Journal Article
. 2021 May; 11(1):10592.
doi: 10.1038/s41598-021-90096-8.

A bispecific antibody agonist of the IL-2 heterodimeric receptor preferentially promotes in vivo expansion of CD8 and NK cells

Katherine E Harris 1 Kyle J Lorentsen 1 Harbani K Malik-Chaudhry 1 Kaitlyn Loughlin 1 Harish Medlari Basappa 1 Sharon Hartstein 1 Ghenima Ahmil 2 Nicole S Allen 1 Brian C Avanzino 1 Aarti Balasubramani 1 Andrew A Boudreau 1 Karen Chang 1 Maria-Cristina Cuturi 2 Laura M Davison 1 Dennis M Ho 1 Suhasini Iyer 1 Udaya S Rangaswamy 1 Preethi Sankaran 1 Ute Schellenberger 1 Roland Buelow 1 Nathan D Trinklein 3 
  • PMID: 34011961
  •     53 References


The use of recombinant interleukin-2 (IL-2) as a therapeutic protein has been limited by significant toxicities despite its demonstrated ability to induce durable tumor-regression in cancer patients. The adverse events and limited efficacy of IL-2 treatment are due to the preferential binding of IL-2 to cells that express the high-affinity, trimeric receptor, IL-2Rαβγ such as endothelial cells and T-regulatory cells, respectively. Here, we describe a novel bispecific heavy-chain only antibody which binds to and activates signaling through the heterodimeric IL-2Rβγ receptor complex that is expressed on resting T-cells and NK cells. By avoiding binding to IL-2Rα, this molecule circumvents the preferential T-reg activation of native IL-2, while maintaining the robust stimulatory effects on T-cells and NK-cells in vitro. In vivo studies in both mice and cynomolgus monkeys confirm the molecule's in vivo biological activity, extended pharmacodynamics due to the Fc portion of the molecule, and enhanced safety profile. Together, these results demonstrate that the bispecific antibody is a safe and effective IL-2R agonist that harnesses the benefits of the IL-2 signaling pathway as a potential anti-cancer therapy.

Improved cancer immunotherapy by a CD25-mimobody conferring selectivity to human interleukin-2.
Natalia Arenas-Ramirez, Chao Zou, +11 authors, Onur Boyman.
Sci Transl Med, 2016 Dec 03; 8(367). PMID: 27903862
Immunogenicity of protein therapeutics.
Anne S De Groot, David W Scott.
Trends Immunol, 2007 Oct 30; 28(11). PMID: 17964218
Highly Cited. Review.
The ratios of CD8+ T cells to CD4+CD25+ FOXP3+ and FOXP3- T cells correlate with poor clinical outcome in human serous ovarian cancer.
Claudia C Preston, Matthew J Maurer, +5 authors, Keith L Knutson.
PLoS One, 2013 Nov 19; 8(11). PMID: 24244610    Free PMC article.
Highly Cited.
Potential induction of anti-PEG antibodies and complement activation toward PEGylated therapeutics.
Johan J F Verhoef, John F Carpenter, Thomas J Anchordoquy, Huub Schellekens.
Drug Discov Today, 2014 Sep 11; 19(12). PMID: 25205349
Multispecific Antibody Development Platform Based on Human Heavy Chain Antibodies.
Starlynn C Clarke, Biao Ma, +19 authors, Shelley Force Aldred.
Front Immunol, 2019 Jan 23; 9. PMID: 30666250    Free PMC article.
IL15 by Continuous Intravenous Infusion to Adult Patients with Solid Tumors in a Phase I Trial Induced Dramatic NK-Cell Subset Expansion.
Kevin C Conlon, E Lake Potter, +15 authors, Thomas A Waldmann.
Clin Cancer Res, 2019 May 31; 25(16). PMID: 31142503    Free PMC article.
A First-in-Human Phase I Study of Subcutaneous Outpatient Recombinant Human IL15 (rhIL15) in Adults with Advanced Solid Tumors.
Jeffrey S Miller, Chihiro Morishima, +12 authors, Kevin C Conlon.
Clin Cancer Res, 2017 Dec 06; 24(7). PMID: 29203590    Free PMC article.
An efficient route to human bispecific IgG.
A M Merchant, Z Zhu, +4 authors, P Carter.
Nat Biotechnol, 1998 Jul 14; 16(7). PMID: 9661204
Highly Cited.
In vivo administration of purified human interleukin 2. II. Half life, immunologic effects, and expansion of peripheral lymphoid cells in vivo with recombinant IL 2.
M T Lotze, Y L Matory, +5 authors, S A Rosenberg.
J Immunol, 1985 Oct 01; 135(4). PMID: 2993418
Highly Cited.
IL-2: the first effective immunotherapy for human cancer.
Steven A Rosenberg.
J Immunol, 2014 Jun 08; 192(12). PMID: 24907378    Free PMC article.
Highly Cited. Review.
89Zr-labeled CEA-targeted IL-2 variant immunocytokine in patients with solid tumors: CEA-mediated tumor accumulation and role of IL-2 receptor-binding.
Emilie M J van Brummelen, Marc C Huisman, +12 authors, C Willemien Menke-van der Houven van Oordt.
Oncotarget, 2018 Jun 07; 9(37). PMID: 29872502    Free PMC article.
The IL-2 cytokine family in cancer immunotherapy.
Geok Choo Sim, Laszlo Radvanyi.
Cytokine Growth Factor Rev, 2014 Sep 10; 25(4). PMID: 25200249
Highly Cited. Review.
NKTR-214, an Engineered Cytokine with Biased IL2 Receptor Binding, Increased Tumor Exposure, and Marked Efficacy in Mouse Tumor Models.
Deborah H Charych, Ute Hoch, +19 authors, Stephen K Doberstein.
Clin Cancer Res, 2016 Feb 03; 22(3). PMID: 26832745
Highly Cited.
High-dose recombinant interleukin 2 therapy for patients with metastatic melanoma: analysis of 270 patients treated between 1985 and 1993.
M B Atkins, M T Lotze, +10 authors, S A Rosenberg.
J Clin Oncol, 1999 Nov 24; 17(7). PMID: 10561265
Highly Cited. Review.
Redistribution, hyperproliferation, activation of natural killer cells and CD8 T cells, and cytokine production during first-in-human clinical trial of recombinant human interleukin-15 in patients with cancer.
Kevin C Conlon, Enrico Lugli, +20 authors, Thomas A Waldmann.
J Clin Oncol, 2014 Nov 19; 33(1). PMID: 25403209    Free PMC article.
Highly Cited.
Improved IL-2 immunotherapy by selective stimulation of IL-2 receptors on lymphocytes and endothelial cells.
Carsten Krieg, Sven Létourneau, Giuseppe Pantaleo, Onur Boyman.
Proc Natl Acad Sci U S A, 2010 Jun 16; 107(26). PMID: 20547866    Free PMC article.
Single-cell quantification of IL-2 response by effector and regulatory T cells reveals critical plasticity in immune response.
Ofer Feinerman, Garrit Jentsch, +6 authors, Grégoire Altan-Bonnet.
Mol Syst Biol, 2010 Dec 02; 6. PMID: 21119631    Free PMC article.
Highly Cited.
gp100 peptide vaccine and interleukin-2 in patients with advanced melanoma.
Douglas J Schwartzentruber, David H Lawson, +23 authors, Patrick Hwu.
N Engl J Med, 2011 Jun 03; 364(22). PMID: 21631324    Free PMC article.
Highly Cited.
Establishment of an IL-2 independent, human T-cell line possessing only the p70 IL-2 receptor.
G Starkebaum, T P Loughran, C A Waters, F W Ruscetti.
Int J Cancer, 1991 Sep 19; 49(2). PMID: 1879969
Therapeutic administration of IL-15 superagonist complex ALT-803 leads to long-term survival and durable antitumor immune response in a murine glioblastoma model.
Dimitrios Mathios, Chul-Kee Park, +6 authors, Michael Lim.
Int J Cancer, 2015 Jul 16; 138(1). PMID: 26174883    Free PMC article.
Sequence-Based Discovery Demonstrates That Fixed Light Chain Human Transgenic Rats Produce a Diverse Repertoire of Antigen-Specific Antibodies.
Katherine E Harris, Shelley Force Aldred, +19 authors, Nathan D Trinklein.
Front Immunol, 2018 May 10; 9. PMID: 29740455    Free PMC article.
Regulatory T-cell responses to low-dose interleukin-2 in HCV-induced vasculitis.
David Saadoun, Michelle Rosenzwajg, +6 authors, David Klatzmann.
N Engl J Med, 2011 Dec 02; 365(22). PMID: 22129253
Highly Cited.
De novo design of potent and selective mimics of IL-2 and IL-15.
Daniel-Adriano Silva, Shawn Yu, +22 authors, David Baker.
Nature, 2019 Jan 11; 565(7738). PMID: 30626941    Free PMC article.
Highly Cited.
The biology of interleukin-2 and interleukin-15: implications for cancer therapy and vaccine design.
Thomas A Waldmann.
Nat Rev Immunol, 2006 Jul 27; 6(8). PMID: 16868550
Highly Cited. Review.
The tumour-targeting human L19-IL2 immunocytokine: preclinical safety studies, phase I clinical trial in patients with solid tumours and expansion into patients with advanced renal cell carcinoma.
Manfred Johannsen, Gianluca Spitaleri, +18 authors, Filippo de Braud.
Eur J Cancer, 2010 Aug 28; 46(16). PMID: 20797845
Antibody-cytokine fusion proteins: A novel class of biopharmaceuticals for the therapy of cancer and of chronic inflammation.
Patrizia Murer, Dario Neri.
N Biotechnol, 2019 Apr 17; 52. PMID: 30991144    Free PMC article.
ALKS 4230: a novel engineered IL-2 fusion protein with an improved cellular selectivity profile for cancer immunotherapy.
Jared E Lopes, Jan L Fisher, +5 authors, Heather C Losey.
J Immunother Cancer, 2020 Apr 23; 8(1). PMID: 32317293    Free PMC article.
Durability of complete responses in patients with metastatic cancer treated with high-dose interleukin-2: identification of the antigens mediating response.
S A Rosenberg, J C Yang, D E White, S M Steinberg.
Ann Surg, 1998 Sep 22; 228(3). PMID: 9742914    Free PMC article.
Highly Cited.
Exploiting a natural conformational switch to engineer an interleukin-2 'superkine'.
Aron M Levin, Darren L Bates, +11 authors, K Christopher Garcia.
Nature, 2012 Mar 27; 484(7395). PMID: 22446627    Free PMC article.
Highly Cited.
Intrachain disulfide bond in the core hinge region of human IgG4.
J W Bloom, M S Madanat, +2 authors, S Y Chan.
Protein Sci, 1997 Feb 01; 6(2). PMID: 9041643    Free PMC article.
Interleukin-2: Biology, Design and Application.
Natalia Arenas-Ramirez, Janine Woytschak, Onur Boyman.
Trends Immunol, 2015 Nov 18; 36(12). PMID: 26572555
Highly Cited. Review.
Prospects of IL-2 in Cancer Immunotherapy.
Hani Choudhry, Nawal Helmi, +7 authors, Mohammad S Jamal.
Biomed Res Int, 2018 Jun 02; 2018. PMID: 29854806    Free PMC article.
'Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization.
J B Ridgway, L G Presta, P Carter.
Protein Eng, 1996 Jul 01; 9(7). PMID: 8844834
Highly Cited.
Efficacy and mechanism-of-action of a novel superagonist interleukin-15: interleukin-15 receptor αSu/Fc fusion complex in syngeneic murine models of multiple myeloma.
Wenxin Xu, Monica Jones, +10 authors, Hing C Wong.
Cancer Res, 2013 May 07; 73(10). PMID: 23644531    Free PMC article.
Highly Cited.
Bempegaldesleukin (NKTR-214) plus Nivolumab in Patients with Advanced Solid Tumors: Phase I Dose-Escalation Study of Safety, Efficacy, and Immune Activation (PIVOT-02).
Adi Diab, Nizar M Tannir, +20 authors, Daniel C Cho.
Cancer Discov, 2020 May 23; 10(8). PMID: 32439653
Fc-dependent depletion of tumor-infiltrating regulatory T cells co-defines the efficacy of anti-CTLA-4 therapy against melanoma.
Tyler R Simpson, Fubin Li, +11 authors, Sergio A Quezada.
J Exp Med, 2013 Jul 31; 210(9). PMID: 23897981    Free PMC article.
Highly Cited.
Durable responses and reversible toxicity of high-dose interleukin-2 treatment of melanoma and renal cancer in a Community Hospital Biotherapy Program.
Roxanne Payne, Lyn Glenn, +6 authors, Brendan D Curti.
J Immunother Cancer, 2014 May 24; 2. PMID: 24855563    Free PMC article.
Interleukin-2 and regulatory T cells in graft-versus-host disease.
John Koreth, Ken-ichi Matsuoka, +15 authors, Robert J Soiffer.
N Engl J Med, 2011 Dec 02; 365(22). PMID: 22129252    Free PMC article.
Highly Cited.
IL-2 therapy promotes suppressive ICOS+ Treg expansion in melanoma patients.
Geok Choo Sim, Natalia Martin-Orozco, +9 authors, Laszlo Radvanyi.
J Clin Invest, 2013 Dec 03; 124(1). PMID: 24292706    Free PMC article.
Highly Cited.
The role of interleukin-2 during homeostasis and activation of the immune system.
Onur Boyman, Jonathan Sprent.
Nat Rev Immunol, 2012 Feb 22; 12(3). PMID: 22343569
Highly Cited. Review.
In vitro characterization of five humanized OKT3 effector function variant antibodies.
D Xu, M L Alegre, +9 authors, R A Zivin.
Cell Immunol, 2000 Mar 16; 200(1). PMID: 10716879
Phase I Trial of ALT-803, A Novel Recombinant IL15 Complex, in Patients with Advanced Solid Tumors.
Kim Margolin, Chihiro Morishima, +14 authors, Marc S Ernstoff.
Clin Cancer Res, 2018 Jul 27; 24(22). PMID: 30045932    Free PMC article.
Highly Cited.
A function for interleukin 2 in Foxp3-expressing regulatory T cells.
Jason D Fontenot, Jeffrey P Rasmussen, Marc A Gavin, Alexander Y Rudensky.
Nat Immunol, 2005 Oct 18; 6(11). PMID: 16227984
Highly Cited.
Elimination of Fc receptor-dependent effector functions of a modified IgG4 monoclonal antibody to human CD4.
M P Reddy, C A Kinney, +12 authors, A Truneh.
J Immunol, 2000 Feb 05; 164(4). PMID: 10657642
Randomized phase III trial of high-dose interleukin-2 versus subcutaneous interleukin-2 and interferon in patients with metastatic renal cell carcinoma.
David F McDermott, Meredith M Regan, +15 authors, Michael B Atkins.
J Clin Oncol, 2004 Dec 31; 23(1). PMID: 15625368
Highly Cited.
High donor FOXP3-positive regulatory T-cell (Treg) content is associated with a low risk of GVHD following HLA-matched allogeneic SCT.
Katayoun Rezvani, Stephan Mielke, +8 authors, A John Barrett.
Blood, 2006 Apr 22; 108(4). PMID: 16627754    Free PMC article.
Highly Cited.
The human IL-15 superagonist N-803 promotes migration of virus-specific CD8+ T and NK cells to B cell follicles but does not reverse latency in ART-suppressed, SHIV-infected macaques.
Gabriela M Webb, Jhomary Molden, +19 authors, Jonah B Sacha.
PLoS Pathog, 2020 Mar 13; 16(3). PMID: 32163523    Free PMC article.
A whole blood in vitro cytokine release assay with aqueous monoclonal antibody presentation for the prediction of therapeutic protein induced cytokine release syndrome in humans.
Babette Wolf, Hannah Morgan, +7 authors, Andrea Kiessling.
Cytokine, 2012 Sep 19; 60(3). PMID: 22986013
The binding affinity of human IgG for its high affinity Fc receptor is determined by multiple amino acids in the CH2 domain and is modulated by the hinge region.
S M Canfield, S L Morrison.
J Exp Med, 1991 Jun 01; 173(6). PMID: 1827828    Free PMC article.
Complete eradication of human B-cell lymphoma xenografts using rituximab in combination with the immunocytokine L19-IL2.
Christoph Schliemann, Alessandro Palumbo, +6 authors, Dario Neri.
Blood, 2008 Nov 14; 113(10). PMID: 19005180
The structure of interleukin-2 complexed with its alpha receptor.
Mathias Rickert, Xinquan Wang, +2 authors, K Christopher Garcia.
Science, 2005 Jun 04; 308(5727). PMID: 15933202
Projecting human pharmacokinetics of therapeutic antibodies from nonclinical data: what have we learned?
Rong Deng, Suhasini Iyer, +3 authors, Saileta Prabhu.
MAbs, 2010 Oct 22; 3(1). PMID: 20962582    Free PMC article.
Highly Cited.
Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy: Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines.
Christian Klein, Inja Waldhauer, +27 authors, Pablo Umaña.
Oncoimmunology, 2017 Apr 14; 6(3). PMID: 28405498    Free PMC article.
Highly Cited.