Plasmodium yoelii: axenic development of the parasite mosquito stages.
Study of the parasite mosquito stages of Plasmodium and its use in the production of sporozoite vaccines against malaria has been hampered by the technical difficulties of in vitro development. Here, we show the complete axenic development of the parasite mosquito stages of Plasmodium yoelii. While we demonstrate that matrigel is not required for parasite development, soluble factors produced and secreted by Drosophila melanogaster S2 cells appear to be crucial for the ookinete to oocyst transition. Parasites cultured axenically are both morphologically and biologically similar to mosquito-derived ookinetes, oocysts, and sporozoites. Axenically derived sporozoites were capable of producing an infection in mice as determined by RT-PCR; however, the parasitemia was significantly much less than that produced by mosquito-derived sporozoites. Our cell free system for development of the mosquito stages of P. yoelii provides a simplified approach to generate sporozoites that may be for biological assays and genetic manipulations.
Porter-Kelley
Johanna M
JM
Department of Microbiology and Immunology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA. porterkelleyj@wssu.edu
Dinglasan
Rhoel R
RR
Alam
Uzma
U
Ndeta
George A
GA
Sacci
John B
JB
Azad
Abdu F
AF
D000818:Animals; D000852:Anopheles / Q000469:parasitology; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000032:analysis / Q000096:biosynthesis; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D003094:Collagen; D017077:Culture Media, Conditioned; D016054:DNA, Protozoan / Q000032:analysis; D004331:Drosophila melanogaster; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D022781:Hepatocytes / Q000469:parasitology; D007797:Laminin; D008288:Malaria / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D018512:Parasitemia / Q000469:parasitology; D010967:Plasmodium yoelii / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017077:Culture Media, Conditioned; D016054:DNA, Protozoan; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen