First evaluation of an influenza viral vector based Brucella abortus vaccine in sheep and goats: Assessment of safety, immunogenicity and protective efficacy against Brucella melitensis infection

ABSTRACT Brucellosis is a zoonotic disease with a worldwide distribution that can be transmitted via intentional or accidental aerosol exposure. In order to engineer superior vaccine strains against Brucella species for use in animals as well as in humans, the possibility of challenge infection via aerosol needs to be considered to properly evaluate vaccine efficacy. In this study, we assessed the use of an aerosol chamber to infect deep lung tissue of mice to elicit systemic infections with either Brucella abortus or B. melitensis at various doses. The results reveal that B. abortus causes a chronic infection of lung tissue in BALB/c mice and peripheral organs at low doses. In contrast, B. melitensis infection diminishes more rapidly, and higher infectious doses are required to obtain infection rates in animals similar to those of B. abortus. Whether this difference translates to severity of human infection remains to be elucidated. Despite these differences, unmarked deletion mutants BAΔasp24 and BMΔasp24 consistently confer superior protection to mice against homologous and heterologous aerosol challenge infection and should be considered viable candidates as vaccine strains against brucellosis.

Download Full-text

Evaluation of Lipopolysaccharides and Polysaccharides of Different Epitopic Structures in the Indirect Enzyme-Linked Immunosorbent Assay for Diagnosis of Brucellosis in Small Ruminants and Cattle

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.6.749-754.1998 ◽

1998 ◽

Vol 5 (6) ◽

pp. 749-754 ◽

Cited By ~ 24

Author(s):

B. Alonso-Urmeneta ◽

C. Marín ◽

V. Aragón ◽

J. M. Blasco ◽

R. Díaz ◽

...

Keyword(s):

Immunosorbent Assay ◽

Brucella Abortus ◽

Diagnostic Performance ◽

Lipid A ◽

Brucella Melitensis ◽

Outer Membrane Proteins ◽

Small Ruminants ◽

Enzyme Linked Immunosorbent Assay ◽

Sheep And Goats ◽

Antibody Levels

ABSTRACT Brucella abortus and Brucella melitensishave surface lipopolysaccharides and polysaccharides carryingB. melitensis-type (M) and B. abortus-type (A) epitopes as well as common (C) epitopes present in all smooth Brucella biotypes. Crude lipopolysaccharides, hydrolytic O polysaccharides, and native hapten polysaccharides of MC or AC specificity were evaluated in indirect enzyme-linked immunosorbent assays with polyclonal, monoclonal, or protein G conjugates by using sera from cattle, sheep, and goats infected with AC, MC, or AMC Brucella biotypes. Regardless of the antigen, the levels of antibodies were lower in goats than in sheep and highest in cattle. The diagnostic performance of the assay was not affected by the absence of lipid A-core epitopes, the presence of contaminating outer membrane proteins, the AC or MC epitopic structure of the absorbed antigen, or the conjugate used. Moreover, with sera from cattle vaccinated with B. abortus S19 (AC) or from sheep and goats vaccinated with B. melitensis Rev 1 (MC), AC and MC antigens showed similar levels of reactivity. The results show that antibodies to the C epitopes largely dominate in infection, and this is consistent with the existence of multiple overlapping C epitopes (V. Weynants, D. Gilson, A. Cloeckaert, A. Tibor, P. A. Denoel, F. Godfroid, J. N. Limet, and J.-J. Letesson, Infect. Immun. 65:1939–1943, 1997) rather than with one or two C epitopes. It is concluded that, by adaptation to the corresponding antibody levels, brucellosis in cattle, sheep, and goats can be diagnosed by immunosorbent assay with a single combination of conjugate and antigen.

Download Full-text

An influenza viral vector Brucella abortus vaccine induces good cross-protection against Brucella melitensis infection in pregnant heifers

Vaccine ◽

10.1016/j.vaccine.2015.06.045 ◽

2015 ◽

Vol 33 (31) ◽

pp. 3619-3623 ◽

Cited By ~ 13

Author(s):

Kaissar Tabynov ◽

Sholpan Ryskeldinova ◽

Abylai Sansyzbay

Keyword(s):

Brucella Abortus ◽

Viral Vector ◽

Brucella Melitensis ◽

Cross Protection

Download Full-text

Identification, geographic distribution and risk factors of Brucella abortus and Brucella melitensis infection in cattle in Algeria

Veterinary Microbiology ◽

10.1016/j.vetmic.2021.109004 ◽

2021 ◽

Vol 254 ◽

pp. 109004

Author(s):

Nedjma Lounes ◽

Falk Melzer ◽

Ashraf E. Sayour ◽

Hassiba Tali Maamar ◽

Kheira Rahal ◽

...

Keyword(s):

Risk Factors ◽

Geographic Distribution ◽

Brucella Abortus ◽

Brucella Melitensis

Download Full-text

Deletion of znuA Virulence Factor Attenuates Brucella abortus and Confers Protection against Wild-Type Challenge

Infection and Immunity ◽

10.1128/iai.01957-05 ◽

2006 ◽

Vol 74 (7) ◽

pp. 3874-3879 ◽

Cited By ~ 58

Author(s):

Xinghong Yang ◽

Todd Becker ◽

Nancy Walters ◽

David W. Pascual

Keyword(s):

Brucella Abortus ◽

Deletion Mutant ◽

Pasteurella Multocida ◽

Brucella Melitensis ◽

Normal Growth ◽

Wild Type ◽

Minimal Growth ◽

M Genome ◽

S19 Vaccine ◽

Knockout Mutation

ABSTRACT znuA is known to be an important factor for survival and normal growth under low Zn2+ concentrations for Escherichia coli, Haemophilus spp., Neisseria gonorrhoeae, and Pasteurella multocida. We hypothesized that the znuA gene present in Brucella melitensis 16 M would be similar to znuA in B. abortus and questioned whether it may also be an important factor for growth and virulence of Brucella abortus. Using the B. melitensis 16 M genome sequence, primers were designed to construct a B. abortus deletion mutant. A znuA knockout mutation in B. abortus 2308 (ΔznuA) was constructed and found to be lethal in low-Zn2+ medium. When used to infect macrophages, ΔznuA B. abortus showed minimal growth. Further study with ΔznuA B. abortus showed that its virulence in BALB/c mice was attenuated, and most of the bacteria were cleared from the spleen within 8 weeks. Protection studies confirmed the ΔznuA mutant as a potential live vaccine, since protection against wild-type B. abortus 2308 challenge was as effective as that obtained with the RB51 or S19 vaccine strain.

Download Full-text

Immunohistochemical Detection of Brucella Melitensis Antigens in Cases of Naturally Occurring Abortions in Sheep

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870802000616 ◽

2008 ◽

Vol 20 (6) ◽

pp. 803-806 ◽

Cited By ~ 16

Author(s):

Fatma Ilhan ◽

Zabit Yener

Keyword(s):

Kupffer Cells ◽

Brucella Melitensis ◽

Immunohistochemical Detection ◽

Zoonotic Pathogen ◽

Sheep And Goats ◽

Naturally Occurring ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Microscopic Studies ◽

Formalin Fixed

Brucella melitensis, a worldwide zoonotic pathogen, is a significant cause of abortion in sheep and goats in some countries. The present study was carried out to determine, by immunohistochemistry, the presence of B. melitensis antigens in 110 naturally occurring aborted sheep fetuses. Sections of lung, liver, kidney, and spleen of each fetus were stained with immunoperoxidase to detect Brucella antigens. Brucella melitensis antigens were detected in 33 of 110 fetuses (30%). In the 33 positive cases, Brucella antigens were found in lung (25 [22.7%]), liver (21 [19%]), spleen (13 [11.8%]), and kidney (6 [5.4%]). Microscopic studies demonstrated that Brucella antigens were mainly located in the cytoplasm of macrophages and neutrophils of the lung, and in the cytoplasm of macrophages in the portal infiltrates and Kupffer cells of the liver. It was concluded that immunohistochemistry in formalin-fixed, paraffin-embedded tissues is a useful tool for the diagnosis of spontaneous ovine abortion caused by B. melitensis.

Download Full-text

Evaluation of Protection Afforded by Brucella abortus and Brucella melitensis Unmarked Deletion Mutants Exhibiting Different Rates of Clearance in BALB/c Mice

Infection and Immunity ◽

10.1128/iai.01787-05 ◽

2006 ◽

Vol 74 (7) ◽

pp. 4048-4057 ◽

Cited By ~ 59

Author(s):

M. M. Kahl-McDonagh ◽

T. A. Ficht

Keyword(s):

Mouse Model ◽

Brucella Abortus ◽

Protective Immunity ◽

Brucella Melitensis ◽

Deletion Mutants ◽

Heterologous Challenge ◽

Current Vaccine ◽

Protection Against Infection ◽

Ideal Vaccine

ABSTRACT Research for novel Brucella vaccines has focused upon the development of live vaccine strains, which have proven more efficacious than killed or subunit vaccines. In an effort to develop improved vaccines, signature-tagged mutant banks were screened to identify mutants attenuated for survival. Mutants selected from these screens exhibited various degrees of attenuation characterized by the rate of clearance, ranging from a failure to grow in macrophages after 24 h of infection to a failure to persist in the mouse model beyond 8 weeks. Ideal vaccine candidates should be safe to the host, while evoking protective immunity. In the present work, we constructed unmarked deletion mutants of three gene candidates, manBA, virB2, and asp24, in both Brucella abortus and Brucella melitensis. The Δasp24 mutants, which persist for extended periods in vivo, are superior to current vaccine strains and to other deletion strains tested in the mouse model against homologous challenge infection after 12, 16, and 20 weeks postvaccination. The Δasp24 mutants also display superior protection compared to ΔmanBA and ΔvirB2 mutants against heterologous challenge in mice. From this study, a direct association between protection against infection and cytokine response was not apparent between all vaccine groups and, therefore, correlates of protective immunity will need to be considered further. A distinct correlation between persistence of the vaccine strain and protection against infection was corroborated.

Download Full-text

Protection of BALB/c Mice against Brucella abortus 544 Challenge by Vaccination with Bacterioferritin or P39 Recombinant Proteins with CpG Oligodeoxynucleotides as Adjuvant

Infection and Immunity ◽

10.1128/iai.69.8.4816-4822.2001 ◽

2001 ◽

Vol 69 (8) ◽

pp. 4816-4822 ◽

Cited By ~ 95

Author(s):

Ayman Al-Mariri ◽

Anne Tibor ◽

Pascal Mertens ◽

Xavier De Bolle ◽

Patrick Michel ◽

...

Keyword(s):

Recombinant Proteins ◽

Brucella Abortus ◽

Mononuclear Cells ◽

Brucella Melitensis ◽

Cpg Odn ◽

Peripheral Blood Mononuclear ◽

Recombinant Antigens ◽

Cpg Oligodeoxynucleotides ◽

Ifn Γ

ABSTRACT The P39 and the bacterioferrin (BFR) antigens of Brucella melitensis 16M were previously identified as T dominant antigens able to induce both delayed-type hypersensivity in sensitized guinea pigs and in vitro gamma interferon (IFN-γ) production by peripheral blood mononuclear cells from infected cattle. Here, we analyzed the potential for these antigens to function as a subunitary vaccine against Brucella abortus infection in BALB/c mice, and we characterized the humoral and cellular immune responses induced. Mice were injected with each of the recombinant proteins alone or adjuvanted with either CpG oligodeoxynucleotides (CpG ODN) or non-CpG ODN. Mice immunized with the recombinant antigens with CpG ODN were the only group demonstrating both significant IFN-γ production and T-cell proliferation in response to either Brucella extract or to the respective antigen. The same conclusion holds true for the antibody response, which was only demonstrated in mice immunized with recombinant antigens mixed with CpG ODN. The antibody titers (both immunoglobulin G1 [IgG1] and IgG2a) induced by P39 immunization were higher than the titers induced by BFR (only IgG2a). Using a B. abortus 544 challenge, the level of protection was analyzed and compared to the protection conferred by one immunization with the vaccine strain B19. Immunization with P39 and CpG ODN gave a level of protection comparable to the one conferred by B19 at 4 weeks postchallenge, and the mice were still significantly protected at 8 weeks postchallenge, although to a lesser extent than the B19-vaccinated group. Intriguingly, no protection was detected after BFR vaccination. All other groups did not demonstrate any protection.

Download Full-text