Characterization of Pv92, a Novel Merozoite Surface Protein of Plasmodium vivax
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The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.
Abstract
The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.
참고문헌 (27)
- World Malaria Report (2015), "WHO", ~ (2015)
- Mueller I, "Development of vaccines for Plasmodium vivax malaria", Vaccine, 33, pp.7489~7495 (2015)
- Herrera S, "An update on the search for a Plasmodium vivax vaccine", Trends Parasitol, 23, pp.122~128 (2007)
- Chen JH, "Immunoproteomics profiling of blood stage Plasmodium vivax infection by high-throughput screening assays", J Proteome Res, 9, pp.6479~6489 (2010)
- Arevalo-Pinzon G, "Conserved high activity binding peptides from the Plasmodium falciparum Pf34 rhoptry protein inhibit merozoites in vitro invasion of red blood cells", Peptides, 31, pp.1987~1994 (2010)
- Cowman AF, "Invasion of red blood cells by malaria parasites", Cell, 124, pp.755~766 (2006)
- Jaskiewicz E, "Proteins involved in invasion of human red blood cells by malaria parasites", Postepy Hig Med Dosw, 64, pp.617~626 (2010)
- Gerloff DL, "Structural models for the protein family characterized by gamete surface protein Pfs230 of Plasmodium falciparum", Proc Natl Acad Sci USA, 102, pp.13598~13603 (2005)
- Sanders PR, "Identification of protein complexes in detergent-resistant membranes of Plasmodium falciparum schizonts", Mol Biochem Parasitol, 154, pp.148~157 (2007)
- Eksi S, "Malaria transmission-blocking antigen, Pfs230, mediates human red blood cell binding to exflagellating male parasites and oocyst production", Mol Microbiol, 61, pp.991~998 (2006)
- Rener J, "Target antigens of transmission-blocking immunity on gametes of Plasmodium falciparum", J Exp Med, 158, pp.976~981 (1983)
- Vermeulen AN, "Plasmodium falciparum transmission blocking monoclonal antibodies recognize monovalently expressed epitopes", Dev Biol Stand, 62, pp.91~97 (1985)
- Quakyi IA, "The 230-kDa gamete surface protein of Plasmodium falciparum is also a target for transmission-blocking antibodies", J Immunol, 139, pp.4213~4217 (1987)
- Chowdhury DR, "A potent malaria transmission blocking vaccine based on codon harmonized full length Pfs48", 45 expressed in Escherichia coli, 2009, pp.4, e6352~
- Williamson KC, "Pfs230: from malaria transmission-blocking vaccine candidate toward function", Parasite Immunol, 25, pp.351~359 (2003)
- Obando-Martinez AZ, "Conserved high activity binding peptides are involved in adhesion of two detergent-resistant membrane-associated merozoite proteins to red blood cells during invasion", J Med Chem, 53, pp.3907~3918 (2010)
- Sanders PR, "Distinct protein classes including novel merozoite surface antigens in Raft-like membranes of Plasmodium falciparum", J Biol Chem, 280, pp.40169~40176 (2005)
- Cheng Y, "Characterization of a novel merozoite surface protein of Plasmodium vivax, Pv41", Acta Trop, 126, pp.222~228 (2013)
- Li J, "Pv12, a 6-Cys antigen of Plasmodium vivax, is localized to the merozoite rhoptry", Parasitol Int, 61, pp.443~449 (2012)
- Tachibana M, "Plasmodium vivax gametocyte proteins, Pvs48", 45 and Pvs47, induce transmission-reducing antibodies by DNA immunization, 2015, pp.33, 1901~1908
- Hostetler JB, "A library of Plasmodium vivax recombinant merozoite proteins reveals new vaccine candidates and protein-protein interactions", PLoS Negl Trop Dis, 9, pp.e0004264~ (2015)
- Schultz J, "SMART, a simple modular architecture research tool: identification of signaling domains", Proc Natl Acad Sci USA, 95, pp.5857~5864 (1998)
- Letunic I, "SMART: recent updates, new developments and status in 2015", Nucleic Acids Res, 43, pp.D257~D260 (2015)
- Neeley ES, "Variable slope normalization of reverse phase protein arrays", Bioinformatics, 25, pp.1384~1389 (2009)
- Chen JH, "Measurement of naturally acquired humoral immune responses against the C-terminal region of the Plasmodium vivax MSP1 protein using protein arrays", Parasitol Res, 109, pp.1259~1266 (2011)
- Tachibana M, "N-terminal prodomain of Pfs230 synthesized using a cell-free system is sufficient to induce complement-dependent malaria transmission-blocking activity", Clin Vaccine Immunol, 18, pp.1343~1350 (2011)
- França CT, "An antibody screen of a Plasmodium vivax antigen library identifies novel merozoite proteins associated with clinical protection", PLoS Negl Trop Dis, 10, pp.e0004639~ (2016)