Scientific publications
Utilisation of cytological samples for multiplex immunofluorescence assay
Laura Garcia Tobar 1 , María Villalba-Esparza 1 2 , Marta Abengozar-Muela 1 3 , Laura Alvarez Gigli 1 3 , José I Echeveste 1 3 , Carlos E de Andrea 1 2 3 , María D Lozano 1 3
Objective: Understanding the immune environment of non-small cell lung cancer (NSCLC) is important for designing effective anticancer immunotherapies. We describe the use of multiplex immunofluorescence (mIF) assays to enable characterisation of the tumour-infiltrating immune cells and their interactions, both across and within immune subtypes.
Methods: Six cytological samples of NSCLC taken by transoesophageal ultrasound-guided fine needle aspiration were tested with an mIF assay designed to detect the expression of key immune cell markers such as CD3, CD8, CD20, CD11b, and CD68. Pan-cytokeratin was used to detect the NSCLC cells. Fluorescence images were acquired on a Vectra-Polaris Automated Quantitative Pathology Imaging System (Akoya Biosciences).
Results: MIF assay was able to reliably detect and quantify the myeloid cell markers CD11b, CD68, CD3+ and CD8+ T cells, and CD20+ B lymphocytes on cytological samples of NSCLC. Whole-tissue analysis and its correlation with the corresponding H&E stains allowed a better understanding of the tissue morphology and the relationship between tumour and stroma compartments. Additionally, a uniform, specific, and correct staining pattern was seen for every immune marker.
Conclusion: The implementation of mIF assay on cytological samples taken with minimally invasive methods seems feasible and can be used to explore the immune environment of NSCLC.
CITATION Cytopathology. 2021 Sep;32(5):611-616. doi: 10.1111/cyt.12979. Epub 2021 Apr 18