ORIGINAL ARTICLE Computer-driven quantitative image analysis in the assessment of tumor cell and T cell features in diffuse large B cell lymphomas Francesco Gaudio 1 & Roberto Tamma 2,3 & Giuseppe Ingravallo 4 & Tommasina Perrone 1 & Filomena Emanuela Laddaga 1 & Mariastella De Candia 1 & Eugenio Maiorano 4 & Domenico Ribatti 2,3 & Giorgina Specchia 1 Received: 17 September 2017 /Accepted: 15 December 2017 # Springer-Verlag GmbH Germany, part of Springer Nature 2017 Abstract Diffuse large B cell lymphoma (DLBCL) is recognized as the most common non-Hodgkin lymphoma subtype. Advanced high- resolution digital scans of pathology slides have enabled the development of computer-based image analysis algorithms that may assist pathologists in quantifying immunohistochemical stains. In this retrospective study, we reviewed data from 29 patients affected by DLBCL. In order to evaluate the number of tumor cells and microenvironment T cells, we performed an analysis of CD20, Ki67, and CD3 counts, assessed with the Positive Pixel Count algorithm embedded in the Aperio ImageScope software. A lower tumor cell count was observed in patients with a non-germinal center immunophenotype, high LDH, splenomegaly and an IPI ≥ 3. A lower number of CD3 was observed in patients with bulky disease, an IPI ≥ 3 and disease stage 3–4. Overall, these data confirm that quantitative analysis of the tumor cells and of the tumor microenvironment by means of computer-driven quanti- tative image analysis may add new information in DLBCL diagnosis. Keywords Diffuse large B cell lymphoma . Image analysis . Tumor cells . T cells Introduction Diffuse large B cell lymphoma (DLBCL) is the most common form of non-Hodgkin lymphoma (NHL), accounting for about 40% of B cell cancers worldwide [1, 2]. DLBCL presents a marked clinical and biological heterogeneity, due to the fact that most of these lymphomas arise from germinal center B cells at different stages of differentiation, in which recurrent genetic alterations contribute to the molecular pathogenesis of the disease [2]. Recent gene expression profiling studies have contributed to unravel the complex biological and clinical heterogeneity of this disease, leading to the definition of a germinal center (GC) B-like DLBCL subgroup, and a non-germinal center (nGC) subgroup, with unique gene expression signatures [3]. In addition to studies focused on the cancer cell and its genetic landscape to identify common features in different subgroups of DLBCL, analysis of the tumor microenviron- ment has become an important aspect in the assessment of progression of this malignancy. Different cellular components of the tumor microenvironment have been investigated in DLBCL, such as mast cells and tumor-associated macro- phages, and several correlations established encompassing prognostic significance, stage-related tumor progression, and differences in treatment outcome [4–7]. Therefore, part of the observed heterogeneity in DLBCL can be attributed to several cellular components of the inflam- matory infiltrate surrounding tumor cells, including lympho- cytes, neutrophils, macrophages, and mast cells. It is well known that the presence of immune and inflammatory cells contributes to modulate tumor growth and invasion. T cells play a major role in the different stages of tumor progression, initially restricting local tumor growth in premalignant lesions through T helper 1 cytotoxic subsets [8, 9]. A shift to the T helper 2 and T regulatory immunosuppressive phenotypes * Francesco Gaudio fragaudio@alice.it 1 Department of Emergency and Organ Transplantation (D.E.T.O.), Hematology Section, University of Bari, Bari, Italy 2 Department of Basic Medical Sciences, Neurosciences and Sensory Organs, University of Bari, Bari, Italy 3 National Cancer Institute BGiovanni Paolo II^, Bari, Italy 4 Department of Emergency and Organ Transplantation (D.E.T.O.), Pathology Section, University of Bari, Bari, Italy Annals of Hematology https://doi.org/10.1007/s00277-017-3212-6