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1.
J Virol ; 87(17): 9538-46, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23785195

RESUMO

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.


Assuntos
Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/fisiologia , Animais , Animais Geneticamente Modificados , Antígenos CD/fisiologia , Antígenos de Diferenciação Mielomonocítica/fisiologia , Técnicas de Inativação de Genes , Interações Hospedeiro-Patógeno/imunologia , Interações Hospedeiro-Patógeno/fisiologia , Macrófagos Alveolares/imunologia , Macrófagos Alveolares/virologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/patogenicidade , Receptores de Superfície Celular/fisiologia , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/deficiência , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/genética , Sus scrofa , Suínos , Ligação Viral , Internalização do Vírus
2.
Stem Cells Dev ; 21(14): 2682-8, 2012 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-22482370

RESUMO

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.


Assuntos
Embrião de Mamíferos/metabolismo , Células-Tronco Neurais/citologia , Pele/citologia , Células-Tronco/citologia , Animais , Animais Geneticamente Modificados , Diferenciação Celular , Linhagem da Célula , Quimera/metabolismo , Transferência Embrionária/métodos , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário , Feminino , Fibroblastos/metabolismo , Fibroblastos/transplante , Camadas Germinativas/citologia , Camadas Germinativas/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Masculino , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/transplante , Transplante de Células-Tronco/métodos , Células-Tronco/metabolismo , Suínos , Transgenes
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