RESUMEN
Previous studies have highlighted the potential physiopathological and diagnostic role of N- and C-terminally truncated amyloid-ß (Aß) peptides in Alzheimer's disease. However, our knowledge about their production remains incomplete, in part due to the lack of very specific and sensitive tools for their detection. We thus developed specific monoclonal antibodies that target either Aß11-x or Aß17-x species, which result from the combined cleavages by ß/γ- or α/γ-secretases, respectively. The presence of Aß peptides truncated at residue 11 and 17 peptides was qualitatively and quantitatively assessed, using surface enhanced laser desorption ionization-time of flight mass spectrometry and xMAP (Multi-Analyte Profiling) immunoassays, in the supernatant of HEK293 cells that overexpress wild type or mutant Aß protein precursor or in which α- and ß-secretase activities had been modulated. Our results show a differential secretion of Aß11-40 and Aß17-40 species by these HEK293 cell lines. Finally, Aß11-40 concentration in human cerebrospinal fluid (measured with the new xMAP immunoassays) from a first pilot study was higher in cerebrospinal fluid samples from patients with Alzheimer's disease than in samples from patients with other types of dementia.
Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/metabolismo , Péptidos beta-Amiloides/metabolismo , Demencia/metabolismo , Fragmentos de Péptidos/metabolismo , Anciano , Péptidos beta-Amiloides/líquido cefalorraquídeo , Humanos , Inmunoensayo , Persona de Mediana Edad , Fragmentos de Péptidos/líquido cefalorraquídeo , Fosforilación , Proyectos Piloto , Proteínas tau/líquido cefalorraquídeoRESUMEN
The development of recombinant capripoxviruses for protective immunization of ruminants against bluetongue virus (BTV) infection is described. Sheep (n=11) and goats (n=4) were immunized with BTV recombinant capripoxviruses (BTV-Cpox) individually expressing four different genes encoding two capsid proteins (VP2 and VP7) and two non-structural proteins (NS1, NS3) of BTV serotype 2 (BTV-2). Seroconversion was observed against NS3, VP7 and VP2 in both species and a lymphoproliferation specific to BTV antigens was also demonstrated in goats. Finally, partial protection of sheep challenged 3 weeks after BTV-Cpox administration with a virulent strain of BTV-2, was observed.
Asunto(s)
Virus de la Lengua Azul/inmunología , Virus de la Lengua Azul/metabolismo , Capripoxvirus/inmunología , Capripoxvirus/metabolismo , Expresión Génica , Proteínas Virales/inmunología , Proteínas Virales/metabolismo , Animales , Virus de la Lengua Azul/genética , Virus de la Lengua Azul/patogenicidad , Capripoxvirus/genética , Células Cultivadas , Chlorocebus aethiops , Vectores Genéticos/genética , Cabras , Ovinos , Tasa de Supervivencia , Proteínas Virales/genética , Vacunas Virales/efectos adversos , Vacunas Virales/inmunologíaRESUMEN
Neural stem cells cultured with fibroblast growth factor 2 (FGF2)/epidermal growth factor (EGF) generate clonal expansions called neurospheres (NS), which are widely used for therapy in animal models. However, their cellular composition is still poorly defined. Here, we report that NS derived from several embryonic and adult central nervous system (CNS) regions are composed mainly of remarkable cells coexpressing radial glia markers (BLBP, RC2, GLAST), oligodendrogenic/neurogenic factors (Mash1, Olig2, Nkx2.2), and markers that in vivo are typical of the oligodendrocyte lineage (NG2, A2B5, PDGFR-alpha). On NS differentiation, the latter remain mostly expressed in neurons, together with Olig2 and Mash1. Using cytometry, we show that in growing NS the small population of multipotential self-renewing NS-forming cells are A2B5(+) and NG2(+). Additionally, we demonstrate that these NS-forming cells in the embryonic spinal cord were initially NG2(-) and rapidly acquired NG2 in vitro. NG2 and Olig2 were found to be rapidly induced by cell culture conditions in spinal cord neural precursor cells. Olig2 expression was also induced in astrocytes and embryonic peripheral nervous system (PNS) cells in culture after EGF/FGF treatment. These data provide new evidence for profound phenotypic modifications in CNS and PNS neural precursor cells induced by culture conditions.