# Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray

Source: https://onco.cc/key-papers/paper-hans-immunohistochemistry-cell-of-origin-dlbcl-blood-2004/  
OnCo record `paper-hans-immunohistochemistry-cell-of-origin-dlbcl-blood-2004` (Key paper). Data CC BY-NC 4.0, attribute "Data from OnCo (onco.cc)"; commercial use needs a licence.

## TL;DR

Three ordinary laboratory stains, available in any hospital, were shown to sort lymphoma into the same two prognostic groups that an expensive gene-expression machine had found.

## Summary

The cell-of-origin classification was a research technique: it needed fresh-frozen tissue and a microarray. Hans and colleagues asked whether routine immunohistochemistry on paraffin-embedded tissue could reproduce it. They built tissue microarray blocks from 152 cases of diffuse large B-cell lymphoma, 142 of which had already been classified by complementary DNA microarray (75 germinal-centre, 41 activated B-cell, 26 type 3), and stained for CD10, BCL6, MUM1, FOXP1, cyclin D2 and BCL2.

BCL6 (p < 0.001) and CD10 (p = 0.019) expression went with better overall survival, MUM1 (p = 0.009) and cyclin D2 (p < 0.001) with worse. Using CD10, BCL6 and MUM1, 64 cases (42 per cent) were classed germinal-centre and 88 (58 per cent) non-germinal-centre. Five-year overall survival was 76 per cent for the germinal-centre group against 34 per cent for the non-germinal-centre group (p < 0.001), similar to what the microarray gave. On multivariate analysis an International Prognostic Index of 3 to 5 and the non-germinal-centre phenotype were independent adverse predictors (p < 0.0001).

The algorithm is the reason the classification exists outside research centres. It is also the reason the classification is noisier than it looks on paper.

## Fields

- Kind: Key paper
- Last checked: 2026-10-01
- Also known as: Hans algorithm; Hans 2004; CD10, BCL6 and MUM1 classifier
- Tags: lymphoma-evidence
- Journal: Blood
- Year: 2004
- DOI: 10.1182/blood-2003-05-1545
- Authors: Hans CP, Weisenburger DD, Greiner TC, et al.
- Findings: A three-antibody algorithm using CD10, BCL6 and MUM1 classified 42 per cent of 152 cases as germinal-centre and 58 per cent as non-germinal-centre.; Five-year overall survival was 76 per cent in the germinal-centre group against 34 per cent in the non-germinal-centre group (p < 0.001), similar to the complementary DNA microarray classification.; BCL6 and CD10 expression were associated with better overall survival; MUM1 and cyclin D2 with worse.; BCL2 and cyclin D2 were adverse predictors within the non-germinal-centre group.; On multivariate analysis, an International Prognostic Index of 3 to 5 and the non-germinal-centre phenotype were independent adverse predictors.
- What it means: The practical form of cell-of-origin classification, used in pathology laboratories worldwide. When a report says germinal-centre or non-germinal-centre, this is almost always the algorithm behind it.
- Caveats: Concordance with gene-expression profiling is imperfect, and estimates of the disagreement rate vary widely between series; the algorithm misclassifies a meaningful minority of cases.; Scoring thresholds for the three stains are not fully standardised between laboratories, so the same tumour can be called differently in two hospitals.; The 152 cases came from the same consortium as the profiling study, so this is a confirmation within a cohort rather than an independent validation.; Trials that selected patients by this algorithm for activated B-cell-directed drugs, such as PHOENIX and ROBUST, were negative, which is part of the evidence that it is too blunt an instrument.

## Sources

- Blood 2004: https://doi.org/10.1182/blood-2003-05-1545
- PubMed: https://pubmed.ncbi.nlm.nih.gov/14504078/
- Europe PMC: https://europepmc.org/article/MED/14504078

## Connected records

- key papers: [A probabilistic classification tool for genetic subtypes of diffuse large B cell lymphoma with therapeutic implications](https://onco.cc/key-papers/paper-wright-lymphgen-genetic-subtypes-dlbcl-cancer-cell-2020/), [Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling](https://onco.cc/key-papers/paper-alizadeh-nature/), [Randomized phase III trial of ibrutinib and R-CHOP in non-germinal centre B-cell diffuse large B-cell lymphoma (PHOENIX)](https://onco.cc/key-papers/paper-phoenix-ibrutinib-r-chop-non-gcb-dlbcl-jco-2019/), [The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma](https://onco.cc/key-papers/paper-rosenwald-molecular-profiling-dlbcl-nejm-2002/)
- roadmaps: [Lymphoma roadmap: from a jaw tumour in Uganda and the first human cancer virus to gene-expression subtypes, PET-adapted chemotherapy, CAR-T cells, bispecific antibodies and the genetics-directed trials now recruiting](https://onco.cc/roadmaps/lymphoma-roadmap/)
- cancers: [Diffuse large B-cell lymphoma](https://onco.cc/cancers/dlbcl/), [Non-Hodgkin lymphoma (all types)](https://onco.cc/cancers/non-hodgkin-lymphoma/)
- fronts: [Diagnostics & Biomarkers](https://onco.cc/fronts/diagnostics/)
- targets: [BCL-2](https://onco.cc/targets/bcl2/), [BCL6](https://onco.cc/targets/bcl6/)
- terms: [Cell of origin (GCB vs ABC)](https://onco.cc/terms/cell-of-origin/), [International Prognostic Index (IPI)](https://onco.cc/terms/ipi-score/)
- bottlenecks: [Biomarkers are not validated or standardised](https://onco.cc/bottlenecks/b-biomarker-validation/), [Most of the world has almost no cancer care](https://onco.cc/bottlenecks/b-global-access/)
- journals: [Blood](https://onco.cc/journals/blood/)

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