BackgroundPD-1/PD-L1 immune checkpoint inhibitors have been approved for monotherapy of metastatic non-small cell lung cancer (mNSCLC) depending on tumor cells' PD-L1 expression. Pleural effusion (PE) is common in mNSCLC. The significance of immunocytochemistry PD-L1 analysis from PE samples is unclear.Aim of the studyTo analyse the sensitivity regarding immunocytochemistry PD-L1 analysis of PE in NSCLC as compared to immunohistochemistry of pleural biopsies.Patients and Methods50 consecutive subjects (17 female, median age 72.5, 7 never-smokers) were enrolled in this prospective controlled two-center study. Inclusion criteria were PE, suspected or known lung cancer, indication for pleural puncture and thoracoscopy, written informed consent. Immunocytochemistry and immunohistochemistry PD-L1 analyses were performed with the Dako-PDL1-IHC-22C3pharmDx assay. Analysis for sensitivity, specificity, positive (PPV) and negative predictive value (NPV) was performed for PD-L1 detection from PE.Results50 subjects underwent pleural puncture and thoracoscopy. Pathologic diagnoses were lung cancer (48), lymphoma (1), mesothelioma (1). Sensitivity, specificity, positive-predictive-value and negative-predictive-value of PD-L1-testing with expression ≥50% defined as positive were 100% (95% confidence interval 46–100%), 63%(36–84%), 45%(18–75%), 100%(66–100%), and with expression ≥1% defined as positive 86%(56–97%), 43%(12–80%), 75%(47–92%), 60%(17–93%).ConclusionPD-L1 analysis in tumor-positive PE samples shows a very high sensitivity and negative-predictive-value, especially regarding PD-L1 expression levels ≥50% (European Medicines Agency approval). Negative results are reliable and help in the decision against a first-line checkpoint inhibitor monotherapy. However, a 1% cut-off level (United States Food and Drug Administration approval) leads to a markedly lower negatve-predictive-value, making other invasive procedures necessary. (NCT02855281)