Antígenos compartidos entre Plasmodium falciparum y Anopheles albimanus

The presence of common antigens between Plasmodium falciparum and Anopheles albimanus was demonstrated. Different groups of rabbits were immunized with: crude extract from female An. albimanus (EAaF), red blood cells infected with Plasmodium falciparum (EPfs), and the SPf66 synthetic malaria vaccine. The rabbit's polyclonal antibodies were evaluated by ELISA, Multiple Antigen Blot Assay (MABA), and immunoblotting. All extracts were immunogenic in rabbits according to these three techniques, when they were evaluated against the homologous antigens. Ten molecules were identified in female mosquitoes and also in P. falciparum antigens by the autologous sera. The electrophoretic pattern by SDS-PAGE was different for the three antigens evaluated. Cross-reactions between An. albimanus and P. falciparum were found by ELISA, MABA, and immunoblotting. Anti-P. falciparum and anti-SPf66 antibodies recognized ten and five components in the EAaF crude extract, respectively. Likewise, immune sera against female An. albimanus identified four molecules in the P. falciparum extract antigen. As far as we know, this is the first work that demonstrates shared antigens between anophelines and malaria parasites. This finding could be useful for diagnosis, vaccines, and the study of physiology of the immune response to malaria.Epítopes de antígenos compartidos entre Plasmodium falciparum y Anopheles albimanus fueron identificados. Diferentes grupos de conejos fueron inmunizados con: extracto crudo de mosquito hembra de An. albimanus (EAaH), glóbulos rojos infectados con P. falciparum (EPfs) y la vacuna antimalárica sintética SPf66. Los anticuerpos policlonales producidos en conejos fueron evaluados por ELISA, inmunoensayo simultáneo de múltiples antígenos (MABA) e Immunoblotting. Todos los extractos resultaron inmunogénicos cuando se evaluaron por ELISA, MABA e Immunoblotting. Diez moléculas fueron identificadas en los mosquitos hembras y diez en los antígenos de P. falciparum por los sueros autólogos. El patrón electroforético por SDS-EGPA fue diferente para los tres antígenos evaluados. La reactividad cruzada de moléculas entre An. albimanus y P. falciparum fue demostrada por ELISA, MABA e Immunoblotting. Anticuerpos anti-P. falciparum y anti-SPf66 reconocieron diez y cinco componentes respectivamente en el extracto crudo de anofelinos (EAaH). Asimismo, sueros inmunes contra An. albimanus hembra identificaron cuatro moléculas en el extracto del antígeno de P. falciparum. Hasta el presente, este es el primer estudio en el que se demuestra la presencia de antígenos compartidos entre anofelinos y los parásitos de malaria. Este hallazgo podría ser de relevancia para el diagnóstico, vacunas e interpretación de la fisiopatología de la respuesta inmunitaria en malaria

Sharing of antigens between Plasmodium falciparum and Anopheles albimanus

The presence of common antigens between Plasmodium falciparum and Anopheles albimanus was demonstrated. Different groups of rabbits were immunized with: crude extract from female An. albimanus (EAaF), red blood cells infected with Plasmodium falciparum (EPfs), and the SPf66 synthetic malaria vaccine. The rabbit's polyclonal antibodies were evaluated by ELISA, Multiple Antigen Blot Assay (MABA), and immunoblotting. All extracts were immunogenic in rabbits according to these three techniques, when they were evaluated against the homologous antigens. Ten molecules were identified in female mosquitoes and also in P. falciparum antigens by the autologous sera. The electrophoretic pattern by SDS-PAGE was different for the three antigens evaluated. Cross-reactions between An. albimanus and P. falciparum were found by ELISA, MABA, and immunoblotting. Anti-P. falciparum and anti-SPf66 antibodies recognized ten and five components in the EAaF crude extract, respectively. Likewise, immune sera against female An. albimanus identified four molecules in the P. falciparum extract antigen. As far as we know, this is the first work that demonstrates shared antigens between anophelines and malaria parasites. This finding could be useful for diagnosis, vaccines, and the study of physiology of the immune response to malaria

Conocimiento, actitudes y prácticas sobre la Malaria en comunidades de Maniapure, Estado Bolívar

Introducción: La Malaria es una enfermedadde importancia epidemiológica, la cual ha tenido unrepunte en los últimos años en el Estado Bolívar. Alser potencialmente mortal, es vital la elaboración deestrategias preventivas basadas en los conocimientosy prácticas comunitarias. Objetivos: Identificar losconocimientos, actitudes y prácticas relacionadoscon la malaria y comparar con hallazgos de otrasáreas endémicas. Métodos: Se realizó un estudioobservacional, descriptivo y de corte transversal enla región de Maniapure del 28 al 31 de agosto del2018. Se aplicó un cuestionario de 30 preguntas derespuestas cerradas. Los resultados fueron presentadossegún su frecuencia absoluta y relativa. Resultados:Se entrevistaron a 77 jefes de familia, de los cuales63,64 % (n=49) refirió tener antecedente de malaria y38,96 % (n=30) conoció a alguien que haya fallecido ensu comunidad por esta. En cuanto a conocimientos,79,22 % (n=61) sabe que la enfermedad se adquierepor la picadura de un mosquito, 51,95 % (n=40) creeque se transmite por el agua y 97,4 % (n=75) asociala fiebre como síntoma principal. A nivel de actitudes yprácticas, el 92,21 % (n=71) acudiría al médico para tratarla enfermedad, 62,34 % (n=29) mantiene los canalesy zanjas limpias y 70,13 % (n=54) usa mosquitero y/orepelente. Conclusión: El área de Maniapure se ubicaen una región endémica, y al tratarse de poblacionessusceptibles con diferente trasfondo cultural, se deben elaborar planes educativos individualizados orientadosa la prevención, y evaluar su impacto en la práctica

Conocimiento, actitudes y prácticas sobre la Malaria en comunidades de Maniapure, Estado Bolívar

Introducción: La Malaria es una enfermedadde importancia epidemiológica, la cual ha tenido unrepunte en los últimos años en el Estado Bolívar. Alser potencialmente mortal, es vital la elaboración deestrategias preventivas basadas en los conocimientosy prácticas comunitarias. Objetivos: Identificar losconocimientos, actitudes y prácticas relacionadoscon la malaria y comparar con hallazgos de otrasáreas endémicas. Métodos: Se realizó un estudioobservacional, descriptivo y de corte transversal enla región de Maniapure del 28 al 31 de agosto del2018. Se aplicó un cuestionario de 30 preguntas derespuestas cerradas. Los resultados fueron presentadossegún su frecuencia absoluta y relativa. Resultados:Se entrevistaron a 77 jefes de familia, de los cuales63,64 % (n=49) refirió tener antecedente de malaria y38,96 % (n=30) conoció a alguien que haya fallecido ensu comunidad por esta. En cuanto a conocimientos,79,22 % (n=61) sabe que la enfermedad se adquierepor la picadura de un mosquito, 51,95 % (n=40) creeque se transmite por el agua y 97,4 % (n=75) asociala fiebre como síntoma principal. A nivel de actitudes yprácticas, el 92,21 % (n=71) acudiría al médico para tratarla enfermedad, 62,34 % (n=29) mantiene los canalesy zanjas limpias y 70,13 % (n=54) usa mosquitero y/orepelente. Conclusión: El área de Maniapure se ubicaen una región endémica, y al tratarse de poblacionessusceptibles con diferente trasfondo cultural, se deben elaborar planes educativos individualizados orientadosa la prevención, y evaluar su impacto en la práctica

Comparison of serological responses in two different populations with pulmonary tuberculosis

Observational studies on the humoural immune responses of the Warao indigenous people from Delta Amacuro, an isolated area, were compared with urban residents of the Venezuelan capital. Mycobacterium tuberculosis -specific reactivities (IgM, IgE, sIgA, IgG and IgG subclasses) were measured by ELISA using PPD and 38-kDa M. tuberculosis antigens. A total of 294 individuals were studied, 162 Warao (indigenous people) and 132 Creole (non-indigenous people). The patient group consisted of 87 Warao patients and 58 Creole patients, while the control group consisted of 75 Warao controls and 74 Creole controls. Combinations among the isotypes studied were performed. The findings showed that for the Warao people, sensitivity to the combination including anti-PPD IgG and IgE was 92.0%, while for the Creole people, sensitivity to the combination including anti-PPD IgG but more so anti-PPD IgG1 and IgG2 was 90.0%. Simple tests were able to show higher specificities, which were population-specific; specificities were anti-PPD IgG3, 100.0% and anti-PPD IgM, 97.4% for the Warao and Creole peoples, respectively. In conclusion, while simple tests reached high specificity, the multi-isotype tests improved sensitivity; the latter shows this approach may be useful in diagnostic testing

Differential NRAMP1gene’s D543N genotype frequency: Increased risk of contracting tuberculosis among Venezuelan populations

NRAMP1 and VDR gene polymorphisms have been variably associated with susceptibility to tuberculosis (TB) amongst populations having different genetic background. NRAMP1 and VDR gene variants’ association with susceptibility to active infection by Mycobacterium tuberculosis (Mtb) was analyzed in the Warao Amerindian population, an ethnic population from Venezuela's Orinoco delta region. Genomic DNA was extracted from individuals with and without TB to evaluate genetic polymorphism by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Four NRAMP1 gene polymorphisms were analyzed: D543N (rs17235409), 3′ UTR (rs17235416), INT4 (rs3731865), and 274C/T (rs2276631), and one VDR gene polymorphism: FokI (rs2228570). The results showed that the genotypes D543N-A/A, 3′UTR-TGTG+/+, INT4-C/C, and 274C/T-T/T of known polymorphism in the NRAMP1 gene, as well as the genotypes FokI-F/f and FokI-f/f in the VDR gene were most often found in indigenous Warao with active TB. Binomial logistic regression was used for evaluating associations between polymorphisms and risk of contracting TB, an association between NRAMP1-D543N-A/A genotype distribution and TB susceptibility was found in Warao Amerindians. Regarding Venezuelan populations having different genetic backgrounds; statistically significant TB associations concerning NRAMP1-D543N-A/A, INT4-C/C and 3′UTR-TGTG+/+ variant genotype distributions in Warao Amerindians (indigenous) compared to Creole (admixed non-indigenous population) individuals were found. In conclusion, the results thus indicated that the association between NRAMP1-D543N-A/A genotype and TB in Warao Amerindians could support such allele's role in host susceptibility to Mtb infection.Incluye referencias bibliográfica

Evaluation of the transcriptional immune biomarkers in peripheral blood from Warao indigenous associate with the infection by Mycobacterium tuberculosis

Abstract INTRODUCTION: Biomarkers are critical tools for finding new approaches for controlling the spread of tuberculosis (TB), including for predicting the development of TB therapeutics, vaccines, and diagnostic tools. METHODS: Expression of immune biomarkers was analyzed in peripheral blood cells stimulated and non-stimulated with M. tuberculosis antigens ESAT-6, CFP10 and TB7.7. in Warao indigenous individuals. These biomarkers may be able to differentiate TB states, such as active tuberculosis (ATB) cases and latent tuberculosis infection (LTBI) from non-infected controls (NIC). A real-time reverse transcription polymerase chain reaction (RT-qPCR) assay was performed on 100 blood samples under non-stimulation or direct ex vivo conditions (NS=50) and stimulation conditions (S=50). RESULTS: The findings are shown as the median and interquartile range (IQR) of relative gene expression levels of IFN-γ, CD14, MMP9, CCR5, CCL11, CXCL9/MIG, and uPAR/PLAUR immune biomarkers. MMP9 levels were significantly higher in the LTBI-NS and LTBI-S groups compared with the NIC-NS and NIC-S groups. However, CCR5 levels were significantly lower in the LTBI-S group compared with both NIC-NS and NIC-S groups. CCL11 levels were significantly lower in the LTBI-S group compared with the NIC-NS group. CONCLUSIONS: Preliminary findings showed that MMP9 immune biomarkers separated LTBI indigenous individuals from NIC indigenous individuals, while CCR5, CCL11, CD14, and IFN-γ did not differentiate TB states from NIC. MMP9 may be useful as a potential biomarker for LTBI and new infected case detection among Warao indigenous individuals at high risk of developing the disease. It may also be used to halt the epidemic, which will require further validation in larger studies

Rerouting of a Follicle-Stimulating Hormone Analog to the Regulated Secretory Pathway

LH and FSH are produced by the same gonadotrope cells of the anterior pituitary but differ in their mode of secretion. This coordinated secretion of LH and FSH is essential for normal follicular development and ovulation in females and for spermatogenesis in males. The structural signals encoded in the LH and FSH subunits that govern the intracellular sorting of LH through the regulated secretory pathway and FSH through the constitutive pathway are largely unknown. Our laboratory recently identified the seven amino acid carboxy tail of LHβ as a sorting signal for LH in GH3 cells. Here we compared the morphological features of GH3 cells expressing an FSH analog containing the heptapeptide (FL7AA) with wild-type FSH using confocal microscopy. These experiments were performed to develop a rerouting model for examining structure-function links between secretion pathways of FSH/LH and their biological action. Both FSH- and LH-expressing cells exhibit a fluorescence pattern of randomly dispersed cytoplasmic puncta. FL7AA expressing cells have more intracellular accumulation compared with wild-type FSH and display a unique halo pattern of fluorescence near the plasma membrane. Such a pattern was not observed in cells expressing FSH or LH. Our results demonstrate that this FSH analog containing the carboxy heptapeptide of LHβ is rerouted to the regulated secretory pathway in GH3 cells. This rerouted gonadotropin provides a unique model to study the trafficking, regulation, and function of LH and FSH

Patterns of selection on Plasmodium falciparum erythrocyte-binding antigens after the colonization of the New World.

Pathogens, which have recently colonized a new host species or new populations of the same host, are interesting models for understanding how populations may evolve in response to novel environments. During its colonization of South America from Africa, Plasmodium falciparum, the main agent of malaria, has been exposed to new conditions in distinctive new human populations (Amerindian and populations of mixed origins) that likely exerted new selective pressures on the parasite's genome. Among the genes that might have experienced strong selective pressures in response to these environmental changes, the eba genes (erythrocyte-binding antigens genes), which are involved in the invasion of the human red blood cells, constitute good candidates. In this study, we analysed, in South America, the polymorphism of three eba genes (eba-140, eba-175, eba-181) and compared it to the polymorphism observed in African populations. The aim was to determine whether these genes faced selective pressures in South America distinct from what they experienced in Africa. Patterns of genetic variability of these genes were compared to the patterns observed at two housekeeping genes (adsl and serca) and 272 SNPs to separate adaptive effects from demographic effects. We show that, conversely to Africa, eba-140 seemed to be under stronger diversifying selection in South America than eba-175. In contrast, eba-181 did not show any sign of departure from neutrality. These changes in the patterns of selection on the eba genes could be the consequence of changes in the host immune response, the host receptor polymorphisms and/or the ability of the parasite to silence or express differentially its invasion proteins