Induction of anti-malarial transmission blocking immunity with a recombinant ookinete surface antigen of Plasmodium berghei produced in silkworm larvae using the baculovirus expression vector system.
We have studied Pbs21, a major ookinete surface protein of Plasmodium berghei, for the development of a model transmission blocking immunogen. In the mouse, recombinant Pbs21 expressed in the Escherichia coli expression system (EcrPbs21) is not as effective in inducing transmission blocking antibodies as native Pbs21 (nPbs21), possibly because of differences in post-translational processing between EcrPbs21 and nPbs21. In an attempt to improve the efficacy of the recombinant molecule, we describe here the use of a baculovirus expression vector system in the silkworm Bombyx mori. Following an injection of recombinant baculovirus containing Pbs21 cDNA, B. mori larvae produced recombinant Pbs21 (BmrPbs21) with a molecular weight indistinguishable from nPbs21. Fifty micrograms of BmrPbs21 could be purified from the hemolymph of each infected larva using affinity chromatography. Immunization of Balb/c mice with BmrPbs21 induced high anti-BmrPbs21 and anti--ookinete antibodies but low anti-EcrPbs21 antibody. In contrast, EcrPbs21 induced high anti--EcrPbs21 antibody but low anti-BmrPbs21 and anti-ookinete antibodies. This suggests that most B-cell epitopes on nPbs21 are conformational and that many of the linear epitopes in EcrPbs21 are not normally exposed in nPbs21. Oocyst formation in Anopheles stephensi mosquitoes, which fed on mice immunized with purified BmrPbs21 and infected with P. berghei, was blocked by 85.5-97.1%. These results suggest that the baculovirus-silkworm system produces useful quantities of recombinant Pbs21 which in limited studies is structurally and immunogenically indistinguishable from the native molecule.
Matsuoka
H
H
Department of Medical Zoology, Mie University School of Medicine, Tsu, Japan.
Kobayashi
J
J
Barker
G C
GC
Miura
K
K
Chinzei
Y
Y
Miyajima
S
S
Ishii
A
A
Sinden
R E
RE
D000818:Animals; D000913:Antibodies, Protozoan / Q000032:analysis; D000953:Antigens, Protozoan / Q000096:biosynthesis* / Q000276:immunology / Q000627:therapeutic use*; D000954:Antigens, Surface / Q000096:biosynthesis / Q000276:immunology / Q000627:therapeutic use; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D012831:Bombyx / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D005260:Female; D005822:Genetic Vectors; D008288:Malaria / Q000276:immunology / Q000517:prevention & control* / Q000635:transmission*; D017780:Malaria Vaccines / Q000096:biosynthesis* / Q000276:immunology / Q000627:therapeutic use*; D051379:Mice; D008807:Mice, Inbred BALB C; D010962:Plasmodium berghei / Q000276:immunology*; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000627:therapeutic use
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D017780:Malaria Vaccines; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C084246:ookinete surface antigen Pbs21, Plasmodium