Dear Sharon, This is a very interesting discussion and a difficult case. However, I believe that I be able to provide a rapid, coherent and scientifically sound approach to diagnosis for this case, and other cases of suspect monoclonality. As you know, approximately 90% of follicular lymphomas display a characteristic bcl2 t(14;18) translocation that can be rapidly and specifically identified using PCR. Now a PCR-based series of tests have been developed and validated that simultaneously identify both bcl2 translocations and clonal rearrangements of the B and T cell antigen receptors. The performance characteristics of Multi-Loci Clonality Testing were recently peer-reviewed and published [Molecular Diagnosis 4(2):101-117, 1999]. New assays using this advanced technology will be presented this year at the annual meeting of the Association for Molecular Pathology (Nov 4-7, St Louis), and the International Society for Laboratory Hematology Meeting (April 12-16, Banff). This technology, which uses a PCR-based approach, will identify clonality and translocations testing as little as 50ng (50 x 10-9g) of genomic DNA, an amount of material easily obtained from blood, bone marrow, fresh (aspirate), frozen or formalin-fixed paraffin embedded tissue, or even a single glass slide. The monoclonality detection rate, determined in a method comparison study with Emory University Medical Center, was determined to be 100%, the sensitivity approximately 1 cell in one hundred normal cells, and the specificity was determined to be 100%. One of the major advantages of this approach is that, in addition to testing for bcl2 translocations, both the immunoglobulin heavy chain gene and T cell receptor gamma chain genes are tested. Therefore, the technology identifies cross-lineage clonal rearrangements that are often missed following lineage assignment and testing for either B cell clonality or T cell clonality. I admit a bias for this approach; it is our company's technology. Turnaround time for the assays is generally one day, and the results can be posted on our SECURE Internet Client Data Access Site. Best regards, Jeffrey Miller >>From: "Nebe, Thomas C." <thomas.nebe@ikc.ma.uni-heidelberg.de> >>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >>Subject: AW: Clinical case, lymphoma >>Date: Fri, 22 Oct 1999 14:59:35 +0200 >>MIME-Version: 1.0 >> >> >>Dear Sharon, >> >>I appreciate the comments from Maryalice and Anna. >> >>We had many of these discussions among collgues here in Germany. For the >>benefit of the patient and clinician: The way to go is to extirpate the >>lymph node in total and send it (or pass on) to a reference pathologist who >>is well trained (belongs to the REAL group). They hate fine needle biopsies >>as the architecture of the lymph node gets lost. Your given immunophenotype >>(by "feeling", missing morphology and conjugation) does not suggest a CLL >>and lymphoma diagnosis by flow is a difficult game even including morpholgy >>and experience. The discussion of your data is complicated by the fact that >>the missing fluorochrome is crucial: PE gives positive signals eg. for CD25 >>or CD11c while FITC conjugates give negative results. >> >>Lymphoma classification and treatment of lymphoma is based upon lymph node >>pathology. The extended marker panel you anticipated we use for a while. >>It helps to exclude B CLL but does not give a safe diagnosis of other B-NHL >>(except HCL, may be Immunocytoma). The so called atypical B-CLL has never >>been defined and even very experienced collegues refused to give a >>presentation on that subject. >> >>SLL from the working formulation is not a well defined clinical entity and >>therefore did not exist in the KIEL classification. >>Harald Stein (Berlin, REAL group) just presented at the german hematologist >>meeting in Jena the new concept of a B1 and B2 CLL, where the latter is CD38 >>positive and forms lymphoma. (The concept behind is based on B cell >>development in the lymph node). This provoqued a lively discussion esp. >>among those who used CD38 in the past. We also found CD38-FITC surface >>positive (dim compared to immunocytoma or plasmocytoma) in those CLL >>patients with spleen and lymph node infiltration (total CLL n=85). Possibly >>cytoplasmic staining resolves the controversial issue. >>I dont want to be bouring but it would be helpful to have a minimum report >>format (combinations / fluorochrome / surface or cytoplasmic / intensity >>compared to normal counterparts) when discussing such cases. >>Best regards >>Thomas Nebe >> >>Dr.med. C. Thomas Nebe >>Universitaetsklinikum Mannheim >>Zentrallabor >>Theodor-Kutzer-Ufer 1-3 >>D-68167 Mannheim >>Tel. +49 621 383-3485 >>FAX +49 621 383-73 3485 >> +49 621 383-3819 >>e-mail: thomas.nebe@ikc.ma.uni-heidelberg.de >> >>Bitte besuchen Sie unsere sehr informativen Webseiten unter >>http://www.ma.uni-heidelberg.de/inst/ikc/ >> >>> -----Ursprüngliche Nachricht----- >>> Von: Gerhard Nebe-von-Caron >>>[SMTP:Gerhard.Nebe-von-Caron@unilever.com] >>> Gesendet am: Mittwoch, 20. Oktober 1999 16:58 >>> An: Nebe, Thomas C. >>> Betreff: FW: Clinical case, lymphoma >>> >>> >>> >>> -----Original Message----- >>> From: Sharon Vogt [SMTP:svogt@bellsouth.net] >>> Sent: Saturday, October 16, 1999 1:14 AM >>> To: Cytometry Mailing List >>> Subject: Clinical case, lymphoma >>> >>> >>> Hi all, >>> >>> We have a clinical case that's interesting to the point of frustration. >>> Any >>> comments? >>> >>> 53 y male, 3 years ago and now (recurrent) lymphoma, B-cell type. >>> Phenotype is >>> similar in both flow cytometric studies: >>> >>> CD19+ CD20+ CD22 dim, CD5 dim before, partial now, CD25+, CD23 partial, >>> CD11c >>> partial, with kappa light chain restriction. Negative for CD10. We do not >>> stock >>> FMC7 (but will soon), CD21 or CD24. >>> >>> By morphology and flow (considering dim CD5 as negative), the diagnosis of >>> follicular center cell lymphoma (noting that 20-30% of these are known not >>> to >>> express CD10) was made. None of us are entirely sure of that diagnosis >>> now, and >>> there appears to be a tad more CD5 expression (recent specimen was FNA of >>> cervical node); it was suggested that this may be a mantle cell lymphoma. >>> >>> OK, simple enough - immunohistochemical stains bcl-2 and cyclin D1 should >>> answer >>> that question. Those are difficult stains to optimize, however, and are >>> affected >>> by variations in fixation. In other words, they didn't help much. CD20 nor >>> kappa >>> light chain expression is remarkable for staining intensity, but I'd favor >>> a CLL >>> based on the rest of the phenotype. The pathologists say small cell, low >>> grade, >>> but not really CLL-like and definitely not PLL. >>> >>> ?? Thanks for any discussion. >>> >>> sharon >>> >>> Sharon F. Vogt >>> svogt@bellsouth.net >>> Dekalb Medical Center >>> Atlanta, GA 30033 >>> >>> >>> >>> >>> << Datei: ATT01392.ATT >> >> > > > >Chris Lena >Laboratory Technician >La Jolla Institute For Allergy and Immunology >phone:(858)678-4536 >fax:(858)678-4595 >cali@liai.org >http://www.liai.org > > _ _ > (_)-(_) > \"/ > =V= > > Jeffrey E. Miller, Ph.D. President & CEO IVS Technologies, LLC 2915 Avenida Valera Carlsbad, CA 92009 (760) 431-6705 - Phone (760) 431-6909 - Fax (877) 431-6705 - Toll free jemiller@invivoscribe.com http://www.invivoscribe.com La Jolla Institute for Allergy & Immunology 10355 Science Center Drive San Diego, California 92121 (619) 678-4529 - direct (619) 678-4595 - Fax
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