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The localization of a heterologous displayed antigen in the baculovirus-budded virion determines the type and strength of induced adaptive immune response


In the search of strategies of presentation of heterologous antigens to elicit humoral or cellular immune responses that modulate and properly potentiate each type of response, researchers have been studying baculovirus (BV) as vaccine vectors with promising results. For some years, several research groups explored different antigen presentation approaches using the BV AcNPV by expressing polypeptides on the surface of budded virions or by de novo synthesis of heterologous antigens by transduction of mammalian cells. In the case of expression on the surface of budded virions, for example, researchers have expressed polypeptides in peplomers as GP64 glycoprotein fusions or distributed throughout the entire surface by fusions to portions of the G protein of vesicular stomatitis virus, VSV. Recently, our group developed the strategy of cross-presentation of antigens by fusions of GP64 to the capsid protein VP39 (capsid display) for the generation of cytotoxic responses. While the different strategies showed to be effective in raising immune responses, the individuality of each analysis makes difficult the comparison of the results. Here, by comparing the different strategies, we show that localization of the model antigen ovalbumin (OVA) strongly determined the quality and intensity of the adaptive response to the heterologous antigen. Furthermore, surface display favored humoral responses, whereas capsid display favored cytotoxic responses. Finally, capsid display showed a much more efficient strategy to activate CD8-mediated responses than transduction. The incorporation of adjuvants in baculovirus formulations dramatically diminished the immunostimulatory properties of baculovirus.

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  • Abe T, Matsuura Y (2010) Host innate immune responses induced by baculovirus in mammals. Curr Gene Ther 10:226–231

    Article  CAS  PubMed  Google Scholar 

  • Abe T, Hemmi H, Miyamoto H, Moriishi K, Tamura S, Takaku H, Akira S, Matsuura Y (2005) Involvement of the Toll-like receptor 9 signaling pathway in the induction of innate immunity by baculovirus. J Virol 79:2847–2858

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Argilaguet JM, Perez-Martin E, Lopez S, Goethe M, Escribano JM, Giesow K, Keil GM, Rodriguez F (2013) BacMam immunization partially protects pigs against sublethal challenge with African swine fever virus. Antivir Res 98:61–65. doi:10.1016/j.antiviral.2013.02.005

    Article  CAS  PubMed  Google Scholar 

  • Boublik Y, Di Bonito P, Jones IM (1995) Eukaryotic virus display: engineering the major surface glycoprotein of the Autographa californica nuclear polyhedrosis virus (AcNPV) for the presentation of foreign proteins on the virus surface. Biotechnology (N Y) 13:1079–1084

    Article  CAS  Google Scholar 

  • Cao Y, Lu Z, Sun P, Fu Y, Tian F, Hao X, Bao H, Liu X, Liu Z (2011) A pseudotype baculovirus expressing the capsid protein of foot-and-mouth disease virus and a T-cell immunogen shows enhanced immunogenicity in mice. Virol J 8:77. doi:10.1186/1743-422X-8-77

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chapple SD, Jones IM (2002) Non-polar distribution of green fluorescent protein on the surface of Autographa californica nucleopolyhedrovirus using a heterologous membrane anchor. J Biotechnol 95:269–275

    Article  CAS  PubMed  Google Scholar 

  • Fang R, Feng H, Nie H, Wang L, Tu P, Song Q, Zhou Y, Zhao J (2010) Construction and immunogenicity of pseudotype baculovirus expressing Toxoplasma gondii SAG1 protein in BALB/c mice model. Vaccine 28:1803–1807. doi:10.1016/j.vaccine.2009.12.005

  • Grabherr R, Ernst W, Oker-Blom C, Jones I (2001) Developments in the use of baculoviruses for the surface display of complex eukaryotic proteins. Trends Biotechnol 19:231–236

    Article  CAS  PubMed  Google Scholar 

  • Hopkins R, Esposito D (2009) A rapid method for titrating baculovirus stocks using the Sf-9 easy titer cell line. BioTechniques 47:785–788. doi:10.2144/000113238

    Article  CAS  PubMed  Google Scholar 

  • Ikawa M, Kominami K, Yoshimura Y, Tanaka K, Nishimune Y, Okabe M (1995) A rapid and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP). FEBS Lett 375:125–128

    Article  CAS  PubMed  Google Scholar 

  • Iyori M, Nakaya H, Inagaki K, Pichyangkul S, Yamamoto DS, Kawasaki M, Kwak K, Mizukoshi M, Goto Y, Matsuoka H, Matsumoto M, Yoshida S (2013) Protective efficacy of baculovirus dual expression system vaccine expressing Plasmodium falciparum circumsporozoite protein. PLoS One 8:e70819. doi:10.1371/journal.pone.0070819

  • Kost TA, Condreay JP (2002) Recombinant baculoviruses as mammalian cell gene-delivery vectors. Trends Biotechnol 20:173–180

    Article  CAS  PubMed  Google Scholar 

  • Kukkonen SP, Airenne KJ, Marjomaki V, Laitinen OH, Lehtolainen P, Kankaanpaa P, Mahonen AJ, Raty JK, Nordlund HR, Oker-Blom C, Kulomaa MS, Yla-Herttuala S (2003) Baculovirus capsid display: a novel tool for transduction imaging. Mol Ther 8:853–862

    Article  CAS  PubMed  Google Scholar 

  • Lin SY, Chung YC, Hu YC (2014) Update on baculovirus as an expression and/or delivery vehicle for vaccine antigens. Expert Rev Vaccines 13:1501–1521. doi:10.1586/14760584.2014.951637

    Article  CAS  PubMed  Google Scholar 

  • Mlambo G, Kumar N, Yoshida S (2010) Functional immunogenicity of baculovirus expressing Pfs25, a human malaria transmission-blocking vaccine candidate antigen. Vaccine 28:7025–7029. doi:10.1016/j.vaccine.2010.08.022

    Article  CAS  PubMed  Google Scholar 

  • Molinari P, Crespo MI, Gravisaco MJ, Taboga O, Moron G (2011) Baculovirus capsid display potentiates OVA cytotoxic and innate immune responses. PLoS One 6:e24108. doi:10.1371/journal.pone.0024108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moron G, Dadaglio G, Leclerc C (2004) New tools for antigen delivery to the MHC class I pathway. Trends Immunol 25:92–97. doi:10.1016/

    Article  CAS  PubMed  Google Scholar 

  • Peralta A, Molinari P, Conte-Grand D, Calamante G, Taboga O (2007) A chimeric baculovirus displaying bovine herpesvirus-1 (BHV-1) glycoprotein D on its surface and their immunological properties. Appl Microbiol Biotechnol 75:407–414

    Article  CAS  PubMed  Google Scholar 

  • Prabakaran M, Meng T, He F, Yunrui T, Qiang J, Lin RT, Kwang J (2011) Subcutaneous immunization with baculovirus surface-displayed hemagglutinin of pandemic H1N1 Influenza A virus induces protective immunity in mice. Clin Vaccine Immunol 18:1582–1585. doi:10.1128/CVI.05114-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rodrigo WW, Block OK, Lane C, Sukupolvi-Petty S, Goncalvez AP, Johnson S, Diamond MS, Lai CJ, Rose RC, Jin X, Schlesinger JJ (2009) Dengue virus neutralization is modulated by IgG antibody subclass and Fcgamma receptor subtype. Virology 394:175–182. doi:10.1016/j.virol.2009.09.024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rohrmann GF (2013) Baculovirus molecular biology. 3rd edn., Bethesda (MD)

  • Thiele F, Tao S, Zhang Y, Muschaweckh A, Zollmann T, Protzer U, Abele R, Drexler I (2015) Modified vaccinia virus Ankara-infected dendritic cells present CD4+ T-cell epitopes by endogenous major histocompatibility complex class II presentation pathways. J Virol 89:2698–2709. doi:10.1128/JVI.03244-14

    Article  PubMed  Google Scholar 

  • Xu XG, Wang ZS, Zhang Q, Li ZC, Zhao HN, Li W, Tong DW, Liu HJ (2011) Baculovirus surface display of E envelope glycoprotein of Japanese encephalitis virus and its immunogenicity of the displayed proteins in mouse and swine models. Vaccine 29:636–643. doi:10.1016/j.vaccine.2010.11.045

    Article  CAS  PubMed  Google Scholar 

  • Yoshida S, Araki H, Yokomine T (2010) Baculovirus-based nasal drop vaccine confers complete protection against malaria by natural boosting of vaccine-induced antibodies in mice. Infect Immun 78:595–602. doi:10.1128/IAI.00877-09

    Article  CAS  PubMed  Google Scholar 

  • Yuseff MI, Pierobon P, Reversat A, Lennon-Dumenil AM (2013) How B cells capture, process and present antigens: a crucial role for cell polarity. Nat Rev Immunol 13:475–486. doi:10.1038/nri3469

    Article  CAS  PubMed  Google Scholar 

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We acknowledge Mr. Silvio Diaz (INTA) for his help with mice experiments and Dr. María José Gravisaco for the assistance with the flow cytometer. We also acknowledge Dr. Julia Sabio y García for English language editing.


This work was supported by the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) (grant number PICT 2012-395) and the Instituto Nacional de Tecnología Agropecuaria (INTA) (grant number PNBIO 1131032).

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Correspondence to Oscar Taboga.

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Eugenia Tavarone and Guido Nicolás Molina contributed equally as first authors. Paula Molinari and Oscar Taboga contributed equally as last authors.

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Tavarone, E., Molina, G.N., Amalfi, S. et al. The localization of a heterologous displayed antigen in the baculovirus-budded virion determines the type and strength of induced adaptive immune response. Appl Microbiol Biotechnol 101, 4175–4184 (2017).

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