ABSTRACT
BACKGROUND: In the last years, plants are being used for the production of a wide variety of biopharmaceuticals, including cytokines, and have the potential to serve as vehicles for mucosal administration of these molecules. We had previously reported the expression of a cytokine, interleukin-12 (IL-12), in transgenic tomato plants and had demonstrated that it retained its biologic activity in vitro. FINDINGS: In this work, we administered crude extracts of IL-12-containing tomato fruits to mice through the intratracheal route, measuring endogenous IL-12 and determining biologic activity by quantification of interferon-gamma (IFN-gamma) in lungs and by histological analysis. IFN-gamma expression in lungs, as well as histological analysis, indicate that tomato-expressed IL-12 retains its biologic activity and, most importantly, its effects are restricted to the site of administration. CONCLUSION: Our results indicate that the functional activity of tomato-expressed IL-12 is comparable to that of commercial recombinant IL-12 when given via the mucosal route. This opens the possibility of using crude extracts prepared from tomatoes expressing IL-12 for certain immunotherapies.
ABSTRACT
Treatment of P388D1, a macrophage-like cell line, with staurosporine triggered apoptosis through the activation of caspase-9 and caspase-3. Unexpected effects of staurosporine on the induction of apoptosis were the activation of caspase-8, and an increase of the levels of TNF-alpha. The increased TNF-alpha levels led to activation of caspase-8 by an autocrine effect via the TNF receptor expressed by the P388D1 macrophages. In contrast, P388D1 macrophages that either had been exposed to UV light or treated with dexamethasone did not undergo apoptosis.
Subject(s)
Apoptosis , Macrophages/drug effects , Protein Kinase Inhibitors/pharmacology , Staurosporine/pharmacology , Animals , Antineoplastic Agents, Hormonal/pharmacology , Caspase 3/drug effects , Caspase 3/metabolism , Caspase 8/drug effects , Caspase 8/metabolism , Caspase 9/drug effects , Caspase 9/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/physiology , DNA Fragmentation/drug effects , Dexamethasone/pharmacology , Macrophages/radiation effects , Mice , Receptors, Tumor Necrosis Factor, Type I/agonists , Receptors, Tumor Necrosis Factor, Type I/metabolism , Signal Transduction/drug effects , Signal Transduction/physiology , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/drug effects , Ultraviolet Rays/adverse effectsABSTRACT
Cell-surface viral proteins most frequently enter the cell through clathrin or caveolae endocytosis. Respiratory syncytial virus antigen internalization by immune cells is via caveolin, however, uptake of paramyxovirus cell membrane proteins by non-immune cells is done through clathrin-coated pits. In this work, the uptake of respiratory syncytial virus cell surface glycoproteins by non-immune human epithelial cells was investigated through indirect immunofluorescence with polyclonal anti-RSV antibody and confocal lasser-scanner microscopy. Clathrin and caveolae internalization pathways were monitored through specific inhibitors monodansylcadaverine (MDC) and methyl-beta-cyclodextrin (MBCD), respectively. Internalization of RSV antigens was inhibited by MDC but not by MBCD, implying that clathrin-mediated endocytosis is the major uptake route of RSV antigens by an epithelial human cell line.
Subject(s)
Clathrin-Coated Vesicles/virology , Endocytosis , Epithelial Cells/virology , Glycoproteins/metabolism , Respiratory Syncytial Viruses/physiology , Viral Proteins/metabolism , Cadaverine/analogs & derivatives , Cadaverine/pharmacology , Cell Line , Enzyme Inhibitors/pharmacology , Fluorescent Antibody Technique, Indirect , Humans , Microscopy, Confocal , beta-Cyclodextrins/pharmacologyABSTRACT
BACKGROUND: The binding of viral-specific antibodies to cell-surface antigens usually results in down modulation of the antigen through redistribution of antigens into patches that subsequently may be internalized by endocytosis or may form caps that can be expelled to the extracellular space. Here, by use of confocal-laser-scanning microscopy we investigated the kinetics of the modulation of respiratory syncytial virus (RSV) antigen by RSV-specific IgG. RSV-infected human epithelial cells (HEp-2) were incubated with anti-RSV polyclonal IgG and, at various incubation times, the RSV-cell-surface-antigen-antibody complexes (RSV Ag-Abs) and intracellular viral proteins were detected by indirect immunoflourescence. RESULTS: Interaction of anti-RSV polyclonal IgG with RSV HEp-2 infected cells induced relocalization and aggregation of viral glycoproteins in the plasma membrane formed patches that subsequently produced caps or were internalized through clathrin-mediated endocytosis participation. Moreover, the concentration of cell surface RSV Ag-Abs and intracellular viral proteins showed a time dependent cyclic variation and that anti-RSV IgG protected HEp-2 cells from viral-induced death. CONCLUSION: The results from this study indicate that interaction between RSV cell surface proteins and specific viral antibodies alter the expression of viral antigens expressed on the cells surface and intracellular viral proteins; furthermore, interfere with viral induced destruction of the cell.
Subject(s)
Antibodies, Viral/immunology , Antigens, Viral/immunology , Epithelial Cells/immunology , Epithelial Cells/virology , Respiratory Syncytial Viruses/immunology , Animals , Cell Line, Tumor , Clathrin/metabolism , Endocytosis/physiology , Epithelial Cells/cytology , Glycoproteins/metabolism , Humans , Immunoglobulin G/immunology , Immunoglobulin G/metabolism , Kinetics , Male , Rabbits , Viral Proteins/metabolismABSTRACT
Aunque el dolor pélvico y la metrorragia son las alteraciones más frecuentemente asociadas al mioma uterino, el hemoperitoneo agudo por rotura de leiomioma subseroso es un hecho excepcional apenas recogido en la literatura médica internacional. Este artículo describe uno de los pocos casos conocidos de abdomen agudo por rotura espontánea de leiomioma uterino subseroso que hemos tenido ocasión de tratar en el Servicio de Ginecología del Hospital Universitario Central de Asturias. La edad de la paciente, la paridad, el tamaño del mioma y ciertas malformaciones vasculares uterinas son considerados factores de riesgo para este cuadro tan insólito, pero además, en nuestro caso, determinadas peculiaridades histológicas del tumor parecen tener cierta relevancia
Although pelvic pain and metrorrhagia are the disorders that are most commonly associated with uterine leiomyoma, acute hemoperitoneum due to rupture of a subserosal leiomyoma is an exceptional event that has scarcely been reported in the international medical literature. This article describes one of the few known cases of acute abdomen caused by spontaneous rupture of a subserosal uterine leiomyoma, which was treated at the Department of Gynecology of the Hospital Universitario Central de Asturias (Spain). Risk factors for this unusual entity are considered to be the patient's age and parity, as well as the size of the myoma and certain uterine vascular malformations; however, in this specific case, some histological peculiarities of the tumor seem to be of relevance
