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Related post: CHECK APPROPRIATE 80X(ES) C (a) HUMAN SUBJECTS D (al) MINORS n (a2) INTERVIEWS a Diamox Cost (b) HUMAN TISSUES D (c) NEITHER SUMMARY OF WORK (200 words or less - underline keywords) Animals have been successfully vaccinated against sexual and asexual malaria parasites . In addition, Plasmodium falciparum , the major malaria of man, has been grown in culture for the production of asexual stages and gametocytes. The present study Diamox Sr will 1) Improve culture conditions for the production of large numbers of gametocytes , gametes and merozoites of P^. falciparum ; 2) Study of the physiology of exf lagellation , fertilization and invasion of red cells by malaria merozoites; 3) Characterize structure , function and immuno - genicity of surface determinants on gametes and merozoites; 4) Evaluate gamete vaccines in model systems for identification of the best antigens and adjuvants . 5) Characterize red cell membrane of infected red cells. 25-65 PHS-6040 (Rev. 10-76) Serial No. ZOl AI 00174-03 LPD Project Description ; Objectives ; 1. Use of cultured Plasmodium falciparum to Diamox Online study the production and maturation of sexual and asexual blood stages of the parasite. The work will include studies on Diamox Sequels controls of differentiation of the parasite stages and the identification and expression of antigens characteristic of each parasite state. 2 . Attempts will be made to measure the natural variation between genetically unrelated isolates of P^. falciparum with respect to various physiological and antigenic properties of the parasites. Variation will be analysed at a physiological and molecular level and also genetically by cross fertilization when it becomes possible to produce gametocytes infective to mosquitoes from cultured material. 3. Diamox Iv Study of structure, function and immunogenicity of antigenic determinants on the surface of extracellular sexual and asexual parasites. This will include the use of monospecific antibodies produced by hybridomas and lectins for parasite component purification. 4. Study of immunogenicity of crude antigenic preparations of sexual and asexual Buy Diamox Online stages in model systems. 5. Study changes in the membrane of malaria infected red cells. Each objective is interrelated with the others: Culture will be needed for production of antigenic material for analysis of both sexual and asexual erythrocytic parasites. Structure and function of parasite deter- minants Diamox Price will be studied in the framework of parasite physiology. Successful immunization with whole parasite preparations will identify sources of more purified immunogenic materials. Methods : 1. Continuous culture of red cell stages of Plasmodium falciparum ; isolation of sexual and asexual stages; purification of merozoites and gametes . 2. Use of model systems: P. gallinaceum -chicken for study of gametes and gamete immunity; P^. knowlesi- rhesus for study of mechanism of invasion and gamete immunity. 3. Use of polyspecific antibody to parasite antigens for immune precipitation; high-resolution gel electrophoresis and electrofocussing separation techniques; i lactoperoxidase surface labelling; labelling of sugars and other components by various methods; general metabolic labelling of all parasite components; and lectin column purification of malarial antigens. 25-66 Serial No. ZOl AI 00174-03 LPD 4. Monospecific antibody produced in hybridomas for analysis of parasite surfaces and isolation of surface determinants. 5. P^. falciparum parasitized red cells with (K+) and without (K-) will be layered over human endothelial cells in culture and the percent adherence measured. 6. Use of membrane feeding to feed malaria infected blood to mosquitoes in the presence of substances, e.g. hybridoma antibodies, to be tested for their effect on infectivity of the parasites to mosquitoes. Results of Research ; Asexual Parasites 1. The majority of animals immune to asexual infection with P. knowlesi whether by immunization with Diamox Sequel antigen in Freund's complete adjuvant OJ^ ^V repeated infection precipitate a particular malarial antigen by crossed- Immunoelectrophoresis. Non- immune animals that were infected only one time or immunized with antigens in other adjuvants (e.g., Freund's incomplete adjuvant or BCG) did not precipitate this antigen. 125 2. I-lactoperoxidase labelled merozoites give a characteristic profile on acrylamide gel electrophoresis. Trypsin-treated merozoites that can no longer attach to or invade red cells lose the high molecular weight labelled proteins (150,000 D and 105,000 D) with the appearance of new bands at 70,000 D and 62,000 D. Thus the high molecular weight proteins are candidates for receptor proteins. 3. Monoclonal antibodies have been developed which bind to the surface of merozoites by immunofluorescence and by ultrastructural studies of ferritin antibody coated merozoites. These antibodies agglutinate merozoites but only one Diamox 500 Mg has an effect on invasion. Three of these hybridomas precipitate from metabolically labbelled schizonts a single glycoprotein with a molecular weight greater than 200,000 D. Pecularly, surface labelled merozoites contain no protein above 150,000 D whereas schizonts contain this 200,000 D protein. It is possible that proteolytic digestion during labelling causes loss of the high molecular weight band
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