Identification and Partial Characterization of the Antigens of Anaplasma Marginale (Florida) and Anaplasma Caudatum (Illinois) (Immunoblotting, Hemoparasite, Anaplasmasis)
Adams, John Howard
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https://hdl.handle.net/2142/71333
Description
Title
Identification and Partial Characterization of the Antigens of Anaplasma Marginale (Florida) and Anaplasma Caudatum (Illinois) (Immunoblotting, Hemoparasite, Anaplasmasis)
Author(s)
Adams, John Howard
Issue Date
1986
Department of Study
Veterinary Medical Science
Discipline
Veterinary Medical Science
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Veterinary Science
Abstract
Shared and unique antigens were identified and partially characterized for two morphologically and antigenically distinct isolates of Anaplasma, A. marginale (Florida) (FAM) and A. caudatum (Illinois) (IAC). Crude CA-like antigens were prepared from the parasitemic blood of each calf, separated by SDS-PAGE, electroblotted onto nitrocellulose, and detected using immune bovine sera and an avidin-biotin-peroxidase assay with biotinylated rabbit anti-goat IgG (cross-reacted with cattle, sheep and deer antibodies). IAC antigens were 200 kilodalton (kd), 100 kd, 96 kd, 75 kd, 47 kd, 43 to 38 kd, and 27 kd, detected using anti-IAC and anti-FAM sera. FAM antigens were 108 kd, 100 kd, 96 kd, 91 kd, 75 kd, 47 kd, 43 to 38 kd, and 27 kd, detected using anti-FAM sera and, except for the 91 kd, with anti-IAC sera. The 91 kd is an FAM isolate-specific antigen. The 108 kd, 100 kd, and 96 kd are group-specific antigens of A. marginale and A. centrale. The 43 to 38 kd is genus-specific. The 96, 91, and 75 kd antigens were not soluble in nonionic detergents but were soluble in SDS and urea. The 108 and 100 kd antigens were totally soluble in nonionic detergents. The 41 and 47 kd antigens were partially soluble in nonionic detergents, with a portion was only solubilized with SDS or urea. A 2-mercaptoethanol-sensitive 200 kd antigen was present in the insoluble fraction from the nonionic detergents. With 2-dimensional electrophoresis, the 41 kd antigen was composed of 4 individual proteins. The Anaplasma antigens had a neutral pI, were heat stable, and lost their antigenicity after treatment with chymotrypsin. The detectable antibody response was highly variable within each animal and between animals. IFA titers did not appear to correlate with the antibody response to any particular antigen. The 96 and 91 kd antigens elicited the most constant antibody response. Detectable antibody to the 108 kd antigen disappeared in the FAM-infected animals after the first or second recrudescent parasitemias.
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