title: The reliability of immunohistochemical analysis of the tumor microenvironment in follicular lymphoma: a validation study from the Lunenburg Lymphoma Biomarker Consortium.
abstract: The cellular microenvironment in follicular lymphoma is of biological and clinical importance. Studies on the clinical significance of non-malignant cell populations have generated conflicting results, which may partly be influenced by poor reproducibility in immunohistochemical marker quantification. In this study, the reproducibility of manual scoring and automated microscopy based on a tissue microarray of 25 follicular lymphomas as compared to flow cytometry is evaluated. The agreement between manual scoring and flow cytometry was moderate for CD3, low for CD4, and moderate to high for CD8, with some laboratories scoring closer to the flow cytometry results. Agreement in manual quantification across the 7 laboratories was low to moderate for CD3, CD4, CD8 and FOXP3 frequencies, moderate for CD21, low for MIB1 and CD68, and high for CD10. Manual scoring of the architectural distribution resulted in moderate agreement for CD3, CD4 and CD8, and low agreement for FOXP3 and CD68. Comparing manual scoring to automated microscopy demonstrated that manual scoring increased the variability in the low and high frequency interval with some laboratories showing a better agreement with automated scores. Manual scoring reliably identified rare architectural patterns of T-cell infiltrates. Automated microscopy analyses for T-cell markers by two different instruments were highly reproducible and provided acceptable agreement with flow cytometry. These validation results provide explanations for the heterogeneous findings on the prognostic value of the microenvironment in follicular lymphoma. We recommend a more objective measurement, such as computer-assisted scoring, in future studies of the prognostic impact of microenvironment in follicular lymphoma patients.
authors: birgitta.sander@ki.se.
authors: Birgitta
authors: B
authors: Sander
authors: Daphne
authors: D
authors: de Jong
authors: Andreas
authors: A
authors: Rosenwald
authors: Wanling
authors: W
authors: Xie
authors: Olga
authors: O
authors: Balagué
authors: Maria
authors: M
authors: Calaminici
authors: Joaquim
authors: J
authors: Carreras
authors: Philippe
authors: P
authors: Gaulard
authors: John
authors: J
authors: Gribben
authors: Anton
authors: A
authors: Hagenbeek
authors: Marie José
authors: MJ
authors: Kersten
authors: Thierry Jo
authors: TJ
authors: Molina
authors: Abigail
authors: A
authors: Lee
authors: Santiago
authors: S
authors: Montes-Moreno
authors: German
authors: G
authors: Ott
authors: John
authors: J
authors: Raemaekers
authors: Gilles
authors: G
authors: Salles
authors: Laurie
authors: L
authors: Sehn
authors: Christoph
authors: C
authors: Thorns
authors: Björn E
authors: BE
authors: Wahlin
authors: Randy D
authors: RD
authors: Gascoyne
authors: Edie
authors: E
authors: Weller
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology / Q000378:metabolism; D008224:Lymphoma, Follicular / Q000276:immunology / Q000378:metabolism* / Q000473:pathology*; D015203:Reproducibility of Results; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D059016:Tumor Microenvironment / Q000276:immunology
chemical_list: D015703:Antigens, CD; D014408:Biomarkers, Tumor; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors