RecombinantBrucella abortusProteins That Induce Proliferation and Gamma-Interferon Secretion by CD4+T Cells fromBrucella-Vaccinated Mice and Delayed-Type Hypersensitivity in Sensitized Guinea Pigs

Oliveira, Sérgio C.; Harms, Jerome S.; Banai, M; Splitter, Gary A.

doi:10.1006/cimm.1996.0241

Cited by 54 publications

(45 citation statements)

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“…The results clearly indicated that rGAPDH induced a Th1-type of immune response in mice, although the levels of IFN-ª and TNF-AE in the rGAPDH-immunised group were lower than in the group vaccinated with strain S19. A similar observation was reported by our group and others when a Brucella recombinant protein was used as antigen, and the less potent T-cell activation stimulus is probably due to the antigenic complexity of B. abortus S19 compared with a single molecule [43]. Immunisation of mice with strain S19 induces protective immunity and splenocytes from such mice produce high levels of IFN-ª.…”

Section: Discussionsupporting

confidence: 83%

Molecular and immunological characterisation of recombinant Brucella abortus glyceraldehyde-3-phosphate-dehydrogenase, a T- and B-cell reactive protein that induces partial protection when co-administered with an interleukin-12-expressing plasmid in a DNA vaccine formulation

Rosinha¹,

Myioshi

Azevedo

et al. 2002

Journal of Medical Microbiology

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To identify antigens of Brucella spp. that are potentially involved in stimulating a protective T-cell-mediated immune response, previous studies identified 10 clones from a Brucella abortus 2308 genomic library with primed lymphocytes as probes. One selected positive clone (182) contained an insert of 1.2 kb which was identified, sequenced and characterised. The deduced amino acid sequence of the open reading frame (ORF) revealed 82% and 81% identity to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) enzymes from Agrobacterium tumefaciens and Xanthobacter flavus, respectively. Southern blot analysis demonstrated that the gap gene is present in only one copy in the Brucella genome. B. abortus GAPDH was then expressed in Escherichia coli as a fusion protein with the maltose-binding protein (MBP). To demonstrate the functional activity of Brucella GAPDH, E. coli gap mutants were transformed with a Brucella pMAL-gap construct. Genetic complementation was achieved and as a result E. coli mutants were able to grow on glucose or other carbon source medium. The humoral and cellular immune responses to the recombinant (r) GAPDH were characterised. In Western blots, sera from naturally infected cattle and sheep showed antibody reactivity against rGAPDH. In response to in-vitro stimulation by rGAPDH, splenocytes from mice vaccinated with rGAPDH or B. abortus S19 were able to produce ª-interferon and tumour necrosis factor-AE but not interleukin (IL)-4. Furthermore, gap associated with murine IL-12 gene in a DNA vaccine formulation partially protected mice against experimental infection.

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Section: Discussionsupporting

confidence: 83%

Molecular and immunological characterisation of recombinant Brucella abortus glyceraldehyde-3-phosphate-dehydrogenase, a T- and B-cell reactive protein that induces partial protection when co-administered with an interleukin-12-expressing plasmid in a DNA vaccine formulation

Rosinha¹,

Myioshi

Azevedo

et al. 2002

Journal of Medical Microbiology

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“…In an effort to create an effective and safe vaccine against B. abortus, several research groups have developed subunit (recombinant proteins) [4][5][6][7][8][9][10][11][12], a DNA [13][14][15][16][17][18], or live vector vaccines (based on bacteria and viruses) [19][20][21][22]. All of these vaccines were safe when tested in animal models (laboratory mice), and some when tested in cattle.…”

Section: Introductionmentioning

confidence: 99%

Safety of the novel vector vaccine against Brucella abortus based on recombinant influenza viruses expressing Brucella L7/L12 and OMP16 proteins, in cattle

Tabynov¹,

Kydyrbayev²,

Ryskeldinova³

et al. 2014

J Vaccines Immunol

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Citation: Tabynov K, Kydyrbayev Z, Ryskeldinova S, Yespembetov B, Syrymkyzy N, et al. (2015) eertechz AbstractThis paper presents the results of a study of the safety of new vector vaccine against B. abortus based on recombinant influenza A subtype H5N1 or H1N1 (viral constructs vaccine formulation) viruses expressing Brucella ribosomal protein L7/L12 and Omp16, in cattle. To increase the effectiveness of the vaccine, adjuvants such as Montanide Gel01 or chitosan were included in its composition. Immunization of cattle (5 animals per group) with the viral constructs vaccine formulation only, or its combination with adjuvants Montanide Gel01 or chitosan, were conducted via the conjunctival method using cross prime (influenza virus subtype H5N1) and booster (influenza virus subtype H1N1) vaccination schedules. Vaccine candidates were evaluated in comparison with the positive (B. abortus S19) and negative (PBS) controls. Comprehensive studies involving thermometry and clinical examination, hematology and biochemical blood analysis, showed that all of the viral constructs vaccine formulation, as well as their combination with adjuvants, compared to the commercial bacterial vaccine B. abortus S19 were completely safe in cattle. Furthermore it is shown that the developed vaccines can effectively differentiate vaccinated animals from infected animals. Brucella L7/L12 or Omp16 proteins. The influenza A virus contains a segmented genome consisting of eight negative-strand RNA fragments. Of these, the smallest fragment (NS) -encoding two proteins: viral nonstructural protein (NS1) and nuclear export protein (Nep) -is a convenient target for genetic manipulation as NS1 is able to tolerate foreign sequences exceeding its own length [25]. Thus, the ORF of NS1 was used for inserting Brucella sequences in this study. The А/Puerto Rico/8/34 (H1N1) strain was used as the backbone for obtaining influenza A virus vectors expressing Brucella L7/L12 or Omp16 sequences in the form of fusion proteins with the N-terminal 124 amino acid residues of NS1.Our previous studies have shown that a bivalent vaccine formulation comprising a mixture of recombinant influenza A virus subtype H5N1 or H1N1 expressing the ribosomal L7/L12 or Omp16 proteins in prime and booster immunization mode (via conjunctival injection) in cattle induced a strong antigen-specific T-cell immune response, and most importantly provided a high level of protectiveness comparable to the commercial B. abortus S19 vaccine and superior to the B. abortus S19 vaccine in combination with Montanide Gel01 adjuvant [24]. Based on this, the next stage of our study was to evaluate the safety of the proposed new live vector vaccine in cattle. Additionally, we evaluated the possibility of differentiating infected animals from vaccinated animals using the developed vaccine.

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“…In attempts to identify immunodominant antigens from B. abortus for use in a subunit vaccine, previous studies reported members of the heat-shock protein family as potential immunogens [13]. GroEL is a wellknown member of the heat-shock protein family that is present in various microbial pathogens and is recognised as an immunodominant target for both humoral and cellular immune responses from different hosts [14,15].…”

Section: Introductionmentioning

confidence: 99%

“…GroEL is a wellknown member of the heat-shock protein family that is present in various microbial pathogens and is recognised as an immunodominant target for both humoral and cellular immune responses from different hosts [14,15]. Interestingly, cattle and mice infected with B. abortus mount a strong antibody and T-cell response to GroEL [13,16]. Regarding protection, the immune response induced by recombinant (r) GroEL protein has been shown to be protective in different infectious disease models [17,18].…”

Section: Introductionmentioning

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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RecombinantBrucella abortusProteins That Induce Proliferation and Gamma-Interferon Secretion by CD4+T Cells fromBrucella-Vaccinated Mice and Delayed-Type Hypersensitivity in Sensitized Guinea Pigs

Cited by 54 publications

References 0 publications

Molecular and immunological characterisation of recombinant Brucella abortus glyceraldehyde-3-phosphate-dehydrogenase, a T- and B-cell reactive protein that induces partial protection when co-administered with an interleukin-12-expressing plasmid in a DNA vaccine formulation

Molecular and immunological characterisation of recombinant Brucella abortus glyceraldehyde-3-phosphate-dehydrogenase, a T- and B-cell reactive protein that induces partial protection when co-administered with an interleukin-12-expressing plasmid in a DNA vaccine formulation

Safety of the novel vector vaccine against Brucella abortus based on recombinant influenza viruses expressing Brucella L7/L12 and OMP16 proteins, in cattle

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

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