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Flow cytometry: Chronic Lymphocytic leukaemia: Difference between revisions

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<gallery mode="nolines" widths="200px" heights="200px" border="1px" >
[[file:CLL.jpg|200px|link=CLL.jpg]] 
file:CLL.jpg|link={{filepath:CLL.jpg}}
 
</gallery>
<span style="size: 90%>:''Typical CLL morphology, note the small/medium size, bland chromatin pattern in a slightly squared nucleus, and the tendency of cytoplasm to gently engage adjacent red cells''</span>
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<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.  
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.  
 
*Atypical features should always be reported and the possibility of alternative diagnoses acknowledged.
However, individual markers may not be expressed in all cases - this can cause diagnostic difficulty particularly in distinguishing CLL from atypical cases of MCL or MZL. If this is the case then the atypical features should 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)]].  
 
Note also that cases with a low lymphocytosis (<1x10<sup>9</sup>) may represent monoclonal B-lymphocytosis with CLL phenotype [[Monoclonal B lymphocytosis|(click here for more)]].  
 
 
[[file:CLLCD45.jpg|300px|link=CLLCD45.jpg]] 
 
<span style="size: 90%>:''Side scatter of CLL cells is typically low, with a broad range and relatively weak expression of CD45 compared with normal lymphocytes. The CLL cells (highlighted in blue) therefore tend to the left of normal T cells (red) - giving the CLL population a rather elongated "slug-like" shape.''




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!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 CLL'''
|-
|-
!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;" |Typically expression of CD45 is less intense than normal T or B cells, with many cases showing variable expression with a wide spread of CD45 intensity.
|-
!colspan="4" style = "font-size:90%; background:#DAE8F2"|Major markers useful in CLL diagnosis  
|-
|-
|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:navy;" |<u>'''CD19'''</u>
|colspan="1" style = "font-size:90%; color:black;" |'''CD19'''
|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%;"|Expression is expected, but less strong than normal B cells
|-
|-
|colspan="1" style = "width: 4%; font-size:90%; color:navy;" |<u>'''κ/λ'''</u>
|colspan="1" style = "width: 4%; font-size:90%; color:black;" |'''κ/λ'''
|colspan="1" style = "width: 8%; background:#00b8e6;"|
|colspan="1" style = "width: 8%; background:#00b8e6;"|
|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%;"|Expect weak restricted κ or λ or absent expression
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''FMC7'''</u>
|colspan="1" style = "font-size:90%; color:black;" |'''CD5'''
|colspan="1" style = "background:#004466;"|
|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:90%; color:black;" |'''CD23'''
|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 = "background:#004466;"|
|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:84%;"|Characteristically positive in CLL and negative in MCL
|-
|colspan="1" style = "font-size:90%; color:black;" |'''FMC7'''
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "background:#66e0ff;"|
|colspan="1" style = "font-size:80%;"|
|colspan="1" style = "font-size:80%;"|
|colspan="1" style = "font-size:80%;"|May be expressed by atypical cases, generally absent
|colspan="1" style = "font-size:80%;"|May be expressed by atypical cases, generally absent
|-
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD5'''</u>
!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:80%;"|Characteristic of CLL, usually less strong than on T cells
|-
|-
|colspan="1" style = "font-size:90%;" |CD10
|colspan="1" style = "font-size:90%;" |CD10
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|colspan="1" style = "font-size:80%;"|wk
|colspan="1" style = "font-size:80%;"|wk
|colspan="1" style = "font-size:84%;"|Expression is expected, but characteristically weak
|colspan="1" style = "font-size:84%;"|Expression is expected, but characteristically weak
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD23'''</u>
|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;" |CD25
|colspan="1" style = "font-size:90%; color:black;" |CD25
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|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%;"|Similar to CD200, may be helpful in some cases
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD79b'''</u>
|colspan="1" style = "background:#004466;"|
|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%;" |CD103
|colspan="1" style = "font-size:90%;" |CD103
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|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%;"|Predominantly expressed at plasma cell differentiation  
|-
|colspan="1" style = "font-size:90%; color:navy;" |<u>'''CD200'''</u>
|colspan="1" style = "background:#004466;"|
|colspan="1" style = "font-size:80%;"|-
|colspan="1" style = "font-size:84%;"|Characteristically positive in CLL and negative in MCL
|-
|-
|}
|}
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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 17:11, 13 July 2023

  • CLL.jpg

Background to flow cytometric diagnosis

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.

  • Atypical features should always be reported and the possibility of alternative diagnoses acknowledged.
  • Cases with a low lymphocytosis (<1x109) may represent monoclonal B-lymphocytosis with CLL phenotype (click here for diagnostic criteria).


Immunophenotype of CLL
CD45 expression
Typically expression of CD45 is less intense than normal T or B cells, with many cases showing variable expression with a wide spread of CD45 intensity.
Major markers useful in CLL diagnosis
Marker Freq Level Comment
CD19 - Expression is expected, but less strong than normal B cells
κ/λ Expect weak restricted κ or λ or absent expression
CD5 - Characteristic of CLL, usually less strong than on T cells
CD23 - Characteristic of CLL, although can rarely be weak or absent
CD79b wk Expression is expected, but characteristically weak
CD200 - Characteristically positive in CLL and negative in MCL
FMC7 May be expressed by atypical cases, generally absent
Other relevant markers
CD10 - Infrequent in CLL, consider FL if detected
CD11C wk Expressed in some cases, but tends to be weak
CD20 wk Expression is expected, but characteristically weak
CD25 wk Expressed by many cases of CLL but characteristically weak
CD38 wk Expressed by some cases of CLL, typically weak
CD43 - Similar to CD200, may be helpful in some cases
CD103 - Not expressed by CLL, consider HCL if found
CD138 - Predominantly expressed at plasma cell differentiation


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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