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1.
J Virol ; 87(17): 9538-46, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23785195

RESUMEN

Surface expression of SIGLEC1, also known as sialoadhesin or CD169, is considered a primary determinant of the permissiveness of porcine alveolar macrophages for infection by porcine reproductive and respiratory syndrome virus (PRRSV). In vitro, the attachment and internalization of PRRSV are dependent on the interaction between sialic acid on the virion surface and the sialic acid binding domain of the SIGLEC1 gene. To test the role of SIGLEC1 in PRRSV infection, a SIGLEC1 gene knockout pig was created by removing part of exon 1 and all of exons 2 and 3 of the SIGLEC1 gene. The resulting knockout ablated SIGLEC1 expression on the surface of alveolar macrophages but had no effect on the expression of CD163, a coreceptor for PRRSV. After infection, PRRSV viremia in SIGLEC1(-/-) pigs followed the same course as in SIGLEC1(-/+) and SIGLEC1(+/+) littermates. The absence of SIGLEC1 had no measurable effect on other aspects of PRRSV infection, including clinical disease course and histopathology. The results demonstrate that the expression of the SIGLEC1 gene is not required for infection of pigs with PRRSV and that the absence of SIGLEC1 does not contribute to the pathogenesis of acute disease.


Asunto(s)
Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Lectina 1 Similar a Ig de Unión al Ácido Siálico/fisiología , Animales , Animales Modificados Genéticamente , Antígenos CD/fisiología , Antígenos de Diferenciación Mielomonocítica/fisiología , Técnicas de Inactivación de Genes , Interacciones Huésped-Patógeno/inmunología , Interacciones Huésped-Patógeno/fisiología , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/virología , Síndrome Respiratorio y de la Reproducción Porcina/inmunología , Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/patogenicidad , Receptores de Superficie Celular/fisiología , Lectina 1 Similar a Ig de Unión al Ácido Siálico/deficiencia , Lectina 1 Similar a Ig de Unión al Ácido Siálico/genética , Sus scrofa , Porcinos , Acoplamiento Viral , Internalización del Virus
2.
Stem Cells Dev ; 21(14): 2682-8, 2012 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-22482370

RESUMEN

Multipotent skin-derived progenitors (SKPs) can be traced back to embryonic neural crest cells and are able to differentiate into both neural and mesodermal progeny in vitro. Neural stem cells (NSCs) are capable of self-renewing and can contribute to neuron and glia in the nervous system. Recently, we derived porcine SKPs and NSCs from the same enhanced green fluorescent protein (EGFP) transgenic fetuses and demonstrated that SKPs could contribute to neural and mesodermal lineages in vivo. However, it remains unclear whether porcine SKPs and NSCs can generate ectoderm and mesoderm lineages or other germ layers in vivo. Embryonic chimeras are a well-established tool for investigating cell lineage determination and cell potency through normal embryonic development. Thus, the purpose of this study was to investigate the in vivo developmental potential of porcine SKPs and fetal brain-derived NSCs by chimera production. Porcine SKPs, NSCs, and fibroblasts were injected into precompact in vitro fertilized embryos (IVF) and then transferred into corresponding surrogates 24 h postinjection. We found that porcine SKPs could incorporate into the early embryos and contribute to various somatic tissues of the 3 germ layers in postnatal chimera, and especially have an endodermal potency. However, this developmental potential is compromised when they differentiate into fibroblasts. In addition, porcine NSCs fail to incorporate into host embryos and contribute to chimeric piglets. Therefore, neural crest-derived SKPs may represent a more primitive state than their counterpart neural stem cells in terms of their contributions to multiple cell lineages.


Asunto(s)
Embrión de Mamíferos/metabolismo , Células-Madre Neurales/citología , Piel/citología , Células Madre/citología , Animales , Animales Modificados Genéticamente , Diferenciación Celular , Linaje de la Célula , Quimera/metabolismo , Transferencia de Embrión/métodos , Embrión de Mamíferos/citología , Desarrollo Embrionario , Femenino , Fibroblastos/metabolismo , Fibroblastos/trasplante , Estratos Germinativos/citología , Estratos Germinativos/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Células-Madre Neurales/metabolismo , Células-Madre Neurales/trasplante , Trasplante de Células Madre/métodos , Células Madre/metabolismo , Porcinos , Transgenes
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