Development of Leishmania mexicana in Lutzomyia longipalpis in the absence of sugar feeding

The leishmaniases are caused by Leishmania parasites and transmitted through the bites of phlebotomine sand flies. During parasite development inside the vector's midgut, promastigotes move towards the stomodeal valve, a mechanism that is crucial for transmission. It has been reported that the sugar meal acquired by sand flies during feeding between bloodmeals is essential for the development and migration of parasites. We demonstrated that the distribution of Leishmania mexicana parasites was affected by the sugar meals obtained by the sand flies. Promastigote migration towards the cardia region seems to be only partially based on the stimuli provided by sugar molecules. In the absence of sugars, significant amounts of parasites developed in the hindgut. In addition, sugar meals were important for the survival of sand flies, especially during blood digestion, presumably supporting their energy requirements.

HSP70 of Leishmania amazonensis alters resistance to different stresses and mitochondrial bioenergetics

The 70 kDa heat shock protein (HSP70) is a molecular chaperone that assists the parasite Leishmania in returning to homeostasis after being subjected to different types of stress during its life cycle. In the present study, we evaluated the effects of HSP70 transfection of L. amazonensis promastigotes (pTEX-HSP70) in terms of morphology, resistance, infectivity and mitochondrial bioenergetics. The pTEX-HSP70 promastigotes showed no ultrastructural morphological changes compared to control parasites. Interestingly, the pTEX-HSP70 promastigotes are resistant to heat shock, H2O2-induced oxidative stress and hyperbaric environments. Regarding the bioenergetics parameters, the pTEX-HSP70 parasites had higher respiratory rates and released less H2O2 than the control parasites. Nevertheless, the infectivity capacity of the parasites did not change, as verified by the infection of murine peritoneal macrophages and human macrophages, as well as the infection of BALB/c mice. Together, these results indicate that the overexpression of HSP70 protects L. amazonensis from stress, but does not interfere with its infective capacity.

Efecto de la Leishmania mexicana, RANTES y TNF alfa en el fenotipo y propiedades migratorias in vitro de células dendríticas esplénicas de ratones BALB/c neonatos y adultos / Dendritic cells of BALB/c mice: phenotypic characteristics and migratory properties

Las células dendríticas (CDs) son esenciales en el desarrollo y regulación la respuesta inmunitaria (RI). Existen controversias en cuanto al potencial de inducción de la RI por las CDs en el período neonatal. Se ha propuesto que la RI específica de un neonato depende de la relación cuantitativa CD/linfocito T, y del momento, etapa neonatal o adulta, del encuentro con el antígeno, lo que parece influir sobre las propiedades fenotípicas y biológicas de las CDs, modificando su comportamiento. Por tal motivo, nos planteamos evaluar el efecto de un antígeno, Leishmania mexicana (L. mexicana) y de las citoquinas TNFa y RANTES sobre las características fenotípicas y propiedades migratorias, in vitro, de las CDs esplénicas provenientes de ratones BALB/c neonatos y adultos, usando citometría de flujo y la cámara de Boyden. Las CDs de ratones neonatos y adultos, en condiciones basales, expresan de manera similar, las moléculas CD40, CD86, CMHII y CD54. Este mismo fenómeno se observó al incubar dichas células con el Ag (L. mexicana) a excepción de la molécula CD40 cuya intensidad de expresión se elevó significativamente (P<0,05) en ambos grupos de estudio. El índice de migración de las CDs en presencia de medio de cultivo condicionado de L. mexicana, RANTES y TNFa fue mayor en adultos que en neonatos. Estos resultados muestran que las CDs neonatales son fenotípicamente similares a las adultas. Ante los mismos estímulos se comportan de manera diferente, sugiriendo la existencia de otros factores, que pudieran explicar la mayor susceptibilidad a infecciones en la etapa neonatal.

Dendritic cells (DCs) are essential in the development and regulation of the immune response (IR). The inherent potential of DCs to induce a specific immune response in the neonatal period is controversial. It has been suggested that the specific IR in neonates depends on the quantitative relation of DC/T lymphocytes, as well as on the neonatal or adult age at which the interaction antigen/DC/T lymphocytes occurs. This suggests that this contact has an influence on the phenotypic and/or biological properties of DCs, which modifies its behavior. Therefore, the effects of Leishmania mexicana (L. mexicana) and of TNFa and RANTES cytokines on immunophenotypical characteristics were evaluated on spleen DCs, from neonate and adult BALB/c mice, by using flow cytometry and in vitro migratory properties with a Boyden Chamber. In basal conditions, neonate and adult DCs express the same molecules (CD40, CD86, MHCII and CD54). When the DCs interact with the antigen L. mexicana, the expression of these molecules are similar in adults and in neonates, with the exception of CD40 whose intensity of expression was raised (P<0,05) in both groups. The rate of migration of the DCs in a culture medium conditioned of L. mexicana, RANTES and TNFa was higher in adults than in newborn mice. These observations suggest that neonatal and adult mice DCs have similar phenotypic characteristics. Under the effect of the same stimulus they respond differently; suggesting that other factors are involved in the higher susceptibility that newborns have to infections.

In vitro initial immune response against Leishmania amazonensis infection is characterized by an increased production of IL-10 and IL-13

The initial encounter of Leishmania with its host's immune system is important in the outcome of infection. Previous studies have shown that PBMCs from healthy volunteers (HV) exposed to Leishmania differ in IFN-γ production. We have expanded such observations evaluating the profile and kinetics of cytokines (IFN-γ, IL-12p70, IL-10, IL-13), chemokines (CCL5, CCL3, CCL4, CXCL10), and chemokine receptors (CCR1,CCR5, CXCR3, CCR4) in vitro L. amazonensis-stimulated of HV's PBMCs. HVs were divided in groups of high (HR) or low (LR) IFN-γ responders. In both groups, HR and LR, after L. amazonensis infection there was a predominance of IL-10 and IL-13 over IFN-γ production, while IL-12 was produced in similar amount. Regarding chemokines, a more striking difference was observed for CCL3 expression that was lower at 12 hours and 48 hours post infection in LR than in HR. Interestingly, a downregulation of CCR5 and a greater expression of CCR4 were found in low IFN-γ responders. These data suggest that early after L. amazonensis infection there is a cytokine milieu dominated by IL-13 and IL-10, and despite of this environment, IFN-γ is produced, supporting the complexity of the response. It is noteworthy that the pattern of immune response is mounted in first hours after Leishmania stimulation, with the definition of the differentiation of Th1 versus Th2 cells. It remains to be determined if such an in vitro difference has an in vivo counterpart in terms of susceptibility to infection.

Effect of a second bloodmeal on the oesophagus colonization by Leishmania mexicana complex in Lutzomyia evansi (Diptera: Psychodidae)

Migration and colonization of the oesophagus by Leishmania mexicana parasites were enhanced after digestion of a second bloodmeal intake in Lutzomyia evansi. This event has epidemiological significance since it affects the infection susceptibility of this sand fly species, which is a proven vector of L. chagasi in Colombian and Venezuelan visceral leishmaniasis foci. Also, it may explain the host seeking behaviour displayed by some partially bloodfed flies found inside houses

La leishmaniasis en México / Leishmaniases in Mexico

La leishmaniasis causa una gran variedad de condiciones clínicas y un cuadro patológico complejo debido a la virulencia variable del parásito y la respuesta inmune del huésped. En la leishmaniasis cutánea causada por L. mexicana frecuentemente se presentan úlceras cutáneas y en ciertos casos, una destrucción parcial o total del pabellón auricular (úlcera de los chicleros). En el noreste de México, las lesiones patológicas se limitan al sitio de la picadura infectante, con edema local, que lleva a la formación de una placa granulomatosa crónica, conocida como leishmaniasis tipo "botón de Oriente". Rara vez la enfermedad toma la forma de nódulos múltiples no ulcerados conocidos como leishmaniasis cutánea difusa; ésta última forma nunca cura espontáneamente y responde mal frente a la quimioterapia. En México, los puntos principales a investigar serán los de establecer la importancia de la leishmaniasis, por medio de un sistema para determinar la incidencia de las varias formas, los aspectos ecológicos, la identificación de los parásitos y una quimioterapia mejor de los casos

Observaciones sobre el parasito leishmania mexicana amazonensis y la infeccion natural que provoca en el flebotomo lutzomyia olmeca nociva / Observations on the parasite leishmania mexicana amazonensis and its natural infection of the sand fly lutzomyia olmeca nociva

In 1984 the authors began a search for vectors of the Leishmania mexicana amazonensis parasite in the Brazilian Amazon city of Manaus by capturing phlebotomines (sandflies) at two periurban collection sites and seeking to isolate the parasite from them. One collection site yielded no positive specimens, but the older yielded 10 sandflies infected with Le m. amazonensis. Six of the positive specimens belonged to the sandfly species Lutzomyia flaviscutellata, and four belonged to the species Lu. olmeca nociva. This is the first time that the latter species has been incriminated as a possible vector of Le. m. amazonensis and the first record of Lu. flaviscutellata being infected with this parasite outside of Para State. The capture of infected phlebotomines at one site but no at another similar nearby site is reminiscent of previous results reported by Lainson and Shaw. Whether or not these results indicate the existence of discrete transmission foci is still unclear