Figure 1

Lung macrophages are efficiently infected with RSV. (A) (i) PMΦ cells and (ii) AMΦ cells were mock-infected or RSV-infected, and at 24 hrs post-infection (hpi) stained with anti-RSV and examined using fluorescence microscopy (objective x 10) (highlighted by white arrows). (B) Mock (M) and RSV-infected (I) PMΦ cell lysates were examined by immunoblotting using anti-N, anti-P, anti-M2-1, anti-F or anti-G. Anti-actin is shown as the loading control. Analysis of mock and RSV-infected HEp2 cells (using same cell numbers) with anti-F and anti-G is shown. Protein bands of the expected sizes for the respective virus proteins are indicated. The 50 kDa F1 subunit, and the O- and N-linked (G(o/n)) and N-linked (G (n)) glycosylated G proteins are indicated. PMΦ cells were infected with RSV and at 24 hpi the cells were fixed, labelled using (C) anti-F or (D) anti-G and examined using florescence scanning confocal microscopy (FSCM) (i) at a focal plane that shows the cell surface staining (inset, highlighted by *) or (ii) in cross-section showing the surface staining (highlighted by white arrow). (E) The AMΦ cells were stained using anti-F and examined at a focal plane that shows the cell surface (inset, highlighted by *). (F) RSV-infected HEp2 cells at 24 hpi stained using anti-F (inset) and examined using FSCM, the virus filaments (VF) on the cell surface are highlighted. (G) (i) Mock and (ii) RSV-infected cells were stained with anti-RSV and anti-HSP90. The presence of virus-induced inclusion bodies and HSP90 staining of these structures is highlighted (white arrows). (H) RSV-infected AMΦ cells were stained using anti-RSV, and the presence of stained inclusion bodies highlighted (white arrow). (I) At 24 hpi RSV-infected PMΦ cells were stained with anti-RSV and anti-LAMP-1. The inclusion bodies (IB) and the LAMP-1 punctate staining pattern (white arrows) are highlighted.