A recent clinical trial in African children demonstrated the potential utility of merozoite surface protein (MSP)-3 as a vaccine against Plasmodium falciparum malaria. The present study evaluated the use of Plasmodium vivax MSP-3 (PvMSP-3) as a target antigen in vaccine formulations against malaria caused by P. vivax. Recombinant proteins representing MSP-3α and MSP-3β of P. vivax were expressed as soluble histidine-tagged bacterial fusions. Antigenicity during natural infection was evaluated by detecting specific antibodies using sera from individuals living in endemic areas of Brazil. A large proportion of infected individuals presented IgG antibodies to PvMSP-3α (68.2%) and at least 1 recombinant protein representing PvMSP-3β (79.1%). In spite of the large responder frequency, reactivity to both antigens was significantly lower than was observed for the immunodominant epitope present on the 19-kDa C-terminal region of PvMSP-1. Immunogenicity of the recombinant proteins was studied in mice in the absence or presence of different adjuvant formulations. PvMSP-3β, but not PvMSP-3α, induced a TLR4-independent humoral immune response in the absence of any adjuvant formulation. The immunogenicity of the recombinant antigens were also tested in formulations containing different adjuvants (Alum, Salmonella enterica flagellin, CpG, Quil A,TiterMax？ and incomplete Freunds adjuvant) and combinations of two adjuvants (Alum plus flagellin, and CpG plus flagellin). Recombinant PvMSP-3α and PvMSP-3β elicited higher antibody titers capable of recognizing P. vivax-infected erythrocytes harvested from malaria patients. Our results confirm that P. vivax MSP-3 antigens are immunogenic during natural infection, and the corresponding recombinant proteins may be useful in elucidating their vaccine potential.
References
[1]
Agnandji ST, Lell B, Soulanoudjingar SS, Fernandes JF, Abossolo BP, et al. (2011) First Results of Phase 3 Trial of RTS,S/AS01 Malaria Vaccine in African Children. N Engl J Med 365: 1863–1875.
[2]
Thera MA, Doumbo OK, Coulibaly D, Laurens MB, Ouattara A, et al. (2011) A field trial to assess a blood-stage malaria vaccine. N Engl J Med 365: 1004–1013.
[3]
Sirima SB, Cousens S, Druilhe P (2011) Protection against malaria by MSP3 candidate vaccine. N Engl J Med 365: 1062–1064.
[4]
Herrera S, Fernández OL, Vera O, Cárdenas W, Ramírez O, et al.. (2011) Phase I safety and immunogenicity trial of Plasmodium vivax CS derived long synthetic peptides adjuvanted with montanide ISA 720 or montanide ISA 51. Am J Trop Med Hyg 84(2 Suppl): 12–20.
[5]
Arévalo-Herrera M, Soto L, Perlaza BL, Céspedes N, Vera O, et al.. (2011) Antibody-mediated and cellular immune responses induced in naive volunteers by vaccination with long synthetic peptides derived from the Plasmodium vivax circumsporozoite protein. Am J Trop Med Hyg 84(2 Suppl): 35–42.
[6]
Wu Y, Ellis RD, Shaffer D, Fontes E, Malkin EM, et al. (2008) Phase 1 trial of malaria transmission blocking vaccine candidates Pfs25 and Pvs25 formulated with montanide ISA 51. PLoS One 3: e2636.
[7]
malERA Consultative Group on Vaccines (2011) A research agenda for malaria eradication: vaccines. PLoS Med 8: e1000398.
[8]
Soares IS, Levitus G, Souza JM, Del Portillo HA, Rodrigues MM (1997) Acquired immune responses to the N- and C-terminal regions of Plasmodium vivax merozoite surface protein 1 in individuals exposed to malaria. Infect Immun 65: 1606–1614.
[9]
Soares IS, Cunha MG, Silva MN, Souza JM, Del Portillo HA, et al. (1999) Longevity of naturally acquired antibody responses to the N- and C-terminal regions of Plasmodium vivax merozoite surface protein 1. Am J Trop Med Hyg 60: 357–363.
[10]
Michon PA, Arevalo-Herrera M, Fraser T, Herrera S, Adams JH (1998) Serologic responses to recombinant Plasmodium vivax Duffy binding protein in a Colombian village. Am J Trop Med Hyg 59: 597–599.
[11]
Rodrigues MH, Cunha MG, Machado RL, Ferreira OC Jr, Rodrigues MM, et al. (2003) Serological detection of Plasmodium vivax malaria using recombinant proteins corresponding to the 19-kDa C-terminal region of the merozoite surface protein-1. Malar J 2: 39.
[12]
Herrera S, Gómez A, Vera O, Vergara J, Valderrama-Aguirre A, et al.. (2005) Antibody response to Plasmodium vivax antigens in Fy-negative individuals from the Colombian Pacific coast. Am J Trop Med Hyg 73(5 Suppl): 44–49.
[13]
Rodrigues MHC, Rodrigues KM, Oliveira TR, Comodo AN, Rodrigues MM, et al. (2005) Antibody response of naturally infected individuals to recombinant Plasmodium vivax apical membrane antigen-1. Int J Parasitol 35: 185–192.
[14]
Morais CG, Soares IS, Carvalho LH, Fontes CJ, Krettli AU, et al. (2005) IgG isotype to C-terminal 19 kDa of Plasmodium vivax merozoite surface protein 1 among subjects with different levels of exposure to malaria in Brazil. Parasitol Res 95: 420–426.
[15]
Morais CG, Soares IS, Carvalho LH, Fontes CJ, Krettli AU, et al. (2006) Antibodies to Plasmodium vivax apical membrane antigen 1: persistence and correlation with malaria transmission intensity. Am J Trop Med Hyg 75: 582–587.
[16]
Barbedo MB, Ricci R, Jimenez MC, Cunha MG, Yazdani SS, et al. (2007) Comparative recognition by human IgG antibodies of recombinant proteins representing three asexual erythrocytic stage vaccine candidates of Plasmodium vivax. Mem Inst Oswaldo Cruz 102: 335–339.
[17]
Ceravolo IP, Sanchez BA, Sousa TN, Guerra BM, Soares IS, et al. (2009) Naturally acquired inhibitory antibodies to Plasmodium vivax Duffy binding protein are short-lived and allele-specific following a single malaria infection. Clin Exp Immunol 156: 502–510.
[18]
Storti-Melo LM, Souza-Neiras WC, Cassiano GC, Taveira LC, Cordeiro AJ, et al.. (2011) Evaluation of the naturally acquired antibody immune response to the Pv200L N-terminal fragment of Plasmodium vivax merozoite surface protein-1 in four areas of the Amazon Region of Brazil. Am J Trop Med Hyg 84(2 Suppl): 58–63.
[19]
Lima-Junior JC, Jiang J, Rodrigues-da-Silva RN, Banic DM, Tran TM, et al. (2011) B cell epitope mapping and characterization of naturally acquired antibodies to the Plasmodium vivax merozoite surface protein-3α (PvMSP-3α) in malaria exposed individuals from Brazilian Amazon. Vaccine 29: 1801–1811.
[20]
Mour？o LC, Morais CG, Bueno LL, Jimenez MC, Soares IS, et al. (2012) Naturally acquired antibodies to Plasmodium vivax blood-stage vaccine candidates (PvMSP-1(19) and PvMSP-3α(359–798)) and their relationship with hematological features in malaria patients from the Brazilian Amazon. Microbes Infect 14: 730–739.
[21]
Cunha MG, Rodrigues MM, Soares IS (2001) Comparison of the immunogenic properties of recombinant proteins representing the Plasmodium vivax vaccine candidate MSP1(19) expressed in distinct bacterial vectors. Vaccine 20: 385–396.
[22]
Herrera S, Bonelo A, Perlaza BL, Valencia AZ, Cifuentes C, et al. (2004) Use of long synthetic peptides to study the antigenicity and immunogenicity of the Plasmodium vivax circumsporozoite protein. Int J Parasitol 34: 1535–1546.
[23]
Arévalo-Herrera M, Castellanos A, Yazdani SS, Shakri AR, Chitnis CE, et al.. (2005) Immunogenicity and protective efficacy of recombinant vaccine based on the receptor-binding domain of the Plasmodium vivax Duffy binding protein in Aotus monkeys. Am J Trop Med Hyg 73(5 Suppl): 25–31.
[24]
Valderrama-Aguirre A, Quintero G, Gómez A, Castellanos A, Pérez Y, et al.. (2005) Antigenicity, immunogenicity, and protective efficacy of Plasmodium vivax MSP1 PV200l: a potential malaria vaccine subunit. Am J Trop Med Hyg 73(5 Suppl): 16–24.
[25]
Rosa DS, Iwai LK, Tzelepis F, Bargieri DY, Medeiros MA, et al. (2006) Immunogenicity of a recombinant protein containing the Plasmodium vivax vaccine candidate MSP1(19) and two human CD4+ T-cell epitopes administered to non-human primates (Callithrix jacchus jacchus). Microbes Infect 8: 2130–2137.
[26]
Bargieri DY, Rosa DS, Braga CJ, Carvalho BO, Costa FT, et al. (2008) New malaria vaccine candidates based on the Plasmodium vivax Merozoite Surface Protein-1 and the TLR-5 agonist Salmonella typhimurium FliC flagellin. Vaccine 26: 6132–6142.
[27]
Bouillet Lé, Dias MO, Dorigo NA, Moura AD, Russell B, et al. (2011) Long-Term Humoral and Cellular Immune Responses Elicited by a Heterologous Plasmodium vivax Apical Membrane Antigen 1 Protein Prime/Adenovirus Boost Immunization Protocol. Infect Immun 79: 3642–3652.
[28]
Galinski MR, Corredor-Medina C, Povoa M, Crosby J, Ingravallo P, et al. (1999) Plasmodium vivax merozoite surface protein-3 contains coiled-coil motifs in an alanine-rich central domain. Mol Biochem Parasitol 101: 131–147.
[29]
Galinski MR, Ingravallo P, Corredor-Medina C, Al-Khedery B, Povoa M, et al. (2001) Plasmodium vivax merozoite surface proteins-3beta and-3gamma share structural similarities with P. vivax merozoite surface protein-3alpha and define a new gene family. Mol Biochem Parasitol 115: 41–53.
[30]
Carlton JM, Adams JH, Silva JC, Bidwell SL, Lorenzi H, et al. (2008) Comparative genomics of the neglected human malaria parasite Plasmodium vivax. Nature 455: 757–763.
[31]
Jimenez MC, Ramos CH, Barbosa JA, Galinski MR, Barnwell JW, et al. (2008) Biophysical characterization of the recombinant merozoite surface protein-3 of Plasmodium vivax. Biochim Biophys Acta 1780: 983–988.
[32]
Singh S, Soe S, Weisman S, Barnwell JW, Pérignon JL, et al. (2009) A conserved multi-gene family induces cross-reactive antibodies effective in defense against Plasmodium falciparum. PLoS One 4: e5410.
[33]
GReferences and further reading may be available for this article. To view references, further reading you must purchase this article.entil F, Bargieri DY, Leite JA, Fran？oso KS, Patricio MB, et al. (2010) A recombinant vaccine based on domain II of Plasmodium vivax Apical Membrane Antigen 1 induces high antibody titres in mice. Vaccine 28: 6183–6190.
[34]
Russell B, Suwanarusk R, Borlon C, Costa FT, Chu CS, et al. (2011) A reliable ex vivo invasion assay of human reticulocytes by Plasmodium vivax. Blood. 118: e74–81.
[35]
Rayner JC, Corredor V, Feldman D, Ingravallo P, Iderabdullah F, et al. (2002) Extensive polymorphism in the Plasmodium vivax merozoite surface coat protein MSP-3alpha is limited to specific domains. Parasitology 125: 393–405.
[36]
Rayner JC, Huber CS, Feldman D, Ingravallo P, Galinski MR, et al. (2004) Plasmodium vivax merozoite surface protein PvMSP-3 beta is radically polymorphic through mutation and large insertions and deletions. Infect Genet Evol 4: 309–319.
[37]
Ribeiro RS, Ladeira L, Rezende AM, Fontes CJ, Carvalho LH, et al. (2011) Analysis of the genetic variability of PvMSP-3α among Plasmodium vivax in Brazilian field isolates. Mem Inst Oswaldo Cruz 106 Suppl 127–33.
[38]
Coler RN, Carter D, Friede M, Reed SG (2009) Adjuvants for malaria vaccines. Parasite Immunol 31: 520–528.
[39]
Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, et al. (2000) A Toll-like receptor recognizes bacterial DNA. Nature 408: 740–745.
[40]
Mullen GE, Ellis RD, Miura K, Malkin E, Nolan C, et al. (2008) Phase1 trial of AMA1-C1/Alhydrogel plus CPG7909: an asexual blood-stage vaccine or Plasmodium falciparum malaria. PLoS One 3: e2940.
[41]
Traore B, Kone Y, Doumbo S, Doumtabe D, Traore A, et al. (2009) The TLR9 agonist CpG fails to enhance the acquisition of Plasmodium falciparum-specific memory B cells in semi-immune adults in Mali. Vaccine 27: 7299–7303.
[42]
Sagara I, Ellis RD, Dicko A, Niambele MB, Kamate B, et al. (2009) A randomized and controlled Phase 1 study of the safety and mmunogenicity of the AMA1-C1/Alhydrogel +CPG 7909 vaccine for Plasmodium falciparum malaria in semi-immune Malian adults. Vaccine 27: 7292–7298.
[43]
Ellis RD, Martin LB, Shaffer D, Long CA, Miura K, et al. (2010) Phase 1 trial of the Plasmodium falciparum blood stage vaccine MSP1(42)-C1/Alhydrogel with and without CPG 7909 in malaria na？ve adults. PLoSOne 5: e8787.
