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[[file:CLL.jpg|200px|link=CLL.jpg]] 




<span style="color: navy>'''Background to flow cytometric diagnosis'''</span>
<span style="color: navy>'''Background to flow cytometric diagnosis'''</span>


Typical CLL can be diagnosed with some confidence using flow cytometry. However, individual markers may not be expressed in all cases - particularly it may be difficult to distinguish atypical CLL from some cases of MCL or MZL.  
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.
*Atypical features should always be reported and the possibility of alternative diagnoses acknowledged.
*Cases with a low lymphocytosis (<1x10<sup>9</sup>) may represent monoclonal B-lymphocytosis with CLL phenotype [[Monoclonal B lymphocytosis|(click here for diagnostic criteria)]].  




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<div style="width: 95%; overflow: auto; border: 1px solid navy; font-size:100%">
{| 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"|Major markers useful in CLL diagnosis
!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 plasma cell identification
|-
|-
|colspan="1" style = "font-size:80%; width: 4%; |'''Marker'''
|colspan="1" style = "font-size:80%; width: 4%; |'''Marker'''
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|colspan="1" style = "font-size:80%; width: 80%; |'''Comment'''
|colspan="1" style = "font-size:80%; width: 80%; |'''Comment'''
|-
|-
|colspan="1" style = "font-size:90%; color:black;" |'''CD19'''
|colspan="1" style = "font-size:90%; color:black;" |CD38
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expression is expected, but less strong than normal B cells
|colspan="1" style = "font-size:84%;"|The relatively strong expression of CD38 is typical of plasma cells
|-
|-
|colspan="1" style = "width: 4%; font-size:90%; color:black;" |'''κ/λ'''
|colspan="1" style = "font-size:90%;" |CD138
|colspan="1" style = "width: 8%; background:#00b8e6;"|
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|var
|colspan="1" style = "font-size:84%;"|Expression is expected although the intensity can vary
|-
|colspan="1" style = "width: 4%; font-size:90%; color:black;" |κ/λ
|colspan="1" style = "width: 8%; background:#E6FAFF;"|
|colspan="1" style = "font-size:80%; width: 8%; |
|colspan="1" style = "font-size:80%; width: 8%; |
|colspan="1" style = "font-size:80%;"|Expect weak restricted κ or λ or absent expression
|colspan="1" style = "font-size:80%;"|Surface light chain expression is lost (internal will be detected in permeabilised cells)
|-
|-
|colspan="1" style = "font-size:90%; color:black;" |'''CD5'''
|colspan="1" style = "font-size:90%; color:black;" |CD19
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|Characteristic of CLL, usually less strong than on T cells
|colspan="1" style = "font-size:84%;"|Expression of CD19 may persist in some cases
|-
|-
|colspan="1" style = "font-size:90%; color:black;" |'''CD23'''
!colspan="4" style = "font-size:90%; background:#DAE8F2"|Other relevant markers
|colspan="1" style = "background:#004466;"|
|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:90%; color:black;" |'''CD79b'''
|colspan="1" style = "font-size:90%; color:black;" |CD5
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|wk
|colspan="1" style = "font-size:84%;"|Expression is expected, but characteristically weak
|-
|colspan="1" style = "font-size:90%; color:black;" |'''CD200'''
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|-
|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:80%;"|Not detected on plasma cells
|-
|colspan="1" style = "font-size:90%; color:black;" |'''FMC7'''
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|
|colspan="1" style = "font-size:80%;"|May be expressed by atypical cases, generally absent
|-
!colspan="4" style = "font-size:90%; background:#DAE8F2"|Other relevant markers
|-
|-
|colspan="1" style = "font-size:90%;" |CD10
|colspan="1" style = "font-size:90%;" |CD10
|colspan="1" style = "background:#00b8e6;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Infrequent in CLL, consider FL if detected
|colspan="1" style = "font-size:84%;"|
|-
|-
|colspan="1" style = "font-size:90%;" |CD11C
|colspan="1" style = "font-size:90%;" |CD11C
|colspan="1" style = "background:#00b8e6;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|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%;"|
|-
|-
|colspan="1" style = "font-size:90%;" |CD20
|colspan="1" style = "font-size:90%;" |CD20
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|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 = "font-size:90%; color:black;" |CD23
|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;" |CD25
|colspan="1" style = "font-size:90%; color:black;" |CD25
|colspan="1" style = "background:#00b8e6;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|wk
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Expressed by many cases of CLL but characteristically weak
|colspan="1" style = "font-size:84%;"|
|-
|-
|colspan="1" style = "font-size:90%; color:black;" |CD38
|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 = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|wk
|colspan="1" style = "font-size:84%;"|Expressed by some cases of CLL, typically weak
|-
|colspan="1" style = "font-size:90%;" |CD43
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|-
|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%;"|Expressed in some cases
|-
|-
|colspan="1" style = "font-size:90%;" |CD103
|colspan="1" style = "font-size:90%;" |CD103
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|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 = "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%;" |CD138
|colspan="1" style = "font-size:90%; color:black;" |FMC7
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|
|colspan="1" style = "font-size:84%;"|Predominantly expressed at plasma cell differentiation
|colspan="1" style = "font-size:80%;"|Not expressed
|-
|-
|}
|}

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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