The concentrations of IL-27 and IL-6 in the MM pleural effusions were assessed with a bead-based multiplexed immunoassay and Luminex technology (MILLIPLEX MAP Human TH17 Magnetic Bead Panel, customized HTH17MAG-8K, Merck Millipore, Darmstadt, Germany), according to the manufacturer’s instructions. Samples were tested both at 1:2 and 1:10 dilutions in Assay Buffer.
The levels of soluble PD-L1 protein were measured by ELISA using the Human PD-L1 [28–8] SimpleStep ELISA Kit (Abcam, ab214565), following datasheet instructions. Both pleural effusions and conditioned media were centrifuged in a microfuge to eliminate debris and used undiluted. Results were processed by the BioTek EL808 absorbance microplate reader and GEN5 Software version 2.03.01.
The levels of soluble PD-L1 protein were measured by ELISA using the Human PD-L1 [28–8] SimpleStep ELISA Kit (Abcam, ab214565), following datasheet instructions. Both pleural effusions and conditioned media were centrifuged in a microfuge to eliminate debris and used undiluted. Results were processed by the BioTek EL808 absorbance microplate reader and GEN5 Software version 2.03.01.