Immunoproteomic and mass spectrometric analysis of Eimeria acervulina antigens recognized by antisera from chickens infected with E. acervulina, E. tenella or E. necatrix

Authors: LIU, JIN - Jiangxi Agricultural University; Tuo, Wenbin; WU, XIANGDONG - Jiangxi Agricultural University; XIONG, JIAMING - Jiangxi Agricultural University; YU, ENCHAO - Jiangxi Agricultural University; YIN, CHAO - Jiangxi Agricultural University; MA, ZHIWU - Jiangxi Agricultural University; LIU, LIHENG - Jiangxi Agricultural University

Submitted to: Parasites & Vectors
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/13/2020
Publication Date: 2/21/2020
Citation: Liu, J., Tuo, W., Wu, X., Xiong, J., Yu, E., Yin, C., Ma, Z., Liu, L. 2020. Immunoproteomic and mass spectrometric analysis of Eimeria acervulina antigens recognized by antisera from chickens infected with E. acervulina, E. tenella or E. necatrix. Veterinary Parasitology. 13:93. https://doi.org/10.1186/s13071-020-3965-y.
DOI: https://doi.org/10.1186/s13071-020-3965-y

Interpretive Summary: Coccidiosis is poultry disease caused by multiple Eimeria species, resulting in large economic losses to the global poultry industry. Due to developing anticoccidial drug resistance and drug residue issues, the objective of the present study was to identify shared antigens which may be used to develop a subunit, multivalent, cross-protective vaccine against most of the economically important Eimeria species. Eimeria sporozoite proteins separated by 2-dimensional electrophoresis were probed with antisera specific to 4 Eimeria parasites using Western blotting. Twenty immunologically common protein spots were identified and their protein sequences were demonstrated by mass spectrometry. The twenty unique and putative Eimeria proteins identified are novel and highly diverse in cellular localizations and biological functions, and none of which were previously reported. Further research on cross-protective potential of these proteins will aid in the development of vaccines against the most common and pathogenic Eimeria parasites.

Technical Abstract: Coccidiosis caused by Eimeria spp results in large economic losses to the global poultry industry. Due to anticoccidial drug resistance developing in the parasite and drug residue in poultry products as a serious concern for human safety, the objective of the present study was to identify common protective antigens which may be used as vaccine candidates in the development of subunit, multivalent, cross-protective vaccines against most of the economically important Eimeria species. Whole sporozoite proteins of Eimeria acervulina were prepared and analyzed by 2-dimensional gel electrophoresis (2-DE) followed by Western blotting using immune sera specific to E. tenella, E. acervulina, E. maxima or E. necatrix. Approximately 620 E. acervulina sporozoite protein spots were demonstrated by 2-DE with silver staining, among which 20 protein spots were recognized by immune sera from all 4 Eimeria species. The protein spots were then excised from the preparative gel and protein ID was performed by MALDI-TOF-MS/MS. The results showed that 20 unique, putative, E. accervulina proteins were identified, which include proteins of enzymatic properties (10) and those which are involved in protein translation, transport and trafficking, and ribosomal biogenesis and function (2 each). There is one protein may be involved in transcription and one heat shock protein. Two proteins contain predicted domains, but with no apparent functionality known. Our research has identified a diverse group of antigens immunologically common to all 4 Eimeria species, none of which was previously characterized and tested as vaccine candidates. Further research on immunogenicity and cross-protective potential of these individual proteins as vaccine candidates will aid in the development of vaccines against the most common and pathogenic Eimeria spp.