Multiplex immunofluorescence staining and image analysis assay for diffuse large B cell lymphoma.

Multiplex immunofluorescence staining and image analysis assay for diffuse large B cell lymphoma. J Immunol Methods. 2019 Nov 26;:112714 Authors: Lee CW, Ren YJ, Marella M, Wang M, Hartke J, Couto SS Abstract With the explosion of immuno-oncology and the approval of many immune checkpoint therapies by regulatory agencies in the last few years, understanding the tumor microenvironment (TME) in the context of patient's immune status has become essential. Among available immune profiling techniques multiplex immunofluorescence (mIF) assays offer the unique advantage of preserving the architectural features of the tumor and revealing the spatial relationships between tumor cells and immune cells. A number of mIF and image analysis assays have been described for solid tumors but most are not sufficiently suitable for lymphoma, in which the lack of clear tumor-stromal boundaries, and high tumor density present significant challenges. Here we describe the development and optimization of a reliable workflow using Akoya Opal staining kit to label and analyze 6 markers per slide in diffuse large B-cell lymphoma (DLBCL). Five panels of 30 markers were developed to characterize infiltrating immune cells and relevant check-point proteins such as PD1, PDL1, ICOS, SIRP-alpha and Lag3 on 70 DLBCL sections. Multiplexed sections were scanned using Akoya multispectral scanner. Image analysis workflow using InForm and Matlab was developed to overcome ch...
Source: Journal of Immunological Methods - Category: Allergy & Immunology Authors: Tags: J Immunol Methods Source Type: research