Neutralization of Human Interleukin 23 by Multivalent Nanobodies Explained by the Structure of Cytokine-Nanobody Complex

Overview

Journal Front Immunol

Specialty Allergy & Immunology

Date 2017 Sep 6

PMID 28871249

Citations 21

Authors

Aline Desmyter

Silvia Spinelli

Carlo Boutton

Michael Saunders

Christophe Blachetot

Hans de Haard

Geertrui Denecker

Maarten Van Roy

Christian Cambillau

Heidi Rommelaere

Affiliations

Soon will be listed here.

Abstract

The heterodimeric cytokine interleukin (IL) 23 comprises the IL12-shared p40 subunit and an IL23-specific subunit, p19. Together with IL12 and IL27, IL23 sits at the apex of the regulatory mechanisms shaping adaptive immune responses. IL23, together with IL17, plays an important role in the development of chronic inflammation and autoimmune inflammatory diseases. In this context, we generated monovalent antihuman IL23 variable heavy chain domain of llama heavy chain antibody (V) domains (Nanobodies) with low nanomolar affinity for human interleukin (hIL) 23. The crystal structure of a quaternary complex assembling hIL23 and several nanobodies against p19 and p40 subunits allowed identification of distinct epitopes and enabled rational design of a multivalent IL23-specific blocking nanobody. Taking advantage of the ease of nanobody formatting, multivalent IL23 nanobodies were assembled with properly designed linkers flanking an antihuman serum albumin nanobody, with improved hIL23 neutralization capacity and , as compared to the monovalent nanobodies. These constructs with long exposure time are excellent candidates for further developments targeting Crohn's disease, rheumatoid arthritis, and psoriasis.

Citing Articles

Redirecting immune signaling with cytokine adaptors.

Abhiraman G, Householder K, Rodriguez G, Glassman C, Saxton R, Breuer C Nat Commun. 2025; 16(1):2432.

PMID: 40069219 PMC: 11897282. DOI: 10.1038/s41467-025-57681-1.

Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF-α and IL-23p19 bioactivities.

Wang J, Kang G, Lu H, de Marco A, Yuan H, Feng Z Clin Transl Med. 2024; 14(3):e1636.

PMID: 38533646 PMC: 10966562. DOI: 10.1002/ctm2.1636.

Nanobody engineering for SARS-CoV-2 neutralization and detection.

Hannula L, Kuivanen S, Lasham J, Kant R, Kareinen L, Bogacheva M Microbiol Spectr. 2024; 12(4):e0419922.

PMID: 38363137 PMC: 10986514. DOI: 10.1128/spectrum.04199-22.

Structures of complete extracellular receptor assemblies mediated by IL-12 and IL-23.

Bloch Y, Felix J, Merceron R, Provost M, Symakani R, De Backer R Nat Struct Mol Biol. 2024; 31(4):591-597.

PMID: 38287195 DOI: 10.1038/s41594-023-01190-6.

Quantitative flow cytometric selection of tau conformational nanobodies specific for pathological aggregates.

Zupancic J, Smith M, Trzeciakiewicz H, Skinner M, Ferris S, Makowski E Front Immunol. 2023; 14:1164080.

PMID: 37622125 PMC: 10445546. DOI: 10.3389/fimmu.2023.1164080.

References

Steyaert J, Kobilka B . Nanobody stabilization of G protein-coupled receptor conformational states. Curr Opin Struct Biol. 2011; 21(4):567-72. PMC: 3166880. DOI: 10.1016/j.sbi.2011.06.011. View

Elsadek B, Kratz F . Impact of albumin on drug delivery--new applications on the horizon. J Control Release. 2011; 157(1):4-28. DOI: 10.1016/j.jconrel.2011.09.069. View

Park Y, Pardon E, Wu M, Steyaert J, Hol W . Crystal structure of a heterodimer of editosome interaction proteins in complex with two copies of a cross-reacting nanobody. Nucleic Acids Res. 2011; 40(4):1828-40. PMC: 3287191. DOI: 10.1093/nar/gkr867. View

Iwakura Y, Ishigame H . The IL-23/IL-17 axis in inflammation. J Clin Invest. 2006; 116(5):1218-22. PMC: 1451213. DOI: 10.1172/JCI28508. View

Spinelli S, Tegoni M, Frenken L, van Vliet C, Cambillau C . Lateral recognition of a dye hapten by a llama VHH domain. J Mol Biol. 2001; 311(1):123-9. DOI: 10.1006/jmbi.2001.4856. View

Muyldermans S, Cambillau C, Wyns L . Recognition of antigens by single-domain antibody fragments: the superfluous luxury of paired domains. Trends Biochem Sci. 2001; 26(4):230-5. DOI: 10.1016/s0968-0004(01)01790-x. View

Roovers R, Laeremans T, Huang L, De Taeye S, Verkleij A, Revets H . Efficient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR Nanobodies. Cancer Immunol Immunother. 2006; 56(3):303-317. PMC: 11030579. DOI: 10.1007/s00262-006-0180-4. View

Aggarwal S, Ghilardi N, Xie M, de Sauvage F, Gurney A . Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17. J Biol Chem. 2002; 278(3):1910-4. DOI: 10.1074/jbc.M207577200. View

Winn M, Ballard C, Cowtan K, Dodson E, Emsley P, Evans P . Overview of the CCP4 suite and current developments. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):235-42. PMC: 3069738. DOI: 10.1107/S0907444910045749. View

10.

McGeachy M, Cua D . Th17 cell differentiation: the long and winding road. Immunity. 2008; 28(4):445-53. DOI: 10.1016/j.immuni.2008.03.001. View

11.

Leslie A . Integration of macromolecular diffraction data. Acta Crystallogr D Biol Crystallogr. 1999; 55(Pt 10):1696-702. DOI: 10.1107/s090744499900846x. View

12.

Spinelli S, Frenken L, Bourgeois D, de Ron L, Bos W, Verrips T . The crystal structure of a llama heavy chain variable domain. Nat Struct Biol. 1996; 3(9):752-7. DOI: 10.1038/nsb0996-752. View

13.

Lupardus P, Garcia K . The structure of interleukin-23 reveals the molecular basis of p40 subunit sharing with interleukin-12. J Mol Biol. 2008; 382(4):931-41. PMC: 2666310. DOI: 10.1016/j.jmb.2008.07.051. View

14.

Yoon C, Johnston S, Tang J, Stahl M, Tobin J, Somers W . Charged residues dominate a unique interlocking topography in the heterodimeric cytokine interleukin-12. EMBO J. 2000; 19(14):3530-41. PMC: 313992. DOI: 10.1093/emboj/19.14.3530. View

15.

Roovers R, van Dongen G, Van Bergen en Henegouwen P . Nanobodies in therapeutic applications. Curr Opin Mol Ther. 2007; 9(4):327-35. View

16.

Furuzawa-Carballeda J, Vargas-Rojas M, Cabral A . Autoimmune inflammation from the Th17 perspective. Autoimmun Rev. 2007; 6(3):169-75. DOI: 10.1016/j.autrev.2006.10.002. View

17.

Bunkoczi G, Echols N, McCoy A, Oeffner R, Adams P, Read R . Phaser.MRage: automated molecular replacement. Acta Crystallogr D Biol Crystallogr. 2013; 69(Pt 11):2276-86. PMC: 3817702. DOI: 10.1107/S0907444913022750. View

18.

Oppmann B, Lesley R, Blom B, Timans J, Xu Y, Hunte B . Novel p19 protein engages IL-12p40 to form a cytokine, IL-23, with biological activities similar as well as distinct from IL-12. Immunity. 2000; 13(5):715-25. DOI: 10.1016/s1074-7613(00)00070-4. View

19.

Blanc E, Roversi P, Vonrhein C, Flensburg C, Lea S, Bricogne G . Refinement of severely incomplete structures with maximum likelihood in BUSTER-TNT. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 12 Pt 1):2210-21. DOI: 10.1107/S0907444904016427. View

20.

Ramamurthy V, Krystek Jr S, Bush A, Wei A, Emanuel S, Das Gupta R . Structures of adnectin/protein complexes reveal an expanded binding footprint. Structure. 2012; 20(2):259-69. DOI: 10.1016/j.str.2011.11.016. View