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Plasmodium berghei circumsporozoite protein encapsulated in oligomannose-coated liposomes confers protection against sporozoite infection in mice.

Terkawi MA, Kuroda Y, Fukumoto S, Tanaka S, Kojima N, Nishikawa Y - Malar. J. (2014)

Bottom Line: The design and development of an effective malaria vaccine against the pre-erythrocytic and erythrocytic-stages of infection present a great challenge.The current results represent the use of an oligomannose-coated liposome-based vaccine against pre-erythrocytic and erythrocytic stages malaria infection.This approach may offer a new vaccination strategy against malaria infection.

View Article: PubMed Central - PubMed

Affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. nisikawa@obihiro.ac.jp.

ABSTRACT

Background: The design and development of an effective malaria vaccine against the pre-erythrocytic and erythrocytic-stages of infection present a great challenge.

Methods: In the present study, protective efficacy of oligomannose-coated liposome (OML)-entrapped merozoite and sporozoite antigens against Plasmodium berghei challenge infection in BALB/c mice was evaluated.

Results: Subcutaneous immunization with truncated merozoite surface protein 1 entrapped with OML (OML-PbMSP1) prolonged survival, but failed to protect the mice from erythrocytic-stage infection, despite the antigen-specific antibody responses induced by the immunization regimen. In contrast, immunization with circumsporozoite protein entrapped with OML (OML-PbCSP) elicited antigen-specific humoral and cellular responses, which correlated with substantial protection against sporozoite challenge infections.

Conclusions: The current results represent the use of an oligomannose-coated liposome-based vaccine against pre-erythrocytic and erythrocytic stages malaria infection. This approach may offer a new vaccination strategy against malaria infection.

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Related in: MedlinePlus

Expression and purification of recombinant proteins. 15% sodium dodecyl sulphate polyacrylamide electrophoresis gel (SDS-PAGE) for recombinant proteins stained with Coomassie blue. Lanes: M, molecular mass marker; lane 1, PbMSP1; lane 2, PbCSP.
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Fig1: Expression and purification of recombinant proteins. 15% sodium dodecyl sulphate polyacrylamide electrophoresis gel (SDS-PAGE) for recombinant proteins stained with Coomassie blue. Lanes: M, molecular mass marker; lane 1, PbMSP1; lane 2, PbCSP.

Mentions: Recombinant proteins comprising truncated regions of PbMSP1 (Leu1609-Ser1768, GenBank accession number: AAC28871) and PbCSP (Asn201-Asn347, GenBank accession number: P23093.1) were produced in Escherichia coli as glutathione S-transferase (GST) fusion proteins. Briefly, the coding regions of the targeted genes were amplified from P. berghei ANKA genomic DNA with specific primer sets designed from the GenBank sequences: these were 5’-AC GGA TCC AGT ATT ACC ACC GAG CAG AA-3’ , which includes a BamHI restriction enzyme site (boldface), and 5’-AG CTC GAG TTA GCT GGA AGA GCT ACA GAA-3’ , which includes a XhoI restriction enzyme site (boldface) for PbMSP; and 5’-AA GGA TCC CAG CCA CAA CCA CAG CCA GGT-3’ , which includes a BamHI restriction enzyme site (boldface), and 5’-GG CTC GAG TTA TGA ACA TTT ATC CAT TTT-3’ , which includes a XhoI site (boldface) for PbCSP. PCR products were digested with the appropriate restriction enzymes and then ligated into a pGEX-4 T1 (GE Healthcare, Buckinghamshire, UK) expression vector, which had been digested with the same set of restriction enzymes. The nucleotide sequences of the PbMSP1- and PbCSP-positive plasmid inserts were determined using an ABI 3100 DNA sequencer (Applied Biosystems, Foster City, CA, USA). Recombinant proteins were expressed in large-scale DH5α strain E. coli cultures (Takara Bio Inc., Osaka, Japan) and purified by Glutathione-Sepharose 4B beads (GE Healthcare). Purified recombinant proteins were treated with thrombin protease (GE Healthcare) to digest the GST tag and then subjected to endotoxin removal using the membrane filter, Acrodisc® Unite (Pall Life Sciences, Ann Arbor, MI, USA). The purity and quantity of the recombinant proteins were tested by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) stained with Coomassie Brilliant Blue R250 (MP Biomedicals Inc., Illkirch-Graffenstaden, France) and by a BCA protein assay kit (Thermo Fisher Scientific, Inc. Rockford, IL, USA). Single bands (20-kDa and 14-kDa) corresponding to each recombinant protein (PbMSP-1 and PbCSP, respectively) were observed with SDS-PAGE (Figure 1). Thereafter, OMLs were prepared for recombinant protein entrapment as described previously [17–21], and the amount of entrapped antigen was measured using a modified Lowry protein assay reagent (Pierce, Rockford, IL, USA).Figure 1


Plasmodium berghei circumsporozoite protein encapsulated in oligomannose-coated liposomes confers protection against sporozoite infection in mice.

Terkawi MA, Kuroda Y, Fukumoto S, Tanaka S, Kojima N, Nishikawa Y - Malar. J. (2014)

Expression and purification of recombinant proteins. 15% sodium dodecyl sulphate polyacrylamide electrophoresis gel (SDS-PAGE) for recombinant proteins stained with Coomassie blue. Lanes: M, molecular mass marker; lane 1, PbMSP1; lane 2, PbCSP.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4232614&req=5

Fig1: Expression and purification of recombinant proteins. 15% sodium dodecyl sulphate polyacrylamide electrophoresis gel (SDS-PAGE) for recombinant proteins stained with Coomassie blue. Lanes: M, molecular mass marker; lane 1, PbMSP1; lane 2, PbCSP.
Mentions: Recombinant proteins comprising truncated regions of PbMSP1 (Leu1609-Ser1768, GenBank accession number: AAC28871) and PbCSP (Asn201-Asn347, GenBank accession number: P23093.1) were produced in Escherichia coli as glutathione S-transferase (GST) fusion proteins. Briefly, the coding regions of the targeted genes were amplified from P. berghei ANKA genomic DNA with specific primer sets designed from the GenBank sequences: these were 5’-AC GGA TCC AGT ATT ACC ACC GAG CAG AA-3’ , which includes a BamHI restriction enzyme site (boldface), and 5’-AG CTC GAG TTA GCT GGA AGA GCT ACA GAA-3’ , which includes a XhoI restriction enzyme site (boldface) for PbMSP; and 5’-AA GGA TCC CAG CCA CAA CCA CAG CCA GGT-3’ , which includes a BamHI restriction enzyme site (boldface), and 5’-GG CTC GAG TTA TGA ACA TTT ATC CAT TTT-3’ , which includes a XhoI site (boldface) for PbCSP. PCR products were digested with the appropriate restriction enzymes and then ligated into a pGEX-4 T1 (GE Healthcare, Buckinghamshire, UK) expression vector, which had been digested with the same set of restriction enzymes. The nucleotide sequences of the PbMSP1- and PbCSP-positive plasmid inserts were determined using an ABI 3100 DNA sequencer (Applied Biosystems, Foster City, CA, USA). Recombinant proteins were expressed in large-scale DH5α strain E. coli cultures (Takara Bio Inc., Osaka, Japan) and purified by Glutathione-Sepharose 4B beads (GE Healthcare). Purified recombinant proteins were treated with thrombin protease (GE Healthcare) to digest the GST tag and then subjected to endotoxin removal using the membrane filter, Acrodisc® Unite (Pall Life Sciences, Ann Arbor, MI, USA). The purity and quantity of the recombinant proteins were tested by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) stained with Coomassie Brilliant Blue R250 (MP Biomedicals Inc., Illkirch-Graffenstaden, France) and by a BCA protein assay kit (Thermo Fisher Scientific, Inc. Rockford, IL, USA). Single bands (20-kDa and 14-kDa) corresponding to each recombinant protein (PbMSP-1 and PbCSP, respectively) were observed with SDS-PAGE (Figure 1). Thereafter, OMLs were prepared for recombinant protein entrapment as described previously [17–21], and the amount of entrapped antigen was measured using a modified Lowry protein assay reagent (Pierce, Rockford, IL, USA).Figure 1

Bottom Line: The design and development of an effective malaria vaccine against the pre-erythrocytic and erythrocytic-stages of infection present a great challenge.The current results represent the use of an oligomannose-coated liposome-based vaccine against pre-erythrocytic and erythrocytic stages malaria infection.This approach may offer a new vaccination strategy against malaria infection.

View Article: PubMed Central - PubMed

Affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. nisikawa@obihiro.ac.jp.

ABSTRACT

Background: The design and development of an effective malaria vaccine against the pre-erythrocytic and erythrocytic-stages of infection present a great challenge.

Methods: In the present study, protective efficacy of oligomannose-coated liposome (OML)-entrapped merozoite and sporozoite antigens against Plasmodium berghei challenge infection in BALB/c mice was evaluated.

Results: Subcutaneous immunization with truncated merozoite surface protein 1 entrapped with OML (OML-PbMSP1) prolonged survival, but failed to protect the mice from erythrocytic-stage infection, despite the antigen-specific antibody responses induced by the immunization regimen. In contrast, immunization with circumsporozoite protein entrapped with OML (OML-PbCSP) elicited antigen-specific humoral and cellular responses, which correlated with substantial protection against sporozoite challenge infections.

Conclusions: The current results represent the use of an oligomannose-coated liposome-based vaccine against pre-erythrocytic and erythrocytic stages malaria infection. This approach may offer a new vaccination strategy against malaria infection.

Show MeSH
Related in: MedlinePlus