Retained memory in tissues T cells (TRM) are a type of T cell genus that is found in peripheral tissues and does not circulate again. The initial T cell response to local infections and inflammatory conditions is represented by them because they are embedded in the tissue.
The expression of CD103, CD69, and CD49a distinguishes TRM from other cell types. TRM has been linked to better prognosis and survival in a number of cancers and is essential to the development and effectiveness of cancer vaccines. Comparisons between TRM and cytotoxic CD8 T cells in the tumor microenvironment are, however, scarce. We created the HISTOPROFILE®-TRM multiplex panel with the goal of phenotyping memory resident T cells in solid tumors and demonstrating the panel’s potential for studying TRM subpopulations in NSCLC FFPE tissue resections. This was achieved using panel design and validation.
After the slides had been identified, the BOND RX® (Leica) slide stainer was used to apply a sequential multiplex protocol using Opal® (Akoya Biosciences) fluorophores. The VECTRA® PolarisTM (Akoya Biosciences) slide scanner was then used to take multispectral images, and unmixed NSCLC images were analyzed using the HALO® (Indica Labs) Highplex module.
Simplex slides were stained for each biomarker in a simplex protocol and compared to a serial section stained with the HISTOPROFILE®-TRM multiplex panel. Staining concordance between the multiplex and simplex slides was determined with HALO® without multispectral deconvolution. The precision of the protocol was determined by intra-run repeatability analysis and inter-run reproducibility analysis (data not shown)