Side-by-side comparison of flow cytometry and immunohistochemistry for detection of calreticulin exposure in the course of immunogenic cell death.

July 8, 2019
Source: Methods Enzymol. 2020;632:15-25. doi: 10.1016/bs.mie.2019.05.025. Epub 2019 Jul 8.

Authors: L. Kasikova, I. Truxova, I. Cremer, C. Sautes-Fridman, O. Kepp, G. Kroemer, R. Spisek, J. Fucikova.

Immunogenic cell death (ICD), a functionally peculiar type of apoptosis, represents a unique way to deliver danger-associated molecular patterns (DAMPs) to the tumor microenvironment. Once emitted by dying cancer cells, DAMPs orchestrate antigen-specific immune responses by acting on both innate and adaptive components of the immune system. Accumulating preclinical and clinical evidence indicates that one of these DAMPs, calreticulin (CALR) represents a novel powerful prognostic biomarker, reflecting the activation of a clinically relevant anticancer immune response in different cancer malignancies. Therefore, the assessment of CALR emission can provide a therapeutic tool for the stratification of cancer patients and the identification of individuals that are intrinsically capable to respond to a particular treatment. Here we describe methods for the quantification of CALR exposure in the tumor microenvironment of cancer patients by flow cytometry and immunohistochemistry.