This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by de Jong, D. Articles by Weller, E. Search for Related Content PubMed PubMed Citation Articles by de Jong, D. Articles by Weller, E. Social Bookmarking                            What's this?

Immunohistochemical Prognostic Markers in Diffuse Large B-Cell Lymphoma: Validation of Tissue Microarray As a Prerequisite for Broad Clinical Applications—A Study From the Lunenburg Lymphoma Biomarker Consortium

Daphne de Jong, Andreas Rosenwald, Mukesh Chhanabhai, Philippe Gaulard, Wolfram Klapper, Abigail Lee, Birgitta Sander, Christoph Thorns, Elias Campo, Thierry Molina, Andrew Norton, Anton Hagenbeek, Sandra Horning, Andrew Lister, John Raemaekers, Randy D. Gascoyne, Gilles Salles, Edie Weller

From the Netherlands Cancer Institute; Academic Medical Center, Amsterdam; University Medical Center Nijmegen, the Netherlands; Institute of Pathology, University of Würzburg, Würzburg; Department of Pathology, Hematopathology Section, University Hospital Schleswig-Holstein, Campus Kiel; University Clinic Schleswig-Holstein, Campus Luebeck, Germany; Department of Pathology & Medical Oncology, British Columbia Cancer Agency, University of British Columbia, Vancouver, Canada; Department of Pathology, Inserm U617, Hôpital Henri Mondor, Créteil; Université Paris-Descartes; AP-HP, Hôtel-Dieu, Paris; Hospices Civils de Lyon & Université Claude Bernard Lyon-1, Lyon, France; CR-UK Medical Oncology Unit, St Bartholomew's Hospital, London, United Kingdom; Karolinska Institutet, Stockholm, Sweden; Hospital Clinic, University of Barcelona, Barcelona, Spain; Stanford University Medical Center, Palo Alto, CA; and Dana-Farber Cancer Institute, Boston, MA

Address reprint requests to Daphne de Jong, MD, PhD, Department of Pathology, the Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, the Netherlands; e-mail: d.d.jong{at}nki.nl

Purpose The results of immunohistochemical class prediction and prognostic stratification of diffuse large B-cell lymphoma (DLBCL) have been remarkably various thus far. Apart from biologic variations, this may be caused by differences in laboratory techniques, scoring definitions, and inter- and intraobserver variations. In this study, an international collaboration of clinical lymphoma research groups from Europe, United States, and Canada concentrated on validation and standardization of immunohistochemistry of the currently potentially interesting prognostic markers in DLBCL.

Patients and Methods Sections of a tissue microarray from 36 patients with DLBCL were stained in eight laboratories with antibodies to CD20, CD5, bcl-2, bcl-6, CD10, HLA-DR, MUM1, and MIB-1 according to local methods. The study was performed in two rounds firstly focused on the evaluation of laboratory staining variation and secondly on the scoring variation.

Results Different laboratory staining techniques resulted in unexpectedly highly variable results and very poor reproducibility in scoring for almost all markers. No single laboratory stood out as uniformly poor or excellent. With elimination of variation due to staining, high agreement was found for CD20, HLA-DR, and CD10. Poor agreement was found for bcl-6 and Ki-67. Optimization of techniques and uniformly agreed on scoring criteria improved reproducibility.

Conclusion This study shows that semiquantitative immunohistochemistry for subclassification of DLBCL is feasible and reproducible, but exhibits varying rates of concordance for different markers. These findings may explain the wide variation of biomarker prognostic impact reported in the literature. Harmonization of techniques and centralized consensus review appears mandatory when using immunohistochemical biomarkers for treatment stratification.

Supported by the van Vlissingen Lymphoma Foundation. In addition, unrestricted grants were received from Genentech, Millennium Pharmaceuticals Inc, Roche International, and Schering AG.

Authors' disclosures of potential conflicts of interest and author contributions are found at the end of this article.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				K S. G. Cunha, A C. Caruso, A S. Goncalves, V G. Bernardo, A R. C. Pires, E C. da Fonseca, P A. S. de Faria, L E. da Silva, M Geller, R S. de Moura-Neto, et al. Validation of tissue microarray technology in malignant peripheral nerve sheath tumours J. Clin. Pathol., July 1, 2009; 62(7): 629 - 633. [Abstract] [Full Text] [PDF]

				K. Dunleavy, S. Pittaluga, M. S. Czuczman, S. S. Dave, G. Wright, N. Grant, M. Shovlin, E. S. Jaffe, J. E. Janik, L. M. Staudt, et al. Differential efficacy of bortezomib plus chemotherapy within molecular subtypes of diffuse large B-cell lymphoma Blood, June 11, 2009; 113(24): 6069 - 6076. [Abstract] [Full Text] [PDF]

				A. J. Garcia-Pineres, A. Hildesheim, L. Dodd, T. J. Kemp, J. Yang, B. Fullmer, C. Harro, D. R. Lowy, R. A. Lempicki, and L. A. Pinto Gene Expression Patterns Induced by HPV-16 L1 Virus-Like Particles in Leukocytes from Vaccine Recipients J. Immunol., February 1, 2009; 182(3): 1706 - 1729. [Abstract] [Full Text] [PDF]

				D. de Jong, A. Koster, A. Hagenbeek, J. Raemaekers, D. Veldhuizen, S. Heisterkamp, J. P. de Boer, and M. van Glabbeke Impact of the tumor microenvironment on prognosis in follicular lymphoma is dependent on specific treatment protocols Haematologica, January 1, 2009; 94(1): 70 - 77. [Abstract] [Full Text] [PDF]

				D. Ennishi, K. Takeuchi, M. Yokoyama, H. Asai, Y. Mishima, Y. Terui, S. Takahashi, H. Komatsu, K. Ikeda, M. Yamaguchi, et al. CD5 expression is potentially predictive of poor outcome among biomarkers in patients with diffuse large B-cell lymphoma receiving rituximab plus CHOP therapy Ann. Onc., November 1, 2008; 19(11): 1921 - 1926. [Abstract] [Full Text] [PDF]

				E. Hartmann, V. Fernandez, V. Moreno, J. Valls, L. Hernandez, F. Bosch, P. Abrisqueta, W. Klapper, M. Dreyling, E. Hoster, et al. Five-Gene Model to Predict Survival in Mantle-Cell Lymphoma Using Frozen or Formalin-Fixed, Paraffin-Embedded Tissue J. Clin. Oncol., October 20, 2008; 26(30): 4966 - 4972. [Abstract] [Full Text] [PDF]

				K. Fu, D. D. Weisenburger, W. W.L. Choi, K. D. Perry, L. M. Smith, X. Shi, C. P. Hans, T. C. Greiner, P. J. Bierman, R. G. Bociek, et al. Addition of Rituximab to Standard Chemotherapy Improves the Survival of Both the Germinal Center B-Cell-Like and Non-Germinal Center B-Cell-Like Subtypes of Diffuse Large B-Cell Lymphoma J. Clin. Oncol., October 1, 2008; 26(28): 4587 - 4594. [Abstract] [Full Text] [PDF]

				N. A. Johnson and R. D. Gascoyne Gene expression signatures in follicular lymphoma: are they ready for the clinic? Haematologica, July 1, 2008; 93(7): 982 - 987. [Full Text] [PDF]

				R. Malumbres, J. Chen, R. Tibshirani, N. A. Johnson, L. H. Sehn, Y. Natkunam, J. Briones, R. Advani, J. M. Connors, G. E. Byrne, et al. Paraffin-based 6-gene model predicts outcome in diffuse large B-cell lymphoma patients treated with R-CHOP Blood, June 15, 2008; 111(12): 5509 - 5514. [Abstract] [Full Text] [PDF]

				W. H. Wilson, K. Dunleavy, S. Pittaluga, U. Hegde, N. Grant, S. M. Steinberg, M. Raffeld, M. Gutierrez, B. A. Chabner, L. Staudt, et al. Phase II Study of Dose-Adjusted EPOCH and Rituximab in Untreated Diffuse Large B-Cell Lymphoma With Analysis of Germinal Center and Post-Germinal Center Biomarkers J. Clin. Oncol., June 1, 2008; 26(16): 2717 - 2724. [Abstract] [Full Text] [PDF]

				J. Jacob, M. Jentsch, D. Kostka, S. Bentink, and R. Spang Detecting hierarchical structure in molecular characteristics of disease using transitive approximations of directed graphs Bioinformatics, April 1, 2008; 24(7): 995 - 1001. [Abstract] [Full Text] [PDF]

				G. A. Salles Clinical Features, Prognosis and Treatment of Follicular Lymphoma Hematology, January 1, 2007; 2007(1): 216 - 225. [Abstract] [Full Text] [PDF]