Effect of in vitro stimulation on immunoregulatory gene expression
. | Foxp3 . | CD25 . | CTLA-4 . | GITR . | PD-1 . | TRF-β . |
|---|---|---|---|---|---|---|
| Mitogen stimulation | ||||||
| ConA | ||||||
| CD4 | 2.7 ± 0.4 | 18.9 ± 2.9* | 6.7 ± 2.1 | 73.1 ± 11.5* | 27.7 ± 7.1* | 1.6 ± 0.6 |
| CD8 | 2.3 ± 0.9 | 23.1 ± 8.6 | 3.2 ± 1.3 | 5.8 ± 2.0 | 28.2 ± 12.0 | 1.6 ± 1.1 |
| LPS | ||||||
| CD4 | 2.9 ± 0.5 | 10.5 ± 1.7* | 97.5 ± 9.7* | 112.0 ± 17.0* | 86.2 ± 14.3* | 6.6 ± 0.9* |
| CD8 | 2.4 ± 1.1 | 60.6 ± 19.8 | 26.6 ± 9.5 | 29.9 ± 12.1 | 6.0 ± 1.9 | 1.3 ± 0.6 |
| Mixed lymphocyte reaction | ||||||
| PBMC | 1.8 ± 0.3 | 26.3 ± 6.7* | 0.5 ± 0.2 | 2.3 ± 0.9 | 3.2 ± 0.8* | 3.7 ± 0.7* |
| Ag stimulation | ||||||
| C-terminal CLIP | ||||||
| CD4 | 226.7 ± 14.8* | 14.8 ± 1.5* | 39.3 ± 5.4* | 880.0 ± 83.0* | 1.8 ± 0.9 | 1.4 ± 0.2 |
| CD8 | 14.8 ± 3.8 | 7.8 ± 2.0 | 1.2 ± 0.4 | 5.9 ± 2.8 | 1.5 ± 0.4 | 1.3 ± 0.3 |
| N-terminal CLIP | ||||||
| CD4 | 28.3 ± 2.6 | 12.0 ± 1.9 | 13.9 ± 2.0 | 223.0 ± 31.0 | 1.2 ± 0.2 | 1.2 ± 0.2 |
| CD8 | 5.2 ± 2.7 | 3.3 ± 1.5 | 0.5 ± 0.2 | 3.4 ± 1.8 | 1.0 ± 0.2 | 0.8 ± 0.2 |
| Class II-binding domain CLIP | ||||||
| CD4 | 23.7 ± 2.0 | 5.3 ± 1.6 | 8.5 ± 1.3 | 118.0 ± 27.0 | 1.1 ± 0.2 | 0.7 ± 0.1 |
| CD8 | 6.2 ± 2.8 | 2.3 ± 0.9 | 0.6 ± 0.3 | 2.2 ± 1.1 | 0.9 ± 0.3 | 0.5 ± 0.1 |
| No peptide | ||||||
| CD4 | 4.1 ± 1.2 | 4.7 ± 1.0 | 15.9 ± 2.5 | 73.1 ± 12.1 | 1.0 ± 0.2 | 0.6 ± 0.2 |
| CD8 | 2.6 ± 1.8 | 1.9 ± 0.9 | 1.0 ± 0.2 | 3.1 ± 1.4 | 1.2 ± 0.3 | 0.6 ± 0.3 |
. | Foxp3 . | CD25 . | CTLA-4 . | GITR . | PD-1 . | TRF-β . |
|---|---|---|---|---|---|---|
| Mitogen stimulation | ||||||
| ConA | ||||||
| CD4 | 2.7 ± 0.4 | 18.9 ± 2.9* | 6.7 ± 2.1 | 73.1 ± 11.5* | 27.7 ± 7.1* | 1.6 ± 0.6 |
| CD8 | 2.3 ± 0.9 | 23.1 ± 8.6 | 3.2 ± 1.3 | 5.8 ± 2.0 | 28.2 ± 12.0 | 1.6 ± 1.1 |
| LPS | ||||||
| CD4 | 2.9 ± 0.5 | 10.5 ± 1.7* | 97.5 ± 9.7* | 112.0 ± 17.0* | 86.2 ± 14.3* | 6.6 ± 0.9* |
| CD8 | 2.4 ± 1.1 | 60.6 ± 19.8 | 26.6 ± 9.5 | 29.9 ± 12.1 | 6.0 ± 1.9 | 1.3 ± 0.6 |
| Mixed lymphocyte reaction | ||||||
| PBMC | 1.8 ± 0.3 | 26.3 ± 6.7* | 0.5 ± 0.2 | 2.3 ± 0.9 | 3.2 ± 0.8* | 3.7 ± 0.7* |
| Ag stimulation | ||||||
| C-terminal CLIP | ||||||
| CD4 | 226.7 ± 14.8* | 14.8 ± 1.5* | 39.3 ± 5.4* | 880.0 ± 83.0* | 1.8 ± 0.9 | 1.4 ± 0.2 |
| CD8 | 14.8 ± 3.8 | 7.8 ± 2.0 | 1.2 ± 0.4 | 5.9 ± 2.8 | 1.5 ± 0.4 | 1.3 ± 0.3 |
| N-terminal CLIP | ||||||
| CD4 | 28.3 ± 2.6 | 12.0 ± 1.9 | 13.9 ± 2.0 | 223.0 ± 31.0 | 1.2 ± 0.2 | 1.2 ± 0.2 |
| CD8 | 5.2 ± 2.7 | 3.3 ± 1.5 | 0.5 ± 0.2 | 3.4 ± 1.8 | 1.0 ± 0.2 | 0.8 ± 0.2 |
| Class II-binding domain CLIP | ||||||
| CD4 | 23.7 ± 2.0 | 5.3 ± 1.6 | 8.5 ± 1.3 | 118.0 ± 27.0 | 1.1 ± 0.2 | 0.7 ± 0.1 |
| CD8 | 6.2 ± 2.8 | 2.3 ± 0.9 | 0.6 ± 0.3 | 2.2 ± 1.1 | 0.9 ± 0.3 | 0.5 ± 0.1 |
| No peptide | ||||||
| CD4 | 4.1 ± 1.2 | 4.7 ± 1.0 | 15.9 ± 2.5 | 73.1 ± 12.1 | 1.0 ± 0.2 | 0.6 ± 0.2 |
| CD8 | 2.6 ± 1.8 | 1.9 ± 0.9 | 1.0 ± 0.2 | 3.1 ± 1.4 | 1.2 ± 0.3 | 0.6 ± 0.3 |
Immunoregulatory gene expression was analyzed following in vitro stimulation of PBMCs from 3 different individuals. CD4+ or CD8+ T cells were incubated for 3, 12, and 24 hours with Concanavalin A (ConA) or lipopolysaccharide (LPS). Time to maximal expression of individual marker transcripts varied. Data presented the maximal transcript levels assessed over 24 hours. In addition, gene expression was evaluated after 5 days of stimulation with MHC unrelated lymphocytes (mixed lymphocyte reaction). CD4+ or CD8+ T cells were also stimulated with Ag-pulsed autologous dendritic cells. Ag peptides used for study included the CLIP N-terminal variant, C-terminal variant, and MHC class II-binding domain variant. Gene expression of Ag-specific T cells was evaluated after 3 days of culture. Fold increases were determined by comparison to unstimulated CD4+ or CD8+ T cells or PBMCs. Data are expressed as mean ± SE.
Significantly greater than control (P < .05)