Immunization of mice with recombinant L7L12 ribosomal protein confers protection against Brucella abortus infection

Oliveira, Sérgio C.; Splitter, Gary A.

doi:10.1016/0264-410x(96)00018-7

Cited by 156 publications

(107 citation statements)

References 20 publications

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“…Until now no satisfactory vaccines against this disease are available and new vaccinal strategies (such as oral live vaccines) should be explored. Recent work demonstrated a protective immunization against Brucella abortus in mice using a ribosomal protein (24). We think that a lactococcal strain secreting this ribosomal protein (or a part of it) would be a promising vector to induce a protective immune response against brucellosis.…”

Section: Discussionmentioning

confidence: 99%

Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

Langella

Loir

1999

Braz J Med Biol Res

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Lactic acid bacteria (LAB) are Gram-positive bacteria and are generally regarded as safe (GRAS) organisms. Therefore, LAB could be used for heterologous protein secretion and they are good potential candidates as antigen delivery vehicles. To develop such live vaccines, a better control of protein secretion is required. We developed an efficient secretion system in the model LAB, Lactococcus lactis. Staphylococcal nuclease (Nuc) was used as the reporter protein. We first observed that the quantity of secreted Nuc correlated with the copy number of the cloning vector. The nuc gene was cloned on a high-copy number cloning vector and no perturbation of the metabolism of the secreting strain was observed. Replacement of nuc native promoter by a strong lactococcal one led to a significant increase of nuc expression. Secretion efficiency (SE) of Nuc in L. lactis was low, i.e., only 60% of the synthesized Nuc was secreted. Insertion of a synthetic propeptide between the signal peptide and the mature moiety of Nuc increased the SE of Nuc. On the basis of these results, we developed a secretion system and we applied it to the construction of an L. lactis strain which secretes a bovine coronavirus (BCV) epitopeprotein fusion (BCV-Nuc). BCV-Nuc was recognized by both anti-BCV and anti-Nuc antibodies. Secretion of this antigenic fusion is the first step towards the development of a novel antigen delivery system based on LAB-secreting strains.

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Section: Discussionmentioning

confidence: 99%

Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

Langella

Loir

1999

Braz J Med Biol Res

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“…As an alternative to the commercially available B. abortus strain 19 vaccine, recombinant Brucella proteins have been studied with limited success [22]. A single molecule may be insuf®cient to confer full protection and also a better antigen delivery system has to be investigated.…”

Section: Discussionmentioning

confidence: 99%

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Leclerq¹,

Harms²,

Rosinha³

et al. 2002

Journal of Medical Microbiology

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The immunogenicity and protective ef®cacy of a DNA vaccine encoding the GroEL heatshock 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 ampli®ed 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 nonsecreted 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-speci®c antiGroEL and high levels of IFN-ã produced by splenic T cells. Gene gun immunisation induced a Th0 type of immune response in mice characterised by a high IgG1/IgG2a ratio, and IL-4 and interferon (IFN)-ã production. Even though a distinct pattern of immune response was generated depending upon the immunisation route used, neither method engendered a signi®cant level of protection with the GroEL DNA vaccine.

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“…Protection was demonstrated against B. abortus by immunising mice with a peptide derived from the sequence of the B. abortus Cu-Zn superoxide-dismutase; however, the whole recombinant protein was not efficacious [52]. More recently a purified recombinant ribosomal protein was shown to be protective in mice, inducing both immune cells and antibodies [53]. This would indicate that immune Oprotective) mechanisms against brucella might involve both internal and outer-membrane antigens.…”

Section: Discussionmentioning

confidence: 99%

Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of Brucella Melitensis (Omp25) Expressed in Escherichia Coli

Bowden

Cloeckaert²,

Zygmunt³

et al. 1998

Journal of Medical Microbiology

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The antibody response specific to the 25-kDa major outer-membrane protein (Omp25) of Brucella melitensis expressed in Escherichia coli was assessed in BALB/c mice. Groups of mice were immunised and boosted either with sonicated E. coli carrying plasmid pAC2533 -E. coli (pAC2533) -expressing the gene coding for Omp25 (omp25 gene) of B. melitensis, or with E. coli carrying plasmid pUCl9 -E. coli (pUC19). One control group received saline. The evolution of antibody responses was investigated by indirect ELISA with whole rough (R) B. melitensis H38 cells as antigen. Serum antibody titres of mice immunised with E. coli (pAC2533) were appreciably higher than those of mice immunised with E. coli (pUC19). The specificity to Omp25 of murine antibodies induced by E. coli (pAC2533) was demonstrated by SDS-PAGE and immunoblotting of five B. melitensis strains. Binding of antibody in E. coli (pAC2533) immune sera to the surface of B. melitensis strains differing in their smooth lipopolysaccharide (S-LPS) expression was also studied by whole-cell ELISA and by flow cytometry. Antibody reactivity to R and smooth-rough (S-R) was much stronger than that to smooth (S) B. melitensis strains, indicating a much better accessibility of Omp25 to antibody on strains lacking or expressing less O-polysaccharide on their surface. The antibodies to Omp25 were predominantly of IgG2a isotype. The capacity of E. coli (pAC2533) to induce protective immune responses against four challenge strains of B. melitensis was further evaluated in mice. Significant reductions in splenic infections, in comparison with mice immunised with E. coli (pUC19) and unimmunised (saline injection) mice, were observed in R B. melitensis B115, S-R B. melitensis EP and S B. melitensis H38 infected mice. Protection against S B. melitensis 16M was not significant. The data from the present study, together with previous results, suggest that humoral immunity against probably conformational, well-exposed epitopes of the Omp25 could contribute to protective mechanisms against B. melitensis infection in mice.

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Immunization of mice with recombinant L7L12 ribosomal protein confers protection against Brucella abortus infection

Cited by 156 publications

References 20 publications

Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

Induction of a Th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene

Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of Brucella Melitensis (Omp25) Expressed in Escherichia Coli

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