Model structure of the immunodominant surface antigen of Eimeria tenella identified as a target for sporozoite-neutralizing monoclonal antibody

Jahn, Doreen; Matros, Andrea; Bakulina, Anastasia; Tiedemann, Jens; Schubert, Ulrike; Giersberg, Martin; Haehnel, Sigrun; Zoufal, Karen; Mock, Hans‐Peter; Kipriyanov, Sergey M.

doi:10.1007/s00436-009-1437-6

Cited by 50 publications

(47 citation statements)

References 49 publications

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“…All assays were performed in triplicate. The deduced percentages of infected cells in the presence/absence of inhibitory antibody were used for the calculation of the inhibition rates as previously described (Jahn et al 2009). …”

Section: Invasion Inhibition Assaymentioning

confidence: 99%

Identification and partial characterization of a serine protease inhibitor (serpin) of Eimeria tenella

et al. 2011

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Serine protease inhibitors (serpins) mediate many biological processes, including immune responses to pathogenic infection. In this study, a member of the serpin superfamily was identified from the common poultry parasite Eimeria tenella by expressed sequence tag analysis and the rapid amplification of cDNA ends technique. The full-length cDNA was 1,918 bp and had an open reading frame of 1,248 bp encoding a polypeptide of 415 amino acids with the theoretical isoelectric point of 5.26 and predicted molecular weight of 45.5 kDa. Real-time quantitative PCR analysis revealed that the serpin gene was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts, and second-generation merozoites). The sequence encoding the mature protein was amplified by PCR, cloned into the pET28(a) vector, and expressed in Escherichia coli. Specific antiserum generated against the recombinant protein was prepared and used to determine invasion inhibition capacity and localization; the results suggested that the serpin may play an important role in invasion and survival of the sporoziotes in the host.

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Section: Invasion Inhibition Assaymentioning

confidence: 99%

Identification and partial characterization of a serine protease inhibitor (serpin) of Eimeria tenella

et al. 2011

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“…As a result, scFvs were generated, in which the two variable regions, VH and VL, are artificially linked by a flexible polypeptide (Fig. 1) [27,28]. This and similar approaches may still be considered as highly convenient once the negative European public perception on biofarming eases its halt in the development of local companies specialized in antibody production.…”

Section: Antibody Fragmentsmentioning

confidence: 99%

Manufacturing antibodies in the plant cell

Orzáez

Granell

Blázquez

2009

Biotechnology Journal

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International audiencePlants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently produce mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being coped with. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production

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“…Invasion inhibition assays were based on invasion of DF-1 cells by E. tenella sporozoites [30], [33]. Freshly isolated sporozoites were irreversibly labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE-Molecular Probes, Beyotime, CN) according to the manufacturer's instruction.…”

Section: Methodsmentioning

confidence: 99%

“…All assays were performed in triplicate. Percentages of infected cells in the presence or absence of inhibitory antibody were used to calculate inhibition rates as previously described [30], [33].…”

Section: Methodsmentioning

confidence: 99%

Molecular Characterization and Analysis of a Novel Protein Disulfide Isomerase-Like Protein of Eimeria tenella

et al. 2014

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Protein disulfide isomerase (PDI) and PDI-like proteins are members of the thioredoxin superfamily. They contain thioredoxin-like domains and catalyze the physiological oxidation, reduction and isomerization of protein disulfide bonds, which are involved in cell function and development in prokaryotes and eukaryotes. In this study, EtPDIL, a novel PDI-like gene of Eimeria tenella, was cloned using rapid amplification of cDNA ends (RACE) according to the expressed sequence tag (EST). The EtPDIL cDNA contained 1129 nucleotides encoding 216 amino acids. The deduced EtPDIL protein belonged to thioredoxin-like superfamily and had a single predicted thioredoxin domain with a non-classical thioredoxin-like motif (SXXC). BLAST analysis showed that the EtPDIL protein was 55–59% identical to PDI-like proteins of other apicomplexan parasites. The transcript and protein levels of EtPDIL at different development stages were investigated by real-time quantitative PCR and western blot. The messenger RNA and protein levels of EtPDIL were higher in sporulated oocysts than in unsporulated oocysts, sporozoites or merozoites. Protein expression was barely detectable in unsporulated oocysts. Western blots showed that rabbit antiserum against recombinant EtPDIL recognized only a native 24 kDa protein from parasites. Immunolocalization with EtPDIL antibody showed that EtPDIL had a disperse distribution in the cytoplasm of whole sporozoites and merozoites. After sporozoites were incubated in complete medium, EtPDIL protein concentrated at the anterior of the sporozoites and appeared on the surface of parasites. Specific staining was more intense and mainly located on the parasite surface after merozoites released from mature schizonts invaded DF-1 cells. After development of parasites in DF-1 cells, staining intensified in trophozoites, immature schizonts and mature schizonts. Antibody inhibition of EtPDIL function reduced the ability of E. tenella to invade DF-1 cells. These results suggested that EtPDIL might be involved in sporulation in external environments and in host cell adhesion, invasion and development of E. tenella.

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Model structure of the immunodominant surface antigen of Eimeria tenella identified as a target for sporozoite-neutralizing monoclonal antibody

Cited by 50 publications

References 49 publications

Identification and partial characterization of a serine protease inhibitor (serpin) of Eimeria tenella

Identification and partial characterization of a serine protease inhibitor (serpin) of Eimeria tenella

Manufacturing antibodies in the plant cell

Molecular Characterization and Analysis of a Novel Protein Disulfide Isomerase-Like Protein of Eimeria tenella

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