Development, expression, and murine testing of a multistage Plasmodium falciparum malaria vaccine candidate
Identifieur interne : 003704 ( Main/Merge ); précédent : 003703; suivant : 003705Development, expression, and murine testing of a multistage Plasmodium falciparum malaria vaccine candidate
Auteurs : Ya Ping Shi [États-Unis] ; Parimal Das [Inde] ; Brian Holloway [États-Unis] ; Venkatachalam Udhayakumar [États-Unis] ; Jon Eric Tongren [États-Unis] ; Francisco Candal [États-Unis] ; Sukla Biswas [États-Unis] ; Raies Ahmad [Inde] ; Seyed E. Hasnain [Inde] ; Altaf A. Lal [États-Unis]Source :
- Vaccine [ 0264-410X ] ; 2000.
English descriptors
- Teeft :
- Adjuvant, Alum, Alum adjuvant formulation, Amino acid sequence, Antibody, Antibody levels, Antibody response, Antibody responses, Antibody titers, Assaya alum control alum vaccine copolymer control copolymer vaccine, Baculovirus, Baculovirus expression vector system, Bevs, Candidate vaccine antigen, Cell control, Cell epitope, Cell epitopes, Cell lysate, Cell lysates, Cell pellets, Circumsporozoite protein, Clone, Codon usage, Control mice, Copolymer, Copolymer adjuvant, Correct sequence, Cytotoxic, Deletion errors, Disease control, Elsevier science, Epitope, Falciparum, Falciparum vaccine, First lane, Full length, Gene, Gene synthesis, Grand island, Human service, Human services, Immune, Immune epitopes, Immune kenyan adults, Immune response, Immune responses, Immunization, Immunization studies, Immunoepidemiologic studies, Individual epitopes, Infectious diseases, Insect cell line control, Insect cells, Linear epitopes, Liver stage, Long oligonucleotides, Malaria, Malaria parasites, Malarial epitopes, Mass markers, Melittin signal sequence, Methylated clone, Mice mice, Model systems, Monoclonal, Monoclonal antibody, Mouse, Mouse antisera, Multistage, Multistage vaccine, National center, Normal mouse serum, Nucleic acids, Nucleotide position, Nucleotide sequencing, Oligonucleotides, Overlap extension, Palo alto, Parasite, Parasite growth, Parasitic diseases, Plasmodium, Plasmodium falciparum, Plasmodium falciparum malaria, Plasmodium falciparum merozoite surface protein, Polymerase, Positive response, Primary immunization, Proc natl acad, Proliferative, Proliferative response, Proliferative responses, Protective immunity, Public health service, Recombinant, Recombinant antigen, Recombinant protein, Recombinant virus, Restriction enzymes, Rpmi media, Second round, Sequence artifacts, Several epitopes, Single band, Spleen cells, Sporozoite, Standard reaction, States department, Stimulation index, Supernatant, Synthetic gene, Synthetic gene approach, Synthetic genes, Synthetic oligonucleotides, Talon histidine metal resin, Terminal primers, Tetanus toxoid, Third round, Vaccine, Vaccine candidate antigens, Vaccine development, Vivo protection studies, Western blot, Wild virus.
Abstract
Abstract: A synthetic gene encoding twelve B cell epitopes, six T-cell proliferative epitopes, and three cytotoxic T lymphocyte (CTL) epitopes from nine stage-specific antigens, representing the sporozoite, liver stage, asexual blood-stage, and sexual-stage antigens of Plasmodium falciparum, was constructed by assembling overlapping oligonucleotides followed by PCR extension and annealing. A three-step PCR protocol using twelve long oligonucleotides was employed to generate a 1053 base-pair synthetic gene, the identity of which was confirmed by sequencing. This synthetic gene, named CDC/NII MAL VAC-1, was cloned, and the recombinant protein was expressed in the Baculovirus Expression Vector System (BEVS). The selection of malarial epitopes for inclusion in this vaccine construct was based on immunoepidemiological studies in malaria endemic area, in vitro, and in vivo protection studies in model systems. The 41 kDa BEVS-expressed recombinant protein reacted with mouse antibodies specific for individual B cell epitopes in the vaccine construct and with sera from clinically immune Kenyan adults. An immunization study in three strains of mice that differ at the H-2 locus demonstrated that the BEVS-expressed recombinant protein is immunogenic; the candidate vaccine antigen induced high titer antibodies, and lymphocyte proliferative and IFN-γ responses. These results demonstrate that individual B and T cell epitopes can be assembled to create synthetic genes that encode proteins capable of eliciting specific antibody and T cell responses.
Url:
DOI: 10.1016/S0264-410X(00)00045-1
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<term>Antibody response</term>
<term>Antibody responses</term>
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<term>Cell pellets</term>
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<front><div type="abstract" xml:lang="en">Abstract: A synthetic gene encoding twelve B cell epitopes, six T-cell proliferative epitopes, and three cytotoxic T lymphocyte (CTL) epitopes from nine stage-specific antigens, representing the sporozoite, liver stage, asexual blood-stage, and sexual-stage antigens of Plasmodium falciparum, was constructed by assembling overlapping oligonucleotides followed by PCR extension and annealing. A three-step PCR protocol using twelve long oligonucleotides was employed to generate a 1053 base-pair synthetic gene, the identity of which was confirmed by sequencing. This synthetic gene, named CDC/NII MAL VAC-1, was cloned, and the recombinant protein was expressed in the Baculovirus Expression Vector System (BEVS). The selection of malarial epitopes for inclusion in this vaccine construct was based on immunoepidemiological studies in malaria endemic area, in vitro, and in vivo protection studies in model systems. The 41 kDa BEVS-expressed recombinant protein reacted with mouse antibodies specific for individual B cell epitopes in the vaccine construct and with sera from clinically immune Kenyan adults. An immunization study in three strains of mice that differ at the H-2 locus demonstrated that the BEVS-expressed recombinant protein is immunogenic; the candidate vaccine antigen induced high titer antibodies, and lymphocyte proliferative and IFN-γ responses. These results demonstrate that individual B and T cell epitopes can be assembled to create synthetic genes that encode proteins capable of eliciting specific antibody and T cell responses.</div>
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