Characterization of stage-specific and cross-reactive antigens from Eimeria acervulina by chicken monoclonal antibodies

The characterization of five chicken monoclonal antibodies (mAbs) that were developed against apical complex antigens of Eimeria acervulina sporozoites is realized and the mAbs reactivity to merozoites belonging to this species is tested. Using immuno-fluorescence assay (IFA), one mAb (HE-4) that recognized apical antigens common to sporozoites of E. acervulina and E. brunetti bound antigens localized on the apical tip of merozoites from all stages of development examined. The mAb 8E-1, reactive with antigens found on the apical tip of all chicken Eimeria sporozoites, also showed binding to antigens common to merozoites from all generations. Another mAb, 8C-3, which identified an antigen shared by sporozoites apical tip and sporocysts wall of E. acervulina reacted very weak and inconstantly with the merozoites from all generations whereas the mAbs 5D-11 and 8D-2 that recognized antigens shared by the sporozoites of E. acervulina and E. maxima (mAb 5D-11) and E. acervulina and E. brunetti (mAb 8D-2) did not react with the merozoites from any generation. Collectively, these results showed that the invasive stages of chicken Eimeria share cross reactive apical complex antigens which are inter-species and inter-generation-specific that might be components of a potential recombinant vaccine

Monoclonal antibodies for the diagnosis of Canine Mastocytoma

Mastocytomas are the most common malignant neoplasm in the dog; they are more aggressive than the mast cell tumors of other species. Therefore, it is imperative to develop a highly sensitive and specific immunoassay for clinical diagnosis of canine mastocytoma. The production and characterization of new mouse monoclonal antibodies (MAb 9-3 and MAb 80) directed against canine mastocytoma are reported here. By immunohistochemistry using fresh frozen tissue of tissue impression smears, we observed that the antigen recognized by MAb 9-3 is expressed exclusively on the surface and cytoplasmic granules of canine mastocytoma\ud but not on the mast cells in normal canine skin. MAb 80 did not compete for binding to mast cells in normal canine skin. Western blot assays performed with canine mastocytoma indicated that MAb 9-3 recognized the 74 kDa band, and MAb 80 recognized the 167 and 248 kDa bands. We studied the immunostaining pattern of impression smears with MAb 9-3 from 36 benign and malignant canine masses, including eight\ud samples of mastocytoma that were positive and other tumor samples that were negative by MAb 9-3. This report for the first time precisely characterizes a monoclonal antibody specific for canine mastocytoma, facilitating clinical and molecular investigation of canine mastocytoma

Characterization of monoclonal antibodies that recognize the Eimeria tenella Microneme Protein MIC2

The apicomplexan pathogens of Eimeria tenella cause coccidiosis, an intestinal disease of chickens that has a major economic impact on the poultry industry. Members of Apicomplexa share an assortment of unique secretory organelles (rhoptries, micronemes, and dense granules) that mediate invasion of host cells and the formation and modification of the parasitophorous vacuole. Among these, microneme protein 2 from E. tenella (EtMIC2) has a putative function in parasite adhesion to the host cell to initiate the invasion process. To investigate the role of EtMIC2 in host parasite interactions, the production and characterization of 12 monoclonal antibodies (mabs) produced\ud against recombinant EtMIC2 proteins is described. In an immunofluorescence assay, all mabs reacted with molecules belonging to the apical complex of sporozoites and merozoites of E. tenella, E. acervulina, and E. maxima. By Western blot analysis, the mabs identified a developmentally\ud regulated protein of 42 kDa corresponding to EtMIC 2 and\ud cross-reacted with proteins in developmental stages of E. acervulina. Collectively, these mabs are useful tools for the detailed investigation of the characterization of EtMIC2-related proteins in Eimeria species