A Comparison of Lipoarabinomannan with other Antigens used in Absorbed Enzyme Immunoassays for the Serological Detection of Cattle Infected with<i>Mycobacterium Paratuberculosis</i>

Sugden, E A; Stilwell, K; Michaelides, Alecos

doi:10.1177/104063879700900413

Cited by 23 publications

(16 citation statements)

References 14 publications

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“…In addition, antibody titers against Map and p34 in sera from infected cattle were highly variable among animals. Individual variations were described in field animals and in sera of Map infected cattle whose results were generally analyzed individually [37,40].…”

Section: Discussionmentioning

confidence: 99%

Bovine IgG1 antibodies againstMycobacterium aviumsubsp.paratuberculosisprotein p34-cx improve association of bacteria and macrophages

et al. 2007

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-Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (Map), is a chronic granulomatous enteric disease in cattle. Among molecular components of Map, protein p34 was identified as specific and immunodominant for bovine B cells. In order to determine if specific antibodies could influence the course of Map pathogenesis, the interaction between bacteria and bovine macrophages was studied. Bovine polyclonal antibodies from 3 calves vaccinated with protein p34-cx, 6 calves vaccinated with heatkilled Map, 8 naturally infected, and 3 healthy calves -as negative controls-were used. Specific anti-Map, -p34-cx and -PPA-3 antibodies were evaluated and isotype characterized. Infected and Map vaccinated animals showed similar IgG1 and IgG2 response against Map whole bacteria. When p34-cx was used as the antigen, mainly IgG1 and IgG3 were detected in infected and only IgG1 in p34-cx vaccinated animals. Bovine polyclonal antibodies from three animals of each category were isolated and affinity purified through Map and p34-cx columns. The effect of these antibodies in association with Map and a transformed bovine peritoneal macrophage's cell line (Bov-Mac) as well as activation of NF-κB transcription factor was studied. Our results show that association of Map significantly increased in vitro after pretreatment with bovine anti-Map or anti-p34-cx antibodies obtained from vaccinated or infected cattle when compared with those of controls. Improved activation of NF-κB was detected in macrophages that ingested Map opsonized with either anti-Map or anti-p34-cx specific antibodies of infected or vaccinated calves, suggesting that both anti-Map and IgG1 anti-p34-cx antibodies support Map-macrophage interactions.Mycobacterium avium subsp. paratuberculosis / p34 / IgG1 / macrophages / cattle

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Section: Discussionmentioning

confidence: 99%

Bovine IgG1 antibodies againstMycobacterium aviumsubsp.paratuberculosisprotein p34-cx improve association of bacteria and macrophages

et al. 2007

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“…LAM-based ELISA assays have demonstrated utility for diagnosis of mycobacterial diseases, including M. avium subsp. paratuberculosis infection (23,26,29,33,(48)(49)(50). It is speculated that selective binding of the lipid portion of LAM to the test plate leaves the more immunogenic carbohydrate portion of the molecule free to react with antibody in the sample.…”

Section: Discussionmentioning

confidence: 99%

Early Induction of Humoral and Cellular Immune Responses during ExperimentalMycobacterium aviumsubsp.paratuberculosisInfection of Calves

et al. 2003

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“…Interestingly, numerous individual cattle were strongly responsive in one assay and not another (8). Although exact details of the antigen preparation methods for commercially available ELISA kits are proprietary, either protoplasmic fractions, other cellular extracts, or lipoarabinomannan are likely the basis of most assays because of the higher antigen yields and ease of preparation (1,4,7,9,21,34,37,46). Based on a literature review, proteins secreted from M. paratuberculosis into the extracellular environment have received little attention as serodiagnostic assay antigen candidates (21).…”

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confidence: 99%

Comparison of the Proteosomes and Antigenicities of Secreted and Cellular Proteins Produced byMycobacterium paratuberculosis

Cho

Collins

2006

Clin Vaccine Immunol

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The protein expression profiles and antigenicities of both culture filtrates (CF) and cellular extracts (CE) of Mycobacterium paratuberculosis were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), one-dimensional electrophoresis (1-DE) and 2-DE immunoblotting, and enzyme-linked immunosorbent assay (ELISA). The CF proteins were harvested from supernatants of stationary-phase liquid cultures and concentrated by size exclusion filtration. The CE proteins were extracted by mechanical disruption of cells using glass beads and a high-speed agitator. Analysis of SDS-PAGE gels showed that the majority of CF proteins had low molecular masses (<50 kDa), whereas CE protein mass ranged more evenly over a broader range up to 100 kDa. By 2-DE, CF proteins had a narrow array of pI values, with most being between pH 4.0 and 5.5; CE proteins spanned pI values from pH 4.0 to 7.0. The antigenicities of CF and CE proteins were first determined by 1-DE and 2-DE immunoblotting with serum from a cow naturally infected with M. paratuberculosis. The serum reacted strongly to more proteins in the CF than the CE. Sera from 444 infected and 412 uninfected cattle were tested by ELISA with CF and CE as solid-phase antigens. Receiver-operator characteristic curve analysis of the ELISA results showed a significantly greater area under the curve for CF compared to CE (P < 0.05). A high degree of variability in protein binding patterns was shown with 1-DE immunoblot analysis with 31 sera from M. paratuberculosis-infected cattle. Collectively, these results indicate that serologic tests for bovine paratuberculosis may be improved by using proteins derived from CF instead of CE. To maximize the diagnostic sensitivity of serologic tests, multiple proteins will be required. Even so, a CF ELISA may not be able to detect all M. paratuberculosis-infected cattle, in particular those in the early stages of infection that have yet to mount an antibody response.

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A Comparison of Lipoarabinomannan with other Antigens used in Absorbed Enzyme Immunoassays for the Serological Detection of Cattle Infected withMycobacterium Paratuberculosis

Abstract: Enzyme-linked immunosorbent assay (ELISA) protocols for the detection of antibodies to Mycobacterium paratuberculosis have employed a variety of antigens. These include crude protoplasmic antigens from M. paratuberculosis strains 18, 3,4

Cited by 23 publications

References 14 publications

Bovine IgG1 antibodies againstMycobacterium aviumsubsp.paratuberculosisprotein p34-cx improve association of bacteria and macrophages

Bovine IgG1 antibodies againstMycobacterium aviumsubsp.paratuberculosisprotein p34-cx improve association of bacteria and macrophages

Early Induction of Humoral and Cellular Immune Responses during ExperimentalMycobacterium aviumsubsp.paratuberculosisInfection of Calves

Comparison of the Proteosomes and Antigenicities of Secreted and Cellular Proteins Produced byMycobacterium paratuberculosis

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