Human IL-2Rɑ subunit binding modulation of IL-2 through a decline in electrostatic interactions: A computational and experimental approach

• Published in: PloS one Vol. 17; no. 2; p. e0264353
• Main Authors: Beig Parikhani, Arezoo, Bagherzadeh, Kowsar, Dehghan, Rada, Biglari, Alireza, Shokrgozar, Mohammad Ali, Riazi Rad, Farhad, Zeinali, Sirous, Talebkhan, Yeganeh, Ajdary, Soheila, Ahangari Cohan, Reza, Behdani, Mahdi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 2022; Public Library of Science (PLoS)
• Subjects: Affinity; Anticancer properties; Antitumor activity; Binding; Biology and Life Sciences; Biotechnology; CD25 antigen; Chemical compounds; Computer applications; Cytokines; Cytotoxicity; Drug dosages; Electrostatic properties; Flow cytometry; Humans; Hypotension; Immunology; Immunotherapy; Interleukin 2; Interleukin 2 receptors; Interleukin-2 - chemistry; Interleukin-2 - metabolism; Interleukin-2 Receptor alpha Subunit - chemistry; Interleukin-2 Receptor alpha Subunit - metabolism; Kidney cancer; Laboratories; Leukocytes, Mononuclear - metabolism; Lymphocytes; Medicine and Health Sciences; Metastasis; Molecular Docking Simulation; Molecular dynamics; Molecular Dynamics Simulation; Monoclonal antibodies; Mutants; Peripheral blood mononuclear cells; Pharmacology; Physical Sciences; Protein Binding; Research and Analysis Methods; Sampling techniques; Side effects; Simulation; Static Electricity; Toxicity; Tumor necrosis factor-TNF; Vascular leak syndrome; Iran
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0264353

Although high-dose IL-2 has clear antitumor effects, severe side effects like severe toxicity and activation of Tregs by binding of IL-2 to high-affinity IL-2R, hypotension, and vascular leak syndrome limit its applications as a therapeutic antitumor agent. Here in this study, a rational computational approach was employed to develop and design novel triple-mutant IL-2 variants with the aim of improving IL-2-based immunotherapy. The affinity of the mutants towards IL-2Rα was further computed with the aid of molecular dynamic simulations and umbrella sampling techniques and the obtained results were compared to those of wild-type IL-2. In vitro experiments by flow cytometry showed that the anti-CD25 mAb was able to bind to PBMC cells even after mutant 2 preincubation, however, the binding strength of the mutant to α-subunit was less than of wtIL-2. Additionally, reduction of IL-2Rα subunit affinity did not significantly disturb IL-2/IL2Rβγc subunits interactions.