Innate lymphoid cells in peripheral blood of patients with American Cutaneous Leishmaniasis.

Innate lymphoid cells (ILCs) are classified by the expression of specific transcription factors: ILC1 depending on T-bet for IFN-γ production; ILC2 depending on GATA3 for IL-5 and IL-13; and ILC3 depending on ROR-γτ and AHR for IL-17 and IL-22. This study aimed to determine circulating ILCs in 23 patients with localized (LCL) = 7, mucocutaneous (MCL) = 10, intermediate (ICL) = 3 and diffuse (DCL) = 3 cutaneous leishmaniasis and 17 healthy controls from endemic area (EC) = 9 and non-endemic area (HC) = 8. Results evidenced a higher proportion of ILC1 in LCL than controls and MCL. ILC2 was higher in DCL compared with controls. ILC3 s were abundant in MCL and DCL concerning controls. A prevalence ratio was calculated to approach cell plasticity: in LCL, the ratio showed a prevalence of ILC1/ILC3 (plasticity 1), in contrast to DCL, and controls, where ILC2/ILC3 (plasticity 3) is prevalent. Also, MCL and ICL showed higher ILC1/ILC2 (plasticity 2). These results suggest that ILC1 and ILC3 in LCL are associated with disease control and regulation of inflammation, while MCL and ICL are related to immunopathology and uncontrolled inflammation. In DCL, ILC2 is associated with the tolerogenic state of these patients.

Células inflamatorias en la secreción nasal y citocinas proinflamatorias Th1, Th2, Th17 y reguladoras en el suero de pacientes con leishmaniasis cutánea Americana / Inflammatory cells in the nasal secretion and serum pro-inflammatory Th1, Th2, Th17 and regulatory cytokines from American cutaneous leishmaniasis patients

En la forma mucocutánea (LCM) y cutánea (LCL) de la leishmaniasis, se genera una respuesta inflamatoria cuyos mediadores (células y citocinas) se han involucrado en la severidad de las úlceras y en el daño tisular observado en estos pacientes, particularmente en los LCM. Por ello, nos propusimos identificar los grupos celulares predominantes en la secreción nasal de pacientes con LCL y LCM, y relacionarlos con citocinas proinflamatorias y reguladoras. Evaluamos en pacientes LCL (n=20), LCM (n=14) y 20 individuos sanos: a) La cuantificación de tipos de leucocitos en "frotis" de secreción nasal, úlceras cutáneas y sangre periférica teñidos con Giemsa empleando microscopía óptica, b) Concentraciones séricas de IL-8, IL-4 e IL-10 por citometría de flujo (CBA array) e IFN-γ, TNF-α e IL-17 por ELISA. El grupo celular predominante en la secreción nasal de pacientes con LCM fueron los neutrófilos (80,7%) y escasos eosinófilos (0,6%), comparados con los LCL y controles, en los que no se observaron estas células. Mientras que los "frotis" de las ulceras de los LCL presentaron 45,3% de neutrófilos y 43% de linfocitos. En contraste, en sangre periférica, de los pacientes se observó un incremento de neutrófilos y linfocitos junto a una frecuencia significativa de monocitos (LCM: 5,3; LCL: 6,3%) y eosinófilos (LCM: 8,2%; LCL: 5,2%). Todo esto sugiere la participación de los neutrófilos en la inmunopatogénesis en la LCM. Adicionalmente, se demostró una mayor (P=0,03) concentración sérica de IL-8 en los pacientes con LCL (18,5ρg/mL) y LCM (18,2ρg/mL) respecto a los individuos sanos, sugiriendo que esta citocina promueve el reclutamiento de neutrófilos al sitio de infección en los LCM, mientras que en los LCL contribuyen junto con los linfocitos T CD4+ de la subpoblación Th1 y productores de IFN-γ, en la activación de mecanismos leishmanicidas.

In mucocutaneous (MCL) and cutaneous (LCL) leishmaniasis, the inflammatory mediators (cytokines and cells) have been associated with ulcers severity and tissue damage observed in these patients, particularly in MCL. Therefore, we decided to identify the predominant cell groups in the nasal secretion of LCL and MCL patients, and related pro-inflammatory and regulatory cytokines. It was evaluated in LCL (n = 20), MCL patients (n = 14) and 20 healthy volunteers: a) Differential leukocyte count by optical microscopy performed in: smear of a runny nose, skin ulcers and peripheral blood dyed with Giemsa, b) serum levels of IL-8, IL-4 and IL-10 using cytometric bead array (CBA) assay and IFN-γ, TNF-α and IL-17 by ELISA. In MCL patients, neutrophils (80.7%) were the most abundant cellular group in nose secretion, followed by a small amount of eosinophils (0.6%) compared to the LCL and controls, where no such cells were observed. In contrast, in peripheral blood from ACL patients were observed an abundant amount of neutrophils and lymphocytes together with a significant frequency of monocytes (MCL:5.3%; LCL: 6.3%) and eosinophils (MCL:8.2%; LCL:5.2%). While the smear from skin ulcers of LCL patients showed 45.3% of neutrophils and 43% lymphocytes. All of these indicate that neutrophils might play a role in the MCL immunopathogenesis. Moreover, an increased serum levels of IL-8 (P=0.03) were found in LCL (18.5ρg/mL) and MCL (18.2ρg/mL) patients, suggesting that this cytokine promotes the recruitment of neutrophils to the infection site in MCL; while in LCL patients may contribute with CD4 + Th1 (IFN-γ) cells in the activation of leishmanicida mechanisms.

Intestinal parasitic infections and atopic dermatitis among Venezuelan Warao Amerindian pre- school children.

BACKGROUND: We evaluated the influence of intestinal parasitic infection on food sensitization associated to the severity of Atopic Dermatitis (AD) in a group of Warao Amerindian pre- school children. METHODS: Feces examinations were performed in fresh stool specimens. Diagnosis of AD was done according to Hannifin and Rajka criteria and SCORAD index. Skin prick tests (SPT) were performed using extracts of cow's milk (CM), hen's egg (HE), Dermatophagoides pteronyssinus and Dermatophagoides farinae . Serum CM and HE-IgE levels (ELISA) were measured. Quantikine (R&D systems) assays were used for the determination of IL-13, TNF-α IL-6, and sCD23 in supernatants of CM- and HE- whole blood stimulated samples. RESULTS: Atopic Dermatitis was reported in 23% of the children. It was significantly (p < 0.0001) associated toward both CM and HE- SPT positivity (p < 0.001). Giardia duodenalis infection (37%) was associated to the presence of AD (p = 0.005) and to a significant increase in the levels of CM stimulated TNF-α (p=0.006), IL-13 (p = 0.01), sCD23 (p = 0.001), CM-IgE (p = 0.012) and CM-SPT (p < 0.0001). Similarly, G. duodenalis infection was particularly associated with the increase on the levels of HE-stimulated TNF-α (p = 0.001), sCD23 (p = 0.001), HE-IgE levels (p = 0.002) and HE-SPT (p = 0.001). CONCLUSION: Gut inflammation caused by G. duodenalis may enhance food allergic reactivity contributing to the manifestation of AD in these children. However, other environmental factors (not considered in this work) as well as an atopic background among the Warao population would also contribute to the presence of AD.

Factores inmunológicos que pueden influir en la presencia de síntomas clínicos en niños con giardiasis / Immunological factors influencing the presence of clinical symptoms in children with giardiasis

El objetivo de este trabajo fue evaluar el perfil de la respuesta inmune según sintomatología intestinal en niños con giardiasis. Se realizó un estudio transversal clínico, inmunológico y parasitológico en 182 escolares (6-7 años) de la región de Barlovento, Estado Miranda. La evaluación clínica fue realizada haciendo énfasis en los síntomas intestinales y evaluación antropométrica. Se realizaron exámenes de heces seriadas. Los niveles de IgA secretora (total y específicos a G. duodenalis) así como los niveles de citocinas fueron determinados mediante ELISA. Los niveles de óxido nítrico usando el método colorimétrico de Griess. Análisis estadístico mediante Med calc Software, Versión 12.4.0. Encontramos que Los niveles plasmáticos de TNF-α y de óxido nítrico (en saliva) fueron más elevados en los niños con síntomas intestinales y giardiasis (P<0.001) mientras que los niveles de TGF-β, IL-10 y los niveles de IgA secretora total y especifica a G. duodenalis fueron más elevados (P<0.0001) en el grupo de asintomáticos. Los niveles de IL-6 fueron más elevados en los niños con giardiasis independientemente de la sintomatología. La co-infeccion con A. lumbricoides se asoció negativamente a la presencia de síntomas intestinales. Los indicadores antropométricos (Talla/edad, peso/Edad y Peso/ Talla) se asociaron positivamente a la presencia de citocinas reguladoras (P<0.005) y niveles de IgA secretora (P<0.0001). Así, Diferencias en el perfil de la respuesta inmune podrían asociarse a síntomas intestinales inducidos por giardiasis. Deficiencias en el estado nutricional se asocian a procesos inflamatorios y a la generación de síntomas mientras que la co-infección con helmintos intestinales favorece la reducción de la sintomatología.

We evaluated the immune profile associated to intestinal symptoms in children with giardiasis. A crosssectional study including clinical, immunological and parasitological aspects in 182 children (6-7 years) from the area of Barlovento, Miranda state, Venezuela was carried out. Clinical evaluation was performed with emphasis on intestinal symptoms and anthropometric parameters were also determined. Serial stool examinations were carried out. Secretory IgA levels (total and specific against G. duodenalis) and cytokine levels were determined by ELISA. Nitric oxide levels were determined using the Griess colorimetric method. Statistical analysis was performed by Med Calc Software, Version 12.4.0. We found that plasma levels of TNF-α and nitric oxide (salivary) were higher in children with intestinal symptoms and giardiasis (P<0.001) while the levels of TGF-β, IL-10 and IgA levels (total and anti-G. duodenalis) were higher (P<0.0001) in the asymptomatic group. The levels of IL-6 were higher in children with giardiasis regardless of symptoms. Coinfection with A. lumbricoides was associated negatively with the presence of intestinal symptoms. Anthropometric indicators (height / age, weight / age and weight / height) were positively associated with the presence of regulatory cytokines (P<0.005) and secretory IgA levels (P<0.0001). Therefore, differences in the immune profile may be associated with intestinal symptoms induced by giardiasis. Co-infection with intestinal helminths helps to reduce symptoms while nutritional deficiencies are associated with inflammatory processes and the generation of symptoms.

[Non invasive diagnostic tools for visceral leishmaniasis: a comparison of the immunoserological tests DAT, rK26 and rK39]. / Herramientas no invasivas en Venezuela: comparación entre las pruebas inmunoserológicas DAT, rK26 y rK39 en el diagnóstico de leishmaniasis visceral.

INTRODUCTION: Human visceral leishmaniasis is a serious public health problem in endemic countries because of its high potential lethality, particularly in children. Rapid diagnosis is essential to early treatment and control of visceral leishmaniasis. OBJECTIVE: The aim was to compare three serodiagnostic tools for human visceral leishmaniasis. MATERIALS AND METHODS: Three methods were compared: the rK39 dipstick (Kalazar detection test, Inbios International Inc.), ELISA rK26 and direct agglutination test (DAT) (KIT Biomedical Research). Fifty serum samples from patients positive for rK39 ELISA were compared from four endemic provinces in Venezuela: Nueva Esparta (Margarita island), Lara, Anzoátegui and Trujillo. Additional serum samples from 17 healthy volunteers and 25 patients with other diseases were included. The rK39 ELISA served as the baseline standard method. Sensitivity, specificity, positive predictive value, negative predictive value and likelihood ratio were calculated for each test. RESULTS: All methods had a positive correlation with rK39 ELISA (p<0.0001). They showed high sensitivity and specificity. The direct agglutination test and the rK39 dipstick showed high sensitivity values, 89.7% (95% CI: 81.34.0-98.2%) and 94.2% (95% CI: 87.7-100%), respectively, and high specificity, 81.0% (95% CI: 80.0-99.5%) and 100%. The rK26 ELISA showed good specificity, 99% (95% CI: 95.2-100%), but a very low sensitivity, 37% (95% CI: 23.4-50.2%). CONCLUSION: Overall results indicated that DAT and rK39 dipstick have the highest specificity and sensitivity. Both are simple, cost-effective and field applicable tests. Therefore, they are recommended for early and accurate diagnosis of visceral leishmaniasis.

Estudio preliminar de Citocinas en suero de perros con Leishmaniasis visceral en una zona endémica Venezolana / Preliminary Study of Cytokines in Sera from Dogs with Visceral Leishmaniasis from a Venezuelan Endemic Area

Diversos estudios en modelos de ratones han demostrado que la inmunidad protectora frente a Leishmania es mediada por linfocitos T, células presentadoras de antígeno y citocinas. Sin embargo, pocos estudios se han realizado para evaluar citocinas en perros infectados en forma natural con Leishmania infantum/chagasi. El perro doméstico es el principal reservorio del parásito, en tal sentido, el objetivo de este trabajo fue determinar las citocinas en suero de 33 perros con Leishmaniasis Visceral Canina (LVC), provenientes del estado Nueva Esparta, Venezuela (foco endémico). Los perros fueron clasificados en sintomáticos o asintomáticos, de acuerdo al análisis de los signos clínicos de la enfermedad, coincidentes con los títulos de anticuerpos contra las kinesinas recombinantes de Leishmania rK39 y rK26. Otros dos grupos incluyeron: 10 perros de la misma zona endémica como grupo control endémico (CE) y 10 perros de la zona no endémica como control sano (CS). Las concentraciones (pg/mL) de las citocinas solubles IFN-γ, TNF-α, IL-10, IL-6, IL-4 e IL-2 se determinaron por citometría de flujo (Kit CBA Hu Th1/Th2, BD TM). Los resultados mostraron concentraciones estadísticamente mayores (p<0,05) de IFN-γ (69,93±7,46), IL-4 (7,51±2,68), TNF-α (3,86±1,46) e IL-2 (39,85±3,84) en el grupo de perros asintomáticos, con respecto a los perros sintomáticos (60,8±10,6; 5,28±0,80; 2,76±0,72 y 36,04±3,61, respectivamente) y los perros sanos (51±14; 4,65±0,2; 3,21±0,89 y 32,65±5,86, respectivamente). Los perros asintomáticos también presentaron mayor concentración de IL-6 (4,9±0,55) que los CS (4,02±0,64) (p<0,01). Estos resultados demuestran que los perros en estado asintomático exhiben mayor proporción de citocinas de activación celular y proinflamatorias. Los resultados señalan a la medición de citocinas séricas como reflejo del estado inmunológico de los caninos en futuros estudios orientados a vacunación o terapia.

Different experimental murine models have shown that protective immunity against Lesihmania depends upon T cells, cytokines, and antigen presenting cells. However, the role of cytokines in naturally-infected hosts like domestic dogs is controversial. Few studies have evaluated cytokines in dogs naturally-infected with Leishmania infantum/chagasi. Since the domestic dog is the main reservoir of the parasite, a study was conducted to determine cytokines in serum of 33 dogs with Canine Visceral Lesihmaniasis from endemic areas of the State of Nueva Esparta, Venezuela. Dogs were classified as symptomatic (SD) and asymptomatic (AD), according to the expression of three or more clinical signs and levels of  antibodies for rK39 and rK26. Ten non-infected, rK39 negative controls were included from an endemic area (EA) and ten dogs from a non-endemic area were used as healthy controls (HC). The following cytokines (pg/mL) were measured in serum by flow cytometry (CBA Hu Th1/Th 2, BD TM kit): IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL-2. Results show a higher  concentration (P<0.05) of  IFN-γ (69.93±7.46), IL-4 (7.51±2.68), TNF-α (3.86±1.46), and IL-2 (39.85±3.84) in AD when compared with SD (60.8±10.6; 5.28±0.80; 2.76± 0.72; and 36.04±3.61, respectively); and HC (51±14; 4.65±0.2; 3.21±0.89, and 32.65±5.86, respectively). The AD also showed higher levels (P<0.01) of IL-6 (4.9±0.55) compared with HC (4.02±0.64). Results show that AD exhibit a higher proportion of cellular activation and proinflammatory cytokines. Results indicate that measuring of serum cytokines could reflect the immunological status in dogs in future clinical trials oriented to either vaccination or therapy.

Herramientas no invasivas en Venezuela: comparación entre las pruebas inmunoserológicas DAT, rK26 y rK39 en el diagnóstico de leishmaniasis visceral / Non invasive diagnostic tools for visceral leishmaniasis: a comparison of the immunoserological tests DAT, rK26 and rK39

Introducción. La leishmaniasis visceral constituye un problema de salud pública en los países en donde es endémica por ser potencialmente letal, principalmente en niños. El diagnóstico rápido es importante en el control de la enfermedad. Objetivo. Comparar las pruebas inmunocromatográficas rK39 (rK39 dipstick, Kalazar detect test, Inbios Internacional Inc.), ELISA rK26 y la prueba de aglutinación directa (Kit Biomedical Research) en relación con la prueba de ELISA rK39, como herramientas serodiagnósticas para la leishmaniasis visceral en Venezuela.Materiales y métodos. Se estudiaron 50 muestras séricas de pacientes positivos por la prueba ELISA rK39, provenientes de diferentes zonas endémicas: Nueva Esparta, Lara, Anzoátegui y Trujillo; se incluyeron 17 muestras de voluntarios sanos y 25 de pacientes con otras enfermedades. Se utilizó la prueba ELISA rK39 como método de referencia, considerándola como patrón de referencia imperfecto, a partir del cual se determinaron los valores de sensibilidad, especificidad, razón de verosimilitud y valores diagnóstico positivo y negativo en las demás pruebas evaluadas. Resultados. Todas las pruebas mostraron una fuerte correlación (p<0,0001) con la ELISA rK39. La aglutinación directa y la prueba inmunocromatográfica rK39 presentaron altos valores de sensibilidad, 89,74% (IC95% 81,34-98,15) y 94,15% (IC95% 87,65-100), respectivamente, y de especificidad, 81% (IC95% 79,96-99,51) y 100% (IC95% 100-100). La prueba ELISA rK26, a pesar de poseer buena especificidad, 99% (IC95% 95,17-100), tuvo baja sensibilidad, 37% (IC95% 23,41-50,15). Conclusión. Las pruebas de aglutinación directa y la prueba inmunocromatográfica rK39 presentaron los mayores valores de sensibilidad y especificidad. Ambas son simples, económicas y fácilmente aplicables. Por ello, son recomendables para efectuar un diagnóstico de leishmaniasis visceral eficaz y precoz en Venezuela.

Introduction. Human visceral leishmaniasis is a serious public health problem in endemic countries because of its high potential lethality, particularly in children. Rapid diagnosis is essential to early treatment and control of visceral leishmaniasis.Objective. The aim was to compare three serodiagnostic tools for human visceral leishmaniasis. Materials and methods. Three methods were compared: the rK39 dipstick (Kalazar detection test, Inbios International Inc.), ELISA rK26 and direct agglutination test (DAT) (KIT Biomedical Research). Fifty serum samples from patients positive for rK39 ELISA were compared from four endemic provinces in Venezuela: Nueva Esparta (Margarita island), Lara, Anzoátegui and Trujillo. Additional serum samples from 17 healthy volunteers and 25 patients with other diseases were included. The rK39 ELISA served as the baseline standard method. Sensitivity, specificity, positive predictive value, negative predictive value and likelihood ratio were calculated for each test. Results. All methods had a positive correlation with rK39 ELISA (p<0.0001). They showed high sensitivity and specificity. The direct agglutination test and the rK39 dipstick showed high sensitivity values, 89.7% (95% CI: 81.34.0-98.2%) and 94.2% (95% CI: 87.7-100%), respectively, and high specificity, 81.0% (95% CI: 80.0-99.5%) and 100%. The rK26 ELISA showed good specificity, 99% (95% CI: 95.2-100%), but a very low sensitivity, 37% (95% CI: 23.4-50.2%). Conclusion. Overall results indicated that DAT and rK39 dipstick have the highest specificity and sensitivity. Both are simple, cost-effective and field applicable tests. Therefore, they are recommended for early and accurate diagnosis of visceral leishmaniasis.

Antibody responses and resistance against Ascaris lumbricoides infection among Venezuelan rural children: the influence of ethnicity.

We studied the production of anti parasite antibodies and the resistance against A. lumbricoides infection among the school population of three different Venezuelan ethnic groups: Warao Amerindians, Afro- Americans and white 'Creole' descendents. The prevalence and intensity of A. lumbricoides were determined before and after 1 year of anthelminthic treatment. Parasitic loads were associated to pre-treatment serum levels of anti A. lumbricoides IgE and IgG4 antibodies. The intensity of the infection and the proportion of re-infected children were significantly lower (p < 0.0001) among the Warao Amerindians, exhibiting higher levels of IgE (p < 0.0001) and lower levels of IgG4 (p < 0.0001) compared to their non-Amerindian counterparts. IgE levels correlated inversely with the proportion of re-infected children, whereas the levels of IgG4 were positively associated. These results suggest that ethnicity may influence the balance of IgE/IgG4 levels, modulating the intensity of A. lumbricoides and the resistance to the infection.

Evaluación de una prueba de aglutinación directa como método alternativo para el diagnóstico de leishmaniasis visceral canina y humana en Venezuela / The direct agglutination test as an alternative method for the diagnosis of canine and human visceral leishmaniasis

Introduction. Visceral leishmaniasis is the most severe clinical form of leishmaniasis and is often fatal without proper treatment. Therefore, early and accurate diagnosis is important, but often difficult in endemic areas. Objective. The aim was to evaluate a direct agglutination test as a potential visceral leishmaniasis diagnostic method in endemic areas of Venezuela Materials and methods. The performance of the direct agglutination test, based on freezedried Leishmania donovani antigen was evaluated under laboratory conditions using serum samples of humans and dogs from several Venezuelan visceral leishmaniasis endemic areas: Nueva Esparta (Margarita Island), Lara, Anzoátegui and Trujillo Status. The study included confirmed visceral leishmaniasis patients (n=30), visceral leishmaniasis suspected subjects (n=4), healthy controls (n=19) and patients with other confirmed diseases (n=20). In addition, 24 serum samples from dogs with confirmed visceral leishmaniasis and 18 healthy control dogs were tested. Results. All serum samples of visceral leishmaniasis patients, either active or recovered, were positive. They showed anti- L. donovani titers above 1:1600. Three out of four suspected visceral leishmaniasis cases were also positive, while serum samples from endemic controls and patients with other diseases had titers lower than 1:800. A sensitivity of 100 percent was obtained for all threshold levels under consideration and 100 percent specificity at the threshold titer of 1:800 (95 percent confidence interval: 91-100 percent). A 93 percent sensitivity (95 percent confidence interval: 76-99 percent) was observed in dog samples, with 100 percent specificity (95 percent confidence interval: 79-100 percent) at the threshold titer of 1:200. Conclusion. The direct agglutination test seems suitable for use in epidemiological studies and for serological diagnosis of human visceral leishmaniasis and canine visceral leishmaniasis.

Introducción. La leishmaniasis visceral es la forma clínica más grave de la leishmaniasis. Ésta puede ser fatal si no se administra el tratamiento adecuado. Por ello, el diagnostico temprano es importante, pero a menudo difícil, en las áreas endémicas. Objetivo. Evaluar el potencial de la prueba de aglutinación directa como un método para el diagnóstico de leishmaniasis visceral en zonas endémicas de Venezuela. Materiales y métodos. La efectividad de la prueba de aglutinación directa con el antígeno congelación-descongelación de Leishmania donovani fue evaluada bajo condiciones de laboratorio usando muestras de sueros de humanos y perros provenientes de diferentes regiones endémicas de leishmaniasis visceral de Venezuela: Nueva Esparta (Isla de Margarita), Estados Lara, Anzoátegui y Trujillo. Se incluyeron pacientes con diagnóstico confirmado de leishmaniasis visceral (n=30), sospecha de leishmaniasis visceral (n=4), voluntarios sanos (n=19) y pacientes con otras enfermedades (n=20). Además, se evaluaron 24 muestras de suero de perros con leishmaniasis visceral y 18 controles. Resultados. Todas las muestras de los pacientes con leishmaniasis visceral activa o curada fueron positivas. Mostraron títulos anti-L. donovani por encima de 1:1.600. Tres de cuatro casos con sospecha de leishmaniasis visceral también resultaron positivos a la prueba, mientras que los sueros controles y los de los pacientes con otras patologías dieron títulos por debajo de 1:800. Se obtuvo una sensibilidad de 100% a todos los puntos de corte considerados y una especificidad de 100% al punto de corte de 1:800 (intervalo de confianza de 95%, IC95%: 90,97%-100%). Las muestras de perros mostraron una sensibilidad de 92,59% (IC95%: 75,69%- 99,09%) y 100% de especificidad (IC95%: 79,42%-100%) al punto de corte de 1:200. Conclusión. En general, nuestros resultados indican que el uso de la prueba de aglutinación directa es apropiado para la realización de estudios epidemiológicos y para el...

An aggregation pheromone modulates lekking behavior in the vector mosquito Aedes aegypti (Diptera: Culicidae).

Males of Aedes aegypti mosquitoes formed swarms in the laboratory, triggered by the onset of the photophase or by the presence of odors from a rat (which is a potential host for females). The swarm attracted both males and females and increased mating activity. The number of copulas per mosquito was positively correlated with the number of mosquitoes in the swarm and with the duration of the swarm. Swarming and mating activity increased with the presence of a host for females. Young sexually immature males, less than 24 h old, flew but did not swarm nor copulate. Observations using an olfactometer showed that swarming males produced a volatile pheromone that stimulates the flying activity of females at a distance. Females also produce a volatile attractant. The results suggest that males, and possibly also females, produce an aggregation pheromone that attracts males and females towards the swarm. The characteristics of the pheromone-mediated swarm may be described as a 3-dimensional lek. Our results suggest that the development of pheromone-based control systems and/or pheromone traps for the monitoring of vector populations is feasible, adding a new tool to combat this vector of several human pathogens.

The direct agglutination test as an alternative method for the diagnosis of canine and human visceral leishmaniasis.

INTRODUCTION: Visceral leishmaniasis is the most severe clinical form of leishmaniasis and is often fatal without proper treatment. Therefore, early and accurate diagnosis is important, but often difficult in endemic areas. OBJECTIVE: The aim was to evaluate a direct agglutination test as a potential visceral leishmaniasis diagnostic method in endemic areas of Venezuela. MATERIALS AND METHODS: The performance of the direct agglutination test, based on freeze-dried Leishmania donovani antigen was evaluated under laboratory conditions using serum samples of humans and dogs from several Venezuelan visceral leishmaniasis endemic areas: Nueva Esparta (Margarita Island), Lara, Anzoátegui and Trujillo Status. The study included confirmed visceral leishmaniasis patients (n=30), visceral leishmaniasis suspected subjects (n=4), healthy controls (n=19) and patients with other confirmed diseases (n=20). In addition, 24 serum samples from dogs with confirmed visceral leishmaniasis and 18 healthy control dogs were tested. RESULTS: All serum samples of visceral leishmaniasis patients, either active or recovered, were positive. They showed anti-L. donovani titers above 1:1600. Three out of four suspected visceral leishmaniasis cases were also positive, while serum samples from endemic controls and patients with other diseases had titers lower than 1:800. A sensitivity of 100% was obtained for all threshold levels under consideration and 100% specificity at the threshold titer of 1:800 (95% confidence interval: 91-100%). A 93% sensitivity (95% confidence interval: 76-99%) was observed in dog samples, with 100% specificity (95% confidence interval: 79-100%) at the threshold titer of 1:200. CONCLUSION: The direct agglutination test seems suitable for use in epidemiological studies and for serological diagnosis of human visceral leishmaniasis and canine visceral leishmaniasis.

Role of the low affinity IgE receptor (CD23) on the IgE response against Ascaris lumbricoides in Warao Amerindian children from Venezuela

El objetivo de este estudio fue investigar los mecanismos inmunitarios frente a helmintos gastrointestinales involucrados en la producción de IgE, particularmente el papel del receptor de baja afinidad para la IgE (CD23) en niños indígenas de la etnia Warao del Delta del Orinoco en Venezuela. Estudiamos un grupo no seleccionado (159) de niños Warao escolares con una alta prevalencia e intensidad de infección por parásitos helmintos, y un grupo control de 70 niños no parasitados de una comunidad rural criolla del mismo Estado. Los niveles totales y específicos de IgE anti-A lumbricoides, determinados por ELISA, fueron extremadamente elevados en los niños Warao (p < 0,001). Sin embargo, solo un reducido grupo de esos niños fue capaz de desarrollar una respuesta de IgE específica clínicamente significativa (> 0,7UI/mL) frente a antígenos de A lumbricoides. El número de células circulantes, T CD4+, B CD20+ CD21+ y la expresión del receptor de baja afinidad para IgE (CD23) determinados por citometría de flujo, fueron significativamente (p < 0,001) más elevados en los niños Warao, particularmente en aquellos con adecuados niveles de IgE específica frente al parásito. Se encontró una alta correlación (p < 0,001) entre los niveles totales y específicos de IgE específica y la expresión del receptor para la IgE (CD23) y una correlación significativa (p < 0,005) entre el número de células circulantes CD20+CD23+ y CD4+. Concluimos entonces que los antígenos de A. lumbricoides podrían estimular la producción de IgE total y específica a través de una vía dependiente del receptor CD23. Sin embargo, los factores ambientales y genéticos que podrían determinar la capacidad de desarrollar respuestas IgE eficientes en la población Warao, requiere de mayor estudio

Role of the low affinity IgE receptor (CD23) on the IgE response against Ascaris lumbricoides in Warao Amerindian children from Venezuela.

The objective of this study was to investigate the immune mechanisms against gastrointestinal helminths involved in the production of IgE, particularly the role of the low affinity IgE receptor (CD23) in Amerindian Warao children from the Orinoco Delta, Venezuela. We studied a group of unselected (n = 159) Warao school children with high prevalence and intensity of helminthic infection and a similar non- parasitized (n = 70) control group of a "creole" community also located at the Delta of the Orinoco. The levels of total and specific anti-Ascaris IgE, determined by ELISA, were extremely higher in the Warao children (p < 0.001) compared to the control group. However, only a reduced group of these children were able to develop a clinically significant (> 0.7 PRU/mL) IgE response against A lumbricoides antigens. Circulating T helper cells (CD3+CD4+), activated B (CD20+, CD20+CD21+) lymphocytes and the expression of the low affinity receptor for IgE (CD23), determined by flow cytometry, were significantly (p < 0.001) elevated in the Warao children, particularly in those with an enhanced capacity to mount efficient specific anti Ascaris IgE responses. Strong correlations (p < 0.001) were found between the number of circulating CD20+CD23+ cells and the level of total and anti-A lumbricoides IgE. Also, a significant (p < 0.005) correlation between CD20+CD23+ and CD4+ circulating cells was observed. We conclude that A lumbricoides antigens may stimulate the production of total and specific IgE through a CD23 dependent pathway. However the different environmental and genetics factors involved in the capacity to develop efficient specific responses among the Warao population still need to be elucidated.

Variations in the serum levels of soluble CD23, nitric oxide and IgE across the spectrum of American cutaneous leishmaniasis.

Serum IgE levels and the expression of low affinity receptor for IgE (Fc epsilon RII/CD23) are increased in cutaneous leishmaniasis. The ligation of CD23 receptor by IgE-anti-IgE immune complexes results in nitric oxide (NO) production, which is a critical leishmanicidal factor. Human monocytes/macrophages express Fc epsilon RII/CD23 after activation with IFN-gamma and IL-4. In the present study, we assessed the relationship between NO, total and Leishmania-specific IgE and soluble CD23 across the clinical spectrum of American cutaneous leishmaniasis (ACL). Sixty-nine ACL patients and 22 endemic controls were studied. NO concentration was estimated using the Greiss method. Soluble CD23, total IgE and anti-Leishmania IgE levels were measured using ELISA. Soluble CD23 was increased in the four patient groups (0.001