Skip to main content
SpringerLink
Log in
Book cover

Natural Antibodies pp 187–196Cite as

Evaluating the Impact of Natural IgM on Adenovirus Type 5 Gene Therapy Vectors

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1643))

Abstract

Natural IgM antibodies have an innate ability to recognize many viruses and viral-based gene therapy vectors. Naive mice have natural IgM antibodies that bind to adenoviruses, and these antibodies can profoundly affect the biodistribution and efficiency of gene delivery by adenovirus type 5 vectors. Here, we present protocols for isolating IgM from mouse serum, for assaying the concentration and adenoviral reactivity of mouse IgM, and for evaluating how natural antibodies and complement can synergize to neutralize adenovirus vectors.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. The Journal of Gene Medicine Gene Therapy Clinical Trials Worldwide (.2016) http://wwwabediacom/wiley/vectorsphp

    Google Scholar 

  2. Parker AL, Waddington SN, Buckley SM, Custers J, Havenga MJ, van Rooijen N et al (2009) Effect of neutralizing sera on factor X-mediated adenovirus serotype 5 gene transfer. J Virol 83(1):479–483. doi:10.1128/JVI.01878-08

    Article  CAS  PubMed  Google Scholar 

  3. Vogels R, Zuijdgeest D, van Rijnsoever R, Hartkoorn E, Damen I, de Bethune MP et al (2003) Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination: efficient human cell infection and bypass of preexisting adenovirus immunity. J Virol 77(15):8263–8271

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Tsai V, Johnson DE, Rahman A, Wen SF, LaFace D, Philopena J et al (2004) Impact of human neutralizing antibodies on antitumor efficacy of an oncolytic adenovirus in a murine model. Clin Cancer Res 10(21):7199–7206. doi:10.1158/1078-0432.CCR-04-0765. 10/21/7199 [pii]

    Article  CAS  PubMed  Google Scholar 

  5. Pichla-Gollon SL, Lin SW, Hensley SE, Lasaro MO, Herkenhoff-Haut L, Drinker M et al (2009) Effect of preexisting immunity on an adenovirus vaccine vector: in vitro neutralization assays fail to predict inhibition by antiviral antibody in vivo. J Virol 83(11):5567–5573. doi:10.1128/JVI.00405-09

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Byrnes AP (2016) Antibodies against adenoviruses. In: Curiel D (ed) Adenovirus vectors for gene therapy, 2nd edn. Elsevier, Amsterdam, pp 367–390

    Google Scholar 

  7. Wolff G, Worgall S, van Rooijen N, Song WR, Harvey BG, Crystal RG (1997) Enhancement of in vivo adenovirus-mediated gene transfer and expression by prior depletion of tissue macrophages in the target organ. J Virol 71(1):624–629

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Tao N, Gao GP, Parr M, Johnston J, Baradet T, Wilson JM et al (2001) Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver. Mol Ther 3(1):28–35. doi:10.1006/mthe.2000.0227

    Article  CAS  PubMed  Google Scholar 

  9. Manickan E, Smith JS, Tian J, Eggerman TL, Lozier JN, Muller J et al (2006) Rapid Kupffer cell death after intravenous injection of adenovirus vectors. Mol Ther 13(1):108–117. doi:10.1016/j.ymthe.2005.08.007

    Article  CAS  PubMed  Google Scholar 

  10. Di Paolo NC, Doronin K, Baldwin LK, Papayannopoulou T, Shayakhmetov DM (2013) The transcription factor IRF3 triggers “defensive suicide” necrosis in response to viral and bacterial pathogens. Cell Rep 3(6):1840–1846. doi:10.1016/j.celrep.2013.05.025. S2211-1247(13)00242-8 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  11. He JQ, Keatschke KJ Jr, Gribling P, Suto E, Lee WP, Diehl L et al (2013) CRIg mediates early Kupffer cell responses to adenovirus. J Leukoc Biol 93:301–306. doi:10.1189/jlb.0612311. jlb.0612311 [pii]

    Article  CAS  PubMed  Google Scholar 

  12. Smith JS, Xu Z, Tian J, Stevenson SC, Byrnes AP (2008) Interaction of systemically delivered adenovirus vectors with Kupffer cells in mouse liver. Hum Gene Ther 19(5):547–554. doi:10.1089/hum.2008.004

    Article  CAS  PubMed  Google Scholar 

  13. Xu ZL, Tian J, Smith JS, Byrnes AP (2008) Clearance of adenovirus by Kupffer cells is mediated by scavenger receptors, natural antibodies, and complement. J Virol 82(23):11705–11713. doi:10.1128/Jvi.01320-08

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Unzu C, Melero I, Morales-Kastresana A, Sampedro A, Serrano-Mendioroz I, Azpilikueta A et al (2014) Innate functions of immunoglobulin M lessen liver gene transfer with helper-dependent adenovirus. PLoS One 9(1):e85432. doi:10.1371/journal.pone.0085432

    Article  PubMed  PubMed Central  Google Scholar 

  15. Qiu Q, Xu Z, Tian J, Moitra R, Gunti S, Notkins AL et al (2015) Impact of natural IgM concentration on gene therapy with adenovirus type 5 vectors. J Virol 89(6):3412–3416. doi:10.1128/JVI.03217-14. JVI.03217-14 [pii]

    Article  CAS  PubMed  Google Scholar 

  16. Khare R, Hillestad ML, Xu Z, Byrnes AP, Barry MA (2013) Circulating antibodies and macrophages as modulators of adenovirus pharmacology. J Virol 87(7):3678–3686. doi:10.1128/JVI.01392-12. JVI.01392-12 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Corte-Real J, Rodo J, Almeida P, Garcia J, Coutinho A, Demengeot J et al (2009) Irf4 is a positional and functional candidate gene for the control of serum IgM levels in the mouse. Genes Immun 10(1):93–99. doi:10.1038/gene.2008.73. gene200873 [pii]

    Article  CAS  PubMed  Google Scholar 

  18. Snoeys J, Mertens G, Lievens J, van Berkel T, Collen D, Biessen EA et al (2006) Lipid emulsions potently increase transgene expression in hepatocytes after adenoviral transfer. Mol Ther 13(1):98–107. doi:10.1016/j.ymthe.2005.06.477

    Article  CAS  PubMed  Google Scholar 

  19. Borsos T, Rapp HJ (1965) Complement fixation on cell surfaces by 19S and 7S antibodies. Science 150(3695):505–506

    Article  CAS  PubMed  Google Scholar 

  20. Xu Z, Qiu Q, Tian J, Smith JS, Conenello GM, Morita T et al (2013) Coagulation factor X shields adenovirus type 5 from attack by natural antibodies and complement. Nat Med 19(4):452–457. doi:10.1038/nm.3107. nm.3107 [pii]

    Article  CAS  PubMed  Google Scholar 

  21. Tian J, Xu Z, Smith JS, Hofherr SE, Barry MA, Byrnes AP (2009) Adenovirus activates complement by distinctly different mechanisms in vitro and in vivo: indirect complement activation by virions in vivo. J Virol 83(11):5648–5658. doi:10.1128/JVI.00082-09

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Waddington SN, McVey JH, Bhella D, Parker AL, Barker K, Atoda H et al (2008) Adenovirus serotype 5 hexon mediates liver gene transfer. Cell 132(3):397–409. doi:10.1016/j.cell.2008.01.016

    Article  CAS  PubMed  Google Scholar 

  23. Doronin K, Flatt JW, Di Paolo NC, Khare R, Kalyuzhniy O, Acchione M et al (2012) Coagulation factor X activates innate immunity to human species C adenovirus. Science 338(6108):795–798. doi:10.1126/science.1226625. science.1226625 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Kalyuzhniy O, Di Paolo NC, Silvestry M, Hofherr SE, Barry MA, Stewart PL et al (2008) Adenovirus serotype 5 hexon is critical for virus infection of hepatocytes in vivo. Proc Natl Acad Sci U S A 105(14):5483–5488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Duffy MR, Doszpoly A, Turner G, Nicklin SA, Baker AH (2016) The relevance of coagulation factor X protection of adenoviruses in human sera. Gene Ther. doi:10.1038/gt.2016.32

    PubMed  PubMed Central  Google Scholar 

  26. Ma J, Duffy MR, Deng L, Dakin RS, Uil T, Custers J et al (2015) Manipulating adenovirus hexon hypervariable loops dictates immune neutralisation and coagulation factor X-dependent cell interaction in vitro and in vivo. PLoS Pathog 11(2):e1004673. doi:10.1371/journal.ppat.1004673. PPATHOGENS-D-14-02650 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  27. Vellekamp G, Porter FW, Sutjipto S, Cutler C, Bondoc L, Liu YH et al (2001) Empty capsids in column-purified recombinant adenovirus preparations. Hum Gene Ther 12(15):1923–1936. doi:10.1089/104303401753153974

    Article  CAS  PubMed  Google Scholar 

  28. Lachmann PJ (2010) Preparing serum for functional complement assays. J Immunol Methods 352(1–2):195–197. doi:10.1016/j.jim.2009.11.003. S0022-1759(09)00339-1 [pii]

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the FDA, including funding from the CBER’s Critical Path program. We thank Mike Havert and Nirjal Bhattarai for reviewing the manuscript.

Author information

Authors and Affiliations

  1. Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD, 20993, USA

    Zhili Xu, Jie Tian, Andrew W. Harmon & Andrew P. Byrnes

Authors
  1. Zhili Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jie Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrew W. Harmon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andrew P. Byrnes
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrew P. Byrnes .

Editor information

Editors and Affiliations

  1. Centre de Recherche des Cordeliers, INSERM, PARIS, Paris, France

    Srinivas V. Kaveri

  2. Centre de Recherche des Cordeliers, INSERM, PARIS, Paris, France

    Jagadeesh Bayry

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Xu, Z., Tian, J., Harmon, A.W., Byrnes, A.P. (2017). Evaluating the Impact of Natural IgM on Adenovirus Type 5 Gene Therapy Vectors. In: Kaveri, S., Bayry, J. (eds) Natural Antibodies. Methods in Molecular Biology, vol 1643. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7180-0_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7180-0_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7179-4

  • Online ISBN: 978-1-4939-7180-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation