Identification of sero-reactive Brucella melitensis cytosoluble proteins which discriminate between antibodies elicited by infection and Rev. 1 vaccination in sheep

Debbarh, Hanane Salih-Alj; Cloeckaert, Axel; Zygmunt, Michel S.; Dubray, Gérard

doi:10.1016/0378-1135(94)00058-5

Cited by 33 publications

(22 citation statements)

References 17 publications

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“…Reported studies have focused on cytosoluble and membrane proteins [4, 5, from most sheep infected with B. melitensis [5] and therefore seemed to be more appropriate than OMPs in the development of such a diagnostic test. As shown in the present study, this CP is expressed in all Brucella species and biovars.…”

Section: Discussionmentioning

confidence: 99%

“…A large number of protein bands ranging in apparent molecular mass from 12 to 100 kDa, of which proteins previously identified as discriminating antibody responses of B. melitensis infected from Rev. 1 vaccinated sheep [5], were recognised by sera of mice infected with both H38 and Rev. 1.…”

Section: Antibody Responses To Cp In Mice After Infectionmentioning

confidence: 99%

“…to be the most interesting regarding specificity and number of infected animals detected [5]. Groups of five 8-week-old female BALB/c mice were inoculated intraperitoneally with lo7 cfu of B.…”

Section: Mice Immunisations and Hybridomasmentioning

confidence: 99%

“…A previous study [5] identified a number of B. melitensis cytosoluble proteins (CP) by immunoblotting, mainly proteins ranging in apparent molecular masses from 10 to 32 kDa, which made distinction possible between antibody responses from B. melitensis Rev. 1 sis.…”

Section: Introductionmentioning

confidence: 99%

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Production and characterisation of monoclonal antibodies to Brucella melitensis cytosoluble proteins that are able to differentiate antibody responses of infected sheep from Rev. 1 vaccinated sheep

Cloeckaert

Debbarh²,

Zygmunt³

et al. 1996

Journal of Medical Microbiology

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Monoclonal antibodies (MAbs) were produced to Brucella melitensis cytosoluble proteins (CP) with apparent molecular masses of 12, 24 and 28kDa (CP12, CP24 and CP28) which were previously shown by immunoblotting to differentiate antibody responses of infected sheep from those of B. melitensis strain Rev. 1 vaccinated sheep. These MAbs were derived from mice infected with virulent smooth (S) B. melitensis strain H38. Most MAbs obtained were directed to CP28, which indicated (as was shown in infected sheep) that this protein was also highly immunogenic in mice. A large number of MAbs that showed reactivity to CP in ELISA but did not show reactivity in immunoblotting of CP were also obtained and might recognise conformational epitopes of these proteins. MAbs were used to localise CP12, CP24 and CP28. None of the MAbs reacted with whole B. melitensis cells in ELISA but showed reactivity with sonicated bacteria in ELISA, which indicated an internal localisation of these proteins. Among several B. melitensis B115 subcellular fractions tested, the anti-CP12 and anti-CP28 MAbs reacted essentially with the CP extract (CPE) in both ELISA and immunoblotting, whereas the anti-CP24 MAbs reacted with both CPE and cell envelope fraction (CEF) -although with lower intensity to the latter fraction. The internal localisation of these proteins was confirmed by immuno-electron microscopy of thin-sectioned B. melitensis B115 or B. melitensis 16M cells. Immunogold labelling was mainly observed in the cytoplasm and, consequently, CP12, CP24 and CP28 are probably cytoplasmic proteins. Immunoblotting of whole cell lysates with the MAbs also showed the presence of these proteins in all Brucella species and biovars, including the vaccine strains B. melitensis Rev. 1 and B. abortus B19. The use of these MAbs should help further study of antibody responses in sheep and other hosts and may be of considerable value for developing new diagnostic tests for ovine brucellosis.

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Section: Discussionmentioning

confidence: 99%

Section: Antibody Responses To Cp In Mice After Infectionmentioning

confidence: 99%

Section: Mice Immunisations and Hybridomasmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Production and characterisation of monoclonal antibodies to Brucella melitensis cytosoluble proteins that are able to differentiate antibody responses of infected sheep from Rev. 1 vaccinated sheep

Cloeckaert

Debbarh²,

Zygmunt³

et al. 1996

Journal of Medical Microbiology

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“…This cross-reactivity hampered the interpretation of many of serological tests used in the diagnosis of brucellosis [48]. The situation is worsened in farm animals vaccinated with the live-attenuated Rev1 vaccine, which makes it difficult to differentiate between vaccinated and infected animals [49]. In humans, the use of serological assays had complicated the clinical differentiation between acute, chronic, or chronic persistent forms.…”

Section: Humoral Response To Brucella Infectionmentioning

confidence: 99%

Host response to Brucella infection: review and future perspective

Elfaki

Alaidan

Alhokail

2015

J Infect Dev Ctries

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Brucellosis is a zoonotic and contagious infectious disease caused by infection with Brucella species. The infecting brucellae are capable of causing a devastating multi-organ disease in humans with serious health complications. The pathogenesis of Brucella infection is influenced largely by host factors, Brucella species/strain, and the ability of invading brucellae to survive and replicate within mononuclear phagocytic cells, preferentially macrophages (M). Consequently, the course of human infection may appear as an acute fatal or progress into chronic debilitating infection with periodical episodes that leads to bacteremia and death. The existence of brucellae inside M represents one of the strategies used by Brucella to evade the host immune response and is responsible for treatment failure in certain human populations treated with anti-Brucella drugs. Moreover, the persistence of brucellae inside M complicates the diagnosis and may affect the host cell signaling pathways with consequent alterations in both innate and adaptive immune responses. Therefore, there is an urgent need to pursue the development of novel drugs and/or vaccine targets against human brucellosis using high throughput technologies in genomics, proteomics, and immunology.

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Mapping and identification of Brucella melitensis proteins by two‐dimensional electrophoresis and microsequencing

et al. 1997

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Two-dimensional (2-D) gel electrophoresis was used to map Brucella melitensis proteins. The 2-D proteins map of B. melitensis B115 revealed 595 silver-stained protein spots separated by both isoelectric point and molecular mass. Twenty-five proteins were identified either by immunoblotting using monoclonal antibodies (MAbs) or by N-terminal microsequencing. The protein spots identified by MAbs were the 89 kDa outer membrane protein, DnaK, bacterioferritin, CP24, and BP26. Some spots were identified by N-terminal microsequencing as proteins whose sequences had been reported previously from Brucella, such as three heat-shock proteins, namely DnaK, GroEL and GroES; bacterioferritin; Cu-Zn superoxide dismutase; and the 50S ribosomal protein L7/L12. Other proteins had amino acid sequences homologous with those of various proteins from other bacteria found in protein databases: ClpP; the 10K-S protein; the ORFU phosphoprotein; succinyl-CoA synthetase alpha sub-unit; an inorganic pyrophosphatase; the Fe and/or Mn superoxide dismutase; the nucleoside diphosphate kinase, an amino acid ABC type transporter, and an electron transfer flavoprotein small subunit. Seven proteins were identified with N-terminal sequences not yet reported in databases. The 2-D map established in this study will be the basis for comparative studies of protein expression in Brucella.

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Identification of sero-reactive Brucella melitensis cytosoluble proteins which discriminate between antibodies elicited by infection and Rev. 1 vaccination in sheep

Cited by 33 publications

References 17 publications

Production and characterisation of monoclonal antibodies to Brucella melitensis cytosoluble proteins that are able to differentiate antibody responses of infected sheep from Rev. 1 vaccinated sheep

Production and characterisation of monoclonal antibodies to Brucella melitensis cytosoluble proteins that are able to differentiate antibody responses of infected sheep from Rev. 1 vaccinated sheep

Host response to Brucella infection: review and future perspective

Mapping and identification of Brucella melitensis proteins by two‐dimensional electrophoresis and microsequencing

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