Immunological properties of recombinant proteins of the transmission blocking vaccine candidate, Pfs48/45, of the human malaria parasite <i>Plasmodium falciparum</i> produced in <i>Escherichia coli</i>

Milek, Richard L.B.; Roeffen, Will; Kocken, Clemens H. M.; Jansen, Josephine; Kaan, Anita; Eling, W.M.C.; Sauerwein, Robert W.; Konings, Ruud N. H.

doi:10.1046/j.1365-3024.1998.00171.x

Cited by 30 publications

(20 citation statements)

References 34 publications

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“…Antibodies-Polyclonal rabbit antiserum K96 was used to detect recombinant Pfs48/45 (12). To detect epitopes of Pfs48/ 45, various mAbs from mouse origin, 32F5 (epitope I) (6) or from rat origin, 85RF45.1 (epitope I), 85RF45.2b (epitope IIb), 85RF45.3 (epitope III), and 85RF45.5 (epitope V) (8) were used.…”

Section: Methodsmentioning

confidence: 99%

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Epitope Analysis of the Malaria Surface Antigen Pfs48/45 Identifies a Subdomain That Elicits Transmission Blocking Antibodies

Outchkourov¹,

Vermunt²,

Jansen³

et al. 2007

Journal of Biological Chemistry

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Pfs48/45, a member of a Plasmodium-specific protein family, displays conformation-dependent epitopes and is an important target for malaria transmission-blocking (TB) immunity. To design a recombinant Pfs48/45-based TB vaccine, we analyzed the conformational TB epitopes of Pfs48/45. The Pfs48/45 protein was found to consist of a C-terminal six-cysteine module recognized by anti-epitope I antibodies, a middle four-cysteine module recognized by anti-epitopes IIb and III, and an N-terminal module recognized by anti-epitope V antibodies. Refolding assays identified that a fragment of 10 cysteines (10C), comprising the middle four-cysteine and the C-terminal six-cysteine modules, possesses superior refolding capacity. The refolded and partially purified 10C conformer elicited antibodies in mice that targeted at least two of the TB epitopes (I and III). The induced antibodies could block the fertilization of Plasmodium falciparum gametes in vivo in a concentration-dependent manner. Our results provide important insight into the structural organization of the Pfs48/45 protein and experimental support for a Pfs48/45-based subunit vaccine.Malaria parasites are spread in the human population by Plasmodium-infected Anopheles mosquitoes. After a blood meal on infected humans, mosquitoes become infected by ingesting a sexual form of the malaria parasite called gametocytes. Subsequent sporogonic development in the mosquito can be prevented by the presence of anti-malaria, transmissionblocking (TB) 3 antibodies in the ingested blood meal (1-3). Pfs48/45 is a TB vaccine candidate that belongs to a family of malaria-specific proteins that contain conserved motifs with four-or six-cysteine residues (4). Pfs48/45 plays a key role in parasite fertilization (5), and monoclonal antibodies (mAbs) against Pfs48/45 prevent fertilization (6). Anti-Pfs48/45 antibodies are present in human sera from endemic areas, and there is evidence that the presence of anti-Pfs48/45 antibodies in natural sera correlates with TB activity (7). The induction of antibodies after natural infection, as observed in the field, creates the highly beneficial potential of vaccine boosting in the endemic setting. At least four different epitopes (I, IIb, III, and V) on Pfs48/45 are targeted by monoclonal antibodies that block or reduce malaria transmission (8, 9). Apart from epitope V, the TB epitopes are sensitive to reducing agents, which indicates the importance of the disulfide bridges and hence the conformational dependence of these epitopes (8, 10).To design a Pfs48/45-based TB vaccine, we aimed to delineate and characterize the TB epitopes of Pfs48/45. Protease digestion of the native protein, expression in Escherichia coli of truncations, cysteine mutations, and refolding assays were evaluated by immunological analysis of the TB epitopes. An N-terminally truncated protein with higher refolding efficiency was identified, partially purified, and tested in mice immunization experiments. The elicited antibodies recognized the native Pfs48/45 protein, competed ...

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Section: Methodsmentioning

confidence: 99%

“…Expression of full-length Pfs48/45 fused to GST (Fig. 2B, 16C) resulted in a protein of ϳ80kDathatwasrecognizedbythepolyclonal,non-conformationdependent K96 sera (12) and by the mAbs recognizing epitopes I, IIb, III, and V (8) (Fig. 2B).…”

Section: Pfs48/45 Domain Organization and Subunit Vaccine Designmentioning

confidence: 99%

Epitope Analysis of the Malaria Surface Antigen Pfs48/45 Identifies a Subdomain That Elicits Transmission Blocking Antibodies

Outchkourov¹,

Vermunt²,

Jansen³

et al. 2007

Journal of Biological Chemistry

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“…[6][7][8][9][10] Recombinant Pfs48/45 protein was highly immunogenic in mice and rabbits, but expression of this protein in the correct conformation has proven difficult because the elicited antibodies failed to block parasite transmission to mosquitoes. 11 Only two paralogous P. vivax antigens, Pvs25 and Pvs28, have been assessed as vaccine candidates. 6 Both Pvs25 and Pvs28 produced in recombinant Saccharomyces cerevisiae and formulated with aluminum hydroxide induced strong transmission-blocking antibody responses in immunized mice.…”

Section: Introductionmentioning

confidence: 99%

Induction of Transmission-Blocking Immunity in Aotus Monkeys by Vaccination With a Plasmodium Vivax Clinical Grade Pvs25 Recombinant Protein

Arévalo‐Herrera

Solarte

Yasnot

et al. 2005

The American Journal of Tropical Medicine and Hygiene

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Abstract. Aotus monkeys were used to determine the immunogenicity of Pvs25 protein expressed in the zygote/ ookinete surface. Animals were immunized in three times with 100 g of Pvs25 formulated in Montanide ISA-720. Antibodies to Pvs25 detected by an enzyme-linked immunosorbent assay appeared by day 30 after the first immunization, with a peak of antibodies levels on day 150. These antibodies were still detectable on day 300. Plasma samples on day 150 from experimental group were able to completely block the development of the parasite in Anopheles albimanus mosquitoes artificially fed with human isolates of Plasmodium vivax. Immunized Aotus monkeys were infected with blood forms of the P. vivax Salvador I strain and no boosting effect of blood infection on titers of antibodies to Pvs25 was observed despite the presence of infective gametocytes. In conclusion, Pvs25 protein formulated in Montanide ISA-720 induces efficient and long-lasting transmission-blocking antibodies that cannot be boosted by parasite infection.

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“…In addition to identifying TBV candidates that are effective and may span different insect vector species, studies on TBV development must include antigenic variability present in field isolates (Kocken et al 1995;Drakeley et al 1996;Duffy & Kaslow 1997;Sattabongkot et al 2003), immunogenicity of such antigens (Kubler-Kielb et al 2007), reactogenicity caused by adjuvants (Saul et al 2007;Wu et al 2008), non-specific responses (Quakyi et al 1987;Tonui et al 2001a), and improper folding of antigens (Kaslow et al 1994;Milek et al 1998a;Milek et al 1998b;Milek et al 2000). Natural antigenic boosting is another important issue that must be dealt with (Arevalo-Herrera et al 2005).…”

Section: Future Of Tbvsmentioning

confidence: 99%

Transgenesis, Paratransgenesis and Transmission Blocking Vaccines to Prevent Insect-Borne Diseases

Ramalho-Ortigão¹,

Coutinho-Abreu²

2012

Integrated Pest Management and Pest Control - Current and Future Tactics

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Immunological properties of recombinant proteins of the transmission blocking vaccine candidate, Pfs48/45, of the human malaria parasite Plasmodium falciparum produced in Escherichia coli

Cited by 30 publications

References 34 publications

Epitope Analysis of the Malaria Surface Antigen Pfs48/45 Identifies a Subdomain That Elicits Transmission Blocking Antibodies

Epitope Analysis of the Malaria Surface Antigen Pfs48/45 Identifies a Subdomain That Elicits Transmission Blocking Antibodies

Induction of Transmission-Blocking Immunity in Aotus Monkeys by Vaccination With a Plasmodium Vivax Clinical Grade Pvs25 Recombinant Protein

Transgenesis, Paratransgenesis and Transmission Blocking Vaccines to Prevent Insect-Borne Diseases

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