Novel regulatory SNPs in the promoter region of the TNFRSF18 gene in a Gabonese population

Parasites are accountable for driving diversity within immune gene families. We identified and investigated regulatory single nucleotide polymorphisms (SNPs) in the promoter regions of the tumor necrosis factor receptor superfamily member 18 (TNFRSF18) gene by direct sequencing in a group of male Gabonese individuals exposed to a wide array of parasitic diseases such as malaria, filariasis and schistosomiasis. Two new promoter variants were identified in 40 individuals. Both novel variants were heterozygous and were linked to SNP #rs3753344 (C/T), which has been described. One of the SNP variants (ss2080581728) was close to the general transcription factor site, the TATA box. We further validated these new promoter variants for their allelic gene expression using transient transfection assays. One new promoter variant with two base changes (C/T - ss2080581728/rs3753344) displayed an altered expression of the marker gene. Both novel variants remained less active at the non-induced state in comparison to the major allele. The allele frequencies observed in this study were consistent with data for other African populations. The detection and analysis of these human immune gene polymorphisms contribute to a better understanding of the interaction between host-parasite and expression of Treg activity.

Novel regulatory SNPs in the promoter region of the TNFRSF18 gene in a Gabonese population.

Parasites are accountable for driving diversity within immune gene families. We identified and investigated regulatory single nucleotide polymorphisms (SNPs) in the promoter regions of the tumor necrosis factor receptor superfamily member 18 (TNFRSF18) gene by direct sequencing in a group of male Gabonese individuals exposed to a wide array of parasitic diseases such as malaria, filariasis and schistosomiasis. Two new promoter variants were identified in 40 individuals. Both novel variants were heterozygous and were linked to SNP #rs3753344 (C/T), which has been described. One of the SNP variants (ss2080581728) was close to the general transcription factor site, the TATA box. We further validated these new promoter variants for their allelic gene expression using transient transfection assays. One new promoter variant with two base changes (C/T - ss2080581728/rs3753344) displayed an altered expression of the marker gene. Both novel variants remained less active at the non-induced state in comparison to the major allele. The allele frequencies observed in this study were consistent with data for other African populations. The detection and analysis of these human immune gene polymorphisms contribute to a better understanding of the interaction between host-parasite and expression of Treg activity.

Prevalence and level of antibodies to the circumsporozoite proteins of human malaria parasites, including a variant of Plasmodium vivax, in the population of two epidemiologically distinct areas in the state of Acre, Brazil.

A seroepidemiological study of the prevalence of antibodies against the repeating epitopes of circumsporozoite (CS) proteins of human malaria parasites was conducted in 2 different areas in the state of Acre, Brazil in 1987 and 1990. In 1987 antibodies against the CS protein of the VK 247 variant Plasmodium vivax as well as antibodies against the CS proteins of P. falciparum and the classic P. vivax were found at relatively high rates in the 2 areas, but significant microepidemiological differences were observed. In 1990, when large scale migration in Amazonia had ceased and control measures were applied in the study areas, the malaria endemicity decreased, as determined by the declining prevalence of anti-sporozoite antibodies against all Plasmodium species, and the small number of individuals with positive blood smears. Antibodies against sporozoites of the variant P. vivax did not cross-react with the CS proteins of the classic P. vivax, nor with antibodies against sporozoites of P. falciparum and P. malariae. Sera containing antibodies against the CS protein of P. malariae were found at a very low frequency, and only in 1987. The anti-CS protein antibody response to all Plasmodium species was age-related.

Levels of the schistosome circulating anodic and cathodic antigens in serum of schistosomiasis patients from Brazil.

Serum levels of 2 schistosome circulating antigens, the circulating anodic antigen (CAA) and the circulating cathodic antigen (CAA), were determined in persons infected with Schistosoma mansoni in Brazil. Sensitive monoclonal antibody-based enzyme-linked immunosorbent assays were used to measure levels of the 2 antigens. The study group consisted of 38 individuals with intestinal schistosomiasis, and 20 persons with the hepatosplenic form of the disease. Age and intensity of infection were comparable for the 2 groups. CAA was detected in 65.5% of all patients' sera and CCA was found in the serum of 82.8% of all patients. CAA levels correlated well with the egg output, as determined by duplicate Kato-Katz smears; CCA was significantly positively correlated with egg output in patients with intestinal schistosomiasis only. Whereas no significant difference was found between CAA titre in patients with intestinal schistosomiasis and those with the hepatosplenic form, a significantly higher CCA titre was found in patients with hepatosplenomegaly compared to patients with intestinal schistosomiasis.

Drug susceptibility of Plasmodium falciparum in the western Amazon region State of Acre, Brazil

Estudos de campo na Amazonia ocidental (Estado do Acre, Brasil) indicam que as 4-aminoquinolinas, assim como a sua combinacao com sulfadoxina-pirimetamina nao podem mais ser recomendadas para o tratamento e profilaxia das infeccoes pelo P. falciparum nesta regiao. A quinina permanece como droga efetiva quando usada corretamente. Entretanto, problemas podem surgir devido aos efeitos colaterais durante a sua aplicacao por periodo de 10 dias. Possibilidades de ultrapassar estes problemas combinando curto espaco de administracao da quinina com outras drogas estao no momento limitadas devido a falta de um composto associado adequado. Por esta razao, a combinacao quinina/clindamicina parece ser a terapeutica mais adequada para a malaria pelo P. falciparum. Nossos estudos in vitro sugerem que a mefloquina e outra alternativa efetiva para o tratamento da malaria falciparum nesta regiao da Amazonia.

Drug susceptibility of Plasmodium falciparum in the western Amazon region, State of Acre, Brazil.

Field studies in the western Amazon region (state of Acre, Brazil) indicate that the 4-aminoquinolines, as well as the combined regimen with sulfadoxine-pyrimethamine, can no longer be recommended for the treatment and prophylaxis of P. falciparum infections in this region. Quinine remains an effective drug when used correctly. However, compliance problems arise due to the often occurring side-effects during a ten day regimen. Prospects of overcoming these constraints by combining a short course of quinine with other drugs are limited, because of the lack of suitable partner compounds. For this reason quinine/clindamycin appears to be a more practical therapy of P. falciparum malaria. In vitro data from this study suggest that mefloquine is another effective alternative for the treatment of falciparum malaria in this Amazon region.

Interaction of rheumatoid factor and Entamoeba histolytica

Testes para citotoxicidade e lise amebiana foram utilizados para deminstrar uma possível interaçäo entre o fator reumatóide e a Entamoeba histolytica. A atividade citotóxica amebiana foi inibida pela IgG antiameba de coelho purificada através de cromatografia. Constatou-se inibiçäo aumentada com IgG antiameba de coelho mais fator reumatóide. A mesma inibiçäo acentuada da atividade citotóxica amebiana pôde ser constatada quando se substituiu o fator reumatóide por sor humano normal, inativado pelo calor, como controle. Cerca de 50% de lise amebiana ocorreu quando as amebas foram misturadas com soro normal humano como fonte de complemento. A lise amebiana aumentou para 60% quando incubadas com soro humano normal, acrescido de anticorpos humanos antiameba. Nenum aumento adicional pode ser obtido pela adiçäo de fator reumatóide. Usando IgG antiameba de coelho em vez de anticorpos humanos, a proporçäo de lise näo aumentou. A incubaçäo de amebas com soro humano normal, IgG antiameba de coelho e fator reumatóide reduziu acentuadamente a lise amebiana. O fator reumatóide näo teve efeito na atividade citotóxica amebiana, nem na lise amebiana mediada pelo complemento in vitro

Interaction of rheumatoid factor and Entamoeba histolytica.

The amoebae's cytotoxicity test and the amoebae's lysis test were used to show possible interactions between rheumatoid factor (RF) and Entamoeba histolytica. Amoebae's cytotoxic activity (ACA) was inhibited by affinity chromatography purified antiamoebae rabbit IgG (RIgG). Enhanced inhibition could be demonstrated with RIgG plus RF. But the same marked inhibition of ACA could be seen when replacing RF by heat inactivated normal human serum as a control. About 50% amoebae's lysis occurred when amoebae were brought together with native normal human serum (NNHS) as a source of complement. Amoebae's lysis increased to 60% when incubated with NHS plus human antiamoebae antibodies. No further augmentation could be obtained by the addition of RF. Using RIgG instead of human antibodies the lysis rate did not increase. Incubation of amoebae, NNHS, RIgG and RF even reduced amoebae's lysis. RF neither has an effect on ACA nor on complement mediated AL in vitro.

Effects of intradermal gamma-interferon in cutaneous leishmaniasis.

The clinical and immunological evolution of lesions in cutaneous leishmaniasis was assessed after treatment with human recombinant gamma interferon (rIFN-gamma). 3 weeks after rIFN-gamma treatment of lesions due to Leishmania braziliensis guyanensis, 12/13 had become smaller compared with 6/13 control lesions; only 4 treated lesions were free of parasites. 9 of 13 L tropica lesions treated with rIFN-gamma resolved completely within 4-8 weeks of treatment. An acute inflammatory reaction around treated lesions was more common in lesions due to L tropica. There were no other local or systemic adverse reactions. Histological and immunohistochemical studies indicate that local application of rIFN-gamma enhances cell-mediated immune responses and thus promotes healing of cutaneous leishmaniasis.

Differences in drug response of Plasmodium falciparum within an area of the Amazon region.

Clinical characteristics of patients with falciparum malaria, as well as sensitivity of Plasmodium falciparum to chloroquine and mefloquine, were investigated in 2 distinct strata within the same geographical area of the Amazon Basin. One stratum was the population living along the road, the other that living along the river, both near Rio Branco, capital of Acre State, Brazil. The clinical features did not differ between the 2 strata. Full in vitro sensitivity of P. falciparum to mefloquine was observed in both areas. However, significant differences in chloroquine sensitivity were observed between the 2 strata. EC50 values for chloroquine were 0.8484 mumol/litre for parasite isolates from the road stratum (E) and 0.4638 mumol/litre for parasite isolates from the river stratum (R). EC90 values were 2.8095 and 1.2549 mumol/litre in strata 'E' and 'R', respectively. Continuous drug pressure over years in area 'E' and relatively low drug pressure in area 'R' were presumably responsible for these differences.

Clindamycin treatment of falciparum malaria in Brazil.

Oral treatment with clindamycin (5 mg/kg twice a day, for five consecutive days) was studied in patients with uncomplicated falciparum malaria in Acre, Brazil, an area with multiresistant Plasmodium falciparum. Parasitaemia ranged between 12 and 79560/microliters of blood admission. Thirty-five out of 44 patients admitted to the study could be followed up for 28 days. Only two patients showed parasitaemia six days after admission, and no asexual parasites were observed by day seven. Twenty-eight days after admission all patients were cured. Of the nine patients withdrawn from the study, five were lost during follow up and four needed different treatment (quinine 15 mg/kg twice a day, for ten days) because clinical symptoms did not improve within 60 h after admission. These patients had experienced their first attack by P. falciparum. In individual cases oral clindamycin can be used as an alternative treatment in semi-immune patients with uncomplicated falciparum malaria from an area where multiresistant parasites frequently occur. However, because of the slow response in all cases described here, and the risk of development of resistance if clindamycin is used alone it cannot be recommended as monotherapy in non-immune patients.

In vitro drug sensitivity of Plasmodium falciparum in Acre, Brazil.

In Acre, the westernmost state of Brazil in the Amazon region, the sensitivity of Plasmodium falciparum to chloroquine, amodiaquine, mefloquine, quinine and sulfadoxine/pyrimethamine was determined in vitro by the Rieckmann microtechnique. The study was performed between January and June 1987; the in vitro parasite responses to all antimalarial drugs were determined according to the recommendations of WHO. Of 83 isolates of P. falciparum, all were sensitive to mefloquine and of 87 isolates of P. falciparum, 84 (97%) were sensitive to quinine. The EC50 for mefloquine was 0.27 mumol/l and for quinine 4.60 mumol/l. In contrast, 65 of 89 (73%) and 70 of 83 (84%) isolates were resistant to amodiaquine and chloroquine, respectively; 11 isolates even grew at 6.4 mumol chloroquine/l. The EC50 for amodiaquine was 0.34 mumol/l and for chloroquine 0.73 mumol/l. Sulfadoxine/pyrimethamine resistance was seen in 23 of 25 (92%) cases. These data clearly indicate that in the western part of the Amazon region the 4-aminoquinolines, as well as sulfadoxine/pyrimethamine, can no longer be recommended for the treatment of P. falciparum infections.