A DNA vaccine coding for the chimera BLSOmp31 induced a better degree of protection against B. ovis and a similar degree of protection against B. melitensis than Rev.1 vaccination

In the present study, we reported an attempt to improve the immunogenicity and protective capacity of the chimera BLSOmp31 using a different antigen delivery: DNA vaccination. Vaccination of BALB/c mice with the DNA vaccine coding for the chimera BLSOmp31 (pCIBLSOmp31) provided the best protection level against Brucella ovis, which was significantly higher than the given by the co-delivery of both plasmids coding for the whole proteins (pcDNABLS+pCIOmp31) and even higher than the control vaccine Rev.1. Moreover, pCIBLSOmp31 induced higher protection against Brucella melitensis than pcDNABLS+pCIOmp31 but similar protection than Rev.1. The chimera induced a strong humoral response against the inserted peptide. It also induced peptide- and BLS-specific cytotoxic T responses. The insertion of this peptide on BLS induced stronger T helper 1 responses specific for the carrier (BLS), thus our results represent a case of synergic strengthening between two Brucella antigens. Hitherto, this is the first indication that a recombinant subunit vaccine elicits greater protection than whole Brucella.     

Efficacy of bp26 and bp26/omp31 B. melitensis Rev.1 deletion mutants against Brucella ovis in rams

Brucella melitensis Rev.1 is the most effective vaccine against B. ovis infection in sheep but induces antibodies interfering with B. melitensis diagnosis. Brucella BP26 and Omp31 proteins are differential diagnostic antigens. Single or double bp26 and omp31 Rev.1 deletion mutants have been proven effective against B. melitensis in sheep. Here, the CGV26 (deleted in bp26 gene) and CGV2631 (deleted in both bp26 and omp31 genes) mutants have been tested for efficacy against B. ovis in rams. Either inoculated subcutaneously or conjunctivally, both mutants conferred significant protection against B. ovis. The protection induced by CGV26 was similar to that of Rev.1 but significantly higher than that conferred by CGV2631. In conclusion, the CGV26 mutant, in association with the adequate diagnostic strategy, could be a useful alternative to Rev.1 for sheep vaccination against B. ovis infections in those countries performing simultaneously B. melitensis and B. ovis eradication campaigns.     

Simultaneous subcutaneous and conjunctival administration of the influenza viral vector based Brucella abortus vaccine to pregnant heifers provides better protection against B. abortus 544 infection than the commercial B. abortus S19 vaccine

In this study, we explored possibility of increasing the protective efficacy of our novel influenza viral vector based B. abortus vaccine (Flu-BA) in pregnant heifers by adapting an innovative method of vaccine delivery. We administered the vaccine concurrently via the conjunctival and subcutaneous routes to pregnant heifers, and these routes were previously tested individually. The Flu-BA vaccination of pregnant heifers (n = 9) against a challenge B. abortus 544 infection provided protection from abortion, infection of heifers and fetuses/calves by 88.8%, 100% and 100%, respectively (alpha = 0.004–0.0007 vs. negative control; n = 7). Our candidate vaccine using this delivery method provided slightly better protection than the commercial B. abortus S19 vaccine in pregnant heifers (n = 8), which provided protection from abortion, infection of heifers and fetuses/calves by 87.5%, 75% and 87.5%, respectively. This improved method of the Flu-BA vaccine administration is highly recommended for the recovery of farms which has high prevalence of brucellosis.     

Charged amino acids in the AMPV fusion protein have more influence on induced protection than deletion of the SH or G genes

Modifications to F, G and SH genes of an avian metapneumovirus (AMPV) field isolate were made by reverse genetics and their virulence and protective capacity were tested in young turkeys. Infection of one-day-old turkeys with a subtype A AMPV neither caused disease nor stimulated detectable protection against subsequent virulent challenge. While serial passage of this virus in tracheal tissue increased virulence, protection stimulated remained moderate. Substitution of the fusion protein from a protective AMPV very minimally increasing virulence but dramatically increased induced protection; and this was associated with five amino acid substitutions all involving charged amino acids which computational analysis predicted to affect protein surface properties but not immunodominant helper T-lymphocyte antigenic sites. When SH or G genes were deleted, viruses caused no disease but still conferred full protection to the majority of turkeys. In the case of the SH deletion, shed virus post-inoculation was undetectable. Partial SH deletions were found to confer protection related to the length of SH open reading frame remaining. Removal of both SH and G genes together produced a virus conferring negligible protection. We conclude that the characteristics of the AMPV fusion protein are important in inducing protection while the SH and G genes under investigation played a lesser role.     

Oral vaccination with liposome-encapsulated recombinant fusion peptide of urease B epitope and cholera toxin B subunit affords prophylactic and therapeutic effects against H. pylori infection in BALB/c mice

A new fusion peptide CtUBE of cholera toxin B subunit and Helicobacter pylori urease B subunit epitope was expressed in Escherichia coli. With this fusion peptide, an oral liposome vaccine against H. pylori infection was prepared and evaluated in BALB/c mice. Based on the results of urease tests, quantitation of culturable bacteria colonies in mice stomachs and histological identification of gastritis, the mice were protected significantly after intragastric vaccination with this CtUBE liposome vaccine, which increased the content levels of specific anti-urease serum IgG and mucosal IgA for both prophylactic and therapeutic vaccination protocols. These results showed that the fusion peptide CtUBE retained immunogenicity and could be used as antigen in the development of an oral vaccine against H. pylori infection.     

Further analysis of protection induced by the MIC3 DNA vaccine against T. gondii: CD4 and CD8 T cells are the major effectors of the MIC3 DNA vaccine-induced protection,both Lectin-like and EGF-like domains of MIC3 conferred protection

The present study was conducted mainly to evaluate the contribution of the cellular and the humoral responses in protection conferred by the MIC3 DNA vaccine (pMIC3i) that was proved as a potent vaccine against toxoplasmosis. We performed the adoptive transfer of CD4⁺ and CD8⁺ T lymphocytes from pMIC3i immunized mice to naive ones and the role of humoral immunity was evaluated by in vitro invasion assays. We also constructed plasmids encoding the EGF-like domains and the Lectin-like domain of MIC3, to define which domains of MIC3 are involved in the protection. Furthermore, the adjuvant effect of the GM-CSF-expressing vector (granulocyte-macrophage colony-stimulating factor) required the precise temporal and spatial codelivery of GM-CSF with antigen, thus, we constructed a bicistronic plasmid expressing MIC3 and GM-CSF. In conclusion, the protection induced by pMIC3i was mainly mediated by CD4⁺ and CD8⁺ T lymphocytes and both EGF and Lectin domains of MIC3 conferred protection. Furthermore, the codelivery of GM-CSF by a bicistronic plasmid appeared to be a most effective way for enhancing the adjuvant properties of GM-CSF.     

Sequence variation in pertussis S1 subunit toxin and pertussis genes in Bordetella pertussis strains used for the whole-cell pertussis vaccine produced in Poland since 1960: efficiency of the DTwP vaccine-induced immunity against currently circulating B. pertussis isolates

The present study indicates that the appearance of the B. pertussis harbouring prn2 gene allele variant (not found among clinical isolates before 1990s) may have been induced by long-term vaccination in Poland with DTP-composed vaccine strains presenting exclusively prn1. However, ptxS1A allele of pertussis toxin subunit S1 encoding gene, predominant in the currently isolated B. pertussis strains, has been found in vaccine strains used for whole-cell pertussis component (wP) production of DTP vaccine in 1960-1978. This outrules the possibility that the appearance of ptxSIA allele might be related to vaccine pressure driven by non-ptxS1A vaccine strains used for long-term immunization with wP. Intranasal challenge animal model testing the efficiency of the clearance of B. pertussis strains harbouring different ptxS1/prn allele gene combinations revealed that currently produced DTwP vaccine may not contain adequate B. pertussis vaccine strains, since isolates with gene variants different from those observed in vaccine strains were eliminated from the lungs of the immunized animals with lower efficiency.