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Flow Cytometry > B-cell Maturation > Flashcards

B-cell Maturation Flashcards

1
Q

What are the stages of B cell development in the bone marrow?

A
  • Progenitor B cell (Pro-B cell)
  • Pre-B cell
  • Immature B-cell
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2
Q

How are the Pro-B cells further subcategorized?

A
  • Pre-pro B
  • Early pro B
  • Late pro B
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3
Q

What is the immunophenotype of Pro-B cells?

A
  • CD34+
  • CD43 +
  • CD117 +
  • CD45+
  • CD79a+
  • CD10+
  • TdT+

Note: CD19 expression is gained by the early Pro-B cells

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4
Q

How are the Pre-B cells further subcategorized?

A
  • Large pre-B
  • Small pre-B
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5
Q

What occurs in the transition from Pro-B cells to Pre-B cells?

A
  • VDJ rearrangement of the heavy chain locus
  • therefore, the Pre- B cell has cytoplasmic u heavy chains
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6
Q

What occurs in the transition from Pre-B cells to Immature B cells?

A
  • Pre-B cells rearrange their immunoglobulin light chains (VJ segments)
  • therefore, Immature B cells have surface IgM+
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7
Q

What happens to the immature B cells?

A
  • they enter the circulation and become mature, naive B cells with surface expression of IgM and IgD
  • T1 immature B cells
  • T2 immature B cells
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8
Q

What is the immunophenotype of the immature B cells?

A
  • CD19+
  • CD43 -
  • CD79a+
  • CD25 -
  • CD117 -
  • CD34+
  • IgM+
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9
Q

What is characteristic of the T1 immature B cells in the bone marrow?

A
  • located in the bone marrow
  • short half life
  • IgM ++, IgD-, CD21-, CD23 -
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10
Q

What is characteristic of the T2 immature B cells ?

A
  • they leave the bone marrow and enter the spleen follicles and acquire IgD, CD21, and CD23
  • they are able to circulate and become mature, naive B cells
  • once they mature they express: CD22, CD24, and CD40
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11
Q

In the spleen, the B cells can be subdivided into Follicular B cells and Marginal zone B cells. What are their immunophenotypes?

A
  • Follicular: CD19+, IgM dim+, IgD bright+, CD21 moderate, CD23+
  • Marginal: CD19+, IgM bright+, IgD dim+, CD21 bright, CD23-
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12
Q

When do the expression of Pax5 and CD79a occur in B cells?

A
  • at the time of heavy chain gene rearrangement
  • transition from Pro-B cell to Pre-B cell
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13
Q

When does the expression of CD20 occur in B cells?

A
  • at the time of light chain gene rearrangement/ immunoglobulin formation
  • in immature B cells
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14
Q

What is the phenotype of Naive B cells (mature)?

A
  • CD19+
  • CD20+
  • CD5+
  • sIgM+
  • CD34 -
  • TdT -
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15
Q

What markers do early hematogones express?

A

(+)
- CD19, TdT, CD34, HLA-DR
- bright CD10 and CD38

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16
Q

What markers are altered as the hematogones transition to intermediate hematogones?

A
  • gradual loss of CD34 and CD10
  • still maintain CD38
17
Q

When does down-regulation of CD38 occur in hematogones?

A
  • down-regulation of CD38 occurs in late hematogones and mature B cells

Note: CD38 is picked up again in germinal centers

18
Q

What is the pattern of expression of HLA-DR in intermediate hematogones?

A
  • HLA-DR becomes brighter
19
Q

When does the loss of HLA-DR occur?

A
  • marks the transition from late hematogones to mature B cell
20
Q

In terms of hematogones, when does CD20 get acquired?

A
  • starts to be acquired in the intermediate stage with bright expression in the late hematogones and then mature B cells.
21
Q

Is there surface light chain expression in B-ALL?

A
  • should be absent in majority of cases
22
Q

What is the pattern of expression of CD9?

A
  • positive heterogeneous expression in hematogones
  • usually a positive tight cluster in B lymphoblasts
23
Q

What can be the pattern of CD20 expression in B-ALL?

A
  • CD20 can be heterogeneously expressed in B-ALL
24
Q

Are myeloid markers ever present in B-ALL?

A
  • yes, but not lineage defining (MPO), or would have to consider an MPAL
  • markers for CD13, CD33 and CD117 can be seen in rare cases
25
Q

Are T cell markers ever seen in B-ALL?

A
  • T cell antigens (CD3, CD5, CD2, and CD7) have been rarely reported

Flow Cytometry (17 decks)

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