Combining Multiplex Immunohistochemistry with Computer-Aided Topographical and Single-Cell Analysis to Investigate the Mechanism of Spontaneous Regression in a Case of Metastatic Congenital Neuroblastoma

This is a description of a novel combination of chromogenic multiplex immunohistochemistry, digital pathology, computer-aided cell detection and topographical analysis of tumor tissue to allow a detailed study of the immune infiltrate. This is applied to a rare clinical case, where a tumor sample is available from an infant with metastatic neuroblastoma at the point of spontaneous regression. This allowed detailed analysis of the immune infiltrate, including spatial distribution and phenotype of lymphoid and myeloid populations, with a distinction between heterogeneous areas within the intra- and extra- tumoral immune microenvironments. The mechanism of spontaneous regression in congenital neuroblastoma is poorly understood, but the data obtained suggested an immune-mediated phenomenon, characterised by an adaptive T cell driven response with a significant delayed-type hypersensitivity (granulomatous) component.