Figure 9. The phagocytic index for RSV was decreased in the AMs from the ethanol-exposed pups but this decrease was blocked by maternal zinc supplements.
Dams were fed the control or ethanol diets ± zinc acetate supplements (100 mg/L) during pregnancy. On post-natal day 10, some pups (± ethanol and ± maternal zinc supplement) were given intranasal injections of saline or RSV (20 μl; each nasal nare; 2 × 105 PFU) and then returned to their respective dams. After 48 h, the pups were sacrificed and the lungs lavaged. After the cells in the lavage were removed by centrifugation and cultured on slides, they were fixed with 3.7% paraformaldehyde, permeabilized with ice-cold methanol, and then incubated with the primary antibody (1 h; a 1:100 dilution; Santa Cruz Biotechnology, Inc., Santa Cruz, CA). After washing the cells, AM were incubated with the secondary antibody (anti-goat IgG; a 1:200 dilution; 45 min). RSV in the cell was determined using confocal fluorescent analysis. Phagocytic index (PI) was calculated as the percentage of cells with internalized fluorescence × the mean RFU per field. Bar heights represent mean PI relative to the control ± S.E.M. from at least 6 separate litters (A). Representative fluorescent images are shown for each condition (B). CTRL = pup AM from control dams; CTRL + Zn = pup AM from control dams with dietary zinc supplements; EtOH = pup AM from ethanol-fed dams; and EtOH + Zn = pup AM from ethanol-fed dams with dietary zinc supplements. *** denotes p ≤ 0.05 when compared to the CTRL group and ** denotes p ≤ 0.01 EtOH + Zn vs. EtOH.