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Flow Cytometry > Population Identification (small Flow Book) > Flashcards

Population Identification (small Flow Book) Flashcards

1
Q

What is the pattern of CD45 expression in hematopoietic cells?

A
  • increasing intensity is seen on most maturing hematopoietic cells.
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2
Q

CD123 is a marker most helpful in identifying which populations?

A
  • Basophils

- Plasmacytoid dendritic cells

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3
Q

What cell types will have the highest level of expression of CD45 ?

A
  • mature lymphocytes
  • monocytes
  • granulocytes

Note: immature granulocytes usually have lower CD45 expression

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4
Q

What is the immunophenotype of the earliest precursor cells within the marrow?

A
  • cells are capable of differentiating along a variety of lineages
  • CD34(++), CD38 (low to negative), CD45, HLA-DR (intermediate)
  • CD13 (intermediate), CD33 (low), CD117 (low), and CD123 (low)
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5
Q

What markers do blasts acquire when they transition to the myeloid lineage ?

A
  • CD13, CD15, and CD33
  • Note:
    • promyelocytes have a high level of expression of CD13, CD33, and CD38
      * the level of CD33 and CD64 is lower than a mature monocyte
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6
Q

What is the pattern of CD13 and CD16 in maturing myeloids ?

A
  • myeloids lose CD13
  • then they acquire CD13 and CD16 together
    • both these antigens are highly expressed on the neutrophils
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7
Q

What are the initial markers that define myeloid maturation from an early precursor?

A
  • begins when the CD34+ precursor starts acquiring CD38 along with:
    • higher levels of CD13, CD33, and CD117
    • with a slight decrease in CD45
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8
Q

What antigens are lost when myeloblasts move into the promyelocyte stage?

A
  • CD34 and HLA-DR are lost
  • CD13 expression increases as CD15 is acquired
  • IMPORTANT:
    • Promyelocytes have levels of CD33 and CD64 that are the highest for myeloid cells
      But should still be less than mature monocytes
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9
Q

What happens to antigen expression as cells move into the myelocyte stage?

A
  • There is a decrease in SSC, CD13 and CD33
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10
Q

What myeloid stage are CD13 and CD16 acquired together?

A
  • Metamyelocytes

- Bands and neutrophils show high levels of CD13 and CD16 with maximal CD45

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11
Q

What antigenic changes can be seen in neutrophils in reactive conditions?

A
  • can upregulate CD64 or downregulate CD16
  • marrow-regeneration can lead to acquisition of CD56
  • Remember
    • PNH causes a loss of CD16
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12
Q

What antigens are obtained during monocytic maturation from the stem cells?

A
  • begins in a similar fashion compared to myeloid maturation
  • CD34+ acquire CD38
  • then there are higher levels of CD13, CD33 and CD117
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13
Q

What markers indicate commitment to the monocytic lineage ?

A
  • high levels of HLA-DR, CD11b, CD33, and CD64

* the expression of these antigens is much higher compared to myeloid precursors

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14
Q

What markers do immature monocytes demonstrate (monoblasts and promonocytes) ?

A
  • they acquire CD15 with low variable expression of CD13
  • Note:
    • with progressive maturation (mature monocytes) express CD13, CD14 and CD45 at high levels
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15
Q

What is the activated monocyte phenotype?

A
  • CD16+
  • slightly decreased levels of CD14, CD33, CD64, and HLA-DR
  • Note:
    • CD14 can be lost in PNH monocytes
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16
Q

How is CD36 used in identifying monocytes ?

A
  • CD36 is another marker that is acquired by promonocytes
  • the issue is that platelets and erythroids can be positive for this as well
    • issue becomes when platelets stick to things then you have non-specific expression
    • also platelets can compete for the anti-CD36 antibody and decrease the intensity on monocytes
17
Q

What antigens are acquired when the stem cell progenitor becomes erythroid?

A
  • CD34+ progenitor acquires CD38

- high levels of CD117 are then acquired

18
Q

What is antigen expression as the erythroid cells mature ?

A
  • CD13 and CD33 are dropped (sometimes low levels are retained)
  • then after that CD38, CD117 and CD45 are lost
  • then CD71 is gained at a high level
  • following this glycophorin and heme are acquired
19
Q

When is the expression of CD71 lost in the erythroid precursor ?

A
  • when the nucleus is lost, then CD71 is no longer expressed

- mature red cells are lysed by ammonium chloride for flow cytometry so evaluation of them is limited

20
Q

How are eosinophils identified by flow cytometry ?

A
  • gate around the high CD45 and SSC

- then subgate the negative CD16 population [eosinophils]

21
Q

Where in the CD45 vs. SSC are basophils and PDCs found?

A
  • “blast gate”

- they are differentiated from blasts by their expression of CD123

22
Q

How are basophils and PDC’s differentiated by flow cytometry ?

A
  • Basophils : negative for HLA-DR and positive for CD38 (bright)
  • PDC: express high levels of HLA-DR
23
Q

Where does B cell development start and what antigens are expressed/lost initially ?

A
  • begins in the bone marrow
  • stem cell (CD34+, CD38-) acquires CD38 and then loses CD13, CD117, and CD123
  • then CD33 is lost after
24
Q

What are the earliest markers expressed by B cell progenitors ?

A
  • CD34, CD79a, TdT and CD10
  • they then acquire CD19
  • these progenitors are also considered to be hematogones
    • hematogones show an inverse relationship with age
    • they can be increased in regenerative marrows, immune mediated cytogenetic, and
      Acquired immune deficiency
25
Q

What are the three stages of hematogone maturation ?

A
  • Stage I:
    * CD19, TdT, CD34 and CD10 (bright)
  • Stage II:
    • decreased CD10, loss of TdT and gradual loss of CD34
  • Stage III:
    • acquisition of CD20 with still some CD10 expression

Loss of CD10 completely means the mature B cell stage has been reached.

26
Q

Where can naive mature B cells be found?

A
  • all compartments (bone marrow, peripheral blood, tissues)

- primary follicles (predominantly), mantle zones of reactive follicles

27
Q

What immunophenotypic changes are seen in B cells when they enter the germinal center and get exposed to antigen?

A
  • Acquire CD10 and have increased CD38

- bright CD20 expression

28
Q

When are CD5 positive, normal B cells encountered?

A
  • these are called transitional B cells (B1)
  • they are a type of naive B cells that expresses CD5
  • more common in children and young adults but can be seen in adults
  • they can be seen in all types of tissues, but frequently in head and neck

Flow Cytometry (17 decks)

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