Clonality analysis in

hematolymphoid malignancies
Dr J ay Mehta
Centre of Excellence : Histopathology
Gene clonality principle
Key feature of functional lymphocytes : rearranged
antigen-receptor genes (unique for each lymphocyte)
When normal lymphocytes differentiate, every cell will
have a slightly different but unique gene rearrangement.
Clonal ( malignant ) lymphocytes, on the other hand, will
contain identical gene rearrangements because they
originated from the same cell
The Gene Clonality assays are designed to identify the
presence of clonal ( malignant) populations of cells
among polyclonal ( normal ) cells, with the aid of PCR
based techniques
a problem case
A 32 year old lady was found to have
cervical lymphadenopathy.
She had palpable splenomegaly, but no
lymphadenopathy at any other site.
Her cervical lymph node was biopsied.
CD 3
CD 5
CD 2
CD 7
CD 4
CD 8
CD 10
CD 23
CD 20
CD 20
CD 30
the dilemma
young lady
single group of lymph nodes
histology :
? PTCL
? T cell rich B cell lymphoma
PCR clonality assay IgH
clonality assay helped
clonal B cell population established
No clonal T cell population seen
final diagnosis : T cell rich B cell non
hodgkin lymphoma
clinical case 10W 5176
22 year old lady presented with history of
fever accompanied by generalized
lymphadenopathy
Her axillary lymph node was biopsied.
CD 30
CD 30
CD 15
CD 15
LCA
CD 20
CD 3
ALK-1
EMA
EBER
Pax-5
differential diagnosis
? Hodgkins lymphoma
? Anaplastic large cell lymphoma
TCR beta assay TCR gamma assay
No clonal T cell population seen
diagnosis
Hodgkins lymphoma
clinical case 09W 11575
35 year old lady presented with generalized
lymphadenopathy
Her cervical lymph node was biopsied
CD 20
CD 3
CD 5
CD 10
CD 23
CD 2
CD 7
CD 4
CD 8
history
So far the thought process favours a T cell
lymphoproliferative process
At this stage we got the history of
retroviral positivity in this patient
P 24
P 24
CD 20
CD 30 EBER
synopsis
The lymphoproliferation was T cell predominant,
but had EBER+ B cell proliferation as well
Difficult to decide in light of retroviral positivity,
if this was reactive / malignant, and if latter,
what lineage
Clonality analysis was resorted to .
TCR beta assay IgH assay
Clonal TCR beta rearrangement confirmed
diagnosis
NHL of T cell type, NOS
Summary
Clonality assessment is warranted for those
cases that remain clinically suspect & / or
histomorphologically unclear.
Clonality assays can be reliably performed on
archival material ( paraffin embedded tissues ),
obviating the need to collect fresh tissues
Imperative that clonality results be interpreted
with full knowledge of the clinical data &
pathology of the hematologic malignancy
( integrated interpretation )
thank you