Utilisation of cytological samples for multiplex immunofluorescence assay.

Area of Science:

• Oncology
• Immunology
• Pathology

Background:

• Understanding the tumor immune microenvironment is crucial for developing effective non-small cell lung cancer (NSCLC) immunotherapies.
• Characterizing tumor-infiltrating immune cells and their interactions is essential for therapeutic advancements.

Purpose of the Study:

• To evaluate the utility of multiplex immunofluorescence (mIF) assays for characterizing immune cells within NSCLC cytological samples.
• To assess the feasibility of using mIF for exploring the immune landscape of NSCLC.

Main Methods:

• Six NSCLC cytological samples were analyzed using a custom mIF assay.
• The assay detected key immune markers (CD3, CD8, CD20, CD11b, CD68) and NSCLC cells (pan-cytokeratin).
• Image acquisition was performed on a Vectra-Polaris Automated Quantitative Pathology Imaging System.

Main Results:

• The mIF assay reliably detected and quantified myeloid cells (CD11b, CD68), T cells (CD3+, CD8+), and B lymphocytes (CD20+) in NSCLC cytological samples.
• Whole-tissue analysis correlated with H&E stains, improving understanding of tissue morphology and tumor-stroma interactions.
• Consistent and specific staining patterns were observed for all immune markers.

Conclusions:

• Multiplex immunofluorescence assays are feasible for analyzing immune cells in minimally invasive NSCLC cytological samples.
• This approach enables comprehensive exploration of the NSCLC immune microenvironment.
• mIF assays hold promise for guiding the development of targeted immunotherapies.