Isolation of Theileria parva schizonts from infected lymphoblastoid cells.
This study set out to develop a rapid method for the isolation of schizonts from Theileria parva-infected bovine and buffalo lymphoblastoid cells. Parasitized lymphoblastoid cells were lysed by treatment with the cytolytic toxins, aerolysin and Ah-1 hemolysin, produced by Aeromonas hydrophila, and the schizonts were separated by Percoll density-gradient centrifugation. Light and electron microscopic examination showed that the isolated schizonts from lymphoblastoid cells infected with T. parva (Muguga) retained their normal morphology and were essentially free from host cell components. The schizonts also retained antigens as recognized by a series of anti-schizont monoclonal antibodies. The concentrations of toxin and Ficoll 400 in the lysis buffer which gave optimal cell lysis varied for 10 different infected cell lines tested.
D000818:Animals; D016782:Apicomplexa / Q000032:analysis / Q000302:isolation & purification* / Q000648:ultrastructure; D001427:Bacterial Toxins; D002020:Buffaloes; D002417:Cattle; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006460:Hemolysin Proteins; D015151:Immunoblotting; D008214:Lymphocytes / Q000469:parasitology*; D008854:Microscopy, Electron; D052899:Pore Forming Cytotoxic Proteins; D011506:Proteins / Q000032:analysis; D013801:Theileriasis / Q000469:parasitology
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D052899:Pore Forming Cytotoxic Proteins; D011506:Proteins; C032676:aerolysin