Article Figures & Data
Figures
- Fig. 1.
Flow cytometric analysis of integrin αvβ3, αvβ1, and αvβ5 expression in A549 cells. Control A549 cells were incubated with FITC-conjugated rabbit anti-mouse IgG (A), integrin αvβ3-specific MAb LM609 (B), integrin β1-specific MAb 6S6 (C), and integrin αvβ5-specific MAb P1F6 (D). The histograms display relative cell numbers as a function of relative fluorescence intensities.
- Fig. 2.
Percent inhibition of HPEV1 infectivity to A549 cells in the presence of LM609 (αvβ3-specific MAb), NK1-M9 (αv-specific MAb), 6S6 (β1-specific MAb), P1F6 (αvβ5-specific MAb), BHA2.1 (α2β1-specific MAb), and isotype control MAb at concentrations of 2.5, 5, 10, and 15 μg. The error bars are calculated from the standard deviation over a number on independent experiments.
- Fig. 3.
Percent inhibition of HPEV1 infectivity to A549 cells in the presence of combinations of MAbs at concentrations of 2.5, 5, 10, and 15 μg. For identities of the MAbs, see the legend to Fig. 2. The error bars are calculated from the standard deviation over a number on independent experiments.
- Fig. 4.
Percent inhibition of HPEV1 infectivity to A549 cells in the presence of different concentrations (10 to 80 μg/ml) of vitronectin, fibronectin, laminin, and a combination of vitronectin and fibronectin. The error bars are calculated from the standard deviation over a number on independent experiments.
- Fig. 5.
Flow cytometric analysis of integrin αvβ3 and αvβ1 expression on CHO-αvβ3, CHO-αvβ1, and CHO-wt cells. Control CHO-αvβ3 (A), CHO-αvβ1 (D), and CHO-wt (G) cells were incubated with FITC-conjugated rabbit anti-mouse IgG. To test integrin expression on CHO-αvβ3 cells, integrin αvβ3-specific MAb LM609 (B) or integrin β1-specific MAb 6S6 (C), followed by FITC-conjugated rabbit anti-mouse IgG, was added to the cells. To CHO-αvβ1 cells, integrin β1-specific MAb 6S6 (E) or integrin αvβ3-specific MAb LM609 (F), followed by FITC-conjugated rabbit anti-mouse IgG, was added. To test integrin expression on CHO-wt cells, specific MAb LM609 (I) or specific 6S6 (H) was added, followed by FITC-conjugated rabbit anti-mouse IgG. The histograms display relative cell numbers as a function of relative fluorescence intensities.
- Fig. 6.
Results of HPEV-1 plaque assay on CHO-αvβ1 (A), CHO-αvβ3 (B), and CHO-wt (C) cells. The plates are representative of a number of independent experiments.
- Fig. 7.
Percent inhibition of HPEV1 binding to CHO-αvβ1 and CHO-αvβ3 cells in the presence of a combination of αv-specific MAb NK1-M9 and β1-specific MAb 6S6 (black bars), in the presence of an isotype control MAb (clear bars), in the presence of αvβ3-specific MAb LM609 (striped bars), and in the presence of an isotype control MAb (black and white bars). The error bars are calculated from the standard deviation over a number of independent experiments.
- Fig. 8.
SDS-PAGE of immunoprecipitated HPEV1 receptor complexes under reducing conditions. Cell surface-biotinylated A549 cells were solubilized in 1% digitonin and immunoprecipitated with HPEV1 virions followed by HPEV1-specific monkey neutralizing serum (G), or in the absence of HPEV1 virions, with HPEV1-specific monkey neutralizing serum alone (E), with an irrelevant antiserum (F), or with normal monkey serum (C). As controls, cell surface-biotinylated CHO-αvβ1 (A), CHO-αvβ3 (D), and CHO-wt (B) were solubilized in 1% digitonin and immunoprecipitated with HPEV1 virions followed by HPEV1-specific monkey neutralizing serum. The membrane from the A549 cell lysate immunoprecipitations was Western blotted with αv-chain-specific MAb VNR139 (M), with β3-chain-specific MAb Y2/51 (N), with β1-chain-specific MAb B3B11 (H), and with rabbit polyclonal sera specific for integrins α2 (I), β4 (J), β5 (K), and α5 (L), followed by either HRP-conjugated goat anti-mouse Ig or HRP-conjugated goat anti-rabbit Ig. The positions of molecular weight markers are shown to the right.
