Plasmodium vivax: Genetic diversity of the apical membrane antigen-1 (AMA-1) in isolates from India

Plasmodium vivax apical membrane antigen-1 (PvAMA-1) is an important vaccine candidate for vivax malaria. However, antigenic variation within PvAMA-1 is a major obstacle to the design of a global protective malaria vaccine. In this study, we analyzed the genetic polymorphism and selection of the PvAMA-1 gene from 152 P. vivax isolates from imported cases to China, collected in the China–Myanmar border (CMB) area in Yunnan Province (YP) during 2009–2011 (n = 71) and 2014–2016 (n = 81), in comparison with PvAMA-1 gene information from Myanmar (n = 73), collected from public data. The overall nucleotide diversity of the PvAMA-1 gene from the 152 YP isolates was 0.007 with 76 haplotypes identified (Hd = 0.958). Results from the population structure suggested three groups among the YP and Myanmar isolates with optimized clusters value of K = 7. In addition, YP (2014–2016) isolates generally lacked some K components that were commonly found in YP (2009–2011) and Myanmar. Meanwhile, PvAMA-1 domain I is found to be the dominant target of positive diversifying selection and most mutation loci were found in this domain. The mutation frequencies of D107N/A, R112K/T, K120R, E145A, E277K, and R438H in PvAMA-1 were more than 70% in the YP isolates. In conclusion, high genetic diversity and positive selection were found in the PvAMA-1 gene from YP isolates, which are significant findings for the design and development of PvAMA-1-based malaria vaccine.

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Analysis of the Plasmodium vivax apical membrane antigen-1 gene from re-emerging Korean isolates

Parasitology Research ◽

10.1007/s00436-002-0777-2 ◽

2003 ◽

Vol 90 (4) ◽

pp. 325-329 ◽

Cited By ~ 20

Author(s):

Joon-Yong Chung ◽

Eui-Hyun Chun ◽

Jin-Ho Chun ◽

Weon-Gyu Kho

Keyword(s):

Plasmodium Vivax ◽

Apical Membrane ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen

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High-Level Expression of Plasmodium vivax Apical Membrane Antigen 1 (AMA-1) in Pichia pastoris: Strong Immunogenicity in Macaca mulatta Immunized with P. vivax AMA-1 and Adjuvant SBAS2

Infection and Immunity ◽

10.1128/iai.67.1.43-49.1999 ◽

1999 ◽

Vol 67 (1) ◽

pp. 43-49 ◽

Cited By ~ 57

Author(s):

Clemens H. M. Kocken ◽

Martin A. Dubbeld ◽

Annemarie Van Der Wel ◽

Jack T. Pronk ◽

Andrew P. Waters ◽

...

Keyword(s):

Pichia Pastoris ◽

Plasmodium Vivax ◽

Apical Membrane ◽

Gel Filtration ◽

Methylotrophic Yeast ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen ◽

High Level

ABSTRACT The apical membrane antigen 1 (AMA-1) family is a promising family of malaria blood-stage vaccine candidates that have induced protection in rodent and nonhuman primate models of malaria. Correct conformation of the protein appears to be essential for the induction of parasite-inhibitory responses, and these responses appear to be primarily antibody mediated. Here we describe for the first time high-level secreted expression (over 50 mg/liter) of thePlasmodium vivax AMA-1 (PV66/AMA-1) ectodomain by using the methylotrophic yeast Pichia pastoris. To prevent nonnative glycosylation, a conservatively mutagenized PV66/AMA-1 gene (PV66Δglyc) lacking N-glycosylation sites was also developed. Expression of the PV66Δglyc ectodomain yielded similar levels of a homogeneous product that was nonglycosylated and was readily purified by ion-exchange and gel filtration chromatographies. Recombinant PV66Δglyc43–487 was reactive with conformation-dependent monoclonal antibodies. With the SBAS2 adjuvant,Pichia-expressed PV66Δglyc43–487 was highly immunogenic in five rhesus monkeys, inducing immunoglobulin G enzyme-linked immunosorbent assay titers in excess of 1:200,000. This group of monkeys had a weak trend showing lower cumulative parasite loads following a Plasmodium cynomolgi infection than in the control group.

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Natural Human Antibody Responses to Plasmodium vivax Apical Membrane Antigen 1 under Low Transmission and Unstable Malaria Conditions in Sri Lanka

Infection and Immunity ◽

10.1128/iai.74.1.798-801.2006 ◽

2006 ◽

Vol 74 (1) ◽

pp. 798-801 ◽

Cited By ~ 17

Author(s):

Thilan Wickramarachchi ◽

Prasad H. Premaratne ◽

K. L. R. Lakshman Perera ◽

Sumith Bandara ◽

Clemens H. M. Kocken ◽

...

Keyword(s):

Sri Lanka ◽

Plasmodium Vivax ◽

Malaria Vaccine ◽

Apical Membrane ◽

Human Antibody ◽

Antibody Prevalence ◽

Low Transmission ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen ◽

Malaria Vaccine Candidate

ABSTRACT Plasmodium vivax apical membrane antigen 1, an important malaria vaccine candidate, was immunogenic during natural malaria infections in Sri Lanka, where low transmission and unstable malaria conditions prevail. Antibody prevalence increased with exposure in areas where malaria was or was not endemic. A marked isotype switch to cytophilic (immunoglobulin G1 [IgG1]/IgG3) antibodies was evident with increasing exposure exclusively in residents from areas of endemicity.

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Identification of a Highly Antigenic Linear B Cell Epitope within Plasmodium vivax Apical Membrane Antigen 1 (AMA-1)

PLoS ONE ◽

10.1371/journal.pone.0021289 ◽

2011 ◽

Vol 6 (6) ◽

pp. e21289 ◽

Cited By ~ 27

Author(s):

Lilian Lacerda Bueno ◽

Francisco Pereira Lobo ◽

Cristiane Guimarães Morais ◽

Luíza Carvalho Mourão ◽

Ricardo Andrez Machado de Ávila ◽

...

Keyword(s):

B Cell ◽

Plasmodium Vivax ◽

Apical Membrane ◽

Cell Epitope ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen ◽

B Cell Epitope ◽

Linear B

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Population genetic structure and polymorphism analysis of gene encoding apical membrane antigen-1 (AMA-1) of Iranian Plasmodium vivax wild isolates

Acta Tropica ◽

10.1016/j.actatropica.2013.02.017 ◽

2013 ◽

Vol 126 (3) ◽

pp. 269-279 ◽

Cited By ~ 20

Author(s):

Sedigheh Zakeri ◽

Hengameh Sadeghi ◽

Akram Abouie Mehrizi ◽

Navid Dinparast Djadid

Keyword(s):

Genetic Structure ◽

Plasmodium Vivax ◽

Population Genetic Structure ◽

Population Genetic ◽

Apical Membrane ◽

Polymorphism Analysis ◽

Gene Encoding ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen

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Invasion-Inhibitory Antibodies Elicited by Immunization with Plasmodium vivax Apical Membrane Antigen-1 Expressed in Pichia pastoris Yeast

Infection and Immunity ◽

10.1128/iai.01169-13 ◽

2013 ◽

Vol 82 (3) ◽

pp. 1296-1307 ◽

Cited By ~ 39

Author(s):

Elaine C. Vicentin ◽

Kátia S. Françoso ◽

Mariana V. Rocha ◽

Dmitri Iourtov ◽

Fernanda L. dos Santos ◽

...

Keyword(s):

Pichia Pastoris ◽

Recombinant Protein ◽

Plasmodium Vivax ◽

Apical Membrane ◽

Content Type ◽

Apical Membrane Antigen 1 ◽

Apical Membrane Antigen ◽

Monophosphoryl Lipid A ◽

Antibody Titers ◽

Inhibitory Antibodies

ABSTRACTIn a recent vaccine trial performed with African children, immunization with a recombinant protein based onPlasmodium falciparumapical membrane antigen 1 (AMA-1) conferred a significant degree of strain-specific resistance against malaria. To contribute to the efforts of generating a vaccine againstPlasmodium vivaxmalaria, we expressed the ectodomain ofP. vivaxAMA-1 (PvAMA-1) as a secreted soluble protein in the methylotrophic yeastPichia pastoris. Recognized by a high percentage of sera from individuals infected byP. vivax, this recombinant protein was found to have maintained its antigenicity. The immunogenicity of this protein was evaluated in mice using immunization protocols that included homologous and heterologous prime-boost strategies with plasmid DNA and recombinant protein. We used the following formulations containing different adjuvants: aluminum salts (Alum),Bordetella pertussismonophosphoryl lipid A (MPLA), flagellin FliC fromSalmonella entericaserovar Typhimurium, saponin Quil A, or incomplete Freund's adjuvant (IFA). The formulations containing the adjuvants Quil A or IFA elicited the highest IgG antibody titers. Significant antibody titers were also obtained using a formulation developed for human use containing MPLA or Alum plus MPLA. Recombinant PvAMA-1 produced under “conditions of good laboratory practice” provided a good yield, high purity, low endotoxin levels, and no microbial contaminants and reproduced the experimental immunizations. Most relevant for vaccine development was the fact that immunization with PvAMA-1 elicited invasion-inhibitory antibodies against different Asian isolates ofP. vivax. Our results show that AMA-1 expressed inP. pastorisis a promising antigen for use in future preclinical and clinical studies.

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