Infectious Plasmodium sporozoites grown in vitro
AssigneeSanaria Inc.
InventorsAbraham G. Eappen, Stephen L Hoffman
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Abstract
The application is directed to in vitro-reared Plasmodium sporozoites of human host range wherein sporogony from gametocyte stage to sporozoite stage is external to mosquitoes, and methods of producing the same. Provided herein are in vitro-reared infectious Plasmodium sporozoites (SPZ) of human host range, particularly P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi, wherein sporogony from gametocyte stage to sporozoite stage is external to mosquitoes, and methods of producing the same.
Claims19
An in vitro-reared mature infectious Plasmodium sporozoites of human host range, wherein sporogony of the sporozoites from gametocyte stage to mature sporozoite stage is external to mosquitoes and absent any mosquito material, and wherein the genome of the in vitro-reared mature infectious Plasmodium sporozoites of human host range comprises a gene alteration or mutation, wherein the gene alternation or mutation disrupts the function of one or more genes, such that the in vitro-reared mature infectious Plasmodium sporozoites retain the ability to infect a host and invade hepatocytes within the liver, but arrest development in liver-stage.
A method of culturing genetically attenuated Plasmodium parasites of human host range in vitro during sporogonic development of said parasites, comprising:
- a. culturing the genetically attenuated Plasmodium parasites of human host range, which are at the gametocyte stage, to zygote stage in the presence of red blood cells in an exflagellation culture medium;
- b. collecting a mixture comprising zygotes, gametes, gametocytes;
- c. culturing said mixture on a substrate in an ookinete culture medium, wherein the genetically attenuated Plasmodium parasites of human host range differentiate to ookinetes;
- d. exchanging said ookinete medium with an oocyst medium;
- e. culturing the genetically attenuated Plasmodium parasites of human host range in the oocyst medium, wherein the genetically attenuated Plasmodium parasites of human host range differentiate to mature sporozoites;
- and
- f. harvesting the genetically attenuated Plasmodium parasites of human host range produced thereby, wherein said genetically attenuated Plasmodium parasites of human host range are at a mature sporozoite stage.
A culture comprising in vitro-reared Plasmodium parasites of human host range and a culture medium, wherein said parasites are at the zygote stage of development and have undergone sporogonic development in vitro.
Dependencies are inferred from the claim text.