Round-robin test for the cell-of-origin classification of diffuse large B-cell lymphoma—a feasibility study using full slide staining
Diffuse large B-cell lymphoma (DLBCL) is subdivided by gene expression analysis (GEP) into two molecular subtypes named germinal center B-cell-like (GCB) and activated B-cell-like (ABC) after their putative cell-of-origin (COO). Determination of the COO is considered mandatory in any new-diagnosed DLBCL, not otherwise specified according to the updated WHO classification. Despite the fact that pathologists are free to choose the method for COO classification, immunohistochemical (IHC) assays are most widely used. However, to the best of our knowledge, no round-robin test to evaluate the interlaboratory variability has been published so far. Eight hematopathology laboratories participated in an interlaboratory test for COO classification of 10 DLBCL tumors using the IHC classifier comprising the expression of CD10, BCL6, and MUM1 (so-called Hans classifier). The results were compared with GEP for COO signature and, in a subset, with results obtained by image analysis. In 7/10 cases (70%), at least seven laboratories assigned a given case to the same COO subtype (one center assessed one sample as not analyzable), which was in agreement with the COO subtype determined by GEP. The results in 3/10 cases (30%) revealed discrepancies between centers and/or between IHC and GEP subtype. Whereas the CD10 staining results were highly reproducible, staining for MUM1 was inconsistent in 50% and for BCL6 in 40% of cases. Image analysis of 16 slides stained for BCL6 (N = 8) and MUM1 (N = 8) of the two cases with the highest disagreement in COO classification were in line with the score of the pathologists in 14/16 stainings analyzed (87.5%). This study describes the first round-robin test for COO subtyping in DLBCL using IHC and demonstrates that COO classification using the Hans classifier yields consistent results among experienced hematopathologists, even when variable staining protocols are used. Data from this small feasibility study need to be validated in larger cohorts.
KeywordsDiffuse large B-cell lymphoma ABC GCB Cell of origin Immunohistochemistry Hans classifier
The authors would like to thank Dana Germer and Charlotte Botz-von Drathen for their excellent technical support.
J.R., R.S., and W.K. developed the research plan. S.R., F.F., A.F., M.L.H., I.O., G.O., K.H., P.M., A.R., H.S., and W.K. generated data. J.R., S.R., M.A., and W.K. analyzed the data. S.R., J.R., and W.K. wrote the manuscript. All authors have read and accepted the final version of the manuscript.
This study was funded by the German Federal Ministry of Education and Research (BMBF) as a part of the MMML-Demonstrators project (grant 031A428D) within the e:Med framework.
Compliance with ethical standards
The use of tissue was in accordance with the guidelines of the internal review board of the medical faculty of the University of Kiel (no. 447/10).
This article does not contain any studies with animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
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