3D image analysis reveals differences of CD30 positive cells and network formation in reactive and malignant human lymphoid tissue (classical Hodgkin Lymphoma)

AIMS: The examination of histological sections is still the gold standard in diagnostic pathology. Important histopathological diagnostic criteria are nuclear shapes and chromatin distribution as well as nucleus-cytoplasm relation and immunohistochemical properties of surface and intracellular proteins. The aim of this investigation was to evaluate the benefits and drawbacks of three-dimensional imaging of CD30(+) cells in classical Hodgkin Lymphoma (cHL) in comparison to CD30(+) lymphoid cells in reactive lymphoid tissues. MATERIALS AND RESULTS: Using immunoflourescence confocal microscopy and computer-based analysis, we compared CD30(+) neoplastic cells in Nodular Sclerosis cHL (NScCHL), Mixed Cellularity cHL (MCcHL), with reactive CD30(+) cells in Adenoids (AD) and Lymphadenitis (LAD). We confirmed that the percentage of CD30(+) cell volume can be calculated. The amount in lymphadenitis was approx. 1.5%, in adenoids around 2%, in MCcHL up to 4,5% whereas the values for NScHL rose to more than 8% of the total cell cytoplasm. In addition, CD30(+) tumour cells (HRS-cells) in cHL had larger volumes, and more protrusions compared to CD30(+) reactive cells. Furthermore, the formation of large cell networks turned out to be a typical characteristic of NScHL. CONCLUSION: In contrast to 2D histology, 3D laser scanning offers a visualisation of complete cells, their network interaction and spatial distribution in the tissue. The possibility to differentiate cells in regards to volume, surface, shape, and cluster formation enables a new view on further diagnostic and biological questions. 3D includes an increased amount of information as a basis of bioinformatical calculations.