The type 2 inflammatory response favors recognition of tumor antigens by IgE in breast cancer.

BACKGROUND: Several studies describe an inverse statistical relationship between the presence of an allergy and development of cancer. However, the immunological mechanism involved in the relationship between these two degenerative diseases has not been explored. AIMS: The main objective of this study was to explore the possibility that the lymphocyte T helper (Th) 2 response, a characteristic of allergy, induces recognition of tumor antigens. METHODS AND RESULTS: Patients with a clinical diagnosis of breast ductal carcinoma were included. Histopathological markers related to proliferation of tumor cells were determined (Her-2-neu, Ki-67, estrogen receptor, and progesterone receptor). IHC was performed using IgE antibodies purified from an allergy patient and from each biopsy donor patient. Serum concentrations of cytokines representative of Th1 and Th2 inflammatory responses were determined. A total of 14 patients with a confirmed diagnosis of breast ductal carcinoma were included. IHC performed on biopsies showed a weak response when using purified IgE antibodies from an allergy patient; however, IHC using the IgE of each patient as the primary antibody showed an intense and highly specific signal. Serum concentrations of cytokines of the Th2 response, that is, IL-4 (130.5 pg/mL (116-135 pg/mL)), IL-5 (202 pg/mL (191-213 pg/mL)), and IL-13 (105.5 pg/mL (98-117 pg/mL)), were significantly higher than those of the Th1 response, that is, IL-6 (86 pg/mL (79-90 pg/mL)) and INF-γ (93 pg/mL (79-99 pg/mL)). CONCLUSION: Purified IgE antibodies specifically recognize tumor cells in breast ductal carcinoma.

Entamoeba histolytica Trophozoites Interact with the c-Met Receptor at the Surface of Liver Origin Cells through the Gal/GalNAc Amoebic Lectin.

Amoebiasis in humans is caused by the protozoan parasite Entamoeba histolytica, which cytotoxic activity has been demonstrated on a wide variety of target cells. The process involves the adherence of the parasite to the cell, and such adherence is mediated by an amoebic surface lectin, known as Gal/GalNAc lectin. It is composed of heavy, intermediate, and light subunits. The carbohydrate recognition domain (CRD) has been identified within a cysteine-rich region in the lectin heavy subunit and has an amino acid sequence identity to the receptor-binding domain of hepatocyte growth factor (HGF). Recombinant CRD has been previously shown to compete with HGF for binding to the c-Met receptor IgG fusion protein. In the present study, we searched for evidence of interaction between the Gal/GalNAc lectin at the surface of trophozoites with the c-Met receptor expressed at the surface of HepG2 in coculture assays. Immunoprecipitation of the coculture lysate indicated interaction of the c-Met with a 60 kDa peptide recognized by antiamoebic lectin antibody. Colocalization of both molecules was detected by fluorescence confocal microscopy. Incubation of HepG2 cells with HGF before coculture with trophozoites prevents the cytotoxic effect caused by the parasites but not their adherence to the cells. Our results point to Gal/GalNAc lectin as a ligand of the c-Met receptor at the surface of HepG2 cells.

Entamoeba histolytica: adhesins and lectins in the trophozoite surface.

Entamoeba histolytica is the causative agent of amebiasis in humans and is responsible for 100,000 deaths annually, making it the third leading cause of death due to a protozoan parasite. Pathogenesis appears to result from the potent cytotoxic activity of the parasite, which kills host cells within minutes. Although the mechanism is unknown, it is well established to be contact-dependent. The life cycle of the parasite alternates with two forms: the resistant cyst and the invasive trophozoite. The adhesive interactions between the parasite and surface glycoconjugates of host cells, as well as those lining the epithelia, are determinants for invasion of human tissues, for its cytotoxic activity, and finally for the outcome of the disease. In this review we present an overview of the information available on the amebic lectins and adhesins that are responsible of those adhesive interactions and we also refer to their effect on the host immune response. Finally, we present some concluding remarks and perspectives in the field.

Hepatocyte production of modulators of extracellular liver matrix in normal and cirrhotic rat liver.

In the present study, we found collagenolytic and gelatinolytic activity in the supernatants of hepatocyte cultures from rats with experimental CCl(4)-induced liver cirrhosis, in levels significantly higher than in comparable supernatants of hepatocyte cultures from normal rats. In addition, we clearly detected the messenger ribonucleic acids (mRNA) of four matrix metalloproteinases (MMP-2, MMP-3, MMP-10, and MMP-13) and of two tissue inhibitors of matrix metalloproteinases (TIMP-1 and TIMP-2) in hepatocytes from both normal and cirrhotic rats by RT-PCR and by in situ hybridization. Finally, we demonstrated MMP-2, MMP-3, and MMP-13 and TIMP-1 and TIMP-2 proteins in the same hepatocyte preparations by immunostaining. We conclude that rat hepatocytes produce the major enzymes and inhibitors involved in liver ECM modulation and therefore suggests that they might participate actively in the pathophysiology of liver cirrhosis in rats.

Desarrollo de un modelo experimental para la caracterización de la respuesta funcional del corioamnios humano

Objetivo: Este estudio fue diseñado para validar y caracterizar un modelo de cultivo de membranas corioamnióticas (MC) humanas que permita mantenerlas íntegras, viables y reproducir su capacidad de respuesta como tejido, que separa al compartimiento materno-fetal ante diversos estímulos asociados al proceso infeccioso. Material y métodos: Se utilizaron MC de mujeres con 37-40 semanas de gestación, sin trabajo de parto activo, ni señales clínicas y/o microbiológicas de infección cervicovaginal. Las MC fueron sujetadas a una placa de cultivo tipo transwell, que permitió la formación de dos compartimentos independientes delimitados por la membranas que se mantuvieron en cultivo 96 h, se evaluó la integridad estructural mediante resistencia eléctrica transepitelial y análisis histológico. La funcionalidad se midió estimulando de manera independiente o cunjunta amnios y/o corion con 500 ng/mL de lipopolisacárido o 5 ng/mL de IL-1β y cuantificando la secreción de TNFα por ELISA y de Metaloproteasa-9 (MMP-9) por zimografía, respectivamente. Resultados: Las diferentes poblaciones celulares de las MC se mantuvieron metabólicamente viables en el cultivo; los parámetros de integridad indicaron que no presentan cambios morfo-estructurales significativos. El estímulo con IL-1β induce secreción diferencial de MMP-9, que en corion alcanzó su máximo a la 4 h y en amnios a las 24 h. La estimulación selectiva con lipopolisacárido indujo la síntesis diferencial de TNFα, siendo el corion el principal productor con aproximadamente 60%. Conclusiones: El sistema de cultivo de MC propuesto, permite estudiar cuantitativa y cualitativamente la contribución de las distintas poblaciones celulares del corioamnios en la respuesta ante el estímulo diferencial con agentes inmunológico-infecciosos.

Objective: This study was designed to validate and characterize a culture model of human choriamniotic membranes (HCM) that keeps their viability, integrity and capacity to reproduce a response to several stimulus associated to an infectious process, as the tissue that separates the fetal and maternal compartment. Material and methods: We use HCM obtained after delivery by elective cesarean section. Women with 37-40 weeks of gestation without evidence of active labor or presence of clinical an microbiological signs of intrauterine/vaginal infection. The membranes were mounted in transwell devices, allowing testing two independent compartments (chorion and amnion) by physically separating the upper and lower chambers. 500 ng/mL of lipopolysaccharide was added to amniotic or chorionic surface and secretions of TNFα was measured in both compartments by specific enzyme-linked immunosorbent assays and Metalloproteinase-9 (MMP-9) secretions after stimulation with 5 ng/mL of IL-1β. Results: The viability test showed that the different cellular populations of the HCM keep their metabolic viability along 96 h of culture. The integrity parameters showed that the stay without morphologic and structural changes. Functional markers showed that membranes responded differentially to IL-1β stimulus; production of MMP-9 in chorion reached its maximum value at 4 h, while amnion reached it at 24 h. The selective stimulation of chorioamnion with lipopolysaccharide induced a differential synthesis of TNFα; the chorion was the principal producer with approximately 60% of total TNFα. Conclusions: The experimental model allows to study qualitatively and quantitatively the contribution of different cellular regions of the HCM, and its response to differential stimulation with immunologic agents.

Secretions of interleukin-1beta and tumor necrosis factor alpha by whole fetal membranes depend on initial interactions of amnion or choriodecidua with lipopolysaccharides or group B streptococci.

The present study evaluated the secretions of interleukin (IL)-1beta and tumor necrosis factor (TNF) alpha by fetal membranes stimulated with group B streptococci (GBS) and lipopolysaccharide (LPS). The aim was to evaluate the initial response of full-thickness membranes to the microbial insult using an in vitro experimental model that allowed testing of the individual contributions of amnion and choriodecidua to stimulation. Full-thickness membranes were obtained after delivery by elective cesarean section from women at 37-40 wk of gestation without evidence of active labor. The membranes were mounted in Transwell devices, physically separating the upper and lower chambers. The LPS (500 ng/ml) or GBS (1 x 10(6) colony-forming units/ml) was added to either the amniotic or choriodecidual surface, and accumulation of IL-1beta and TNFalpha were measured in both compartments using a specific ELISA. Fetal membranes followed different patterns of secretion of proinflammatory cytokines that depended on the side to which the stimulus was added or the nature of the stimulus itself. The TNFalpha was secreted by amnion and choriodecidua in the presence of LPS or GBS, and stimulation with GBS induced a greater synthesis of IL-1beta than did stimulation with LPS. Choriodecidual tissue was more responsive than amniotic tissue, and this response tended to be higher even when the stimulation was only on the amniotic side. However, the amnion plays an active role in recognizing LPS or GBS, contributing a significant amount of TNFalpha. Thus, cooperative and bidirectional communications occur between amnion and choriodecidua in response to bacterial products, which include intermembranous cytokine traffic and signaling between tissues.

Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters.

Amebic cysteine protease 2 (EhCP2) was purified from ethyl ether extracts of axenically grown trophozoites of Entamoeba histolytica strain HM1-IMSS. The purification procedure involved molecular filtration and electroelution. Sequence analysis of the purified product revealed EhCP2 and ubiquitin(s). Electrophoretic migration patterns, isoelectric point determination and Western blot studies failed to reveal other EhCP molecules. Polyclonal antibodies against the purified EhCP2 prepared in rabbits either stabilized or enhanced the enzyme activity in a dose-response manner. Purified EhCP2 was enclosed within inert resin microspheres (22-44 microm in diameter) and injected into the portal vein of normal hamsters. In the liver, the microspheres caused mild acute inflammation and occasional minimal necrosis of short duration. Sections of the liver were immunohistochemically stained with the anti-EhCP2 antibody and the microspheres were positive for only a very short period (1 h) after injection. Sections of experimental acute (1 day, 5 days) amebic liver abscess produced in hamsters were also stained with the anti-EhCP2 antibody; and amebas were intensely positive but no staining was observed at any time in the surrounding necrotic structures. It is suggested that EhCP2 plays either a minor or no role in the causation of tissue damage in experimental acute liver amebiasis.

Mechanism of ammonium translocation in rat liver mitochondria: finger-printing of the translocator

Nosotros hemos estudiado si el amoníaco atraviesa la membrana interna de la mitocondria de hígado de rata como una especie cargada (NH+4) o descargada (NH3). El hinchamiento pasivo de mitocondrias suspendidas en acetato de amonio y/o sodio, mostró que: el hinchamiento depende de la concentración de amonio de la misma forma en que depende de la de sodio (a concentración de acetato constante). Las curvas alcanzan una meseta a 115 mM de amonio y 100 mM de sodio. Un cambio de dos unidades de pH induce un cambio de dos órdenes de magnitud en la concentración de NH3, en tanto que la de NH+4 permanece casi constante. Sin embargo, la magnitud y la velocidad iniciales de hinchamiento no cambian significativamente. Los resultados se discuten en términos que el NH+4 es translocado como una especie cargada vía un sistema de transporte. El sistema de transporte se caracterizó estudiando el hinchamiento de mitocondrias de hígado de rata suspendidas en acetatos de cationes alcalinos, de amonio y de cationes nitrogenados: el patrón de selectividad del translocador es NH+4>Na+>Li+>K+>Rb+>Cs+> (Secuencia X de Eisenman). Para cationes nitrogenados homólogos la permeabilidad disminuye a medida que el peso molecular aumenta o la movilidad en solución disminuye. Los iones con un radio de Ladd menor que 3.7-3.8 A y área por debajo de 15 A2 son permeables, los que tienen un mayor radio o área no lo son. Cuando la cadena de los cationes nitrogenados monosustituidos se alarga, la permeabilidad decrece, pasa por un mínimo cuando el nC = 4 y luego aumenta. Los cationes nitrogenados que no forman puentes de hidrógeno no entran a la mitocondria; los que tienen uno, dos o tres protones para donar tienen permeabilidades semejantes, y aquellos con uno, dos o tres oxígenos aceptores de protones aumentan su permeabilidad casi linealmente. La formamidina, acetamidina, TEA, derivados de la guanidina, N,N-diciclohexilcarbodiimida y N-etil maleimida no fueron capaces de inhibir el hinchamiento pasivo inducido por sodio en la mitocondria

Magnesium and octylguanidinium inhibition of monovalent cation translocation in mitochondria

Nosotros hemos estudiado los efectos inhibitorios de varios cationes (OG+, Mg2+, Ca2+, Ba2+, Mn2+, La3+) sobre las vías"uniport" y de intercambio para el Na+ y el K+ en mitocondrias de hígado de rata. El hinchamiento de las mitocondrias suspendidas en acetato de sodio o de potasio indica que: 1. El influjo pasivo de Na+ a mitocondrias inhibidas no se ve afectado por OG+ (en el rango micromolar), y concentraciones milimolares de cationes polivalentes sólo inducen una inhibición pobre, siendo la secuencia La3+>Mn2+>Ca2+>Mg2+>Sr2+ = 0. 2. El influjo activo de sodio se inhibe en un 50% con Mg2+ 60 micronM o con OG+ 90 micronM. El La3+, Mn2+, Ca2+ y Sr2+ también inhiben el influjo de sodio (rango milimolar). Se necesitan 10 mM de Mg2+ o 35 micronM de OG+ para inhibir en un 50% el influjo activo de potasio. 3. La salida de cationes alcalinos de mitocondrias parcialmente hinchadas e inhibidas se bloquea en un 50% con 2 mM de Mg2+ o 105 micronM OG+ siempre que el sodio sea el principal catión permeable de la solución. Se necesitan 3 mM de Mg2+ o 38 micronM de OG+ para inhibir en un 50% a las mitocondrias suspendidas en acetato de potasio. 4. La salida de cationes alcalinos de mitocondrias activas parcialmente hinchadas suspendidas en acetato de sodio se ve favorecida por concentraciones superiores a 0.1-0.2 Mg2+ o 50-100 micronM OG+. Estos datos se ajustan a un mecanismo que involucra a un translocador que introduce Na+ en un proceso dependiente de energía y que es sensible a Mg2+ y OG+ y un intercambiador catión/H+, insensible a Mg2+ y OG+, que trabajan en balance dinámico