Skip to main content
SpringerLink
Log in

Aptamers that bind to the human complement component receptor hC5aR1 interfere with hC5aR1 interaction to its hC5a ligand

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The complement system plays an important role in inflammation and immunity. In this system, a potent inflammatory ligand is C5a, which initiates its effects by activating its core receptor C5aR1. Thus, compounds that interfere with the C5a–C5aR1 interaction could alleviate some inflammatory conditions. Consequently, several ligands that bind to either C5a or C5aR1 have previously been isolated and evaluated. In the present study, two RNA aptamers, aptamer 1 and aptamer 9, that specifically bind to hC5aR1 with much higher affinity than antibodies were isolated. These two aptamers were tested for their ability to interfere with the cognate ligand of hC5aR1, C5a, using a chemotaxis assay. Both aptamer 1 and 9 interfered with the C5a interaction, suggesting that the aptamers recognized the extracellular domain of hC5aR1 responsible for hC5a ligand binding. Considering the higher affinity of aptamers to the hC5aR1 and their interference with hC5a ligand binding, further study is warranted to explore not only their applications in the diagnosis of inflammatory diseases but also their usefulness in modulating hC5a and hC5aR1 interactions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Mastellos DC, Deangelis RA, Lambris JD (2013) Complement-triggered pathways orchestrate regenerative responses throughout phylogenesis. Semin Immunol 25:29–38

    Article  CAS  Google Scholar 

  2. Gerard NP, Gerard C (1991) The chemotactic receptor for human C5a anaphylatoxin. Nature 349:614–617

    Article  CAS  Google Scholar 

  3. Boulay F, Mery L, Tardif M, Brouchon L, Vignais P (1991) Expression cloning of a receptor for C5a anaphylatoxin on differentiated HL-60 cells. Biochemistry 30:2993–2999

    Article  CAS  Google Scholar 

  4. Chenoweth DE, Goodman MG (1983) The C5a receptor of neutrophils and macrophages. Agents Actions Suppl 12:252–273

    CAS  PubMed  Google Scholar 

  5. Chenoweth DE, Hugli TE (1978) Demonstration of specific C5a receptor on intact human polymorphonuclear leukocytes. Proc Natl Acad Sci USA 75:3943–3947

    Article  CAS  Google Scholar 

  6. Gerard NP, Hodges MK, Drazen JM, Weller PF, Gerard C (1989) Characterization of a receptor for C5a anaphylatoxin on human eosinophils. J Biol Chem 264:1760–1766

    CAS  PubMed  Google Scholar 

  7. Werfel T, Oppermann M, Begemann G, Gotze O, Zwirner J (1997) C5a receptors are detectable on mast cells in normal human skin and in psoriatic plaques but not in weal and flare reactions or in uticaria pigmentosa by immunohistochemistry. Arch Dermatol Res 289:83–86

    Article  CAS  Google Scholar 

  8. Morelli A, Larregina A, Chuluyan I, Kolkowski E, Fainboim L (1996) Expression and modulation of C5a receptor (CD88) on skin dendritic cells. Chemotactic effect of C5a on skin migratory dendritic cells. Immunology 89:126–134

    Article  CAS  Google Scholar 

  9. El-Naggar AK, Van-Epps DE, Williams RC (1980) Human-B and T-lymphocyte locomotion in response to casein, C5a, and f-met-leu-phe. Cell Immunol 56:365–373

    Article  CAS  Google Scholar 

  10. Fayyazi A, Scheel O, Werfel T, Schweyer S, Oppermann M, Otze O, Radzun HJ, Zwirner J (2000) The C5a receptor is expressed in normal renal proximal tubular but not in normal pulmonary or hepatic epithelial cells. Immunology 99:38–45

    Article  CAS  Google Scholar 

  11. Wetsel RA (1995) Expression of the complement C5a anaphylatoxin receptor (C5aR) on non-myeloid cells. Immunol Lett 44:183–187

    Article  CAS  Google Scholar 

  12. DeMartino JA, Van Riper G, Siciliano SJ, Molineaux CJ, Konteatis ZD, Rosen H, Springer MS (1994) The amino terminus of the human C5a receptor is required for high affinity C5a binding and for receptor activation by C5a but not C5a analogs. J Biol Chem 269:14446–14450

    CAS  PubMed  Google Scholar 

  13. Chen Z, Zhang X, Gonnella NC, Pellas TC, Boyar WC, Ni F (1998) Residues 21–30 within the extracellular N-terminal region of the C5a receptor represent a binding domain for the C5a anaphylatoxin. J Biol Chem 273:10411–10419

    Article  CAS  Google Scholar 

  14. Higginbottom A, Cain SA, Woodruff TM, Proctor LM, Madala PK, Tyndall JD, Taylor SM, Fairlie DP, Monk PN (2005) Comparative agonist/antagonist responses in mutant human C5a receptors define the ligand binding site. J Biol Chem 280:17831–17840

    Article  CAS  Google Scholar 

  15. Buck E, Bourne H, Wells JA (2005) Site-specific disulfide capture of agonist and antagonist peptides on the C5a receptor. J Biol Chem 280:4009–4012

    Article  CAS  Google Scholar 

  16. Morgan BP, Harris CL (2015) Complement, a target for therapy in inflammatory and degenerative diseases. Nat Rev Drug Discov 14:857–877

    Article  CAS  Google Scholar 

  17. Brennan FH, Lee JD, Ruitenberg MJ, Woodruff TM (2016) Therapeutic targeting of complement to modify disease course and improve outcomes in neurological conditions. Semin Immunol 28:292–308

    Article  CAS  Google Scholar 

  18. Woodruff TM, Nanadakumar KS, Tedesco F (2011) Inhibiting the C5-C5a receptor axis. Mol Immunol 48:1631–1642

    Article  CAS  Google Scholar 

  19. Hawksworth OA, Li XX, Coulthard LG, Wolvetang EJ, Woodruff TM (2017) New Concepts on the therapeutic control of complement analphylatoxin receptors. Mol Immunol. https://doi.org/10.1016/j.molimm.2017.05.015

    Article  PubMed  Google Scholar 

  20. Li R, Coulthard LM, Wu MCL, Taylor SM, Woodruff TM (2013) C5L2: a controversial receptor of complement anaphylatoxin C5a. FASEB J 27:855–864

    Article  CAS  Google Scholar 

  21. Konteatis ZD, Siciliano SJ, Van Riper G, Molineaux CJ, Pandya S, Fischer P, Rosen H, Mumford RA, Springer MS (1994) Development of C5a receptor antagonists. Differential loss of functional responses. J Immunol 153:4200–4205

    CAS  PubMed  Google Scholar 

  22. Woodruff TM, Pollitt S, Proctor LM, Stocks SZ, Manthey HD, Williams HM, Mahadevan IB, Shiels IA, Taylor SM (2005) Increased potency of a novel complement factor 5a receptor antagonist in a rat model of inflammatory bowel disease. J Pharmacol Exp Ther 314:811–817

    Article  CAS  Google Scholar 

  23. Wagner F, Lange CF, Nowak M, Ignatenko S (2014) FRI0315 first human dose of the anti-C5a receptor-targeting, human monoclonal antibody NNC0215-0384 in patients with rheumatoid arthritis: a phase 1, randomised, double-blind, single-dose, dose escalation trial. Ann Rheum Dis 73:499–499

    Google Scholar 

  24. Gopinath SCB, Hayashi K, Kumar PKR (2012) Aptamers that bind to the gD protein of herpes simplex virus 1 and efficiently inhibits viral entry. J Virol 86:6732–6744

    Article  CAS  Google Scholar 

  25. Wolter O, Mayer G (2017) Aptamers as valuable molecular tools in neurosciences. J Neurosci 37:2517–2523

    Article  CAS  Google Scholar 

  26. Veedu RN, Ed (2016) Aptamer selection methodologies. In: Aptamers: tools for nanotherapy and molecular imaging. Pan Stanford Publishing Pte. Ltd, Singapore, pp 49–74

    Google Scholar 

  27. Suenaga E, Kumar PKR (2014) An aptamer that binds efficiently to the hemagglutinins of highly pathogenic avian influenza viruses (H5N1 and H5N7) and inhibits hemagglutinin-glycan interactions. Acta Biomater 10:1314–1323

    Article  CAS  Google Scholar 

  28. Yadavalli T, Agelidis A, Jaishankar D, Mangano K, Thakkar N, Kumar PKR, Shukla D (2017) Targeting herpes simplex virus-1 gD by a DNA aptamer can be an effictive new strategy to curb viral infection. Mol Ther 9:365–378

    CAS  Google Scholar 

  29. Kanwar JR, Roy K, Maremanda NG, Subramanian K, Veedu RN, Bawa R, Kanwar RK (2015) Nucleic acid-based aptamers: applications, development and cinical trials. Curr Med Chem 22:2539–2557

    Article  CAS  Google Scholar 

  30. Sullenger BA, Nair S (2016) From the RNA world to the clinic. Science 352:1417–1420

    Article  CAS  Google Scholar 

  31. Suvorova ES, Gripentrog JM, Miettinen HM (2005) Different endocytosis pathways of the C5a receptor and N-formyl peptide receptor. Traffic 6:100–115

    Article  CAS  Google Scholar 

  32. Kumar PKR, Machida K, Urvil PT, Kakiuchi N, Vishnuvardhan D, Shimotohno K, Taira K, Nishikawa S (1997) Isolation of RNA aptamers specific to the NS3 protein of hepatitis C virus from a pool of completely random RNA. Virology 237:270–282

    Article  CAS  Google Scholar 

  33. Gopinath SCB, Kumar PKR (2013) Aptamers that bind to the hemagglutinins of recent pathogenic influenza virus H1N1 and efficiently inhibit hemagglutination. Acta Biomater 9:8932–8941

    Article  CAS  Google Scholar 

  34. Yasuhara N, Kumar PKR (2016) Apatmers that bind specifically to human KPNA2 (importin-α1) and efficiently interfere with nuclear transport. J Biochem 160:259–268

    Article  CAS  Google Scholar 

  35. Rasband WS (1997–2016) ImageJ. U. S. National Institutes of Health, Bethesda. https://imagej.nih.gov/ij/. Accessed 15 May 2017

  36. Karlsson R, Katsamba PS, Nordin H, Pol E, Myszka DG (2006) Analyzing a kinetic titration series using affinity biosensors. Anal Biochem 349:136–147

    Article  CAS  Google Scholar 

  37. Trutnau HH (2006) New multi-step kinetics using common affinity biosensors saves time and sample at full access to kinetics and concentration. J Biotechnol 124:191–195

    Article  CAS  Google Scholar 

  38. Yatime L, Maasch C, Hoehlig K, Klussmann S, Andersen GR, Vater A (2015) Structural basis for the targeting of complement anaphylatixin C5a using a mized L-RNA/L-DNA aptamer. Nat Commun 6:6481. https://doi.org/10.1038/ncomms7481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Lee H, Whitfeld PL, Mackay CR (2008) Receptors for complement C5a. The importance of C5aR and the enigmatic role of C5L2. Immunol Cell Biol 86:153–160

    Article  CAS  Google Scholar 

  40. Lee H, Zahra D, Vogelzang A, Newton R, Thatcher J, Quan A, So T, Zirner J, Koentgen F, Padkaer SB, Mackay F, Whitefeld PL, Mackay CR (2006) Human C5aR knock-in mice facilitate the production and assessment of anti-inflammatory monoclonal antibodies. Nat Biotechnol 24:1279–1284

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Author would like to thank Drs. Yoshio Kato, and Hiroshi Mizuno for their assistance in acquiring fluorescent images and discussion, respectively during the course of this study.

Author information

Authors and Affiliations

  1. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Penmetcha K. R. Kumar

Authors
  1. Penmetcha K. R. Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

PKRK designed, performed all the experiments, analyzed, and wrote the manuscript.

Corresponding author

Correspondence to Penmetcha K. R. Kumar.

Ethics declarations

Conflict of interest

The author declares no conflict of interest. The funding agencies had no influence on the experimental design and the conclusions.

Ethical approval

No animal and human clinical trials are involved in this study.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 27 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kumar, P.K.R. Aptamers that bind to the human complement component receptor hC5aR1 interfere with hC5aR1 interaction to its hC5a ligand. Mol Biol Rep 45, 851–864 (2018). https://doi.org/10.1007/s11033-018-4231-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-018-4231-7

Keywords

Search

Navigation