Expression of the Plasmodium berghei ookinete protein Pbs21 in a baculovirus-insect cell system produces an efficient transmission blocking immunogen.
A surface protein of Plasmodium berghei ookinetes, Pbs21, was expressed in a baculovirus-insect cell system in cell culture and in Heliothis virescens larvae. Groups of BALB/c mice received two intraperitoneal inoculations of either i) Tris-buffer or homogenized H. virescens larvae infected with wild-type baculovirus; ii) enriched, homogenized ookinetes, or iii) homogenized H. virescens larvae expressing recombinant Pbs21 (rPbs21). All animals immunized with ookinetes or with rPbs21 had high titres of antibodies (IgG isotype) that bound to native Pbs21. The large majority of antibodies in immune sera of both groups recognized the antigen under non-reducing but not under reducing conditions. The predominant IgG-subclasses in mice immunized with ookinetes was IgG1 and in mice immunized with rPbs21, the subclasses were IgG1 and IgG2a. Immunization with rPbs21 reduced the infectivity of P.berghei to mosquitoes by 91% compared to a 99% reduction following immunization with ookinetes. This preliminary data indicate that rPbs21 expressed in this eukaryotic system induces a transmission-blocking immunity, which is more effective than that achieved using rPbs21 expressed in Escherichia coli (Matsuoka et al. 1994).
Margos
G
G
Department of Biology, Imperial College of Science, Technology and Medicine, London, UK.
Kurtenbach
K
K
Posnett
E
E
Barker
G C
GC
Matsuoka
H
H
Paton
M G
MG
Sinden
R E
RE
D000818:Animals; D000852:Anopheles / Q000276:immunology; D000913:Antibodies, Protozoan / Q000032:analysis / Q000096:biosynthesis*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D004175:Diptera / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis; D007303:Insect Vectors / Q000276:immunology; D008288:Malaria / Q000517:prevention & control* / Q000635:transmission; D051379:Mice; D008807:Mice, Inbred BALB C; D010962:Plasmodium berghei / Q000235:genetics / Q000276:immunology*; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D014162:Transfection / Q000235:genetics
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D007074:Immunoglobulin G; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C084246:ookinete surface antigen Pbs21, Plasmodium