RESUMEN
DNA vaccination holds great promise for the prevention and treatment of cancer and infectious diseases. However, the clinical ability of DNA vaccines is still controversial due to the limited immune response initially observed in humans. We hypothesized that electroporation of a plasmid encoding the HIV-1 Gag viral capsid protein would enhance cancer DNA vaccine potency. DNA electroporation used to deliver plasmids in vivo, induced type I interferons, thereby supporting the activation of innate immunity. The coadministration of ovalbumin (OVA) and HIV-1 Gag encoding plasmids modulated the adaptive immune response. This strategy favored antigen-specific Th1 immunity, delayed B16F10-OVA tumor growth and improved mouse survival in both prophylactic and therapeutic vaccination approaches. Similarly, a prophylactic DNA immunization against the melanoma-associated antigen gp100 was enhanced by the codelivery of the HIV-1 Gag plasmid. The adjuvant effect was not driven by the formation of HIV-1 Gag virus-like particles. This work highlights the ability of both electroporation and the HIV-1 Gag plasmid to stimulate innate immunity for enhancing cancer DNA vaccine immunogenicity and demonstrates interesting tracks for the design of new translational genetic adjuvants to overcome the current limitations of DNA vaccines in humans.
Asunto(s)
Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/inmunología , Plásmidos/genética , Vacunas de ADN/genética , Vacunas de ADN/inmunología , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/inmunología , Animales , Línea Celular , Proliferación Celular , Modelos Animales de Enfermedad , Humanos , Interferón Tipo I/biosíntesis , Melanoma Experimental/inmunología , Melanoma Experimental/metabolismo , Melanoma Experimental/patología , Melanoma Experimental/terapia , Ratones , Plásmidos/administración & dosificación , Modelos de Riesgos Proporcionales , Células TH1/inmunología , Células TH1/metabolismo , Transfección , Resultado del Tratamiento , Carga Tumoral , Vacunas de Partículas Similares a Virus/genética , Vacunas de Partículas Similares a Virus/inmunología , Vacunas de Partículas Similares a Virus/ultraestructuraRESUMEN
The functionalization of poly(allylamine hydrochloride)/poly(acrylic acid) (PAH/PAA) polyelectrolyte multilayers by silanes reacted from the gas phase is studied depending on reaction time and temperature, pH of multilayer assembly, and nature of the reacting silane group. Whereas monochlorosilanes only diffuse in the multilayer and graft in limited amount, trichloro- and triethoxysilanes form rapidly a continuous gel layer on the surface of the multilayer, with a thickness of ca. 10-20 nm. The reactivity is lower in the strongly paired regime of the multilayers (neutral assembly conditions) but otherwise is not affected by the pH of multilayer assembly. Silanization considerably broadens the range of possible functionalities for (PAH/PAA) multilayers: hydrophobicity, surface-initiated polymerization, and grafting of fluorescent probes by the formation of disulfide bridges are demonstrated. Conversely, our results also broaden the range of substrates that can be functionalized by silanes, using (PAH/PAA) multilayers as ubiquitous anchoring layers.
RESUMEN
CONTEXT: Pendred syndrome is caused by mutations in the gene coding for pendrin, an apical Cl-/I- exchanger. OBJECTIVE: To analyze intrathyroidal compensatory mechanisms when pendrin is lacking, we investigated the thyroid of a patient with Pendred syndrome. The expression of proteins involved in thyroid hormone synthesis, markers of oxidative stress (OS), cell proliferation, apoptosis, and antioxidant enzymes were analyzed. RESULTS: Three morphological zones were identified: nearly normal follicles with iodine-rich thyroglobulin in the colloid (zone 1.a), small follicles without iodine-rich thyroglobulin in lumina (zone 1.b), and destroyed follicles (zone 2). In zones 1.a, dual oxidase (Duox) and thyroid peroxidase (TPO) were localized at the apical pole, OS and cell apoptosis were absent, but ClC-5 expression was strongly increased. In zones 1.b, Duox and TPO were aberrantly present and increased in the cytosol and associated with high OS, apoptosis, cell proliferation, and increased expression of peroxiredoxin-5, catalase, and dehalogenase-1 but moderate ClC-5 expression. CONCLUSION: In conclusion, the absence of pendrin is accompanied by increased ClC-5 expression that may transiently compensate for apical iodide efflux. In more affected follicles, Duox and TPO are relocated in the cytosol, leading to abnormal intracellular thyroid hormone synthesis, which results in cell destruction presumably because intracellular OS cannot be buffered by antioxidant defenses.
Asunto(s)
Proteínas de Transporte de Membrana/genética , Glándula Tiroides/metabolismo , Apoptosis/genética , Western Blotting , Proliferación Celular , Canales de Cloruro/genética , Canales de Cloruro/metabolismo , Oxidasas Duales , Femenino , Humanos , Inmunohistoquímica , Yoduro Peroxidasa/genética , Yoduro Peroxidasa/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Persona de Mediana Edad , NADPH Oxidasas/genética , NADPH Oxidasas/metabolismo , Estrés Oxidativo/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transportadores de Sulfato , Pruebas de Función de la Tiroides , Glándula Tiroides/fisiopatologíaRESUMEN
The adhesion of poly(dimethylsiloxane) (PDMS) rubber is largely improved by oxygen plasma surface treatment. The thickness of the silica-like surface layer is characterized by performing transmission electron microscopy imagery on microtome slices of welded plasma treated surfaces. The specific double layer contrast can be considered as equal to twice the thickness of the silica-like layer. The thickness measurements combined with strain-induced elastic buckling instability analysis gives an estimate of the elastic modulus of the silica-like layer equal to 1.5 GPa.