IMEX   05356
INSTITUTO DE MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Infection of Endothelial and Mesothelial Cells with Andes Virus and NK Response
Autor/es:
SCHIERLOH P.; MARTINEZ V; LANDONI V.I; FERNANDEZ G.C.; SASIAIN M.C.; PADULA P
Lugar:
Beijin
Reunión:
Congreso; IX International Conference on HFRS HPS & Hantaviruses; 2013
Institución organizadora:
International Society for Hantaviruses (HFRS and HPS)
Resumen:
In the early phase of the infection pathogenic hantaviruses escape induction of innate antiviral responses, specifically innate interferon system. The rol of NK cells in hantavirus infection remains to be investigated. Lung microvascular endothelium is considered to be the primary site for hantavirus replication. To determine whether two human pulmonary cell types support Andes virus (ANDV) replication and contribute to the earliest induction of the antiviral response, human microvascular pulmonary endothelial cell line HMEC-1 and cultured primary human mesothelial cells were mock infected or infected with ANDV. Infection of cells was confirmed by immunofluorescence and cell morphology was analyzed by confocal microscopy. Expression of cell surface markers related to NK cell activation was analyzed by flow cytometry. Viral RNA, infectious particles and cytokine and chemokine responses were quantified in culture media of both cell types. Infected cells were identified by the characteristically intra-cytoplasmatic pattern of staining for viral nucleoprotein (NP). The infection caused morphological changes in HMEC-1 and mesothelial cells. By day 4 post infection (p.i.) more than 20% of cells were positive for NP staining in HMEC-1 infected at a multiplicity of infection of 0.1. At the same day viral RNA in culture media increased drastically to reach 2,5xE+9 S segment molecules per ml. The kinetics of infection in HMEC-1 was similar to that seen in Vero E6 cells, although in mesothelial cells the titer reached up to 4 days p.i. was lower. Expression of IL-1 and IL-8 and major histocompatibility complex class I (MHC-I), ICAM and CD80/CD86 surface markers were evaluated in each cell type. ANDV infection in HMEC-1 and mesothelial cells significantly increased MHC-1 expression in cell surface and IL-8 secretion. However, ANDV seems to be unable to alter the pattern induced by some added inflammatory mediators such as TNF-alfa and IFN-gamma. NK cells were co-cultured with ANDV-infected or mock-infected HMEC-1 cells to study the direct effect of this interaction in the activation of NK cells. Preliminary data suggested that there were no evident differences in responses between infected and non infected co-cultures. HMEC-1 and mesothelial cells supported ANDV replication and both of them could serve as a model to study distinct roles in modulation of the immune system during hantavirus infection.