The concentrations of IL-6, sIL-6R, sgp130, sCD163 (R&D Systems, Minneapolis, MN, USA), and soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK; eBioscience, Vienna, Austria) were determined using commercially available ELISA kits (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions. The mean minimum detection limit was 0.039 pg/mL for IL-6 High Sensitivity, 6.5 pg/mL for sIL-6R, 0.08 ng/mL for sgp130, 9.7 pg/mL for sTWEAK, and 0.177 ng/mL for sCD163. ELISA assays recognized both free cytokines and their complexes.
The chemokines’ concentration analysis was conducted in patients treated with CRT with multiplex technique of the commercially available panel of 40 inflammatory chemokines (Human Chemokine Array Q1, RayBiotech Inc., Peachtree Corners, GA, USA), as previously described [17 (link)]. This method is based on the specific reaction of antibodies with the chemokine molecules. The visualization of the chemokine–antibody–biotin complex is performed by laser scanner (GenePix 4100A, Axon Instruments Inc., Foster City, CA, USA).
The chemokines’ concentration analysis was conducted in patients treated with CRT with multiplex technique of the commercially available panel of 40 inflammatory chemokines (Human Chemokine Array Q1, RayBiotech Inc., Peachtree Corners, GA, USA), as previously described [17 (link)]. This method is based on the specific reaction of antibodies with the chemokine molecules. The visualization of the chemokine–antibody–biotin complex is performed by laser scanner (GenePix 4100A, Axon Instruments Inc., Foster City, CA, USA).
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