We have found the viability of "large cell lymphoma-cells" to be a
really low percentage. When the pathologist suspects "large cell",
our paneling procedure immediately becomes Stat! Unfortunately, no
matter how fast we respond, the viability is usually less than 40%
in
our disaggregated mix of cells -- and often less then 20%. We feel
it
is the large cells which are "going away". We try to gate around
the
area in which the large cells would be, and on occaision we are
able
to find a monoclonal population.
Kathy Altig
Providence Med Ctr.
Portland, OR
______________________________ Reply Separator
_________________________________
Subject: Re[2]: large cell lymphoma
Author: ,mann0002@mc.duke.edu [SMTP:mann0002@mc.duke.edu] at PHSOR
Date: 10/9/97 3:30 PM
We have had the identical experience and likewise have not seen evidence
of a
large cell population by light scatter characteristics.
Karen
Karen Mann, MD PhD
Duke University Medical Center
Durham, NC
________________________________________________________________________
_______
Subject: Re: large cell lymphoma
From: mkornste@hsc.vcu.edu at internet
Date: 10/09/97 6:50 AM
I have also been surprised by unremarkable flow data
from an FNA of an obvious large cell lymphoma (phenotyped as B cell by
immunocytochemistry). I like the idea that the cells were
fragile. No population of large cells by forward scatter were apparent.
/Michael Kornstein, MD, Medical College of Virginia, Richmond
On Wed, 8 Oct 1997, Brent Dorsett wrote:
>
>
> >Viki Mosiman
> >Robert H Lurie Cancer Center
> >Flow Cytometry Core Facility
> >Northwestern University Medical School
> >Chicago, IL
> >vlm646@nwu.edu
> >
> >I am interested in an opinion on the best way to detect Large-cell
> >lymphoma(B Cell type) in lymph nodes.
> >
> >We currently disaggregate nodes then surface stain using
> >45/19/kappa/lambda. Although
> >we often have success ,there are cases where clearly the node is a
> >malignant B-cell lymphoma by morphology in Surgical Pathology but we
see
> >no evidence in Flow. We have tried blocking with fetal calf and
mouse
> >serum. We have tried different gating strategies such as 45ss
combined
> >with 19ss gating or combined with fs. We have tried different
staining
> >combinations -19/5/kappa/lambda-45/19/10/kappa&45/19/10/lambda. We
have
> >tried live dead assays(7AAD) combined with surface staining in case a
node
> >was necrotic. Someone even suggested injecting nodes with PBS to
express
> >cells instead of mincing and grinding lymph nodes(they thought this
might
> >be a more gentle process).
> >
> >We don't seem to have the same problem with other malignant lymphomas
in
> >lymph nodes,just with the diagnosis of Large-cell lymphoma ( B-Cell
type).
> >Any suggestions would be greatly appreciated. Thanks for your help!
> >
> >Clinical Flow Lab
> >Northwestern Memorial Hospital
> >
> >
> Viki,
>
> Recently we have seen several cases where the malignant population in
large
> cell lymphomas was extremely difficult to demonstrate in flow. One, a
lymph
> node, was easily demonstrated by immunohistology to consist of large
numbers
> of B cells infiltrated with reactive T cells. In flow cytometry after
the
> usual mechanical disaggregation, suspesion in RPMI 1640 + 5% FCS,
filtering
> through 100 micro nylon mesh and washing almost no malignant B cells
could
> be detected. The second was a FNA of a mass in the liver collected
into RPMI
> 1640. This too immunostained positively as large B-cells, but in flow
was
> composed of 99% small to medium T-cells. Cell viability was good in
both
> preparations but there appeared to be substantil very small debris.
I'm
> guessing that these large cells were particularly fragile -- although
the
> appearance of the FNA cells in histology seemed normal. I would be
> interested in the experience of other with large cell lymphomas.
>
> Brent Dorsett
> Special Pathology Laboratory
> Lenox Hill Hospital
> NYC
>
>
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Subject: Re: large cell lymphoma
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