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ICCS 2012
CD2+,CD3+,CD4-, CD5+, CD7+, CD8+, CD16+, CD56-,
TCR gamma/delta+. Diagnostic populations were also found
in blood, however additional TCR gamma/delta subsets
were unexpectedly detected in both cases with unique
phenotypes, possibly arising from immune responses, and
appeared less sensitive to chemotherapy treatment. TCR
gamma/delta markers are not always included in routine
lymphoma screens. Small numbers of these cells in the
blood are easily overlooked and appropriate follow up
markers not always examined. It is recommended that TCR
surface markers be included in all T cell lymphoma screening
panels as recommended by EuroFlow. We suggest cases
of HSTL may be under diagnosed due to incomplete
immunophenotype testing and distinction between TCR
gamma/delta subsets needs to be made during treatment
monitoring.
P26
APPLICATION OF EIGHT-COLOUR FLOW CYTOMETRY
(FC) IN THE ASSESSMENT OF PLASMA CELL MYELOMA
(PCM) AT PETER MACCALLUM CANCER CENTRE
(PMCC).
Vuong Van Nguyen
1
, Peter Gambell
1
, David Westerman
1,2
,
Neil Came
1,2
1
Pathology Department, Peter MacCallum Cancer Centre,
2
University of Melbourne
Aim:
The utility of our 8-colour/3-tube FC method (table
1) for differentiating normal and abnormal bone marrow
(BM) plasma cells (nPC/aPC) at diagnosis and for stringent
complete response assessment (sCRa) of PCM was
compared to our superseded 4-colour/5-tube protocol
(italics).
Methods:
Consecutive BM aspirate samples
were analysed on a BD-FACSCanto-II to define aPC (with
or without 5% nPC/total PCs) at diagnosis, aPC during
treatment (Rx), and aPC at 0.01% sensitivity for sCRa in
patients with PCM. Comparison of sCRa by 4-colour BD-
FACSCalibur (same gating strategy) was performed in the
same number of consecutive samples analysed immediately
prior to commencement of 8-colour FC. Undetectable
aPC, or of any PC, defined sCR provided that >100000
events (averaged/tube) were analysed and that CD45/
SSC confirmed representative BM cellularity.
Results:
From October 2011-July 2012, 85 eligible BM samples
were assessed by 8-colour FC; 69 for PCM (30 sCRa, 27
inter-therapy, 12 diagnostic) and 16 miscellaneous. 100%
sCRa samples were suitable by 8-colour vs 22/30 (73%)
by 4-colour (p=0.005, Fishers); both 100% sensitivity; 86%
vs 27% specificity; median events analysed (average/tube)
463426 (100000-733955) vs 164343 (27292-500000). An
aPC phenotype was defined in 67/85 (79%) cases (table 2).
Table 3 highlights likelihood of each antigen dichotomizing
nPC/aPC in the same sample.
Conclusion:
Eight-colour
FC of the BM in PCM for sCRa at PMCC is superior to
previous 4-colour method. Additional markers (see CD200)
and reliable analysis of more BM cells per tube provides
improved specificity for defining sCR and adds clinically
relevant prognostic information to guide therapy.
P27
IMPACT OF GALLIOSTM ACQUISITION SETTINGS AND
INFINICYTTM ANALYSIS SETTINGS ON DATA ANALYSIS
AND INTERPRETATION (LESSONS LEARNED)
Melanie O’Donahue, Susan Schmitt
1
Michael McPherson,
2
Esther Alvaro,
3
Marla Dorman
Fluorescent compensation can be tricky, especially when
data is acquired and analyzed with different software
platforms. A growing trend in flow is to have the technical
component (staining and acquisition) and the professional
POSTER ABSTRACTS