Fig. 1.

Flow cytometry and ELISA results indicate that EVLP successfully delivers RTX to both lung tissues and lymph nodes. A) The rejected lungs are placed within a sterile dome and cannulated on the pulmonary artery and the left atrial cuff. Oxygenated perfusate exits the lungs through the atrial cannula into the reservoir. The perfusate is then passaged through a centrifugal pump and propelled into the deoxygenator and heat exchanger. The perfusate is mixed with a gaseous mixture (86% N₂, 8% CO₂, and 6% O₂) and heated to normothermia (37 °C). Lastly, the perfusate will proceed through a leukocyte filter prior to reentering the lung through the pulmonary artery cannula. The 500 mg of RTX is administered into the hard-shell reservoir and circulated. B) The experimental timeline illustrating time of sample collection. C) Lymphocytes were differentiated from total cells based on size and complexity, doublet cells excluded and the proportions of live CD45+, CD3-, CD20+ cells were taken. These CD20+ frequencies were used in ratios of CD20+ cells post over pre-EVLP observed in lymph nodes (n = 8 with RTX or n = 7 on EVLP alone) and in lung tissue (n = 8 with RTX or n = 8 on EVLP alone). Displayed are the individual ratios for each perfusion with their median and associated maximum and minimum values illustrated by the error bars. Mann–Whitney U statistical analysis was conducted with significance defined at p < 0.05. D) Tissues that were collected and frozen following perfusions were thawed and dissociated. The resulting lysates (n = 3 with RTX or n = 3 on EVLP alone) were run on a quantitative RTX ELISA. (Abbreviations: Ex-vivo lung perfusion – EVLP, Rituximab – RTX).