title: Interleukin-2 activity can be fine tuned with engineered receptor signaling clamps.
abstract: Interleukin-2 (IL-2) regulates lymphocyte function by signaling through heterodimerization of the IL-2Rβ and γc receptor subunits. IL-2 is of considerable therapeutic interest, but harnessing its actions in a controllable manner remains a challenge. Previously, we have engineered an IL-2 "superkine" with enhanced affinity for IL-2Rβ. Here, we describe next-generation IL-2 variants that function as "receptor signaling clamps." They retained high affinity for IL-2Rβ, inhibiting binding of endogenous IL-2, but their interaction with γc was weakened, attenuating IL-2Rβ-γc heterodimerization. These IL-2 analogs acted as partial agonists and differentially affected lymphocytes poised at distinct activation thresholds. Moreover, one variant, H9-RETR, antagonized IL-2 and IL-15 better than blocking antibodies against IL-2Rα or IL-2Rβ. Furthermore, this mutein prolonged survival in a model of graft-versus-host disease and blocked spontaneous proliferation of smoldering adult T cell leukemia (ATL) T cells. This receptor-clamping approach might be a general mechanism-based strategy for engineering cytokine partial agonists for therapeutic immunomodulation.
authors.affiliation: Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892-1674, USA.
authors.forename: Suman
authors.initials: S
authors.lastname: Mitra
authors.affiliation: Howard Hughes Medical Institute and Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Ring
authors.affiliation: Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
authors.forename: Shoba
authors.initials: S
authors.lastname: Amarnath
authors.affiliation: Howard Hughes Medical Institute and Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Jamie B
authors.initials: JB
authors.lastname: Spangler
authors.affiliation: Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892-1674, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: Lymphoid Malignancies Branch, National Cancer Institute, NIH, Bethesda, MD 20892-1374, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Ju
authors.affiliation: Howard Hughes Medical Institute and Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Fischer
authors.affiliation: Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892-1674, USA.
authors.forename: Jangsuk
authors.initials: J
authors.lastname: Oh
authors.affiliation: Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892-1674, USA.
authors.forename: Rosanne
authors.initials: R
authors.lastname: Spolski
authors.affiliation: Department of Medicine, Division of Oncology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Kipp
authors.initials: K
authors.lastname: Weiskopf
authors.affiliation: Department of Medicine, Division of Oncology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Holbrook
authors.initials: H
authors.lastname: Kohrt
authors.affiliation: Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
authors.forename: Jason E
authors.initials: JE
authors.lastname: Foley
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, NIH, Rockville, MD 20852, USA.
authors.forename: Sumati
authors.initials: S
authors.lastname: Rajagopalan
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, NIH, Rockville, MD 20852, USA.
authors.forename: Eric O
authors.initials: EO
authors.lastname: Long
authors.affiliation: Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Fowler
authors.affiliation: Lymphoid Malignancies Branch, National Cancer Institute, NIH, Bethesda, MD 20892-1374, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Waldmann
authors.affiliation: Howard Hughes Medical Institute and Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: kcgarcia@stanford.edu.
authors.forename: K Christopher
authors.initials: KC
authors.lastname: Garcia
authors.affiliation: Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892-1674, USA. Electronic address: wjl@helix.nih.gov.
authors.forename: Warren J
authors.initials: WJ
authors.lastname: Leonard
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D005260:Female; D005786:Gene Expression Regulation; D006086:Graft vs Host Disease; D006801:Humans; D007376:Interleukin-2 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000276:immunology / Q000503:physiopathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D015375:Receptors, Interleukin-2 / Q000737:chemistry / Q000378:metabolism*; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology*; D016019:Survival Analysis
chemical_list: D007376:Interleukin-2; D015375:Receptors, Interleukin-2; D050799:STAT5 Transcription Factor