TABLE 3.
Phenotypic analysis of cell surface TLR9+ cells on LPS-stimulated ex vivo human PBMCa
| Treatment | Donor | % Total TLR9+ (R1 gate) | % Isotype control (R1 gate) | % TLR9+ MHC II+ CD19+ (R2 gate) | % TLR9+ MHC II+ CD19− (R2 gate) | % TLR9+ MHC II+ CD11c+ (R2 gate) | % TLR9+ MHC IIlow CD123bright (R2 gate) |
|---|---|---|---|---|---|---|---|
| Ex vivo | 8 | 14.8 | 1.4 | 2.4 | 92.1 | 93.4 | 2.5 |
| 9 | 19.7 | 1.2 | 3.5 | 83.7 | 83.4 | 2.9 | |
| 10 | 13.6 | 1.3 | 4.9 | 84.6 | 87.6 | 2.5 | |
| Avg ± SD | 16.0 ± 3.2 | 1.3 ± 0.1 | 3.6 ± 1.3 | 86.8 ± 4.6 | 88.1 ± 5.0 | 2.6 ± 0.2 | |
| Medium, 24 h | 8 | 18.5 | 2.1 | 1.0 | 97.5 | 97.9 | 1.8 |
| 9 | 16.3 | 1.0 | 5.1 | 92.8 | 92.5 | 2.9 | |
| 10 | 16.5 | 1.0 | 2.5 | 96.0 | 92.7 | 0.7 | |
| Avg ± SD | 17.1 ± 1.2 | 1.4 ± 0.6 | 2.9 ± 2.1 | 95.4 ± 2.4 | 94.4 ± 3.1 | 1.8 ± 1.1 | |
| LPS, 24 h | 8 | 19.1 | 2.1 | 5.5 | 70.7 | 81.3 | 31.3 |
| 9 | 7.0 | 1.0 | 22.4 | 68.2 | 81.2 | 14.0 | |
| 10 | 7.1 | 1.6 | 12.3 | 78.5 | 81.7 | 26.1 | |
| Avg ± SD | 11.1 ± 7.0 | 1.6 ± 0.6 | 13.4 ± 8.5 | 72.5 ± 5.4 | 81.4 ± 0.3 | 23.8 ± 8.9 | |
| Medium, 48 h | 8 | 3.2 | 1.0 | 4.3 | 89.2 | 85.7 | 1.9 |
| 9 | 8.3 | 1.0 | 6.9 | 91.9 | 96.7 | 0.7 | |
| 10 | 7.8 | 1.0 | 7.8 | 88.7 | 83.7 | 1.5 | |
| Avg ± SD | 6.4 ± 2.8 | 1.0 ± 0.0 | 6.3 ± 1.8 | 89.9 ± 1.7 | 88.7 ± 7.0 | 1.4 ± 0.6 | |
| LPS, 48 h | 8 | 23.1 | 1.0 | 4.1 | 68.7 | 65.3 | 66.0 |
| 9 | 12.8 | 3.1 | 16.2 | 76.5 | 72.1 | 66.5 | |
| 10 | 10.7 | 1.4 | 7.8 | 82.5 | 76.8 | 38.5 | |
| Avg ± SD | 15.5 ± 6.6 | 1.8 ± 1.1 | 9.4 ± 6.2 | 75.9 ± 6.9 | 71.4 ± 5.8 | 57.0 ± 16.0 |
a
Phenotypic analysis, with or without 24 or 48 h of LPS stimulation, is based on flow cytometric analysis of live-gated PBMC (R1 gate) and TLR9+ cells (R2 gate) obtained from three independent donors. The raw data for ex vivo cell surface TLR9 expression for donors 8 to 10 are also presented in Table 2.