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.
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
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
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
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
InstitutionalAuthorNameCDC (1999) ArticleTitleCenters for Disease Control and Prevention: withdrawal of rotavirus vaccine recommendation MMWR Morb Mortal Wkly Rep 43 1007
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
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
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
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
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
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
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
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
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
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
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
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
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
LM Houdebine (2000) ArticleTitleTransgenic animal bioreactors Transgenic Res 4–5 305–320
LM Houdebine (2002) ArticleTitleAntibody manufacture in transgenic animals and comparisons with other systems Curr Opin Biotechnol 6 625–629
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
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
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
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
M Labbe A Charpilienne SE Crawford MK Estes J Cohen (1991) ArticleTitleExpression of rotavirus VP2 produces empty corelike particles J Virol 6 2946–2952
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
JK Ma PM Drake P Christou (2003) ArticleTitleThe production of recombinant pharmaceutical proteins in plants Nat Rev Genet 10 794–805
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
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
CM O’Neal SE Crawford MK Estes ME Conner (1997) ArticleTitleRotavirus virus-like particles administered mucosally induce protective immunity J Virol 11 8707–8717
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
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
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
L Roberts (2004) ArticleTitleVaccines Rotavirus vaccines’ second chance Science 5692 1890–1893
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
PH Seeburg (1982) ArticleTitleThe human growth hormone gene family: nucleotide sequences show recent divergence and predict a new polypeptide hormone DNA 3 239–249
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 on “Vaccination 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
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
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
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
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
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
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
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
Author information
Authors and Affiliations
Corresponding author
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11248-005-1771-0