Browsing by Author "Cowan, K.M."
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Item Detection of Antibody to Theileria Parva Schizonts and Cell Surf Ace Membrane Antigens of Infected L Ymphoblastoid Cells by Immunoperoxidase Techniques(1984) Cowan, K.M.; Dolan, T.T.; Teale, A.J.; Youngs, A.S.; Stagg, D.A.; Groocock, C.M.; US Department of Agriculture and Corporative Research British Overseas Development Administration KARI Veterinary Research Department Muguga KikuyuPeroxidase-labeled antibody procedures were described for detecting bovine antibodies reactive with intracellular Theileria parva schizonts and cell surface membrane antigens of infected lymphoblastoid cells. Indirect tests were performed where the reacting bovine antibodies were localized with affinity purified rabbit-anti-bovine IgG coupled to horseradish peroxidase. A 4- to 8-fold increase in sensitivity for detecting bovine antibodies was obtained with unlabeled rabbit-anti-bovine IgG which in turn was detected with peroxidase labeled goat-anti-rabbit IgG. The T. parva infected cells used as antigen were attached to poly-I-lysine treated glass slides and all reaction steps were performed on the slides. The intracellular schizonts and cell surface staining reactions were dependent upon the status of the cells; acetone-fixed cells were required for schizont reactions and viable unfixed cells for cell surface membrane reactions. Sera from cattle stimulated in various ways with T. parva were examined by the techniques. Cattle infected by stabilate inoculation or inoculated with infected autologous lymphoblastoid cells developed relatively high levels of antibody to schizonts, but no detectable antibody to cell . surface membrane antigens. This would indicate that parasite antigens do not occur on the surface of infected lymphoblasts. Cattle inoculated with infected allogeneic lymphoblasts developed low-levels of antibody to schizonts and readily demonstrable antibody to cell surface antigens. The immunoperoxidase procedures have certain advantages over immunofluorescence in that light microscopy is used; therefore, the reactions do not fade which permits a more detailed examination and provides a relatively permanent record, the preparations can be counterstained, and the reagents may be used for immunoelectron- microscopy. The procedures could provide suitable alternatives to immunofluorescence methods for East Coast fever investigations and other systems having intracellular and/or cell surface membrane antigens.Item Detection of Antibody to Theileria Parva Schizonts and Cell Surface Membrane Antigens of Infected(Elsevier, 1984) Cowan, K.M.; Dolan, T.T.; Teale, A.J.; Young, A.S.; Stagg, D.D.; Groocock, C.M.;Peroxidase-labeled antibody procedures were described for detecting bovine anti-bodies reactive with intracellular Theileria parva schizonts and cell surfacesmembrane antigens of infected lymphoblastoid cells. Indirect tests were performed where the reacting bovine antibodies were localized with affinity purified rabbit-anti-bovine IgG coupled to horseradish peroxidase. A 4- to 8-fold increase in sensitivity for detecting bovine anti-bodies was obtained with unlabeled rabbit-anti-bovine IgG which in turn was detected with peroxidase labeled goat-anti-rabbit IgG. The T. parva infected cells used as antigen were attached to poly-l-lysine treated glass slides and all reaction steps were performed on the slides. The intracellular schizonts and cell surface staining reactions were dependent upon the status of the cells; acetone-fixed cells were required for schizont reactions and viable unfixed cells for cell surface membrane reactions. Sera from cattle stimulated in various ways with T. parva were examined by the techniques. Cattle infected by stabilate inoculation or inoculated with infected autologous lymphoblastoid cells developed relatively high levels of antibody to schizonts, but no detectable antibody to cell surface membrane antigens. This would indicate that parasite antigens do not occur on the surface of infected lymphoblasts. Cattle inoculated with infected allogeneic lymphoblasts developed low-levels of antibody to schizonts and readily demonstrable antibody to cell surface antigens. The immunoperoxidase procedures have certain advantages over immunofluorescence in that light microscopy is used; therefore, the reactions do not fade which permits a more detailed examination and provides a relatively permanent record, the preparations can be counterstained, and the reagents may be used for immunoelectron-microscopy. The procedures could provide suitable alternatives to immunofluorescence methods for East Coast fever investigations and other systems having intracellular and/or cell surface membrane antigens.Item Effect of Merthiolatelon Agar Gel Diffusion Prescription Reactions with Foot-and-Mouth Disease Virus(1966) Cowan, K.M.suspension of foot and mouth disease virus obtained from infected cattle guinea pigs and tissue cultures have been reported to give two precipitin bands by agar gel immuno diffusion tec techiniques .Item Surface Membrane Components of Theileria Spp Piroplasms and Associated Complement Fixation Activity(1982) Allsopp, B.A.; Cowan, K.M.; Veterinary Research Department, Kenya Agricultural Research Institute, PO Box 32, Kikuyu, KenyaTheileria parva (parva) piroplasms were examined for complement fixation (CF) activity with sera from cattle immune to T parva (parva) (Muguga). The piroplasms were found to be highly antigenic, and the antigens responsible for this CF reactivity were completely insoluble in aqueous media. The antigens were traced through a variety of fractionation procedures and the fractions examined by electrophoresis in the presence of sodium docedyl sulphate. Antigenically active preparations were found always to contain two polypeptides having molecular weights of 21, 500 and 34, 000. When intact piroplasms were surface labelled with iodine 125 these same two polypeptides were (he most heavily labelled, which suggested that the antigens responsible for the CF reactivity were plasma-membrane bound. Electrophoretic analysis of fractions containing plasma membrane components from piroplasms of three species of Theileria (T parva [parva], T mutans and T taurotragi) enabled interspecific differentiation to be made. Three stocks of T parva (parva) could not be differentiated by this means; such different stocks, from various locations within East Africa, appeared to be identical chemically.Item A third antigenic component associated with foot-and-mouth disease infection(1966) Cowan, K.M.; Graves, J.H. ; Plum Island Animal Disease Laboratory; Animal Disease and Parasite Research Division; Agricultural Research Service; U. S. Department of Agriculture, Greenport, Long Island, New York 11944A third antigen is produced in tissue cultures of baby hamster kidney (BHK) cells and tissues of guinea pigs and cattle infected with type A foot-and-mouth disease virus (FMDV). This antigen is distinct from the recognized 140 Sand 12 S antigenic components of FMDY. It does not appear to be a virus constituent but is produced as a consequence of the infection; therefore, it has been termed the "virus infection associated" (VIA) antigen. Guinea pigs and cattle infected with Fl\IDV produce antibody to VIA antigen that can be demonstrated by agar gel precipitin techniques; whereas animals immunized with inactivated preparations of Fl\IDY do not. This further suggests it to be an infection-associated material.
