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Isolating And Immunostaining Lymphocytes and Dendritic Cells from Murine Peyer's Patches
Published on: March 17, 2013
Subcellular immunolocalization of porcine circovirus type 2 (PCV2) in lymph nodes from pigs with post-weaning
C Rodríguez-Cariño1, A Sánchez-Chardi, J Segalés
1Centre de Recerca en Sanitat Animal (CReSA), UAB-IRTA and Departament de Sanitat i Anatomia Animals, Facultat de Veterinària, Campus de la Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain. carolina.carino@cresa.uab.cat
Abstract:
Post-weaning multisystemic wasting syndrome (PMWS) is one of the most significant porcine diseases worldwide. The causative agent is porcine circovirus type 2 (PCV2), the smallest virus known to infect animals. Data related to the structural and ultrastructural aspects of this infectious disease are sparse and there is little knowledge of the subcellular localization of PCV2 and its replication in the tissues of pigs naturally affected by PMWS. The present study describes the cellular localization of PCV2 in the lymph nodes of pigs affected by PMWS by application of immunolabelling techniques for light and transmission electron microscopy (TEM). PCV2 particles were exclusively detected in histiocytes. Ultrastructural alterations including marked dilatation of rough endoplasmic reticulum and swelling of mitochondria were associated with PCV2-labelled intracytoplasmic inclusions (ICIs) with recognizable virions. Within the ICIs icosahedral virus-like particles were specifically labelled with a PCV2 capsid antibody, whereas particles with a granular appearance were not labelled. Colocalization studies with confocal microscopy and double immunolabelling with TEM indicated a close relationship between virus and the mitochondria, suggesting that these organelles may play an important role in the replication of PCV2. The present findings further support the hypothesis that virus replicates within the histiocytes of lymph nodes.
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