Applications of in Vitro Cultivation Systems for Plasmodium Falciparum in the Identification and Partial Characterization of Potentially Protective Exoantigens
Fajfar Whetstone, Carol Jo T.
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https://hdl.handle.net/2142/71340
Description
Title
Applications of in Vitro Cultivation Systems for Plasmodium Falciparum in the Identification and Partial Characterization of Potentially Protective Exoantigens
Author(s)
Fajfar Whetstone, Carol Jo T.
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)
Health Sciences, Immunology
Abstract
Plasmodium falciparum is the most widespread and pathogenic of four species of parasitic protozoa which cause malaria in humans. Comprehensive studies on the immune response to malaria, and development of effective insecticides, drugs and vaccines are of paramount importance for controlling this dreaded disease.
The asexual blood phase of the parasite life cycle is responsible for the clinical manifestations of malaria, and thus has been a primary focus of vaccine research. It has been shown that parasite antigens (exoantigens) are naturally released into the culture medium as the parasite grows in vitro. Immunization of experimental monkeys with crude or partially purified culture medium can induce protection against acute falciparum malaria.
Parasite exoantigens have been isolated from supernatant fluids of P. falciparum organisms in in vitro systems of human and rabbit/human origin. These antigens are proteins and/or glycoproteins in nature, and are susceptible to heating at 56(DEGREES)C for 30 min. Anion-exchange DEAE-cellulose chromatography has been used to partially purify fractions of supernatants from Indochina I cultures (human and rabbit/human systems) which contain parasite antigens. Crude and partially purified antigens derived from supernatants of Indochina I-infected cultures (human and rabbit/human systems) were immunogenic in mice and rabbits, and were partially protective in a monkey (human system). Target antigens identified in crude and partially purified supernatants of Indochina I-infected cultures include proteins with apparent molecular weights of 79 KD, 40 KD, 29 KD, 20 KD, and 15 KD (human system), and 48 KD, 30 KD, and 27 KD (rabbit/human system). The rabbit/human system of P. falciparum parasites was successively applied in development of a neutralization assay as an in vitro correlate of an antigen's protective potential in vivo. Lastly, the Geneve/SGE-1 strain of P. falciparum was adapted to in vitro growth in a squirrel monkey system as a prerequisite for in vivo adaptation in Bolivian squirrel monkeys. This allowed its use as a challenge strain in experiments designed to assess homologous and heterologous strain immunity.
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