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Title: Characterization of monoclonal antibodies that recognize the Eimeria tenella microneme protein MIC2

Author
item SASAI, KAZUMI - JAPAN
item Fetterer, Raymond
item Lillehoj, Hyun
item MATSUURA, SATOMI - JAPAN
item CONSTANTINOIU, CONSTANTIN - JAPAN
item MATSUBAYASHI, MAKOTO - JAPAN
item HIROYUKI, TANI - JAPAN
item EIICHIROK, BABA - JAPAN

Submitted to: Journal of Parasitology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/1/2008
Publication Date: 4/1/2008
Citation: Sasai, K., Fetterer, R.H., Lillehoj, H.S., Matsuura, S., Constantinoiu, C.C., Matsubayashi, M., Hiroyuki, T., Eiichirok, B. 2008. Characterization of monoclonal antibodies that recognize the Eimeria tenella microneme protein MIC2. Jrnl. of Parasitology. Available: http://www.ncbi.nlm.nih.gov/sites/entrez PMID: 18576850.

Interpretive Summary: Poultry coccidiosis is the result of several different species of a protozoan intestinal parasite which causes considerable annual losses to the poultry industry. The primary control for the disease is through application of medications in the feed as birds are raised in confinement housing. The controls by medications are becoming less effective because of increased resistance to the drugs and less desirable due to concerns about drugs possibly remaining in the meat and within the environment. New control methods are needed. Coccidian parasites are intracellular organisms and possess a precise an unique method to invade host cells which requires the release of a specific proteins called micronemes. The current research developed and characterized monoclonal antibodies to a recombinant microneme protein, EtMIC2. Using staining techniques these antibodies were shown to bind to the anterior of the three species of coccidian parasites suggesting localization of the EtMIC2 protein in this area. In addition, protein analysis indicated that the antibodies recognized EtMIC2 in homogenates of developmental stages of the parasite. These results indicate that the EtMIC2 protein is conserved in three species of coccidia and the monoclonal antibodies developed in this study can be used to further investigate this important class of proteins.

Technical Abstract: The Apicomplexan pathogens of the species Eimeria cause coccidiosis, an intestinal disease of chickens, which has a major economic impact on the poultry industry. Members of the phylum Apicomplexa share an assortment of unique secretory organelles (rhoptries, micronemes and dense granules) that mediate invasion of host cells and formation and modification of the parasitophorous vacuole. Among these, microneme protein 2 from Eimeria 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 against recombinant EtMIC2 proteins is described. All mabs reacted with molecules belonging to the apical complex of sporozoites and merozoites of E. tenella, E. acervulina and E. maxima in immunofluorescence assay. By Western blot analysis, the mabs identified a developmentally regulated protein of 42 kDa corresponding to EtMIC 2 and cross-reacted with proteins in developmental stages of E. acervulina. Collectively, these mabs are useful tools for the detail investigation of the characterization of EtMIC2 related proteins in Eimeria species.