Table 1.
Surface phenotype of the microsphere+ DCs within draining and non-draining secondary lymphoid organs following a subcutaneous injection of 0.2μm latex microspheres with 1α25(OH)2D3 or LPS.
| Secondary lymphoid organs | Additions to microsphere inoculuma | Phenotype of microsphere+ cellsb | |
|---|---|---|---|
| CD8α−33D1+ (%) | CD8α+DEC205+ (%) | ||
| Popliteal LN (draining) | None | 68.1±3.2 | 12.2±1.3 |
| 1α25(OH)2D3 | 21.3± 3.5c | 31.1±2.8c | |
| LPS | 47.5±4.3c | 16.3±0.9 | |
| Axillary LN (non-draining) | None | N/Dd | N/D |
| 1α25(OH)2D3 | 98.5±0.3 | 1.9±0.2 | |
| LPS | 95.4±1.7 | 2.1±0.3 | |
| PP (non-draining, mucosal) | None | N/D | N/D |
| 1α25(OH)2D3 | 97.8±1.1 | 2.0±0.1 | |
| LPS | 96.5±1.5 | 1.8±0.3 | |
Green 0.2μm fluorescent microspheres (50μl of a 0.25% solution) were injected into the hind footpads of C3H/HeN mice (3 per group) in the presence of 0.1μg 1α25(OH)2D3 or 10μg LPS. The injection of microspheres alone served as the control.
Single cell suspensions were prepared from multiple secondary lymphoid organs after 48 hours. Cells were stained with antibodies directed against CD8α, DEC205 and 33D1 and analyzed by FACScan. Phenotype analysis was done on microsphere+ cells. At least 1000 microsphere+ events were analyzed.
Differences in the percentages of LPS or 1α25(OH)2D3-exposed microsphere+ DCs in the draining LNs of experimental mice and the percentage of microsphere+ DCs in the draining LNs of mice that received microspheres only were statistically significant (p<0.05-0.002)
N/D – not detected