Dinámica y especialización funcional de las células dendríticas y macrófagos pulmonares durante reacciones alérgicas

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 17-12-2015Esta tesis tiene embargado el acceso al texto completo hasta el 17-06-2017We have analyzed the kinetics of the lung monocyte-DC-macrophage system, as well as the contribution of Ly-6Chigh monocytes to the generation of these populations, using a lung allergy model against House Dust Mite (HDM). The dynamics of lung populations in C57BL/6 and in mice deficient in CCR2, a chemokine that controls Ly-6Chigh monocytes egression from bone marrow, indicate that recruited monocytes differentiate in situ into moDCs during HDM-induced allergy. Additionally, we have observed that monocyte blockade correlates to a higher allergic reaction. These data suggest that monocytes and/or monocyte-derived cells could play a role in the negative regulation of HDM-induced allergy. Furthermore, we have studied alveolar populations during the allergic response. Our data suggest that alveolar macrophages undergo an apoptotic process that leads to a massive disappearance of this cell population during a strong allergic reaction to HDM, paralleled by the peak of alveolar eosinophilia. Once the allergic response dampens, the alveolar macrophage population recovers, but the newly formed alveolar macrophage population differs phenotipically from the pre-existing population. This transient alveolar macrophage disappearance does not occur during Klebsiella pneumoniae infection, suggesting a link between eosinophil induced damage and alveolar macrophages disappearanc

A liquid crystal of ascorbyl palmitate, used as vaccine platform, provides sustained release of antigen and has intrinsic pro-inflammatory and adjuvant activities which are dependent on MyD88 adaptor protein

Modern subunit vaccines require the development of new adjuvant strategies. Recently,32 we showed that CpG-ODN formulated with a liquid crystal nanostructure formed by self33assembly of 6-O-ascorbyl palmitate (Coa-ASC16) is an attractive system for promoting an34 antigen-specific immune response to weak antigens. Here, we showed that after35 subcutaneous injection of mice with near-infrared fluorescent dye-labeled OVA antigen36 formulated with Coa-ASC16, the dye-OVA was retained at the injection site for a longer37 period than when soluble dye-OVA was administered. Coa-ASC16 alone elicited a local38 inflammation, but how this material triggers this response has not been described yet.39 Although it is known that some materials used as a platform are not immunologically inert,40 very few studies have directly focused on this topic. In this study, we explored the41 underlying mechanisms concerning the interaction between Coa-ASC16 and the immune42 system and we found that the whole inflammatory response elicited by Coa-ASC1643 (leukocyte recruitment and IL-1β, IL-6 and IL-12 production) was dependent on the MyD8844 protein. TLR2, TLR4, TLR7 and NLRP3-inflammasome signaling were not required for45 induction of this inflammatory response. Coa-ASC16 induced local release of self-DNA,46 and in TLR9-deficient mice IL-6 production was absent. In addition, Coa-ASC16 revealed47 an intrinsic adjuvant activity which was affected by MyD88 and IL-6 absence. Taken48 together these results indicate that Coa-ASC16 used as a vaccine platform is effective due49 to the combination of the controlled release of antigen and its intrinsic pro-inflammatory50 activity. Understanding how Coa-ASC16 works might have significant implications for51 rational vaccine design.Fil: Sánchez Vallecillo, María Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Minguito de la Escalera, María. Consejo Superior de Investigaciones Científicas. Centro Nacional de Biotecnología. Departamento de Inmunología y Oncología; EspañaFil: Aguirre, María Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Ullio Gamboa, Gabriela Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; Argentina. Consejo Superior de Investigaciones Científicas. Centro Nacional de Biotecnología. Departamento de Inmunología y Oncología; EspañaFil: Palma, Santiago Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Gonzalez Cintado, Leticia. Consejo Superior de Investigaciones Científicas. Centro Nacional de Biotecnología. Departamento de Inmunología y Oncología; EspañaFil: Chiodetti, Ana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Soldano, Germán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Moron, Victor Gabriel. Consejo Superior de Investigaciones Científicas. Centro Nacional de Biotecnología. Departamento de Inmunología y Oncología; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Allemandi, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Ardavín, Carlos. Consejo Superior de Investigaciones Científicas. Centro Nacional de Biotecnología. Departamento de Inmunología y Oncología; EspañaFil: Pistoresi, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Maletto, Belkys Angélica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Airway allergy causes alveolar macrophage death, profound alveolar disorganization and surfactant dysfunction

Respiratory disorders caused by allergy have been associated to bronchiolar inflammation leading to life-threatening airway narrowing. However, whether airway allergy causes alveolar dysfunction contributing to the pathology of allergic asthma remains unaddressed. To explore whether airway allergy causes alveolar dysfunction that might contribute to the pathology of allergic asthma, alveolar structural and functional alterations were analyzed during house dust mite (HDM)-induced airway allergy in mice, by flow cytometry, light and electron microscopy, monocyte transfer experiments, assessment of intra-alveolarly-located cells, analysis of alveolar macrophage regeneration in Cx3cr1cre:R26-yfp chimeras, analysis of surfactant-associated proteins, and study of lung surfactant biophysical properties by captive bubble surfactometry. Our results demonstrate that HDM-induced airway allergic reactions caused severe alveolar dysfunction, leading to alveolar macrophage death, pneumocyte hypertrophy and surfactant dysfunction. SP-B/C proteins were reduced in allergic lung surfactant, that displayed a reduced efficiency to form surface-active films, increasing the risk of atelectasis. Original alveolar macrophages were replaced by monocyte-derived alveolar macrophages, that persisted at least two months after the resolution of allergy. Monocyte to alveolar macrophage transition occurred through an intermediate stage of pre-alveolar macrophage and was paralleled with translocation into the alveolar space, Siglec-F upregulation, and downregulation of CX3CR1. These data support that the severe respiratory disorders caused by asthmatic reactions not only result from bronchiolar inflammation, but additionally from alveolar dysfunction compromising an efficient gas exchange.Ministerio de Economía y CompetitividadMinisterio de Ciencia e InnovaciónComunidad de MadridDepto. de Bioquímica y Biología MolecularFac. de Ciencias QuímicasTRUEpu