Induction of a th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene.
The immunogenicity and protective efficacy of a DNA vaccine encoding the GroEL heat-shock gene from Brucella abortus was tested in BALB/c mice immunised by intramuscular (i.m.) needle injection or epidermally by gene gun. The Brucella GroEL gene was amplified by PCR and cloned into two different mammalian expression vectors pCMV-link and pCMV-tPA. The D17 cell line was transfected with both constructs and GroEL transcripts were detected by Northern blot. To determine the level of protein synthesised, transfected cell lysates were then submitted to Western blot. The non-secreted form of the recombinant GroEL produced by the pCMV-link construct was detected in much greater amount than the secreted form of the protein produced by the pCMV-tPA construct. After immunisation, a strong anti-GroEL IgG response was detected in mice vaccinated by i.m. injection or gene gun only when the pCMV-link/ GroEL plasmid was used. Regarding the pattern of immune response induced, i.m. needle injection raised a predominantly Th1 response with mostly IgG2a-specific anti-GroEL and high levels of IFN-gamma produced by splenic T cells. Gene gun immunisation induced a ThO type of immune response in mice characterised by a high IgG1/IgG2a ratio, and IL-4 and interferon (IFN)-gamma production. Even though a distinct pattern of immune response was generated depending upon the immunisation route used, neither method engendered a significant level of protection with the GroEL DNA vaccine.
Leclerq
Sophie
S
Harms
Jerome S
JS
Department of Biochemistry and Immunology, Federal University of Minas Gerais, ICB-UFMG, Belo Horizonte-MG 30161-970, Brazil and *Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA.
Rosinha
Gracia M S
GMS
Azevedo
Vasco
V
Oliveira
Sergio C
SC
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002004:Brucella Vaccine / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D002003:Brucella abortus / Q000235:genetics / Q000276:immunology*; D002006:Brucellosis / Q000517:prevention & control; D002460:Cell Line; D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology*; D004269:DNA, Bacterial / Q000276:immunology; D004195:Disease Models, Animal; D005784:Gene Amplification; D005822:Genetic Vectors; D006360:Heat-Shock Proteins; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis; D007273:Injections, Intramuscular; D007371:Interferon-gamma / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology*; D014162:Transfection; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*
D002004:Brucella Vaccine; D018834:Chaperonin 60; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D019444:Vaccines, DNA; D015847:Interleukin-4; D007371:Interferon-gamma