Table 1.
Neutralization titers of H7 Monoclonal Antibodies (mAbs).
Table 2.
Hemagglutination inhibition titers of H7 Monoclonal Antibodies (mAbs).
Fig 1.
Comparison of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA.
(A) Alignment of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA amino acid residues 130 to 310. Predicted antigenic sites A (pink), B (orange), C (purple), D (yellow), and E (blue) are highlighted. Glycosylation site N141 is marked in a square. (B) Overlapping of A/Netherlands/219/2003 HA (grey, PDB: 4DJ6) and A/Shanghai/02/2013 HA (green, PDB: 4LN6) crystal structures. Only monomers are shown. Predicted antigenic sites A (pink), B (orange), C (purple), D (yellow), and E (blue) are labeled. RBS: receptor binding site. (C) Western blot comparison of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA treated with and without PNGase F. Wild type (WT) HA and its mutants were detected with goat antisera against A/Netherlands/219/2003 HA (Top panel). The p24 in HA-pseudoviruses, which served as loading controls, were detected with mouse mAb (183-H12-5C) against HIV-1 p24 Gag (Bottom panel). Data shown are representative of two independent experiments.
Fig 2.
Threonine 143 in HA blocks neutralizing antibodies binding to antigenic site.
Immunoprecipitation of A/Shanghai/02/2013 HA (A) and A/Netherlands/219/2003 HA (B) with mAb 5A6. The relative levels of HA shown by Western blots were detected with goat antisera against A/Netherlands/219/2003 HA. Percentages of the HA signal intensities, normalized to the 1/10 volume of the input used for the immunoprecipitation (control), are indicated below each blot. Con: control; IP: immunoprecipitation. Data shown are representative of two independent experiments.
Table 3.
ELISA end-point titers of H7 Monoclonal Antibodies (mAbs).
Fig 3.
Threonine 143 in HA enhances H7 pseudovirus infectivity.
(A) Comparison of WT and mutation on residue 143. (B) Comparison of WT and mutations on residues 183, 195 and 198. The infectivity of pseudoviruses bearing wild type (WT) HA and its mutants is compared (Top panels in A and B). Data are shown as mean and standard deviation of three independent experiments. P<0.001 of T-test were shown. The levels of mature HA (HA1) and p24 in HA-pseudoviruses shown by Western blotting were detected with goat antisera against A/Netherlands/219/2003 HA and mouse mAb (183-H12-5C) against HIV-1 p24 Gag, respectively (Bottom panels in A and B). NL: A/Netherlands/219/2003 HA-pseudoviruses; SH: A/Shanghai/02/2013 HA-pseudoviruses.