Cloning of a Brucella melitensis group 3 antigen gene encoding Omp28, a protein recognized by the humoral immune response during human brucellosis

Lindler, Luther E.; Hadfield, Ted L.; Tall, Ben D.; Snellings, Norma J.; Rubin, Fran A.; Verg, Lillian L. Van De; Hoover, David L.; Warren, Richard L.

doi:10.1128/iai.64.7.2490-2499.1996

Cited by 64 publications

(30 citation statements)

References 37 publications

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“…This was particularly evident with the two major OMPs (OMP25 and OMP36). These results contradict previously published data which described these antigens as major antibody targets in various animal species (6,14,26).…”

Section: Discussioncontrasting

confidence: 90%

“…(31) and other pathogens (1). Lindler et al (26) also reported that cattle do not produce any detectable antibody response against Omp28, while this antigen does elicit significant antibody responses in goats, rabbits, mice, and humans. Alternatively, such differences could be related to the prevalence of the disease in the region where the sera were collected.…”

Section: Discussionmentioning

confidence: 99%

“…Among these are cytoplasmic antigens: the Cu-Zn superoxide dismutase (29), an 18-kDa protein (19), and an antigen called A2 (37), which could correspond to the Brucella bacterioferritin (15), as well as purified membrane proteins (OMP89 [25] and the peptidoglycan-linked lipoprotein [20]). Recently, a protein useful for the serologic diagnosis of brucellosis was cloned independently by two groups and described either as an OMP belonging to Brucella group 3 antigen (26) or as a periplasmic protein (BP26) (28).…”

mentioning

confidence: 99%

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Humoral immune responses of Brucella-infected cattle, sheep, and goats to eight purified recombinant Brucella proteins in an indirect enzyme-linked immunosorbent assay

Letesson¹,

Tibor²,

Eynde³

et al. 1997

Clin Diagn Lab Immunol

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Brucellosis research is currently focused on the identification of nonlipopolysaccharide (LPS) antigens which could potentially be useful for the specific serologic diagnosis of brucellosis as well as for vaccinal prophylaxis. On the basis of previous reports, we selected eight Brucella proteins (OMP36, OMP25, OMP19, OMP16, OMP10, p17, p15, and p39) as candidate antigens to be further evaluated. The genes encoding these proteins were cloned, sequenced, and overexpressed in Escherichia coli. The recombinant proteins were purified with a polyhistidine tag and metal chelate affinity chromatography and evaluated in an indirect enzyme-linked immunosorbent assay (iELISA). The specificity of the iELISA was determined with sera from healthy cattle, sheep, and goats and ranged from 95 to 99%, depending on the recombinant antigen and the species tested. Sera from experimentally infected, and from naturally infected, animals were used to evaluate the sensitivity of the iELISA. The antiprotein antibody response was often delayed when compared to the anti-smooth LPS (S-LPS) response and was limited to animals which developed an active brucellosis infection (experimentally infected pregnant animals and sheep and goats from areas where brucellosis is still endemic). Among the recombinant antigens, the three cytoplasmic proteins (p17, p15, and p39) gave the most useful results. More than 80% of the animals positive in S-LPS serology were also positive with one of these cytoplasmic proteins alone or a combination of two of them. None of the recombinant antigens detected experimentally infected nonpregnant cows and sheep or naturally infected cattle. This study is a first step towards the development of a multiprotein diagnostic reagent for brucellosis.

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Section: Discussioncontrasting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Humoral immune responses of Brucella-infected cattle, sheep, and goats to eight purified recombinant Brucella proteins in an indirect enzyme-linked immunosorbent assay

Letesson¹,

Tibor²,

Eynde³

et al. 1997

Clin Diagn Lab Immunol

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“…Subunit vaccines (including rOmpA) have also been noted to induce IgG1 over IgG2a in other bacteria, including Pasteurella multocida [9], Burkholderia cenocepacia [25], and Acinetobacter baumanni [21]. A great deal of attention has been focused on the Omps as a non-LPS group of immunogens [22]. However, the roles of Omps in survival and virulence during infection remain unknown [11].…”

Section: Discussionmentioning

confidence: 99%

Immune Modulation of Recombinant OmpA against Brucella abortus 544 Infection in Mice

Simborio

Reyes

Hop

et al. 2016

Journal of Microbiology and Biotechnology

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Brucellosis affects a wide range of host species, including humans and many livestock animals. Chronic infections of the disease make antibiotic treatment costly, and the current vaccine used in livestock has not been approved for human use. This study investigated the possible use of the Brucella abortus outer membrane protein A (OmpA) as a candidate subunit vaccine in an infected mouse model. The ompA gene was cloned and overexpressed, and the recombinant OmpA (rOmpA) protein fused to maltose binding protein (MBP) was purified in Escherichia coli. Immunogenicity was verified through western blotting, and mice were immunized and challenged to evaluate its protective effect. Mice treated with rOmpA exhibited induced humoral and host cell-mediated responses, with a significant increase in immunoglobulin G (IgG1 and IgG2a) and cytokine levels, especially TNF-α and IL-12, compared with the control groups treated with either MBP or PBS. In conclusion, rOmpA should be highly considered as a future subunit vaccine for brucellosis, and further studies regarding rOmpA and its protective ability are suggested.

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“…In present study, we selected a QD immunochromatography technology for labeling an outer membrane proteins (OMPs) in samples. Lindler et al identi ed one non-LPS group of immunogens with OMPs for vaccine and diagnostic purposes [20]. This labeling technology forms hundreds or even thousands of particles by encapsulating or connecting to other materials to form nanoparticles, and it has their own unique nature of good light stability , long uorescence lifetime, wide excitation spectrum and a narrow emission spectrum, good biocompatibility , size-tunable, indicated that the QD uorescent microspheres (QDFM) have potential as a new labeling technology immunoassay [21,22].…”

Section: Introductionmentioning

confidence: 99%

A quantum dot fluorescent microsphere based immunochromatographic strip for detection of brucellosis

Kong

Wang

et al. 2020

Preprint

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Abstract Background: Brucellosis is a serious zoonosis disease that frequently causes significant economic loss in animal husbandry and threatens human health. Therefore, we established a rapid, accurate, simple and sensitive fluorescent immunochromatographic test strip (ICTS) based on quantum dots to detect serum antibodies of brucellosis.Result: The test strips were successfully prepared by quantum dot fluorescent microspheres (QDFM) as tracers, which were covalently coupled to an outer membrane protein of Brucella, OMP22. The outer membrane protein OMP28 and monoclonal antibodies of OMP22 against Brucella were separately dispensed onto a nitrocellulose membrane as test and quality control lines, respectively. Antibodies specific for brucellosis was detected qualitatively using the ratio of the fluorescence signals of the test and control lines (HT/HC). The threshold of assay was determined as a HT/HC value of 0.0492. The repeatability was excellent with an overall average CV of 8.78%. Under optimum conditions, the limit of detection was 1.05 ng/mL (1:512 dilution). With regard to the detection of brucellosis in 150 clinical samples, the total coincidence rate of ICTS and Rose Bengal plate test (RBPT) was 97.3%, the coincidence rate of positive samples was 98.8%, the coincidence rate of negative samples was 95.3%, and no cross reaction with the sera of other related diseaseswas observed.Conclusion: In our present study, the QDFM has promising application for on-site screening of brucellosis owing to its high detection speed, high sensitivity high specificity and low cost.

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Cloning of a Brucella melitensis group 3 antigen gene encoding Omp28, a protein recognized by the humoral immune response during human brucellosis

Cited by 64 publications

References 37 publications

Humoral immune responses of Brucella-infected cattle, sheep, and goats to eight purified recombinant Brucella proteins in an indirect enzyme-linked immunosorbent assay

Humoral immune responses of Brucella-infected cattle, sheep, and goats to eight purified recombinant Brucella proteins in an indirect enzyme-linked immunosorbent assay

Immune Modulation of Recombinant OmpA against Brucella abortus 544 Infection in Mice

A quantum dot fluorescent microsphere based immunochromatographic strip for detection of brucellosis

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