| الوصف: |
(A) maBel/09 at a MOI of 0.01 was pre-treated with (i) mock serum, (ii) anti-NA serum, (iii) anti-HA serum, (iv) IgG1 isotype control, (v) Oseltamivir, (vi) monoclonal antibody HCA-2, (vii) N1-C4, or (viii) N1-7D3 and added to HAE cells (Lonza) for 1 h. All sera were RDE treated and heat-inactivated. The sera were used at 1:100 dilution, monoclonal antibodies at 10 μg/ml, and Oseltamivir at 24 μM. The inoculum was washed away and anti-sera, monoclonal antibodies, or Oseltamivir were re-added. At 0, 8, 24, 36, and 46 h post inoculation, the presence of virus released at the air-liquid interface was determined by TCID 50 . The average viral titers from triplicate HAE cultures ± SD is shown. The data is representative of two independent experiments. *p < 0.05, ***p < 0.001, ****p<0.0001 two-way ANOVA in comparison to either mock sera or isotype control. (B) NA-specific polyclonal sera and N1-C4 restrict virus egress from the surface of HAE cells. maBel/09 at a MOI of 1 was incubated with buffer alone, Oseltamivir (24 μM), anti-NA serum (1:100 final concentration), or N1-C4 (10 μg/ml final concentration) and added to washed HAE cells (Lonza). The virus was allowed to bind for 1 h, after which the inoculum was removed by washing. Buffer, Oseltamivir, anti-NA serum, or N1-C4 was re-added for a further 24 h. The cells were fixed and processed for TEM. Shown are representative images taken with a JEM1010 TEM microscope at 10,000x magnification. Scale bar: 1 μm. |