Kappa and lambda light chain mRNA in situ hybridization compared to flow cytometry and immunohistochemistry in B cell lymphomas

Persistent Link:
http://hdl.handle.net/10150/610134
Title:
Kappa and lambda light chain mRNA in situ hybridization compared to flow cytometry and immunohistochemistry in B cell lymphomas
Author:
Rimsza, Lisa; Day, William; McGinn, Sarah; Pedata, Anne; Natkunam, Yasodha; Warnke, Roger; Cook, James; Marafioti, Teresa; Grogan, Thomas
Affiliation:
Department of Pathology, University of Arizona, Tucson, AZ, USA; Ventana Medical Systems, Inc., Tucson, Arizona, USA; Department of Pathology, Stanford University College of Medicine, Stanford, CA, USA; Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, OH, USA; Department of Pathology, University College Hospital London, London, UK
Issue Date:
2014
Publisher:
BioMed Central
Citation:
Rimsza et al. Diagnostic Pathology 2014, 9:144 http://www.diagnosticpathology.org/content/9/1/144
Journal:
Diagnostic Pathology
Rights:
© 2014 Rimsza et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
Collection Information:
This item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.
Abstract:
BACKGROUND:Detection of B cell clonality is useful for assisting in the diagnosis of B cell lymphomas. Clonality assessment can be accomplished through evaluation of KAPPA and LAMBDA light chain expression. Currently, only slide based methods are available for the majority of patient biopsies and do not detect light chain protein or mRNA in many B-cell lymphomas. Herein we evaluated a new method, known as colorimetric in situ hybridization (CISH), with improved sensitivity and multiplexing capacity, for its usefulness in clonality detection in mature B cell malignancies.METHODS:The KAPPA and LAMBDA ISH was performed on a Ventana Benchmark XT utilizing two color chromogenetic detection. The probes comprised 2 haptenated riboprobes each approximately 500 base pairs long directed against the conserved regions of either KAPPA or LAMBDA mRNA. The dual colors consisted of silver deposition (black) for KAPPA light chain and a novel (pink) chromogen for LAMBDA light chain. Following optimization, CISH allowed visualization of mRNA in benign B cells in reactive tissues including germinal center, mantle zone, and post-germinal center cells. We then identified 79 cases of B cell lymphoma with formalin-fixed paraffin-embedded (FFPE) biopsies including: follicular (36 cases), mantle cell (6 cases), marginal zone (12 cases), lymphoplasmacytic (6 cases), small lymphocytic (4 cases), and diffuse large B cell (15 cases), which were selected on the basis of either prior flow cytometry or immunohistochemistry (IHC) results to serve as the predicate, "gold standard," comparator.RESULTS:39/79 (49.4%) cases were classified as KAPPA and 29/79 (36.7%) as LAMBDA light chain restricted; while 9/79 (11.3%) cases were classified as indeterminate. Of the 70 cases with KAPPA or LAMBDA light chain restricted CISH, 69/70 (98.6%) were concordant with the reference method, while 1/70 (1.4%) was discordant.CONCLUSIONS:Optimized CISH detected lower levels of mRNA than can be visualized with current slide based methods, making clonality assessment in FFPE biopsies possible for mature B cell neoplasms. In this preliminary study, CISH was highly accurate compared to flow cytometry or IHC. CISH offers the possibility of wider applicability of light chain ISH and is likely to become a useful diagnostic tool.Virtual Slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1430491067123856
EISSN:
1746-1596
DOI:
10.1186/1746-1596-9-144
Keywords:
Clonality; B cell lymphoma; Kappa; Lambda; In situ hybridization; Light chains
Version:
Final published version
Additional Links:
http://www.diagnosticpathology.org/content/9/1/144

Full metadata record

DC FieldValue Language
dc.contributor.authorRimsza, Lisaen
dc.contributor.authorDay, Williamen
dc.contributor.authorMcGinn, Sarahen
dc.contributor.authorPedata, Anneen
dc.contributor.authorNatkunam, Yasodhaen
dc.contributor.authorWarnke, Rogeren
dc.contributor.authorCook, Jamesen
dc.contributor.authorMarafioti, Teresaen
dc.contributor.authorGrogan, Thomasen
dc.date.accessioned2016-05-20T08:59:19Z-
dc.date.available2016-05-20T08:59:19Z-
dc.date.issued2014en
dc.identifier.citationRimsza et al. Diagnostic Pathology 2014, 9:144 http://www.diagnosticpathology.org/content/9/1/144en
dc.identifier.doi10.1186/1746-1596-9-144en
dc.identifier.urihttp://hdl.handle.net/10150/610134-
dc.description.abstractBACKGROUND:Detection of B cell clonality is useful for assisting in the diagnosis of B cell lymphomas. Clonality assessment can be accomplished through evaluation of KAPPA and LAMBDA light chain expression. Currently, only slide based methods are available for the majority of patient biopsies and do not detect light chain protein or mRNA in many B-cell lymphomas. Herein we evaluated a new method, known as colorimetric in situ hybridization (CISH), with improved sensitivity and multiplexing capacity, for its usefulness in clonality detection in mature B cell malignancies.METHODS:The KAPPA and LAMBDA ISH was performed on a Ventana Benchmark XT utilizing two color chromogenetic detection. The probes comprised 2 haptenated riboprobes each approximately 500 base pairs long directed against the conserved regions of either KAPPA or LAMBDA mRNA. The dual colors consisted of silver deposition (black) for KAPPA light chain and a novel (pink) chromogen for LAMBDA light chain. Following optimization, CISH allowed visualization of mRNA in benign B cells in reactive tissues including germinal center, mantle zone, and post-germinal center cells. We then identified 79 cases of B cell lymphoma with formalin-fixed paraffin-embedded (FFPE) biopsies including: follicular (36 cases), mantle cell (6 cases), marginal zone (12 cases), lymphoplasmacytic (6 cases), small lymphocytic (4 cases), and diffuse large B cell (15 cases), which were selected on the basis of either prior flow cytometry or immunohistochemistry (IHC) results to serve as the predicate, "gold standard," comparator.RESULTS:39/79 (49.4%) cases were classified as KAPPA and 29/79 (36.7%) as LAMBDA light chain restricteden
dc.description.abstractwhile 9/79 (11.3%) cases were classified as indeterminate. Of the 70 cases with KAPPA or LAMBDA light chain restricted CISH, 69/70 (98.6%) were concordant with the reference method, while 1/70 (1.4%) was discordant.CONCLUSIONS:Optimized CISH detected lower levels of mRNA than can be visualized with current slide based methods, making clonality assessment in FFPE biopsies possible for mature B cell neoplasms. In this preliminary study, CISH was highly accurate compared to flow cytometry or IHC. CISH offers the possibility of wider applicability of light chain ISH and is likely to become a useful diagnostic tool.Virtual Slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1430491067123856en
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.urlhttp://www.diagnosticpathology.org/content/9/1/144en
dc.rights© 2014 Rimsza et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)en
dc.subjectClonalityen
dc.subjectB cell lymphomaen
dc.subjectKappaen
dc.subjectLambdaen
dc.subjectIn situ hybridizationen
dc.subjectLight chainsen
dc.titleKappa and lambda light chain mRNA in situ hybridization compared to flow cytometry and immunohistochemistry in B cell lymphomasen
dc.typeArticleen
dc.identifier.eissn1746-1596en
dc.contributor.departmentDepartment of Pathology, University of Arizona, Tucson, AZ, USAen
dc.contributor.departmentVentana Medical Systems, Inc., Tucson, Arizona, USAen
dc.contributor.departmentDepartment of Pathology, Stanford University College of Medicine, Stanford, CA, USAen
dc.contributor.departmentRobert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, OH, USAen
dc.contributor.departmentDepartment of Pathology, University College Hospital London, London, UKen
dc.identifier.journalDiagnostic Pathologyen
dc.description.collectioninformationThis item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.en
dc.eprint.versionFinal published versionen
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