RESUMO
BACKGROUND: The human malaria parasite Plasmodium vivax infects red blood cells through a key pathway that requires interaction between Duffy binding protein II (DBPII) and its receptor on reticulocytes, the Duffy antigen/receptor for chemokines (DARC). A high proportion of P. vivax-exposed individuals fail to develop antibodies that inhibit DBPII-DARC interaction, and genetic factors that modulate this humoral immune response are poorly characterized. Here, we investigate if DBPII responsiveness could be HLA class II-linked. METHODOLOGY/PRINCIPAL FINDINGS: A community-based open cohort study was carried out in an agricultural settlement of the Brazilian Amazon, in which 336 unrelated volunteers were genotyped for HLA class II (DRB1, DQA1 and DQB1 loci), and their DBPII immune responses were monitored over time (baseline, 6 and 12 months) by conventional serology (DBPII IgG ELISA-detected) and functional assays (inhibition of DBPII-erythrocyte binding). The results demonstrated an increased susceptibility of the DRB1*13:01 carriers to develop and sustain an anti-DBPII IgG response, while individuals with the haplotype DRB1*14:02-DQA1*05:03-DQB1*03:01 were persistent non-responders. HLA class II gene polymorphisms also influenced the functional properties of DBPII antibodies (BIAbs, binding inhibitory antibodies), with three alleles (DRB1*07:01, DQA1*02:01 and DQB1*02:02) comprising a single haplotype linked with the presence and persistence of the BIAbs response. Modelling the structural effects of the HLA-DRB1 variants revealed a number of differences in the peptide-binding groove, which is likely to lead to altered antigen binding and presentation profiles, and hence may explain the differences in subject responses. CONCLUSIONS/SIGNIFICANCE: The current study confirms the heritability of the DBPII antibody response, with genetic variation in HLA class II genes influencing both the development and persistence of IgG antibody responses. Cellular studies to increase knowledge of the binding affinities of DBPII peptides for class II molecules linked with good or poor antibody responses might lead to the development of strategies for controlling the type of helper T cells activated in response to DBPII.
Assuntos
Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Cadeias HLA-DRB1/imunologia , Malária Vivax/genética , Malária Vivax/imunologia , Plasmodium vivax/imunologia , Proteínas de Protozoários/imunologia , Receptores de Superfície Celular/imunologia , Adulto , Alelos , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/metabolismo , Brasil/epidemiologia , Proteínas de Transporte/genética , Estudos de Coortes , Sistema do Grupo Sanguíneo Duffy/imunologia , Ensaio de Imunoadsorção Enzimática , Eritrócitos/parasitologia , Feminino , Variação Genética , Genótipo , Cadeias HLA-DRB1/genética , Haplótipos , Humanos , Imunoglobulina G/imunologia , Malária Vivax/epidemiologia , Malária Vivax/parasitologia , Masculino , Pessoa de Meia-Idade , Plasmodium vivax/química , Plasmodium vivax/genética , Polimorfismo GenéticoRESUMO
Plasmodium simium is a parasite from New World monkeys that is most closely related to the human malaria parasite Plasmodium vivax; it also naturally infects humans. The blood-stage infection of P. vivax depends on Duffy binding protein II (PvDBPII) and its cognate receptor on erythrocytes, the Duffy antigen receptor for chemokines (hDARC), but there is no information on the P. simium erythrocytic invasion pathway. The genes encoding P. simium DBP (PsDBPII) and simian DARC (sDARC) were sequenced from Southern brown howler monkeys (Alouatta guariba clamitans) naturally infected with P. simium because P. simium may also depend on the DBPII/DARC interaction. The sequences of DBP binding domains from P. vivax and P. simium were highly similar. However, the genetic variability of PsDBPII was lower than that of PvDBPII. Phylogenetic analyses demonstrated that these genes were strictly related and clustered in the same clade of the evolutionary tree. DARC from A. clamitans was also sequenced and contained three new non-synonymous substitutions. None of these substitutions were located in the N-terminal domain of DARC, which interacts directly with DBPII. The interaction between sDARC and PvDBPII was evaluated using a cytoadherence assay of COS7 cells expressing PvDBPII on their surfaces. Inhibitory binding assays in vitro demonstrated that antibodies from monkey sera blocked the interaction between COS-7 cells expressing PvDBPII and hDARC-positive erythrocytes. Taken together, phylogenetic analyses reinforced the hypothesis that the host switch from humans to monkeys may have occurred very recently in evolution, which sheds light on the evolutionary history of new world plasmodia. Further invasion studies would confirm whether P. simium depends on DBP/DARC to trigger internalization into red blood cells.
Assuntos
Sistema do Grupo Sanguíneo Duffy/genética , Eritrócitos/parasitologia , Plasmodium vivax/genética , Plasmodium/genética , Alouatta , Animais , Anticorpos Antiprotozoários/imunologia , Células COS , Chlorocebus aethiops , Sistema do Grupo Sanguíneo Duffy/imunologia , Eritrócitos/imunologia , Evolução Molecular , Variação Genética , Haplótipos , Humanos , Filogenia , Plasmodium/imunologia , Plasmodium vivax/imunologia , Polimorfismo de Nucleotídeo Único , Conformação Proteica , Proteínas de Protozoários/genética , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Análise de Sequência de DNARESUMO
The development of pre-erythrocytic Plasmodium vivax vaccines is hindered by the lack of in vitro culture systems or experimental rodent models. To help bypass these roadblocks, we exploited the fact that naturally exposed Fy- individuals who lack the Duffy blood antigen (Fy) receptor are less likely to develop blood-stage infections; therefore, they preferentially develop immune responses to pre-erythrocytic-stage parasites, whereas Fy+ individuals experience both liver- and blood-stage infections and develop immune responses to both pre-erythrocytic and erythrocytic parasites. We screened 60 endemic sera from P. vivax-exposed Fy+ or Fy- donors against a protein microarray containing 91 P. vivax proteins with P. falciparum orthologs that were up-regulated in sporozoites. Antibodies against 10 P. vivax antigens were identified in sera from P. vivax-exposed individuals but not unexposed controls. This technology has promising implications in the discovery of potential vaccine candidates against P. vivax malaria.
Assuntos
Antígenos de Protozoários/isolamento & purificação , Sistema do Grupo Sanguíneo Duffy/imunologia , Eritrócitos/parasitologia , Malária Vivax/parasitologia , Plasmodium vivax/imunologia , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Formação de Anticorpos , Antígenos de Protozoários/genética , Colômbia , Biologia Computacional , Eritrócitos/imunologia , Perfilação da Expressão Gênica , Ensaios de Triagem em Larga Escala , Humanos , Fígado/parasitologia , Fígado/patologia , Vacinas Antimaláricas/imunologia , Plasmodium vivax/genética , Plasmodium vivax/crescimento & desenvolvimento , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Esporozoítos/imunologia , Esporozoítos/metabolismo , Regulação para CimaRESUMO
We present evidence for Plasmodium vivax infection among Duffy blood group-negative inhabitants of Brazil. The P. vivax identification was determined by both genotypic and non-genotypic screening tests. The Duffy blood group was genotyped by PCR/RFLP and phenotyped using a microtyping kit. We detected two homozygous FY*B-33 carriers infected by P. vivax, whose circumsporozoite protein genotypes were VK210 and/or P. vivax-like. Additional efforts are necessary in order to clarify the evidence that P. vivax is being transmitted among Duffy blood group-negative patients from the Brazilian Amazon region.