathy
Bone marrow involved in less than half of cases
Morphology
Cells appear as normal germinal center B cells
Follicular pattern – closely packed follicles
Follicles contain either cleaved centrocytes or large centroblasts
Also exists a diffuse pattern that will stain in a follicular pattern
Grading Follicular Lymphoma
Grade 1 : 0 to 5 centroblasts per high power field
Grade 2 : 6 to 15 centroblasts per high power field
Grade 3 : > 15 centroblasts per high power field
Immunophenotype
CD19, CD20, CD22, CD79a, CD10, BCL-2, BCL-6
Molecular Genetics
Translocation t(14;18)(q32;q21)
BCL-2 rearrangement leading to BCL-2 overexpression and inhibition of apoptosis
Chronic Lymphocytic Leukemia
Chronic Lymphocytic Leukemia
Small Lymphocytic Lymphoma
Chronic Lymphocytic Leukemia and Small Lymphocytic Lymphoma are the same disease but differ in the extent of malignant white blood cells in the peripheral blood (lymphocytosis).
Chronic Lymphocytic Leukemia
4000 per mm3
Diagnostic requirement for Chronic Lymphocytic Leukemia is an absolute lymphocyte count of greater than 4000 per mm3.
Most common leukemia in Western world
Less common in Asia
Average age is 60
Males more common than females
Patients often asymptomatic
Nonspecific symptoms: fatigue, anorexia, weight loss
Half of cases have lymphadenopathy
Half of cases have hepatosplenomegaly
1/10 of patients have autoantibodies to red blood cells and platelets
Chronic Lymphocytic Leukemia often transforms to a more aggressive lymphoid malignancy including:
Prolymphocytic transformation (25% of patients)
Diffuse large B cell lymphoma (10% of patients)
Transformation to one of these leads to a survival of less than 1 year
Morphology
Diffuse effacement of lymph node by small lymphocytes
Irregular nuclei
Condensed chromatin
Scant cytoplasm
Large cells, prolymphocytes, admixed with small lymphocytes – often form aggregates termed proliferation centers containing many mitotically active cells
Proliferation centers are diagnostic for Small Lymphocytic Lymphoma / Chronic Lymphocytic Leukemia
Peripheral Blood in Chronic Lymphocytic Leukemia
Increased numbers of small, round lymphocytes
Scant cytoplasm
Smudge cells – fragile cells that have ruptured during making of smear
Bone Marrow in Chronic Lymphocytic Leukemia
Always involved by small lymphocytes
Interstitial infiltrates
Non-paratrabecular aggregates
Spleen in Chronic Lymphocytic Leukemia
Involvement of both white and red pulp
Liver
Involvement of hepatic portal tracts
Immunophenotype
Pan B cell markers CD19, CD20
CD5 (T cell marker)
CD23
Surface immunoglobulin (IgM, IgD)
Molecular Genetics
Deletions of 13q12-14
Lymphoma Protocol
Lymphoma Protocol
Lymphoid tissue once received from the surgeon should be examined grossly with touch preparations and/or frozen section being done.
The lymph node is cut in perpendicular sections and submitted into different tubes for specific studies to aid in diagnosis.
Cut lymph node perpendicularly along long axis.
1. Wet touch preparations fixed in formalin or 95% alcohol for hematoxylin and eosin staining
2. Air dried touch preparations (left alone to air dry) for cytogenetics (FISH) or cytochemistry or Giemsa staining
3. Snap freeze (rapidly freezing, in –70 degrees, liquid nitrogen) tissue, small amount (e.g. 0.5 x 0.2 cm), for molecular analysis including genetics, also for immunohistochemistry
4. Fresh tissue in saline for flow cytometry (also small amount of tissue e.g. 0.5 x 0.2 cm)
5. B5 fixative and tissue (e.g. one slice of tissue if have a lot, if not 0.5 x 0.2 cm should suffice). B5 provides high nuclear detail, good for Hodgkin’s lymphoma. Usually done when the patient is young and suspecting Hodgkin’s lymphoma.
B5 is toxic to the environment as it has high levels of mercury. It is also very expensive and molecular studies are not effectively done on B5 fixed tissue.
6. Remaining tissue (all) into formalin (10% neutral buffered) for hematoxylin and eosin staining and immunohistochemistry
Optional (depends on your suspicion)
7. Thin shaved tissue in glutaraldehyde for electron microscopy (must put tissue QUICKLY into glutaraldehyde)
8. Sterile tissue, fresh, for microbiology (e.g. sent for cultures). Can also be used for cytogenetics (e.g. FISH)
Posted in Lymphoma
Tagged , Lymphoma pathology protocol, Lymphoma protocol, Protocol lymphoma
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Small Cell Lymphoma
Small Cell Lymphoma and Chronic Lymphocytic Leukemia
Appearance in lymph node
Diffuse effacement of lymph node architecture by population of small cells (6-12 um in diameter)
Cells contain small, round, slightly irregular nuclei with condensed chromatin and scant cytoplasm
These cells are mixed with variable numbers of larger cells known as prolymphocytes
Prolymphocytes gather to form loose aggregates known as proliferation centres (which contain large numbers of mitotically active cells) and are pathognomonic for CLL/SLL
Appearance in bone marrow
Observed in all cases of CLL and most cases of SLL
Small lymphocytes are seen to form interstitial infiltrates and/or non-paratrabecular aggregates
Small Cell Lymphoma Immunohistochemistry
CD19/20 +
CD5+
CD21, CD23+
CD43+
CD10-
Cyclin D1-
IgM, or IgM/IgD +
Either kappa or lambda restricted
Prognosis
Patients present over age 50 (median age: 60 y.o.), often asymptomatic
When symptomatic, symptoms are non-specific (fatigue, weight loss, anorexia)
Course and prognosis is variable and dependent on clinical stage
Median survival is between 4-6 years, however some patients with minimal disease survive over 10 years
Presence of 11q and 17p deletions portends a poor prognosis
Transformation to more aggressive neoplasms can occur (eg. prolymphocytic transformation [15-30% of patients] or transformation to DLBCL [10% of patients]) with most patients surviving less than 1 year
Differential diagnosis
Mantle cell lymphoma
Follicular cell lymphoma
Marginal zone lymphoma
Reactive lymph node hyperplasia
Posted in Lymphoma
Tagged , Chronic lymphocytic leukemia, cll, sll, Small Cell Lymphoma
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Burkitt
Burkitt lymphoma
Burkitt Lymphoma Pathogenesis
Burkitt lymphoma is caused by the translocation of c-myc (on chromosome 8 ) to immunoglobulin gene locus allowing for upregulation of c-myc (increased transcription and translation). Translocation occurs to IgH locus t(8;14) or light chain locus t(2;8) or t(8;22). This is thought to occur during errors in class switching or during attempted V(D)J joining, or breaks generated during somatic hypermutation of Ig genes.
Additional mutation of p53, MDM, or p16 are needed for malignant transformation.
Burkitt Lymphoma Histology
Tissues involved by Burkitt lymphoma are effaced by a diffuse proliferation of intermediate-sized cells (10 to 25 um diameter; similar to nuclear size of benign macrophages within tumor), containing round or oval nuclei with coarse chromatin, several nucleoli, and a moderate amount of faintly basophilic to amphophilic cytoplasm. There is a high mitotic index and high rate of apoptotic cell death within the tumor. This is a requirement. Tingible body macrophages are diffusely scattered among the tumor cells imparting a “starry sky” appearance to the neoplasm.
Classification in the Working Formulation and the REAL classification
Working Formulation
- Classified under high grade lymphoma (small, noncleaved cells)
REAL Classification
- Classified as peripheral B-cell neoplasm
References:
Robbins Basic Pathology 7th ed, edited by Vinay Kumar, Ramzi S. Cotran, and Stanley J. Robbins, 873 pp, Philadelphia, Pa, Sounders, 2003.
Posted in Lymphoma
Tagged burkitt, burkitt lymphoma, Cancer, high grade lymphoma, lyphoma, peripheral b cell
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Lymphoma Mutations
Lymphoma Mutations
Mantle cell lymphoma mutation:
cyclin D1 translocation [t(11;14]
Follicular lymphoma mutation:
translocation of bcl-2 to IgH heavy chain of immunoglobulin[t(14;18)]
Chronic Myelogenous Leukemia (CML) mutation:
creation of fusion protein bcr-abl from translocation [t(9;22)]
References:
Robbins Basic Pathology 7th ed, edited by Vinay Kumar, Ramzi S. Cotran, and Stanley J. Robbins, 873 pp, Philadelphia, Pa, Sounders, 2003.