Table 1.

Effects of Opsonizing Antibodies or rHSP-D on Neutrophil Apoptosis Induced by E coli

Neutrophil Treatment Apoptotic Neutrophils (%) Fas Antigen Expression
Control buffer  5 ± 1  5 ± 0.4 
E coli alone  26 ± 5* 10 ± 2* 
E coli + 18 μg/mL IgG  36 ± 8*, 16 ± 2*, 
E coli + 36 μg/mL IgG  38 ± 8*, 19 ± 1*, 
36 μg/mL IgG alone  4 ± 1 4.5 ± 1  
E coli + 0.13 μg/mL rHSP-D 23 ± 4* 10 ± 2* 
E coli + 0.26 μg/mL rHSP-D  30 ± 6* 12.5 ± 2*, 
E coli + 1 μg/mL rHSP-D  28 ± 4* 12 ± 2* 
E coli + 2 μg/mL rHSP-D  24 ± 1* 10 ± 2* 
Neutrophil Treatment Apoptotic Neutrophils (%) Fas Antigen Expression
Control buffer  5 ± 1  5 ± 0.4 
E coli alone  26 ± 5* 10 ± 2* 
E coli + 18 μg/mL IgG  36 ± 8*, 16 ± 2*, 
E coli + 36 μg/mL IgG  38 ± 8*, 19 ± 1*, 
36 μg/mL IgG alone  4 ± 1 4.5 ± 1  
E coli + 0.13 μg/mL rHSP-D 23 ± 4* 10 ± 2* 
E coli + 0.26 μg/mL rHSP-D  30 ± 6* 12.5 ± 2*, 
E coli + 1 μg/mL rHSP-D  28 ± 4* 12 ± 2* 
E coli + 2 μg/mL rHSP-D  24 ± 1* 10 ± 2* 

Results represent mean ± SEM for five experiments (ie, 5 individual blood donors). Percentages of apoptotic neutrophils were assessed using PI as described in Fig 1. Fas antigen expression was assessed by flow cytometry using polyclonal rabbit anti-Fas antibodies. Fas antigen results are expressed in fluorescence units (ie, mean channel fluorescence of 5,000 cells). All assays were done after 20 hours of incubation of neutrophils with control buffer, E colialone, or E coli that had been preincubated with either opsonizing IgG or rHSP-D as indicated. Opsonizing IgG alone (in absence of E coli) did not cause any increase in percent of apoptotic cells compared with control (n = 3; data not shown).

*

P ≤ .05 as compared with neutrophil treated with buffer alone.

Indicates P ≤ .05 as compared with neutrophils treated withE coli alone (ie, no opsonizing agent).

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