Transgenic Research

, Volume 14, Issue 6, pp 833–844 | Cite as

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

  • Eric Soler
  • Agnès Le Saux
  • Frédéric Guinut
  • Bruno Passet
  • Ruxandra Cohen
  • Christine Merle
  • Annie Charpilienne
  • Cynthia Fourgeux
  • Véronique Sorel
  • Antoine Piriou
  • Isabelle Schwartz-Cornil
  • Jean Cohen
  • Louis-Marie Houdebine
Article

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.

Keywords

bioreactor milk rotavirus VP2 VP6 

Abbreviations

RV

rotavirus

Tg

transgenic

; VLP

virus-like particles

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References

  1. Attal, J, Theron, MC, Kann, G, Bolifraud, P, Puissant, C, Houdebine, LM 2000The stimulation of gene expression by the R region from HTLV-1 and BLVJ Biotechnol2–3179189Google Scholar
  2. Bertolotti-Ciarlet, A, Ciarlet, M, Crawford, SE, Conner, ME, Estes, MK 2003Immunogenicity and protective efficacy of rotavirus 2/6-virus-like particles produced by a dual baculovirus expression vector and administered intramuscularly, intranasally, or orally to miceVaccine25–2638853900Google Scholar
  3. Bijvoet, AG,  et al. 1999Human acid alpha-glucosidase from rabbit milk has therapeutic effect in mice with glycogen storage disease type IIHum Mol Genet1221452153Google Scholar
  4. Burns, JW, Siadat-Pajouh, M, Krishnaney, AA, Greenberg, HB 1996Protective effect of rotavirus VP6-specific IgA monoclonal antibodies that lack neutralizing activityScience5258104107Google Scholar
  5. CDC1999Centers for Disease Control and Prevention: withdrawal of rotavirus vaccine recommendationMMWR Morb Mortal Wkly Rep431007Google Scholar
  6. Charpilienne, A, Lepault, J, Rey, F, Cohen, J 2002Identification of rotavirus VP6 residues located at the interface with VP2 that are essential for capsid assembly and transcriptase activityJ Virol1578227831Google Scholar
  7. Charpilienne, A, Nejmeddine, M, Berois, M, Parez, N, Neumann, E, Hewat, E,  et al. 2001Individual rotavirus-like particles containing 120 molecules of fluorescent protein are visible in living cellsJ Biol Chem312936129367Google Scholar
  8. Choi, AH, Basu, M, McNeal, MM, Bean, JA, Clements, JD, Ward, RL 2004Intranasal administration of an Escherichia coli-expressed codon-optimized rotavirus VP6 protein induces protection in miceProtein Expr Purif2205216Google Scholar
  9. Choi, AH, Basu, M, McNeal, MM, Flint, J, VanCott, JL, Clements, JD,  et al. 2000Functional mapping of protective domains and epitopes in the rotavirus VP6 proteinJ Virol241157411580Google Scholar
  10. Ciarlet, M, Crawford, SE, Barone, C, Bertolotti-Ciarlet, A, Ramig, RF, Estes, MK,  et al. 1998Subunit rotavirus vaccine administered parenterally to rabbits induces active protective immunityJ Virol1192339246Google Scholar
  11. Coste, A, Sirard, JC, Johansen, K, Cohen, J, Kraehenbuhl, JP 2000Nasal immunization of mice with virus-like particles protects offspring against rotavirus diarrheaJ Virol1989668971Google Scholar
  12. Emslie, KR, Molloy, MP, Barardi, CR, Jardine, D, Wilkins, MR, Bellamy, AR,  et al. 2000Serotype classification and characterisation of the rotavirus SA11 VP6 protein using mass spectrometry and two-dimensional gel electrophoresisFunct Integr Genom11224Google Scholar
  13. Estes MK (1996) Rotaviruses and their replication. Fields Virol:1625–1650.Google Scholar
  14. Fan, J, Watanabe, T 2003Transgenic rabbits as therapeutic protein bioreactors and human disease modelsPharmacol Ther3261282Google Scholar
  15. Feng, N, Burns, JW, Bracy, L, Greenberg, HB 1994Comparison of mucosal and systemic humoral immune responses and subsequent protection in mice orally inoculated with a homologous or a heterologous rotavirusJ Virol1277667773Google Scholar
  16. Fromantin, C, Jamot, B, Cohen, J, Piroth, L, Pothier, P, Kohli, E 2001Rotavirus 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 responseJ Virol221101011016Google Scholar
  17. Garcia-Diaz, A, Lopez-Andujar, P, Diaz, JR, Montava, R, Barcelo, CT, Ribes, JM,  et al. 2004Nasal immunization of mice with a rotavirus DNA vaccine that induces protective intestinal IgA antibodiesVaccine4489498Google Scholar
  18. Gavel, Y, von Heijne, G 1990Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineeringProtein Eng5433442Google Scholar
  19. Glass, RI, Bresee, JS, Parashar, UD, Jiang, B, Gentsch, J 2004The future of rotavirus vaccines: a major setback leads to new opportunitiesLancet942015471550Google Scholar
  20. Houdebine, LM 2000Transgenic animal bioreactorsTransgenic Res4–5305320Google Scholar
  21. Houdebine, LM 2002Antibody manufacture in transgenic animals and comparisons with other systemsCurr Opin Biotechnol6625629Google Scholar
  22. Houdebine, LM, Attal, J, Vilotte, JL 2002Pinkert, Carl A. eds. Vector Design for Transgene Expression Transgenic Animal Technology, 2nd ednAcademic PressNew york419458Google Scholar
  23. Kim, TG, Langridge, WH 2003Assembly of cholera toxin B subunit full-length rotavirus NSP4 fusion protein oligomers in transgenic potatoPlant Cell Rep9884890Google Scholar
  24. Kim, TG, Langridge, WH 2004Synthesis of an HIV-1 Tat transduction domain-rotavirus enterotoxin fusion protein in transgenic potatoPlant Cell Rep6382387Google Scholar
  25. Koles, K, Berkel, PH, Mannesse, ML, Zoetemelk, R, Vliegenthart, JF, Kamerling, JP 2004Influence of lactation parameters on the N-glycosylation of recombinant human C1 inhibitor isolated from the milk of transgenic rabbitsGlycobiology11979986Google Scholar
  26. Labbe, M, Charpilienne, A, Crawford, SE, Estes, MK, Cohen, J 1991Expression of rotavirus VP2 produces empty corelike particlesJ Virol629462952Google Scholar
  27. Lepault, J, Petitpas, I, Erk, I, Navaza, J, Bigot, D, Dona, M,  et al. 2001Structural polymorphism of the major capsid protein of rotavirusEMBO J714981507Google Scholar
  28. Ma, JK, Drake, PM, Christou, P 2003The production of recombinant pharmaceutical proteins in plantsNat Rev Genet10794805Google Scholar
  29. Mathieu, M, Petitpas, I, Navaza, J, Lepault, J, Kohli, E, Pothier, P,  et al. 2001Atomic structure of the major capsid protein of rotavirus: implications for the architecture of the virionEMBO J714851497Google Scholar
  30. Matsumura, T, Itchoda, N, Tsunemitsu, H 2002Production of immunogenic VP6 protein of bovine group A rotavirus in transgenic potato plantsArch Virol612631270Google Scholar
  31. O’Neal, CM, Crawford, SE, Estes, MK, Conner, ME 1997Rotavirus virus-like particles administered mucosally induce protective immunityJ Virol1187078717Google Scholar
  32. Recillas-Targa, F, Pikaart, MJ, Burgess-Beusse, B, Bell, AC, Litt, MD, West, AG,  et al. 2002Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activitiesProc Natl Acad Sci USA1068836888Google Scholar
  33. Rival-Gervier, S, Pantano, T, Viglietta, C, Maeder, C, Prince, S, Attal, J, Jolivet, G, Houdebine, LM 2003The insulator effect of the 5′HS4 region from the beta-globin chicken locus on the rabbit WAP gene promoter activity in transgenic miceTransgenic Res6723730Google Scholar
  34. Rival-Gervier, S, Viglietta, C, Maeder, C, Attal, J, Houdebine, LM 2002Position-independent and tissue-specific expression of porcine whey acidic protein gene from a bacterial artificial chromosome in transgenic miceMol Reprod Dev2161167Google Scholar
  35. Roberts, L 2004Vaccines Rotavirus vaccines’ second chanceScience569218901893Google Scholar
  36. Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual.Google Scholar
  37. Schwartz-Cornil, I, Benureau, Y, Greenberg, H, Hendrickson, BA, Cohen, J 2002Heterologous protection induced by the inner capsid proteins of rotavirus requires transcytosis of mucosal immunoglobulinsJ Virol1681108117Google Scholar
  38. Seeburg, PH 1982The human growth hormone gene family: nucleotide sequences show recent divergence and predict a new polypeptide hormoneDNA3239249Google Scholar
  39. 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 04290589.3Google Scholar
  40. 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, JamaicaGoogle Scholar
  41. Stowers, AW,  et al. 2002A recombinant vaccine expressed in the milk of transgenic mice protects Aotus monkeys from a lethal challenge with Plasmodium falciparumProc Natl Acad Sci USA1339344Google Scholar
  42. Taboit-Dameron, F, Malassagne, B, Viglietta, C, Puissant, C, Leroux-Coyau, M,  et al. 1999Association 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 rabbitsTransgenic Res3223235Google Scholar
  43. Tosser, G, Labbe, M, Bremont, M, Cohen, J 1992Expression of the major capsid protein VP6 of group C rotavirus and synthesis of chimeric single-shelled particles by using recombinant baculovirusesJ Virol1058255831Google Scholar
  44. Vesikari, T, Karvonen, A, Korhonen, T, Espo, M, Lebacq, E, Forster, J,  et al. 2004Safety and immunogenicity of RIX4414 live attenuated human rotavirus vaccine in adults, toddlers and previously uninfected infantsVaccine21–2228362842Google Scholar
  45. Wigdorovitz, A,  et al. 2004Protective lactogenic immunity conferred by an edible peptide vaccine to bovine rotavirus produced in transgenic plantsJ Gen VirolPt 718251832Google Scholar
  46. Wu, YZ,  et al. 2003Oral immunization with rotavirus VP7 expressed in transgenic potatoes induced high titers of mucosal neutralizing IgAVirology2337342Google Scholar
  47. Yang, K, Wang, S, Chang, KO, Lu, S, Saif, LJ, Greenberg, HB,  et al. 2001Immune responses and protection obtained with rotavirus VP6 DNA vaccines given by intramuscular injectionVaccine23–2432853291Google Scholar
  48. Yu, J, Langridge, W 2003Expression of rotavirus capsid protein VP6 in transgenic potato and its oral immunogenicity in miceTransgenic Res2163169Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Eric Soler
    • 1
    • 2
  • Agnès Le Saux
    • 2
  • Frédéric Guinut
    • 2
  • Bruno Passet
    • 2
  • Ruxandra Cohen
    • 2
  • Christine Merle
    • 2
  • Annie Charpilienne
    • 3
  • Cynthia Fourgeux
    • 4
  • Véronique Sorel
    • 2
  • Antoine Piriou
    • 2
  • Isabelle Schwartz-Cornil
    • 4
  • Jean Cohen
    • 3
  • Louis-Marie Houdebine
    • 1
  1. 1.Biologie du Développement et de la ReproductionINRAJouy-en-JosasFrance
  2. 2.Bioprotein TechnologiesParisFrance
  3. 3.Virologie Moléculaire & StructuraleCNRS-INRAGif-sur-YvetteFrance
  4. 4.Virologie et Immunologie MoléculairesINRAJouy-en-JosasFrance

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