Identification of common surface antigens among<i>Babesia bigemina</i>isolates by using monoclonal antibodies

Millán, Julio Vicente Figueroa; Buening, G. M.; Kinden, Darrell A.; Green, T. J.

doi:10.1017/s0031182000061163

Cited by 31 publications

(26 citation statements)

References 40 publications

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“…1B). This could be similar to a non-speci®c band of 53 kDa described by Figueroa et al (1990). c43 serum also reacted faintly with 51-and 48-kDa proteins in B. bigemina (Mexican) but with no proteins in B. bovis (Fig.…”

Section: Culturesupporting

confidence: 74%

In vitro cultivation of an African strain of Babesia bigemina , its characterisation and infectivity in cattle

Posnett

Metaferia

Wiliamson

et al. 1998

Parasitology Research

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An African (Kenyan) strain of Babesia bigemina, Muguga (B(2-1)), was inoculated into a calf from a stabilate and blood from the calf was used to establish the parasite in vitro. The strain has been cultured continuously for 20 months, initially in bovine erythrocytes with 60% adult bovine serum, later, with 50%. Cultures were incubated at 37 degrees C in RPMI 1640 medium with a gas mixture of 1% O2, 5% CO2, 94% N2. Adaptation in vitro was demonstrated when serum from a calf which had recovered from infection with B(2-1) bound to proteins of Mr 46 kDa, 49 kDa, 52 kDa, 61 kDa and 72 kDa on Western blots of B(2-1) antigens from cattle blood but did not recognise the 49 kDa or 52 kDa antigens from in-vitro-derived parasites. These proteins were considered specific for B(2-1), as they were not recognised by the same serum on profiles of a Mexican isolate of B. bigemina or an African isolate of B. bovis (Kwanyange). After 9 months of in vitro culture, a stabilate of the cultured parasite was injected into two splenectomised calves and one intact calf. The calves experienced a drop in packed cell volume and low parasitaemias but recovered spontaneously. Two of these animals, one splenectomised and one intact, were challenged with virulent B(2-1) and experienced only mild babesiosis, in contrast to a previously uninfected calf also challenged with B(2-1), which had to be euthanised after 5 days with severe babesiosis.

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Section: Culturesupporting

confidence: 74%

In vitro cultivation of an African strain of Babesia bigemina , its characterisation and infectivity in cattle

Posnett

Metaferia

Wiliamson

et al. 1998

Parasitology Research

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“…Posttranslational modification of proteins, through the addition of carbohydrate moieties, has been shown to affect the apparent molecular weight of glycoproteins analyzed by SDS-PAGE. Glycosylation contributes as much as 50 to 80% of the molecular weight of mature glycoproteins in mammals (5,8,28) and up to 40% in invertebrates (5,16). In addition, it has been shown that the glycosylation profile can influence MAb binding to glycoproteins (5,28).…”

Section: Discussionmentioning

confidence: 99%

Monoclonal Antibody MG96 Completely Blocks Plasmodium yoelii Developmentin Anophelesstephensi

et al. 2003

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In spite of research efforts to develop vaccines against the causative agent of human malaria, Plasmodium falciparum, effective control remains elusive. The predominant vaccine strategy focuses on targeting parasite blood stages in the vertebrate host. An alternative approach has been the development of transmissionblocking vaccines (TBVs). TBVs target antigens on parasite sexual stages that persist within the insect vector, anopheline mosquitoes, or target mosquito midgut proteins that are presumed to mediate parasite development. By blocking parasite development within the insect vector, TBVs effectively disrupt transmission and the resultant cascade of secondary infections. Using a mosquito midgut-specific mouse monoclonal antibody (MG96), we have partially characterized membrane-bound midgut glycoproteins in Anopheles gambiae and Anopheles stephensi. These proteins are present on the microvilli of midgut epithelial cells in both blood-fed and unfed mosquitoes, suggesting that the expression of the protein is not induced as a result of blood feeding. MG96 exhibits a dose-dependent blocking effect against Plasmodium yoelii development in An. stephensi. We achieved 100% blocking of parasite development in the mosquito midgut. Preliminary deglycosylation assays indicate that the epitope recognized by MG96 is a complex oligosaccharide. Future investigation of the carbohydrate epitope as well as gene identification should provide valuable insight into the possible mechanisms of ookinete attachment and invasion of mosquito midgut epithelial cells.

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“…The JG-29 biological clone of the Mexico strain was obtained by limiting dilution, as previously described (36,37). The Puerto Rico, St. Croix, Argentina S1A, and Texcoco strains were isolated from infected cattle in their respective locations (9) and provided as cyropreserved stabilates (37) by Gerald Buening (University of Missouri, Columbia, Mo.) and Ignacio Echaide (Instituto Nacional de Tecnología Agropecuaria, Rafaela, Argentina).…”

Section: Methodsmentioning

confidence: 99%

Molecular Basis for Variable Expression of Merozoite Surface Antigen gp45 among American Isolates ofBabesia bigemina

2001

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Immunization with the merozoite surface glycoprotein gp45 induces protection against challenge using the homologous Babesia bigemina strain. However, gp45 B-cell epitopes are highly polymorphic among B. bigemina strains isolated from different geographical locations within North and South America. The molecular basis for this polymorphism was investigated using the JG-29 biological clone of a Mexico strain of B. bigemina and comparison with the Puerto Rico, St. Croix, and Texcoco strains. The molecular size and antibody reactivity of gp45 expressed by the JG-29 clone were identical to those of the parental Mexico strain. gp45 cDNA and the genomic locus encompassing gp45 were cloned and sequenced from JG-29. The locus sequence and Southern blot data were consistent with a single gp45 copy in the JG-29 genome. The JG-29 cDNA expressed the full-length protein recognized by the gp45-specific monoclonal antibody 14/1.3.2. The genomes of the Puerto Rico and St. Croix strains of B. bigemina were shown to lack a closely related gp45-like gene by PCR using multiple primer sets and by Southern blots using both full-length and region-specific gp45 probes. This genomic difference was confirmed using unpassaged isolates from a 1999 disease outbreak in Puerto Rico. In contrast, the Texcoco strain retains a gp45 gene, encoding an open reading frame identical to that of JG-29. However, the Texcoco gp45 gene is not transcribed. These two mechanisms, lack of a closely related gp45-like gene and failure to transcribe gp45, result in generation of antigenic polymorphism among B. bigemina strains, and the latter mechanism is unique compared to prior mechanisms of antigenic polymorphism identified in babesial parasites.

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Identification of common surface antigens amongBabesia bigeminaisolates by using monoclonal antibodies

Cited by 31 publications

References 40 publications

In vitro cultivation of an African strain of Babesia bigemina , its characterisation and infectivity in cattle

In vitro cultivation of an African strain of Babesia bigemina , its characterisation and infectivity in cattle

Monoclonal Antibody MG96 Completely Blocks Plasmodium yoelii Developmentin Anophelesstephensi

Molecular Basis for Variable Expression of Merozoite Surface Antigen gp45 among American Isolates ofBabesia bigemina

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