Skip to main content
Log in

Production of Two Vaccinating Recombinant Rotavirus Proteins in the Milk of Transgenic Rabbits

  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Rotaviruses are the main cause of infantile viral gastroenteritis worldwide leading to approximately 500,000 deaths each year mostly in the developing world. For unknown reasons, live attenuated viruses used in classical vaccine strategies were shown to be responsible for intussusception (a bowel obstruction). New strategies allowing production of safe recombinant non-replicating rotavirus candidate vaccine are thus clearly needed. In this study we utilized transgenic rabbit milk as a source of rotavirus antigens. Individual transgenic rabbit lines were able to produce several hundreds of micrograms per ml of secreted recombinant VP2 and VP6 proteins in their milk. Viral proteins expressed in our model were immunogenic and were shown to induce a significant reduction in viral antigen shedding after challenge with virulent rotavirus in the adult mouse model. To our knowledge, this is the first report of transgenic mammal bioreactors allowing the rapid co-production of two recombinant viral proteins in milk to be used as a vaccine.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

RV:

rotavirus

Tg:

transgenic

; VLP:

virus-like particles

References

  • J Attal MC Theron G Kann P Bolifraud C Puissant LM Houdebine (2000) ArticleTitleThe stimulation of gene expression by the R region from HTLV-1 and BLV J Biotechnol 2–3 179–189

    Google Scholar 

  • A Bertolotti-Ciarlet M Ciarlet SE Crawford ME Conner MK Estes (2003) ArticleTitleImmunogenicity and protective efficacy of rotavirus 2/6-virus-like particles produced by a dual baculovirus expression vector and administered intramuscularly, intranasally, or orally to mice Vaccine 25–26 3885–3900

    Google Scholar 

  • AG Bijvoet et al. (1999) ArticleTitleHuman acid alpha-glucosidase from rabbit milk has therapeutic effect in mice with glycogen storage disease type II Hum Mol Genet 12 2145–2153

    Google Scholar 

  • JW Burns M Siadat-Pajouh AA Krishnaney HB Greenberg (1996) ArticleTitleProtective effect of rotavirus VP6-specific IgA monoclonal antibodies that lack neutralizing activity Science 5258 104–107

    Google Scholar 

  • InstitutionalAuthorNameCDC (1999) ArticleTitleCenters for Disease Control and Prevention: withdrawal of rotavirus vaccine recommendation MMWR Morb Mortal Wkly Rep 43 1007

    Google Scholar 

  • A Charpilienne J Lepault F Rey J Cohen (2002) ArticleTitleIdentification of rotavirus VP6 residues located at the interface with VP2 that are essential for capsid assembly and transcriptase activity J Virol 15 7822–7831

    Google Scholar 

  • A Charpilienne M Nejmeddine M Berois N Parez E Neumann E Hewat et al. (2001) ArticleTitleIndividual rotavirus-like particles containing 120 molecules of fluorescent protein are visible in living cells J Biol Chem 31 29361–29367

    Google Scholar 

  • AH Choi M Basu MM McNeal JA Bean JD Clements RL Ward (2004) ArticleTitleIntranasal administration of an Escherichia coli-expressed codon-optimized rotavirus VP6 protein induces protection in mice Protein Expr Purif 2 205–216

    Google Scholar 

  • AH Choi M Basu MM McNeal J Flint JL VanCott JD Clements et al. (2000) ArticleTitleFunctional mapping of protective domains and epitopes in the rotavirus VP6 protein J Virol 24 11574–11580

    Google Scholar 

  • M Ciarlet SE Crawford C Barone A Bertolotti-Ciarlet RF Ramig MK Estes et al. (1998) ArticleTitleSubunit rotavirus vaccine administered parenterally to rabbits induces active protective immunity J Virol 11 9233–9246

    Google Scholar 

  • A Coste JC Sirard K Johansen J Cohen JP Kraehenbuhl (2000) ArticleTitleNasal immunization of mice with virus-like particles protects offspring against rotavirus diarrhea J Virol 19 8966–8971

    Google Scholar 

  • KR Emslie MP Molloy CR Barardi D Jardine MR Wilkins AR Bellamy et al. (2000) ArticleTitleSerotype classification and characterisation of the rotavirus SA11 VP6 protein using mass spectrometry and two-dimensional gel electrophoresis Funct Integr Genom 1 12–24 Occurrence Handle1:CAS:528:DC%2BD3cXktVGhtL0%3D

    CAS  Google Scholar 

  • Estes MK (1996) Rotaviruses and their replication. Fields Virol:1625–1650.

  • J Fan T Watanabe (2003) ArticleTitleTransgenic rabbits as therapeutic protein bioreactors and human disease models Pharmacol Ther 3 261–282

    Google Scholar 

  • N Feng JW Burns L Bracy HB Greenberg (1994) ArticleTitleComparison of mucosal and systemic humoral immune responses and subsequent protection in mice orally inoculated with a homologous or a heterologous rotavirus J Virol 12 7766–7773

    Google Scholar 

  • C Fromantin B Jamot J Cohen L Piroth P Pothier E Kohli (2001) ArticleTitleRotavirus 2/6 virus-like particles administered intranasally in mice, with or without the mucosal adjuvants cholera toxin and Escherichia coli heat-labile toxin, induce a Th1/Th2-like immune response J Virol 22 11010–11016

    Google Scholar 

  • A Garcia-Diaz P Lopez-Andujar JR Diaz R Montava CT Barcelo JM Ribes et al. (2004) ArticleTitleNasal immunization of mice with a rotavirus DNA vaccine that induces protective intestinal IgA antibodies Vaccine 4 489–498

    Google Scholar 

  • Y Gavel G von Heijne (1990) ArticleTitleSequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering Protein Eng 5 433–442

    Google Scholar 

  • RI Glass JS Bresee UD Parashar B Jiang J Gentsch (2004) ArticleTitleThe future of rotavirus vaccines: a major setback leads to new opportunities Lancet 9420 1547–1550

    Google Scholar 

  • LM Houdebine (2000) ArticleTitleTransgenic animal bioreactors Transgenic Res 4–5 305–320

    Google Scholar 

  • LM Houdebine (2002) ArticleTitleAntibody manufacture in transgenic animals and comparisons with other systems Curr Opin Biotechnol 6 625–629

    Google Scholar 

  • LM Houdebine J Attal JL Vilotte (2002) NoChapterTitle Carl A. Pinkert (Eds) Vector Design for Transgene Expression Transgenic Animal Technology, 2nd edn Academic Press New york 419–458

    Google Scholar 

  • TG Kim WH Langridge (2003) ArticleTitleAssembly of cholera toxin B subunit full-length rotavirus NSP4 fusion protein oligomers in transgenic potato Plant Cell Rep 9 884–890

    Google Scholar 

  • TG Kim WH Langridge (2004) ArticleTitleSynthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potato Plant Cell Rep 6 382–387

    Google Scholar 

  • K Koles PH Berkel Particlevan ML Mannesse R Zoetemelk JF Vliegenthart JP Kamerling (2004) ArticleTitleInfluence of lactation parameters on the N-glycosylation of recombinant human C1 inhibitor isolated from the milk of transgenic rabbits Glycobiology 11 979–986

    Google Scholar 

  • M Labbe A Charpilienne SE Crawford MK Estes J Cohen (1991) ArticleTitleExpression of rotavirus VP2 produces empty corelike particles J Virol 6 2946–2952

    Google Scholar 

  • J Lepault I Petitpas I Erk J Navaza D Bigot M Dona et al. (2001) ArticleTitleStructural polymorphism of the major capsid protein of rotavirus EMBO J 7 1498–1507

    Google Scholar 

  • JK Ma PM Drake P Christou (2003) ArticleTitleThe production of recombinant pharmaceutical proteins in plants Nat Rev Genet 10 794–805

    Google Scholar 

  • M Mathieu I Petitpas J Navaza J Lepault E Kohli P Pothier et al. (2001) ArticleTitleAtomic structure of the major capsid protein of rotavirus: implications for the architecture of the virion EMBO J 7 1485–1497

    Google Scholar 

  • T Matsumura N Itchoda H Tsunemitsu (2002) ArticleTitleProduction of immunogenic VP6 protein of bovine group A rotavirus in transgenic potato plants Arch Virol 6 1263–1270

    Google Scholar 

  • CM O’Neal SE Crawford MK Estes ME Conner (1997) ArticleTitleRotavirus virus-like particles administered mucosally induce protective immunity J Virol 11 8707–8717

    Google Scholar 

  • F Recillas-Targa MJ Pikaart B Burgess-Beusse AC Bell MD Litt AG West et al. (2002) ArticleTitlePosition-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities Proc Natl Acad Sci USA 10 6883–6888

    Google Scholar 

  • S Rival-Gervier T Pantano C Viglietta C Maeder S Prince J Attal G Jolivet LM Houdebine (2003) ArticleTitleThe insulator effect of the 5′HS4 region from the beta-globin chicken locus on the rabbit WAP gene promoter activity in transgenic mice Transgenic Res 6 723–730

    Google Scholar 

  • S Rival-Gervier C Viglietta C Maeder J Attal LM Houdebine (2002) ArticleTitlePosition-independent and tissue-specific expression of porcine whey acidic protein gene from a bacterial artificial chromosome in transgenic mice Mol Reprod Dev 2 161–167

    Google Scholar 

  • L Roberts (2004) ArticleTitleVaccines Rotavirus vaccines’ second chance Science 5692 1890–1893

    Google Scholar 

  • Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual.

  • I Schwartz-Cornil Y Benureau H Greenberg BA Hendrickson J Cohen (2002) ArticleTitleHeterologous protection induced by the inner capsid proteins of rotavirus requires transcytosis of mucosal immunoglobulins J Virol 16 8110–8117

    Google Scholar 

  • PH Seeburg (1982) ArticleTitleThe human growth hormone gene family: nucleotide sequences show recent divergence and predict a new polypeptide hormone DNA 3 239–249

    Google Scholar 

  • Soler E, Fourgeux C, Parez N, Garbarg-Chenon A, Schwartz-Cornil I, Cohen J et al. (2004a) Preparation of recombinant rotavirus proteins in milk of transgenic non-human mammals. European Patent Reference (EPO) EP 04290589.3

  • Soler E et al. (2004b) Preparation of recombinant rotavirus 2/6 virus-like particles in milk of transgenic mice. International Meeting onVaccination Against Enteric Diseases”. Montego Bay, Jamaica

  • AW Stowers et al. (2002) ArticleTitleA recombinant vaccine expressed in the milk of transgenic mice protects Aotus monkeys from a lethal challenge with Plasmodium falciparum Proc Natl Acad Sci USA 1 339–344

    Google Scholar 

  • F Taboit-Dameron B Malassagne C Viglietta C Puissant M Leroux-Coyau et al. (1999) ArticleTitleAssociation of the 5′HS4 sequence of the chicken beta-globin locus control region with human EF1 alpha gene promoter induces ubiquitous and high expression of human CD55 and CD59 cDNAs in transgenic rabbits Transgenic Res 3 223–235

    Google Scholar 

  • G Tosser M Labbe M Bremont J Cohen (1992) ArticleTitleExpression of the major capsid protein VP6 of group C rotavirus and synthesis of chimeric single-shelled particles by using recombinant baculoviruses J Virol 10 5825–5831

    Google Scholar 

  • T Vesikari A Karvonen T Korhonen M Espo E Lebacq J Forster et al. (2004) ArticleTitleSafety and immunogenicity of RIX4414 live attenuated human rotavirus vaccine in adults, toddlers and previously uninfected infants Vaccine 21–22 2836–2842

    Google Scholar 

  • A Wigdorovitz et al. (2004) ArticleTitleProtective lactogenic immunity conferred by an edible peptide vaccine to bovine rotavirus produced in transgenic plants J Gen Virol Pt 7 1825–1832

    Google Scholar 

  • YZ Wu et al. (2003) ArticleTitleOral immunization with rotavirus VP7 expressed in transgenic potatoes induced high titers of mucosal neutralizing IgA Virology 2 337–342

    Google Scholar 

  • K Yang S Wang KO Chang S Lu LJ Saif HB Greenberg et al. (2001) ArticleTitleImmune responses and protection obtained with rotavirus VP6 DNA vaccines given by intramuscular injection Vaccine 23–24 3285–3291

    Google Scholar 

  • J Yu W Langridge (2003) ArticleTitleExpression of rotavirus capsid protein VP6 in transgenic potato and its oral immunogenicity in mice Transgenic Res 2 163–169

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Eric Soler.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Soler, E., Saux, A.L., Guinut, F. et al. Production of Two Vaccinating Recombinant Rotavirus Proteins in the Milk of Transgenic Rabbits. Transgenic Res 14, 833–844 (2005). https://doi.org/10.1007/s11248-005-1771-0

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-005-1771-0

Keywords

Navigation