Re: kappa/lambda
Brian Smith (smith@beaker.med.yale.edu)
Tue, 25 Apr 95 07:28:25 -0400
>To: William Ainsley Baughman <wab@gwis2.circ.gwu.edu>
>From: "Brian Smith" <smith@beaker.med.yale.edu> (Brian Smith)
>Subject: Re: kappa/lambda
>
>Bill-
>
>Our standard analysis for lymphocytosis/lymphoma includes a CD19 "clonal
excess analysis" using PE-CD19 and FITC-Fab2-kappa (Dako) plus CD19/lambda -
than we analyze on the FACScan using the K-S stats - based on our titrations
and about 50 normal controls a D(s) of 7 or more is outside the reference
range. The rest of the standard panel includes CD25, CD23, CD20, CD22,
CD10, and IgM/IgD as well as a few T cell markers. For patients with
suspected hairy cell we use the CD19 clonal if there are a lot of hairy cell
suspects - if they are post 2CDA or there are sparse funny-looking cells we
will use PE-CD11c versus FITC k/l and gate on the CD11c very bright. Rarely
we will tricolor it using PerCP-CD19 as well. Normally, we see probably
5-15% of the B cells showing some CD11c staining, but always in about the
first decade over control - the hairy cells in our experience are almost
always 2-3 decades away from control. We will only call the CD11c brights
c/w hairy cell if we can also demonstrate clonality. There are a couple of
papers in Blood from about 1-2 years ago on CD11c+ CLL (a very frequent
finding as well, at least for us) - they may have discussed a "reference
range" in there but I can't pull the papers out immediately.
>
>We have also had reasonable luck focusing on the clonality of small numbers
of CD5+ B cells either by tri-color (FITC-kappa/lambda, PE-CD5, PerCP-CD19)
or by gating in a CD5 vs k/l clonal on the CD5 intermediate cells. We have
gotten our plasma cell analysis to a point where we are reasonably
comfortable using a fix/perm k/l labelling and focussing on very bright (4th
decade) CD38+ cells - the backgrounds are much higher than for surface but
the signal is also generally quite acceptable. We always run a positive
control cell line as well.
>
>Hope this is helpful.
>
>-Brian
>
>>Hi,
>>What cocktails have you used? We are interested in CD5+, CD20+,-, or +/-
>>in combination with kappa or lambda. We have been exploring cytoplasmic k
>>or l with CD45(-), CD38(++). This works well with Caltag fix-perm
>>reagents although the background is up a little. We are a clinical lab
>>and offer a hairy cell panel which includes CD11c+CD20, CD25+CD20 as well
>>as CD19, CD20+CD5, CD10 etc. We have some experience with hairy cells
>>with the frequency of <.5% of LCA+ cells. I believe that there are a
>>small frequency of CD11c+ B cells normally found in people. Do you know
>>of any publications or have any data you are willing to share? We are
>>using FACScans with our own cocktails. These instruments do very well in
>>3 color.
>>Thanks
>>Bill Baughman
>>American Medical Labs., Inc.
>>Chantilly, VA.
>>703-802-7070 x5807
>>
>>
>>
>
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
