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Background  Antibodies have an essential role in the acquired immune response against blood stage  P. falciparum  infection. Although several antigens have been identified as important antibody targets, it is still elusive which antigens have to be recognized for clinical protection. Herein, we analyzed antibodies from plasmas from symptomatic or asymptomatic individuals living in the same geographic area in the Western Amazon, measuring their recognition of multiple merozoite antigens.    Methods  Specific fragments of genes encoding merozoite proteins AMA1 and members of MSP and EBL families from circulating  P. falciparum  field isolates present in asymptomatic and symptomatic patients were amplified by PCR. After cloning and expression of different versions of the antigens as recombinant GST-fusion peptides, we tested the reactivity of patients’ plasmas by ELISA and the presence of IgG subclasses in the most reactive plasmas.    Results  11 out of 24 recombinant antigens were recognized by plasmas from either symptomatic or asymptomatic infections. Antibodies to MSP9 (X 2  DF=1  = 9.26/ p  = 0.0047) and MSP5 (X 2  DF=1  = 8.29/ p  = 0.0069) were more prevalent in asymptomatic individuals whereas the opposite was observed for MSP1 block 2-MAD20 (X 2  DF=1  = 6.41/ p  = 0.0206, Fisher’s exact test). Plasmas from asymptomatic individuals reacted more intensely against MSP4 (U = 210.5,  p  < 0.03), MSP5 (U = 212,  p  < 0.004), MSP9 (U = 189.5,  p  < 0.002) and EBA175 (U = 197, p < 0.014, Mann-Whitney’s U test). IgG1 and IgG3 were predominant for all antigens, but some patients also presented with IgG2 and IgG4. The recognition of MSP5 (OR = 0.112, IC 95%  = 0.021-0.585) and MSP9 (OR = 0.125, IC 95%  = 0.030-0.529, cross tab analysis) predicted 8.9 and 8 times less chances, respectively, to present symptoms. Higher antibody levels against MSP5 and EBA175 were associated by odds ratios of 9.4 (IC 95%  = 1.29-69.25) and 5.7 (IC 95%  = 1.12-29.62, logistic regression), respectively, with an asymptomatic status.    Conclusions  Merozoite antigens were targets of cytophilic antibodies and antibodies against MSP5, MSP9 and EBA175 were independently associated with decreased symptoms.

bmc infectious diseases

Natural antibody response to Plasmodium falciparum merozoite antigens MSP5, MSP9 and EBA175 is associated to clinical protection in the Brazilian Amazon