Isolation and purification of the schizont stage of Theileria annulata from host leukocytes through novel biochemical techniques.

Document Type: Research Article

Authors

1 Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN

2 Department of Chemistry, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN

3 Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, I.R. IRAN

4 Immunology Section, Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN

Abstract

The intracellular protozoan parasite, Theileria annulata, induces uncontrolled proliferation and transformation in bovine B lymphocytes and monocytes in blood circulation and lymph nodes of host cells. This uncontrolled replication happens in macroschizont stage of life cycle of the parasites. The development of a rapid and efficient technique is likely to necessitate for isolation of purified schizonts from host cells. This is necessary for isolation of highly purified RNA, protein or glycoproteins of schizonts from host cells. This study aimed to isolate the purified schizont based on the aerolysin–nocodazole technique. Aerolysin which was purified from gram-negative pathogen aeromonas hydrophila has an ability to form discrete channels and unstable eukaryotic cell membrane in 0C• and low concentration. Nocodazole used for parasite separation from microtubule network of infected lymphocytes and monocytes. In purified schizonts no large nucleus of host cell visualized in giemsa and DAPI staining. The isolated schizonts were free and intact from host cells. Intact T.annulata schizonts obtained from this study are suitable for purification of RNAs, proteins, glycolipids and glycoproteins of schizonts free of host cell debris.

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