Effects of IAV, E coli, or the Combination of IAV and E coli on Neutrophil Expression of Fas Antigen
| Antibody . | Time (h) . | Control Buffer . | IAV . | E coli . | IAV + E coli . |
|---|---|---|---|---|---|
| CD95 | 3 | 36 ± 2 | 41 ± 3* | 39 ± 2 | 46 ± 4* |
| Isotype control | 3 | 4.4 ± 0.4 | 5.6 ± 0.3* | 7.3 ± 1* | 13.2 ± 2* |
| CD95 | 20 | 26 ± 3 | 35 ± 5* | 33 ± 4* | 40 ± 3*,† |
| Isotype control | 20 | 5 ± 1 | 7 ± 1* | 11 ± 2* | 16 ± 2*,† |
| Rabbit anti-Fas | 20 | 10 ± 1 | 16 ± 2* | 15 ± 2* | 49 ± 15*,† |
| Rabbit anti-Fas + blocker | 20 | 9.4 ± 1 | 11 ± 2 | 9 ± 2 | 26 ± 8 |
| Antibody . | Time (h) . | Control Buffer . | IAV . | E coli . | IAV + E coli . |
|---|---|---|---|---|---|
| CD95 | 3 | 36 ± 2 | 41 ± 3* | 39 ± 2 | 46 ± 4* |
| Isotype control | 3 | 4.4 ± 0.4 | 5.6 ± 0.3* | 7.3 ± 1* | 13.2 ± 2* |
| CD95 | 20 | 26 ± 3 | 35 ± 5* | 33 ± 4* | 40 ± 3*,† |
| Isotype control | 20 | 5 ± 1 | 7 ± 1* | 11 ± 2* | 16 ± 2*,† |
| Rabbit anti-Fas | 20 | 10 ± 1 | 16 ± 2* | 15 ± 2* | 49 ± 15*,† |
| Rabbit anti-Fas + blocker | 20 | 9.4 ± 1 | 11 ± 2 | 9 ± 2 | 26 ± 8 |
Results shown are mean ± SEM of three or more experiments. Expression of Fas antigen was assessed by flow cytometry using monoclonal antibody CD95 or polyclonal rabbit anti-Fas (C-20) antibodies. Values given are mean channel fluorescence for 5,000 cells. Controls included a mouse monoclonal of the same isotype as monoclonal antibody CD95, rabbit anti-Fas that had been preincubated with specific peptide blocker, and nonspecific rabbit IgG. Neutrophils were incubated with either control buffer, IAV, E coli, or IAV and E coli for 3 or 20 hours as indicated. IAV and/or E coli did not cause any increase in binding of nonspecific rabbit IgG (data not shown).
Significant increase (P ≤ .05) as compared with neutrophil treated with buffer alone.
P ≤ .05 as compared with neutrophils treated with IAV orE coli alone (ie, no opsonizing agent).