Immunochemical Methods for Identification ofBabesia bovisAntigens Expressed on the Erythrocyte Surface

Allred, David R.

doi:10.1006/meth.1997.0510

Cited by 10 publications

(11 citation statements)

References 27 publications

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“…The procedure used to detect the IRBC surface reactivity of bovine sera and mAbs has been described in several publications (3,14,17,32). In live-cell IFAs using rabbit sera as the primary Ab, reactions were amplified with monoclonal anti-rabbit Igs (clone RG-16, Sigma, St. Louis, MO) and the immune complexes detected with FITC-conjugated goat anti-mouse IgG.…”

Section: Live-cell Immunofluorescence Assays (Live-cell Ifa)mentioning

confidence: 99%

“…The details of surface-specific immunoprecipitations with bovine serum, and conventional immunoprecipitations with mAbs are described elsewhere (3,14,17,32). L- [ 35 S]methionine-labeled IRBCs were used as the Ag source, since mature bovine erythrocytes fail to metabolically incorporate this label into protein.…”

Section: Surface-specific and Conventional Immunoprecipitationsmentioning

confidence: 99%

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Selection of Babesia bovis-Infected Erythrocytes for Adhesion to Endothelial Cells Coselects for Altered Variant Erythrocyte Surface Antigen Isoforms

O’Connor

Allred

2000

The Journal of Immunology

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Sequestration of Babesia bovis-infected erythrocytes (IRBCs) in the host microvasculature is thought to constitute an important mechanism of immune evasion. Since Ig is considered to be important for protection from disease, an in vitro assay of B. bovis sequestration was used to explore the ability of anti-B. bovis Ig to interfere with IRBC cytoadhesion, and to identify IRBC surface Ags acting as endothelial cell receptors. Bovine infection sera reactive with the IRBC surface inhibited and even reversed the binding of IRBCs to bovine brain capillary endothelial cells (BBECs). This activity is at least partially attributable to serum IgG. IgG isolated from inhibitory serum captured the variant erythrocyte surface ag 1 (VESA1) in surface-specific immunoprecipitations of B. bovis-IRBCs. Selection for the cytoadhesive phenotype concurrently selected for antigenic and structural changes in the VESA1 Ag. In addition, the anti-VESA1 mAb, 4D9.1G1, proved capable of effectively inhibiting and reversing binding of adhesive, mAb-reactive parasites to BBECs, and by immunoelectron microscopy localized VESA1 to the external tips of the IRBC membrane knobs. These data are consistent with a link between antigenic variation and cytoadherence in B. bovis and suggest that the VESA1 Ag acts as an endothelial cell ligand on the B. bovis-IRBC.

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Section: Live-cell Immunofluorescence Assays (Live-cell Ifa)mentioning

confidence: 99%

Section: Surface-specific and Conventional Immunoprecipitationsmentioning

confidence: 99%

Selection of Babesia bovis-Infected Erythrocytes for Adhesion to Endothelial Cells Coselects for Altered Variant Erythrocyte Surface Antigen Isoforms

O’Connor

Allred

2000

The Journal of Immunology

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“…In this assay, IRBC are metabolically labelled with L-[ 35 S]methionine, incubated with imm une sera or antibodies, and washed to remove unbound antibodies. The IRBC are then disrupted in Nonidet P-40 and the solubilized immune complexes are captured with protein G-Sepharose Allred, 1997). Because mature bovine erythrocytes do not synthesise nascent polypeptides, any amino-acid labels incorporated into polypeptides must be of parasite origin.…”

Section: Identification Of Surfac E Antigens Of Infected Erythro Cytesmentioning

confidence: 99%

“…Attem pts were made to clarify the situation with regard to antigenic variation in B. bovis, and to identify the antigens involved. The techniques used to achieve these aims are described in detail elsewhere (Allred et al, , 1994Allred, 1997).…”

Section: Detection Of Antigenic Vari-ation By B Bovis and Identifi-mentioning

confidence: 99%

“…Anchorage is indicated by the stable lateral distribution of VESA1 on the IRBC surface. For example, preparation of IRBC for live-cell immuno¯uorescence involves incubation with a primary antibody, cross-linking by a secondary antibody, and visualization by incubation with a¯uores-cently labelled tertiary antibody (Allred et al, 1994;Allred, 1997). Despite these manipulations and cross-linking of surface components, the labelling patterns observed are indistinguishable whether all manipulations are performed at 0± 4°C or at room temperature (O'Connor et al, 1997).…”

Section: Detection Of Antigenic Vari-ation By B Bovis and Identifi-mentioning

confidence: 99%

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Antigenic variation in Babesia bovis : how similar is it to that in Plasmodium falciparum

Allred

1998

Annals of Tropical Medicine And Parasitology

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Despite significant differences in some aspects of their life-cycles, the Apicomplexan parasites Babesia bovis and Plasmodium falciparum share many parallels. Significant among these are participation in rapid, clonal antigenic variation, and cyto-adherence and sequestration in the deep vasculature. Antigenic variation has long been thought to be primarily a mechanism of escape from antibody-mediated mechanisms of the host's immune system. In each species, the components demonstrated to participate in antigenic variation are parasite-derived proteins expressed on the infected erythrocyte's surface. Recently, the malarial component PfEMP1 has been found to be a multifunctional protein that is not only subject to antigenic variation, but also participates in cyto-adherence and rosetting (adhesion to uninfected erythrocytes). In the present review, the antigens elaborated on the surface of an erythrocyte infected with B. bovis, for immune evasion via antigenic variation, are described, and compared and contrasted with those from P. falciparum. The significance of the similarities between B. bovis and P. falciparum, and the potential for contributions to be made to our understanding of malaria through the study of babesiosis are discussed.

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Universal primers suitable to assess population dynamics reveal apparent mutually exclusive transcription of the Babesia bovis ves1α gene

Zupanska

Drummond

Swetnam

et al. 2009

Molecular and Biochemical Parasitology

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Babesia bovis is an intraerythrocytic hemoparasite of widespread distribution, which adversely affects livestock production in many regions of the world. This parasite establishes persistent infections of long duration, at least in part through rapid antigenic variation of the VESA1 protein on the infected-erythrocyte surface. To understand the dynamics of in vivo antigenic variation among the parasite population it is necessary to have sensitive and broadly applicable tools enabling monitoring of variation events in parasite antigen genes. To address this need for B. bovis, "universal" primers for the polymerase chain reaction have been designed for the ves1α gene, spanning from exon 2 to near the 3′ end of cysteine-lysine-rich domain (CKRD) sequences in exon 3. These primers robustly amplified this segment, with minimal bias, from essentially the entire repertoire of fulllength ves1α sequences in the B. bovis Mexico isolate genome, and are equivalently present in other isolates. On purified genomic DNA, this primer set can achieve a sensitivity of 10 genome equivalents or less. When applied to the amplification of cDNA derived from the B. bovis C9.1 clonal line evidence consistent with mutually exclusive transcription of the ves1α gene was obtained, concomitant with detection of numerous mutational events among members of the parasite population. These characteristics of the primers will facilitate the application of polymerase chain reaction-based methodologies to the study of B. bovis population and antigenic switching dynamics.

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Immunochemical Methods for Identification ofBabesia bovisAntigens Expressed on the Erythrocyte Surface

Cited by 10 publications

References 27 publications

Selection of Babesia bovis-Infected Erythrocytes for Adhesion to Endothelial Cells Coselects for Altered Variant Erythrocyte Surface Antigen Isoforms

Selection of Babesia bovis-Infected Erythrocytes for Adhesion to Endothelial Cells Coselects for Altered Variant Erythrocyte Surface Antigen Isoforms

Antigenic variation in Babesia bovis : how similar is it to that in Plasmodium falciparum

Universal primers suitable to assess population dynamics reveal apparent mutually exclusive transcription of the Babesia bovis ves1α gene

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