Vaccine for, diagnostic assay for and method of treating parasit

Chemistry: molecular biology and microbiology – Vector – per se

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536 231, C07H 2102, C12N 1500

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060279346

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
The present invention relates generally to the use of a novel vaccine to treat and novel assays to diagnose a host of hemoflagellate protozoal infections, including infections arising from a variety of African trypanosomes.
2. Description of the Related Art
The World Health Organization estimates that at least 37 million people are afflicted annually with a hemoflagellate protozoal infection, some fatally. As many, if not more, domestic protein producing animals are also similarly afflicted. A vaccine has not been produced prior to now because (1) the parasite's ability to change it's surface coat, in the case of African trypanosomes, (2) the ability to induce immunosuppression in all trypanosomes and Leishmania, and (3) the ability of the parasite to hide intracellularly at some point in the infection. Previously, in African trypanosomes, the only immunity that could be obtained was to a single isolate from a single strain. This immunity was ineffective against any other isolate from the same strain or any other strain or species of African trypanosomes.
Olenick et al., Infect. Immun., 56: 92 (1988), report that a mixture of proteins of approximately 80, 74, 40 and 25 kDa, originally derived from the flagellar pocket of Trypanosoma brucei rhodesiense gives protection across variants of Trypanosoma brucei rhodesiense. However, it is unclear from Olenick et al. whether the antigens are common to all African trypanosomes or, indeed, to all hemoflagellate protozoa. In this regard, the mere fact that Olenick et al. achieved immunoprotection across variants of Trypanosoma brucei rhodesiense does not alone suggest that the antigens involved are common to other species of African trypanosomes or to other hemoflagellate protozoa.
Possibly, a similar protein gives immunoprotection across species of African trypanosomes. See, Powell, Med. J. Zambia, 10: 32 (1976); Powell et al., Med. J. Zambia, 12: 67 (1978); and Powell, Experientia, 34: 1450 (1978). Ruiz et al., Immunol. Lett., 12: 1 (1986), report the flagellar antigens in Trypanosoma cruzi to be immunoprotective against Trypanosoma cruzi, but antibodies to this antigen also give antibodies to mammalian cardiac tissue (see U.S. Pat. No. 4,298,590). This flagellar pocket site on the parasite, in African trypanosomes, is also reported to be involved in receptor-mediated endocytosis. See, Coppens et al., Proc. Natl. Acad. Sci. USA, 85: 6753 (1988); Webster, Eur. J. Cell Biol., 49: 295 (1989); and Webster et al., Eur. J. Cell Biol., 49: 303 (1989).
Published patent application WO 92/22325 teaches that a unique protein antigen originally derived from Trypanosoma brucei rhodesiense organisms can be used to induce immunoprotection against all African trypanosomes, Trypanosoma cruzi, all species of Leishmania, and other parasitic hemoflagellate protozoa. However, sequence and other data that would facilitate the preparation of the antigen by recombinant means are not disclosed. Preparation by recombinant means is necessary to produce the vaccine in sufficient quantities to meet current needs and in a cost effective manner.


SUMMARY OF THE INVENTION

It was an object of the present invention to provide a vaccine generally useful to treat hemoflagellate protozoal infections.
It was another object of the present invention to provide a method for immunoprotection against hemoflagellate protozoal infections generally.
It was another object of the present invention to provide a means for diagnosing hemoflagellate protozoal infections in patients suffering therefrom.
It was a further object to provide such a vaccine and such methods relatively inexpensively.
These and other objects have now been met with the present invention wherein it has now been discovered that an immunodominant fragment of a previously described antigen derived from the flagellar pocket of Trypanosoma brucei rhodesiense gives immunoprotection across the genus of hemoflagellate protozoa. The nucleotide and amino acid sequence of the inventive immunodomi

REFERENCES:
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Boehringer Mannheim Chemicals Catalog, see p. 557, 1991.
Kumar et al. PNAS 87:1337-1341, Feb. 1990.
Lazar et al. Mol. Cell. Biol. 8(3):1247-52, Mar. 1988.
Parsons, J.A. (ed) "Peptide Hormones", published by University Park Press, see Chapter 1, pp. 1-7 by Rudinger, Jun. 1976.
Burgess et al. J. Cell Biol. 111:2129-2137, Nov. 1990.
Infection and Immunity, "A Flagellar Pocket Membrane Fraction from Tryponosoma Brucei rhodesinse: Immunogold Localization and Nonvariant Immunoprotection" Olenick et al.; Jan. 1988, pp. 92-98.
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"Experimental Immunity Against Trypanosomiasis", C. Powell; Jan. 1978, p. 1450.
Immunology Letters, 12; Ruiz et al.; "Immunoprotection of Mice Against Trupanosoma Cruzi with a Lyophilized Flagellar Fraction of the Parasite Plus Adjuvant", 1986, pp. 1-4.
Proc. Natl. Acad. Sci. USA; Coppens et al.; "Receptors for the Host Low Density Lipoprotiens on the Hemoflagellate Trypanosoma Brucei: Purification and Involvement in the Growth of the Parasite", Sep. 1988, pp. 6753-6757.
CRC Press, Inc.; E.T. Maggio; "Immunochemistry Techniques", 1987, pp. 167-179.
Journal of Biological Chemistry, vol. 267, No. 6, Feb. 25, 1992; J. Schatzle et al.; Molecular Cloning and Characterization of the Structural Gene Coding for the Developmentally Regulated Lysosomal Enzyme, .alpha.-mannosidase, in Dictyostekuym Discoideum:, pp. 4000-40007.
European Journal of Cell Biology, 49; 1989; "Endocytosis by African Trypanosomes. I. Three-dimensional Structure of the Endocytic Organelles in Trypanosoma Brucei and T. Congolense"; pp. 295-302.
European Journal of Cell Biology, 49; 1989; "Endocytosis by African Trypanosomes. II. Occurrence in Different Life-cycle Stages and Intracellular Sorting"; pp. 303-310.

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