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<span style="color: navy>'''Background'''</span>


Typical CLL can be diagnosed with some confidence using flow cytometry. CLL cells express low levels of B-cell receptor associated molecules (e.g. CD19, CD79B and sIg) together with CD5 and CD23 in typical cases. However, atypical cases are regularly encoutered and can cause diagnosic difficulty.  
<span style="color: navy>'''Background to flow cytometric diagnosis'''</span>
 
Differentiation to plasma cells involves (in mast cases) the loss of typical pan B-cell markers and surface light chains. The expression of CD45 also differs both within and between cases making standard gating strategies difficult to apply. Cells are most often identified through their co-expression of CD138 with a relatively high expression of CD38, together with the absence of typical B-cell features.




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{| class="wikitable" style="color:black; background-color:#ffffff;" cellpadding="5"
{| class="wikitable" style="color:black; background-color:#ffffff;" cellpadding="5"
!colspan="4" style = "font-size:110%; color: navy; text-align:center; background:#DEE0DE;"|'''Immunophenotype of CLL'''
!colspan="4" style = "font-size:110%; color: navy; text-align:center; background:#DEE0DE;"|'''Immunophenotype of plasma cells'''
|-
!colspan="4" style = "font-size:90%; background:#DAE8F2"|'''CD45''' expression
|-
|colspan="4" style = "font-size:90%; color:black;" |There is no consistent pattern of expression of CD45 although it is most often weak, but may be variable and is occasionally strong.
|-
|-
!colspan="4" style = "font-size:90%; background:#DAE8F2"|'''Major markers useful in CLL diagnosis'''
!colspan="4" style = "font-size:90%; background:#DAE8F2"|Major markers useful in plasma cell identification
|-
|-
|colspan="1" style = "width: 4%; font-size:90%; font-color:navy;" |'''κ/λ'''
|colspan="1" style = "font-size:80%; width: 4%; |'''Marker'''
|colspan="1" style = "width: 8%; background:#00b8e6;"|
|colspan="1" style = "font-size:80%; width: 8%; |'''Freq'''
|colspan="1" style = "width: 5%; |wk/ abs
|colspan="1" style = "font-size:80%; width: 8%; |'''Level'''
|colspan="1" style = "font-size:85%;"|Expect weak or absent restricted κ or λ expression
|colspan="1" style = "font-size:80%; width: 80%; |'''Comment'''
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''FMC7'''
|colspan="1" style = "font-size:90%; color:black;" |CD38
|colspan="1" style = "width: 6%; background:#66e0ff;"|
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "width: 4%; |wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|May be expressed by atypical cases, generally absent
|colspan="1" style = "font-size:84%;"|The relatively strong expression of CD38 is typical of plasma cells
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''CD5'''
|colspan="1" style = "font-size:90%;" |CD138
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Mod
|colspan="1" style = "font-size:80%;"|var
|colspan="1" style = "font-size:84%;"|Characteristic of CLL, usually less strong than on T cells
|colspan="1" style = "font-size:84%;"|Expression is expected although the intensity can vary
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |CD10
|colspan="1" style = "width: 4%; font-size:90%; color:black;" |κ/λ
|colspan="1" style = "width: 6%; background:#00b8e6;"|
|colspan="1" style = "width: 8%; background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|N/A
|colspan="1" style = "font-size:80%; width: 8%; |
|colspan="1" style = "font-size:84%;"|Rare in CLL, consider FL if detected
|colspan="1" style = "font-size:80%;"|Surface light chain expression is lost (internal will be detected in permeabilised cells)
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |CD11C
|colspan="1" style = "font-size:90%; color:black;" |CD19
|colspan="1" style = "width: 6%; background:#00b8e6;"|
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expressed in some cases, but tends to be weak
|colspan="1" style = "font-size:84%;"|Expression of CD19 may persist in some cases  
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''CD19'''
!colspan="4" style = "font-size:90%; background:#DAE8F2"|Other relevant markers
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk/ mod
|colspan="1" style = "font-size:84%;"|Expression is expected, but less strong than normal B cells
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |CD20
|colspan="1" style = "font-size:90%; color:black;" |CD5
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expression is expected, but characteristically weak
|colspan="1" style = "font-size:80%;"|Not detected on plasma cells
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''CD23'''
|colspan="1" style = "font-size:90%;" |CD10
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk/ Mod
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Characteristic of CLL, although can rarely be weak or absent
|colspan="1" style = "font-size:84%;"|
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |CD43
|colspan="1" style = "font-size:90%;" |CD11C
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk/ Mod
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Similar to CD200, may be helpful in some cases
|colspan="1" style = "font-size:84%;"|
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''CD79A'''
|colspan="1" style = "font-size:90%;" |CD20
|colspan="1" style = "width: 6%; background:#004466;"|
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expression is expected, but characteristically weak
|colspan="1" style = "font-size:84%;"|Expression retained in some cases
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |CD103
|colspan="1" style = "font-size:90%; color:black;" |CD23
|colspan="1" style = "width: 6%; background:#E6FAFF;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|N/A
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Not expressed by CLL, consider HCL if found
|colspan="1" style = "font-size:84%;"|
|-
|-
|colspan="1" style = "width: 6%; font-size:90%;" |'''CD200'''
|colspan="1" style = "font-size:90%; color:black;" |CD25
|colspan="1" style = "width: 6%; background:#66e0ff;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "width: 4%; font-size:90%;"|Wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Characteristically positive in CLL and negative in MCL
|colspan="1" style = "font-size:84%;"|
|-
|colspan="1" style = "font-size:90%;" |CD43
|colspan="1" style = "background:#006699;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Generally not expressed by LPL
|-
|colspan="1" style = "font-size:90%; color:black;" |CD79b
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expressed in some cases
|-
|colspan="1" style = "font-size:90%;" |CD103
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|
|-
|colspan="1" style = "font-size:90%; color:black;" |CD200
|colspan="1" style = "background:#006699;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Detected frequently|-|}
|-
|colspan="1" style = "font-size:90%; color:black;" |FMC7
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|
|colspan="1" style = "font-size:80%;"|Not expressed
|-
|-
|}
|}
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<span style="color: navy>'''Pitfalls and atypical cases'''</span>
<span style="color:navy;font-size:80% ">'''Key to table: '''</span>
 
<span style="font-size:80% ">Key to colour code for expression frequency [[Key_to_marker_expression_table|Click for link]]</span>
 
<span style="font-size:80% ">Key to expression strength code and use [[Expression strength key|Click for link]]</span>
 
 
 
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Latest revision as of 18:09, 14 July 2023

  • PC.jpg


Background to flow cytometric diagnosis

Differentiation to plasma cells involves (in mast cases) the loss of typical pan B-cell markers and surface light chains. The expression of CD45 also differs both within and between cases making standard gating strategies difficult to apply. Cells are most often identified through their co-expression of CD138 with a relatively high expression of CD38, together with the absence of typical B-cell features.


Immunophenotype of plasma cells
CD45 expression
There is no consistent pattern of expression of CD45 although it is most often weak, but may be variable and is occasionally strong.
Major markers useful in plasma cell identification
Marker Freq Level Comment
CD38 - The relatively strong expression of CD38 is typical of plasma cells
CD138 var Expression is expected although the intensity can vary
κ/λ Surface light chain expression is lost (internal will be detected in permeabilised cells)
CD19 - Expression of CD19 may persist in some cases
Other relevant markers
CD5 - Not detected on plasma cells
CD10 -
CD11C -
CD20 - Expression retained in some cases
CD23 -
CD25 -
CD43 - Generally not expressed by LPL
CD79b - Expressed in some cases
CD103 -
CD200 - Detected frequently|-|}
FMC7 Not expressed


Key to table:

Key to colour code for expression frequency Click for link

Key to expression strength code and use Click for link


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