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file:PC.jpg|link={{filepath:PC.jpg}}
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[[file:HCL.jpg|200px|link=HCL.jpg]] 
<span style="size: 90%>:''Typical HLL morphology, these medium sized cells have a distinctive "partly open" chromatin in an oval or bean-shaped nucleus, cytoplasm is blue/grey with typical hairy projections''




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


HCL expresses typical and unusual markers that together with morphology can allow diagnosed with some confidence using flow cytometry.  
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.
 
However, no marker is unique to HCL and unusual marker patterns can lead to diagnostic difficulty. If this is the case then the atypical features should be reported, since histopathology and molecular testing can each be used to further evaluate these cases.




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<div style="width: 95%; overflow: auto; border: 1px solid navy; font-size:100%">
<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 = "width: 4%; font-size:90%; color:navy;" |<u>'''κ/λ'''</u>
|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%; width: 8%; |Stg
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|Expect weak restricted κ or λ or absent expression
|colspan="1" style = "font-size:84%;"|The relatively strong expression of CD38 is typical of plasma cells
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''FMC7'''</u>
|colspan="1" style = "font-size:90%;" |CD138
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|Stg
|colspan="1" style = "font-size:80%;"|var
|colspan="1" style = "font-size:80%;"|May be expressed by atypical cases, generally absent
|colspan="1" style = "font-size:84%;"|Expression is expected although the intensity can vary
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD5'''</u>
|colspan="1" style = "width: 4%; font-size:90%; color:black;" |κ/λ
|colspan="1" style = "background:#00b8e6;"|
|colspan="1" style = "width: 8%; background:#E6FAFF;"|
|colspan="1" style = "font-size:80%; width: 8%; |
|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;" |CD19
|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="4" style = "font-size:90%; background:#DAE8F2"|Other relevant markers
|-
|colspan="1" style = "font-size:90%; color:black;" |CD5
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:80%;"|Not detected on plasma cells
|-
|-
|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:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|Stg
|colspan="1" style = "font-size:84%;"|Expressed in some cases, but tends to be weak
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD19'''</u>
|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%;"|
|-
|-
|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:navy;" |<u>'''CD23'''</u>
|colspan="1" style = "font-size:90%; color:black;" |CD23
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "font-size:80%;"|-
|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 = "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 = "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 = "font-size:90%;" |CD43
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#006699;"|
|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%;"|Generally not expressed by LPL
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD79b'''</u>
|colspan="1" style = "font-size:90%; color:black;" |CD79b
|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%;"|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%;" |CD138
|colspan="1" style = "font-size:90%; color:black;" |CD200
|colspan="1" style = "background:#E6FAFF;"|
|colspan="1" style = "background:#006699;"|
|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:84%;"|Detected frequently|-|}
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD200'''</u>
|colspan="1" style = "font-size:90%; color:black;" |FMC7
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "background:#E6FAFF;"|
|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 expressed
|-
|-
|}
|}
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<span style="color:navy;font-size:80% ">'''Key to table: '''</span>
<span style="color:navy;font-size:80% ">'''Key to table: '''</span>
<span style="font-size:80% ">Markers most useful in initial diagnosis are <span style=color:navy><u>underlined in blue text</u></span></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 colour code for expression frequency [[Key_to_marker_expression_table|Click for link]]</span>

Latest revision as of 18:09, 14 July 2023




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