High-level expression of the malaria blood-stage vaccine candidate Plasmodium falciparum apical membrane antigen 1 and induction of antibodies that inhibit erythrocyte invasion

被引:165
作者
Kocken, CHM
Withers-Martinez, C
Dubbeld, MA
van der Wel, A
Hackett, F
Blackman, MJ
Thomas, AW
机构
[1] BPRC, Dept Parasitol, NL-2288 GJ Rijswijk, Netherlands
[2] Natl Inst Med Res, Div Prot Struct, London NW7 1AA, England
[3] Natl Inst Med Res, Div Parasitol, London NW7 1AA, England
关键词
D O I
10.1128/IAI.70.8.4471-4476.2002
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Apical membrane antigen I (AMA-1) is a highly promising malaria blood-stage vaccine candidate that has induced protection in rodent and nonhuman primate models of malaria. Authentic conformation of the protein appears to be essential for the induction of parasite-inhibitory antibody responses. Here we have developed a synthetic gene with adapted codon usage to allow expression of Plasmodium falciparum FVO strain AMA-1 (PfAMA-1) in Pichia pastoris. In addition, potential N-glycosylation sites were changed, exploiting the lack of conservation of these sites in Plasmodium, to obtain high-level secretion of a homogeneous product, suitable for scale-up according to current good manufacturing procedures. Purified PfAMA-1 displayed authentic antigenic properties, indicating that the amino acid changes had no deleterious effect on the conformation of the protein. High-titer antibodies, raised in rabbits, reacted strongly with homologous and heterologous P.falciparum by immunofluorescence. In addition, purified immunoglobulin G from immunized animals strongly inhibited invasion of red blood cells by homologous and, to a somewhat lesser extent, heterologous P.falciparum.
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收藏
页码:4471 / 4476
页数:6
相关论文
共 25 条
  • [1] Immunisation with recombinant AMA-1 protects mice against infection with Plasmodium chabaudi
    Anders, RF
    Crewther, PE
    Edwards, S
    Margetts, M
    Matthew, MLSM
    Pollock, B
    Pye, D
    [J]. VACCINE, 1998, 16 (2-3) : 240 - 247
  • [2] [Anonymous], 1988, Antibodies: A Laboratory Manual
  • [3] PROTECTIVE IMMUNITY INDUCED IN SQUIRREL-MONKEYS WITH RECOMBINANT APICAL MEMBRANE ANTIGEN-1 OF PLASMODIUM FRAGILE
    COLLINS, WE
    PYE, D
    CREWTHER, PE
    VANDENBERG, KL
    GALLAND, GG
    SULZER, AJ
    KEMP, DJ
    EDWARDS, SJ
    COPPEL, RL
    SULLIVAN, JS
    MORRIS, CL
    ANDERS, RF
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1994, 51 (06) : 711 - 719
  • [4] Protective immune responses to apical membrane antigen 1 of Plasmodium chabaudi involve recognition of strain-specific epitopes
    Crewther, PE
    Matthew, MLSM
    Flegg, RH
    Anders, RF
    [J]. INFECTION AND IMMUNITY, 1996, 64 (08) : 3310 - 3317
  • [5] DEANS JA, 1982, CLIN EXP IMMUNOL, V49, P297
  • [6] VACCINATION TRIALS IN RHESUS-MONKEYS WITH A MINOR, INVARIANT, PLASMODIUM-KNOWLESI 66 KD MEROZOITE ANTIGEN
    DEANS, JA
    KNIGHT, AM
    JEAN, WC
    WATERS, AP
    COHEN, S
    MITCHELL, GH
    [J]. PARASITE IMMUNOLOGY, 1988, 10 (05) : 535 - 552
  • [7] STRUCTURAL STUDIES ON A PUTATIVE PROTECTIVE PLASMODIUM-KNOWLESI MEROZOITE ANTIGEN
    DEANS, JA
    JEAN, WC
    [J]. MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1987, 26 (1-2) : 155 - 166
  • [8] Garrity RR, 1997, J IMMUNOL, V159, P279
  • [9] The disulfide bond structure of Plasmodium apical membrane antigen-1
    Hodder, AN
    Crewther, PE
    Matthew, MLSM
    Reid, GE
    Moritz, RL
    Simpson, RJ
    Anders, RF
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (46) : 29446 - 29452
  • [10] Specificity of the protective antibody response to apical membrane antigen 1
    Hodder, AN
    Crewther, PE
    Anders, RF
    [J]. INFECTION AND IMMUNITY, 2001, 69 (05) : 3286 - 3294