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Flow Cytometry > CLL and Exfoliating Lymphoma (PFC) > Flashcards

CLL and Exfoliating Lymphoma (PFC) Flashcards

1
Q

What is the normal count

for peripheral blood lymphocytes and normal composition ?

A
  • absolute levels 0.75-3.6 x 10^9/L
  • T cells normally account for 80-90%
    • identified by sCD3, which means that has matured beyond precursor T cell stage
    • cytoplasmic CD3 expression is the gold standard marker of T cell lineage
    • normal peripheral blood CD4/CD8 ratio is ~2:1
      • normal in bone marrow is 1:2
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2
Q

What is the normal T cell maturation/

immunophenotype pattern ?

A
  • express CD2, CD5, CD7
  • co-expression of CD4/CD8 is seen in thymic maturation/migration
    • can be seen in peripheral blood at low levels

IMP: should NOT have CD4 or CD8 negative lymphocytes in the peripheral blood

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3
Q

What is the normal amount of NK

cells within the peripheral blood ?

A
  • NK cells : 654 x 10^9/L
  • these cells express CD2 but not CD3
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4
Q

What is the normal proportion of B cells in

the peripheral blood ?

A
  • 10-20% of peripheral blood lymphocytes
  • cytoplasmic CD79a is the gold standard used by some to identify B cells
    • CD19 is usually reliable but can be aberrantly expressed by some non-B cell malignancies
  • blood B cells usually express CD22 and FMC7 as well as HLA-DR
    • IMP: nomrla, non-activated T cells do not express HLA-DR
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5
Q

What cells are positive for

CD38, CD23, CD5 and FMC7

normally ?

A
  • CD38
    • activated B cells, plasma cells, T and NK cells
  • CD23
    • activated, mature B cells expressing IgM or IgD
  • CD5
    • small subset of normal B cells and T cells
  • FMC7
    • mature B cells
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6
Q

What cells are normally positive for

CD10, HLA-DR, Kappa and Lambda ?

A
  • CD10
    • pre B lymphocytes and germinal center B cells
  • HLA-DR
    • Normal B cells and activated T cells
  • Kappa and Lambda
    • normal polyclonal mature B cells
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7
Q

What cells are normally positive for

CD11c and CD103 ?

A
  • CD11c
    • granulocytes, macrophages
    • NK cells
    • Dendritic cells
  • CD103
    • intraepithelial T cells
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8
Q

What cells are positive for

CD25 and CD123 ?

A
  • CD25
    • activated B and T cells
    • macrophages
  • CD123
    • plasmacytoid monocytes
    • PDCs
    • Basophils
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9
Q

Where do CLL cells likely arise from ?

A
  • either arise from:
    • Naive pre-germinal center cells (unmutated CLL)
    • or
    • post-germinal center memory B cell (mutated CLL)
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10
Q

In CLL, what do increased

prolymphocytes show by cytogenetics ?

A
  • proportional increases in prolymphocytes are associated with cytogenetic abnormalities
    • often trisomy 12 and del17p
  • also have progressive disease with refractoriness to therapy
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11
Q

FMC7 is considered to be

an epitope of which marker ?

A
  • CD20 molecule
  • FMC7 is absent or dim in CLL
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12
Q

What does CD38 expression in CLL

correlate with ?

A
  • unmutated IgVH gene or Zap70 expression
  • associated with poor prognosis

IMP: CD38 expression may vary in any individual patient cells over time

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13
Q

What marker when positive, helps differentiate

a CLL with abnormal immunophenotype vs. Mantle cell ?

A
  • CD200
    • universally positive in CLL
    • negative in Mantle cell lymphoma
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14
Q

Detection of what mutation

likely reflects refractoriness to Fludarabine therapy ?

A
  • del 17p (loss of TP53 function)
  • refractory to Fludarabine therapy
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15
Q

What marker usually is a good way to differentiate

Mantle cell lymphoma from CLL….but not always ?

A
  • CD23
  • has been reported to be positive in 25% of Mantle cell lymphoma patients

Note: rare cases of MCL have also been shown to be CD10 positive (frequently blastoid morphology)

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16
Q

What cytogenetic and morphologic

findings in Mantle cell are associated with

rapid disease progression ?

A
  • rearrangements of MYC gene (8q24) and TP53 at 17p13
    • often seeni n blastoid morphology and with progression
17
Q

How can splenic margin present ?

A
  • often appear in the blood in low number
  • can express CD5
  • mild lymphocytosis, splenomegaly and cytopenias
    • cytopenias due to hypersplenism and sometimes autoimmune phenomena
  • 1/3 of patients have a small paraprotein, often associated with hepatitis C infection
  • bone marrow involvement is variable
    • interstitial or nodular pattern
    • **affinity for the sinusoids
  • liver and spleen
    • involves the sinusoids and red pulp
18
Q

When splenic marginal zone acquires

a TP53 mutation on chromsome 17p, what

morphology is typically seen ?

A
  • larger cells with prominent nucleoli
  • voluminous cytoplasm
  • must differentiate from HCL and B-PLL
19
Q

What is the immunophenotype of

splenic marginal zone lymphoma ?

A
  • positive:
    • pan B cell antigens
    • light chain and CD20 expression is moderate to strong
    • CD5 (~20%) of cases
      • often seen in cases with more peripheral blood involvement
  • negative:
    • CD103, CD123, CD25
  • positive or negative
    • CD23 and CD11c
20
Q

What are the most common

cytogenetic findings in splenic marginal zone lymphoma ?

A
  • del 7q and gain of 3q
  • TP53 does occur in more aggressive disease
21
Q

What is the typical presentation of

B-PLL ?

A
  • middle aged to elderly males with significant lymphocytosis
  • anemia, thrombocytopenia and splenomegaly
  • lymphoadenopathy is very rare
    • if present should question the diagnosis
22
Q

What is the immunophenotype of

B-PLL ?

A
  • CD5 + in 20-30% of cases
  • CD23 + in 10-20% of cases
  • pan-B antigens positive with CD20 and light chain strong MFI
  • FMC7 and CD11c positive
    • therefore should not confuse with CLL
23
Q

What is the characteristic immunohenotype

of LPL ?

A
  • variable degrees of surface and cytoplasmic light chain expression
  • positive for mature pan-B cells
  • associated plasma cells may be restricted but do not show a malignant phenotype
  • Other markers seen:
    • weak CD23 and CD5 +
    • loss of FMC7
    • variable expression of CD38
    • CD11c and CD25 can be +
24
Q

What is a feature of follicular lymphoma

by flow cytometry that can help differentiate it from

background B cells ?

A
  • generally will have a dim CD19
25
Q

What is the typical immunophenotype of

Burkitt Lymphoma cells ?

A
  • mature precursor or early mature phenotype
  • pan B cell markers are positive
    • CD19, CD20, CD79a, CD22, FMC7
  • early surface immunoglobulin is often weak in intensity
  • express CD10
    • pre-GC origin
  • CD38 is usually bright
    • help differentiate it from true BL
26
Q

What is a marker that can be often

seen on HCL ?

A
  • CD10
  • if the rest of the immunophenotype fits and CD10 is + it can still be HCL
27
Q

What immunophenotype when seen in a B cell

LPD should prompt the addition of

the hairy cell tubes?

A
  • strong surface Ig
  • strong CD20
  • Negative
    • CD5
    • CD10
  • especially if the patient has cytopenias and splenomegaly
28
Q

What are the additional antibodies that should be

run to assess for HCL ?

A
  • CD11c
  • CD103
  • CD123
  • CD25
29
Q

What markers are HCL-v

typically negative for ?

A
  • NEG
    • CD25 and CD123
  • May be positive or negative
    • CD11c
    • CD103

Flow Cytometry (17 decks)

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