Frequency of TNFA

Polymorphisms in cytokine genes can alter the production of these proteins and consequently affect the immune response. The trihybrid heterogeneity of the Brazilian population is characterized as a condition for the use of ancestry informative markers. The objective of this study was to evaluate the frequency of -1031T>C, -308G>A and -238G>A TNFA, +874 A>T IFNG and -819C>T, and -592C>A IL10 gene polymorphisms and their association with malaria vivax and genomic ancestry. Samples from 90 vivax malaria-infected individuals and 51 noninfected individuals from northern Brazil were evaluated. Genotyping was carried out by using ASO-PCR or PCR/RFLP. The genomic ancestry of the individuals was classified using 48 insertion/deletion polymorphism biallelic markers. There were no differences in the proportions of African, European, and Native American ancestry between men and women. No significant association was observed for the allele and genotype frequencies of the 6 SNPs between malaria-infected and noninfected individuals. However, there was a trend toward decreasing the frequency of individuals carrying the TNF-308A allele with the increasing proportion of European ancestry. No ethnic-specific SNPs were identified, and there was no allelic or genotype association with susceptibility or resistance to vivax malaria. Understanding the genomic mechanisms by which ancestry influences this association is critical and requires further study

Polimorfismos dos Genes CD40, CD40L e BLYS, associados na co-estimulação dos Linfócitos B, em indivíduos naturalmente infectados pelo Plasmodium vivax na Amazônia Brasileira

Plasmodium vivax é a espécie mais prevalente de malária no Brasil. O processo co-evolutivo parasita-hospedeiro pode ser visto como uma ferramenta, na qual trocas genéticas adaptativas podem influenciar na diversidade da população. Objetivo: Investigar polimorfismos de genes envolvidos na resposta imune humoral visando identificar possíveis associações com a malária. Material e Métodos: a amostra foi constiuída por 103 pacientes com malária vivax não complicada e como grupo controle 97 indíviduos não-maláricos. A identificação dos SNPs –726T>C no gene CD40L, -1 C>T no gene CD40 e -871C>T no gene BLYS foram efetuadas pelo método de PCR-RFLP. As frequências genotípicas, alélicas e de indivíduos portadores de cada alelo foram estimadas por contagem direta. Também foram comparadas as frequências genotípicas observadas com as esperadas segundo o teorema de Hardy e Weinberg. Resultados: As freqüências genotípicas e alélicas para esses SNPs não diferiram estatisticamente entre pacientes e indivíduos do grupo controle. A combinação dos genótipos entre os genes CD40 e BLYS e entre CD40L e BLYS não revelou interação gênica na população estudada. Não foi observada associação entre resposta imune humoral e parasitemia nos indivíduos maláricos com os polimorfismos dos genes investigados. Ambos os genes se encontram em equilíbrio de Hardy e Weinberg. Conclusões: Os resultados deste estudo sugerem que as variantes genéticas analisadas nos genes CD40, CD40L e BLYS não afetam a funcionalidade das moléculas de modo que possa interferir na susceptibilidade a doença, mas estas variantes podem influenciar o curso clínico em vez de simplesmente aumentar ou diminuir a susceptibilidadePlasmodium vivax is the most prevalent malaria species in Brazil. The parasite-host coevolutionary process can be viewed as an ‘arms race’, in which adaptive genetic changes in one are eventually matched by alterations in the other. Objectives: following the candidate gene approach we analyzed the CD40, CD40L and BLYS genes that participate in B-cell co-stimulation, for associations with P. vivax malaria. Methods: the study sample included 97 patients and 103 controls. We extracted DNA using the extraction and purification commercial kit and identified the following SNPs: -1C>T in the CD40 gene, –726T>C in the CD40L gene and the -871C>T in the BLyS gene using PCR-RFLP. We analyzed the genotype and allele frequencies by direct counting. We also compared the observed with the expected genotype frequencies using the Hardy-Weinberg Equilibrium. Results: The allele and genotype frequencies for these SNPs did not differ statistically between patient and control groups. Gene-gene interactions were not observed between the CD40 and BLYS and between the CD40L and BLYS genes. Overall, the genes were in Hardy-Weinberg Equilibrium. Significant differences were not observed among the frequencies of antibody responses against P. vivax sporozoite and erythrocytic antigens and the CD40 and BLYS genotypes Conclusions: the results of this study show that, although the investigated CD40, CD40L and BLYS alleles differ functionally, this variation does not alter the functionality of the molecules in a way that would interfere in susceptibility to the disease. Significance: The variants of these genes may influence the clinical course rather than simply increase or decrease susceptibilit

Frequency of TNFA, INFG, and IL10 Gene Polymorphisms and Their Association with Malaria Vivax and Genomic Ancestry

Polymorphisms in cytokine genes can alter the production of these proteins and consequently affect the immune response. The trihybrid heterogeneity of the Brazilian population is characterized as a condition for the use of ancestry informative markers. The objective of this study was to evaluate the frequency of -1031T>C, -308G>A and -238G>A TNFA, +874 A>T IFNG and -819C>T, and -592C>A IL10 gene polymorphisms and their association with malaria vivax and genomic ancestry. Samples from 90 vivax malaria-infected individuals and 51 noninfected individuals from northern Brazil were evaluated. Genotyping was carried out by using ASO-PCR or PCR/RFLP. The genomic ancestry of the individuals was classified using 48 insertion/deletion polymorphism biallelic markers. There were no differences in the proportions of African, European, and Native American ancestry between men and women. No significant association was observed for the allele and genotype frequencies of the 6 SNPs between malaria-infected and noninfected individuals. However, there was a trend toward decreasing the frequency of individuals carrying the TNF-308A allele with the increasing proportion of European ancestry. No ethnic-specific SNPs were identified, and there was no allelic or genotype association with susceptibility or resistance to vivax malaria. Understanding the genomic mechanisms by which ancestry influences this association is critical and requires further study

Os polimorfismos do gene da citocina não estão associados aos níveis de anticorpos IgG anti-PvDBP, anti-PvAMA-1 ou anti-PvMSP-119 em uma área endêmica da malária da Amazônia brasileira

This study was financed by the Brazilian National Council for Scientific and Technological Development (Conselho Nacional de Desenvolvimento Cientí‑ fico e Tecnológico) process number 472135/2012, the Pará State Research Support Foundation (Fundação de Amparo à Pesquisa do Estado do Pará) and CAPPES. We thank Dr. Irene Soares and Luzia Carvalho for their help in serological analysis. This project was funded by CNPq and Capes.São José do Rio Preto Medical School. Department of Dermatology, Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil.São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil.Federal University of Sergipe. Department of Biology. Laboratory of Molecular Genetics and Biotechnology. São Cristóvão, SE, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Microbiology and Immunology. Belém, PA, Brazil.Universidade de Campinas. Department of Genetics, Evolution and Bioagents. Laboratory of Tropical Diseases. Campinas, SP, Brazil.São José do Rio Preto Medical School. Department of Dermatology, Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil / São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica em Malária. Ananindeua, PA, Brasil.The immune response against Plasmodium vivax immunogenic epitopes is regulated by pro- and anti-inflammatory cytokines that determine antibody levels and class switching. Cytokine gene polymorphisms may be responsible for changes in the humoral immune response against malaria. The aim of this study was to evaluate whether polymorphisms in the TNFA, IFNG and IL10 genes would alter the levels of anti-PvAMA1, PvDBP and -PvMSP119 IgG antibodies in patients with vivax malaria. Methods: Samples from 90 vivax malaria-infected and 51 uninfected subjects from an endemic area of the Brazilian Amazon were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to identify polymorphisms of the genes TNFA (-1031T > C, -308G > A, -238G > A), IFNG (+874T > A) and IL10 (-819C > T, -592C > A). The levels of total IgG against PvAMA1, PvDBP and PvMSP-119 were determined using an enzyme-linked immunosorbent assay (ELISA). Associations between the polymorphisms and the antibody response were assessed by means of logistic regression models. Results: No significant differences were found in the levels of IgG antibodies against the PvAMA-1, PvDBP or PvMSP119 proteins in relation to the studied polymorphisms. Conclusions: Although no associations were found among the evaluated genotypes and alleles and anti-merozoite IgG class P. vivax antibody levels, this study helps elucidate the immunogenic profile involved in the humoral immune response in malaria

Immunogenetic markers associated with a naturally acquired humoral immune response against an N-terminal antigen of Plasmodium vivax merozoite surface protein 1 (PvMSP-1)

São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil / São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil.São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil.São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil / São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil.Federal University of Sergipe. Department of Biology. São Cristóvão, SE, Brazil.Oswaldo Cruz Foundation. Leônidas and Maria Deane Institute. Manaus, AM, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Oswaldo Cruz Foundation. Leônidas and Maria Deane Institute. Manaus, AM, Brazil.São Paulo State University. Department of Biology. São José do Rio Preto, SP, Brazil / São José do Rio Preto Medical School. Department of Skin. Infectious and Parasitic Diseases. São José do Rio Preto, SP, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisa Básica de Malária. Belém, PA, Brasil.Background: Humoral immune responses against proteins of asexual blood-stage malaria parasites have been associated with clinical immunity. However, variations in the antibody-driven responses may be associated with a genetic component of the human host. The objective of the present study was to evaluate the influence of co-stimulatory molecule gene polymorphisms of the immune system on the magnitude of the humoral immune response against a Plasmodium vivax vaccine candidate antigen. Methods: Polymorphisms in the CD28, CTLA4, ICOS, CD40, CD86 and BLYS genes of 178 subjects infected with P. vivax in an endemic area of the Brazilian Amazon were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The levels of IgM, total IgG and IgG subclasses specific for ICB2-5, i.e., the N-terminal portion of P. vivax merozoite surface protein 1 (PvMSP-1), were determined by enzyme-linked immuno assay. The associations between the polymorphisms and the antibody response were assessed by means of logistic regression models. Results: After correcting for multiple testing, the IgG1 levels were significantly higher in individuals recessive for the single nucleotide polymorphism rs3116496 in CD28 (p = 0.00004). Furthermore, the interaction between CD28 rs35593994 and BLYS rs9514828 had an influence on the IgM levels (p = 0.0009). Conclusions: The results of the present study support the hypothesis that polymorphisms in the genes of co-stimulatory components of the immune system can contribute to a natural antibody-driven response against P. vivax antigens

Polymorphism in the IL-1β promoter is associated with IgG antibody response to circumsporozoite protein repeats of Plasmodium vivax

Conselho Nacional para o Desenvolvimento Científico e Tecnológico (CNPq) ; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil (CAPES – DS and PDSE) – Finance Code 001São Paulo State University. Graduate Program in Biosciences. São José do Rio Preto, SP, Brazil.Lisboa University. Tropical Medicine and Hygiene Institut. Global Health and Tropical Medicine. Lisboa, Portugal.Federal University of Sergipe. Department of Biology. Aracajú, SE, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Imunogenética da Malária Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil / Universidade Federal do Pará. Belém, PA, Brasil.Fluminense Federal University. Center of Microorganisms Investigation. Niterói, RJ, Brazil.Fluminense Federal University. Center of Microorganisms Investigation. Niterói, RJ, Brazil.Fluminense Federal University. Center of Microorganisms Investigation. Niterói, RJ, Brazil.Fundação Oswaldo Cruz. Oswaldo Cruz Institute. Laboratory of Malaria Research. Rio de Janeiro, RJ, Brazil.São Paulo State University. Graduate Program in Biosciences. São José do Rio Preto, SP, Brazil.Fundação Oswaldo Cruz. Oswaldo Cruz Institute. Laboratory of Immunoparasitology. Rio de Janeiro, RJ, Brazil.São Paulo State University. Graduate Program in Biosciences. São José do Rio Preto, SP, Brazil / Fluminense Federal University. Center of Microorganisms Investigation. Niterói, RJ, Brazil.BACKGROUND: It is well established that infection by Plasmodium vivax is a result of host-parasite interactions. In the present study, association with the IL1/IL2 cytokine profiles, anticircumsporozoite protein antibody levels and parasitic loads was evaluated in individuals naturally infected with P. vivax in an endemic area of the Brazilian Amazon. METHODS: Molecular diagnosis of P. vivax and variants was performed using the PCR-RFLP method and IL1B -511C>T, IL2 -330T>G and IL2+114T>G polymorphisms were identified using PCR-RFLP and allele-specific PCR. IL-1β and IL-2 cytokine levels were detected by flow cytometry and circumsporozoite protein (CSP) antibodies were measured by ELISA. RESULTS: Three variants of P. vivax CSP were identified and VK247 was found to be the most frequent. However, the prevalence and magnitude of IgG antibodies were higher for the VK210 variant. Furthermore, the antibody response to the CSP variants was not associated with the presence of the variant in the infection. Significant differences were observed between the single nucleotide polymorphism (SNP) -511T>C in the IL1B gene and levels of antibodies to the VK247 and P. vivax-like variants, but there were no associations between SNPs in IL1 and IL2 genes and their plasma products. CONCLUSIONS: Individuals with the rs16944 CC genotype in the IL1β gene have higher antibody levels to the CSP of P. vivax of VK247 and P. vivax-like variants

Humoral immune responses against the malaria vaccine candidate antigen Plasmodium vivax AMA-1 and IL-4 gene polymorphisms in individuals living in an endemic area of the Brazilian Amazon

Background: Several studies have recently demonstrated that the immune responses against malaria is governed by different factors, including the genetic components of the host. The IL-4 gene appears to be a strong candidate factor because of its role in the regulation of the Th2 response. The present study investigated the role of IL-4 polymorphisms in the development of IgG antibodies against PvAMA-1 and the IL-4 levels in individuals infected with Plasmodium vivax in a malaria endemic area in the Brazilian Amazon.Methods: The study sample included 83 patients who were diagnosed with P. vivax infection using thick smear and confirmed by nested-PCR. The IL-4 590 C>T and IL-4 33 C>T polymorphisms were genotyped by PCR-RFLP, and the intron 3 VNTR was genotyped by PCR. A standardised ELISA protocol was used to measure the total IgG against PvAMA-1. The cytokine/chemokine levels were measured using a Milliplex multiplex assay (Millipore). All of the subjects were genotyped with 48 ancestry informative markers to determine the proportions of African, European and Amerindian ancestry using STRUCTURE software.Results: Of the 83 patients, 60 (73%) produced IgG antibodies against PvAMA-1. A significant decrease in the percentage of respondents was observed among the primo-infected individuals. No significant differences were observed in the frequencies of genotypes and haplotypes among individuals who were positive or negative for IgG antibodies against PvAMA-1. Furthermore, no significant correlation was observed between the IL-4 polymorphisms, antibody levels, IL-4 levels, and parasitemia.Conclusions: This study indicated that the polymorphisms identified in the IL-4 gene are not likely to play a role in the regulation of the antibody response against PvAMA-1 and IL-4 production in vivax malaria. (C) 2015 Elsevier Ltd. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq