Combination of anti-CD28 mAb and rapamycin inhibits donor CD4+ cell expansion and improves early B-cell reconstitution
. | Spleen-derived CD4+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived CD4+Ly5.1-, total cells per mouse × 106 (%) . | Spleen-derived CD8+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived CD8+Ly5.1-, total cells per mouse × 106 (%) . | Spleen-derived B220+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived B220+Ly5.1-, total cells per mouse × 106 (%) . |
|---|---|---|---|---|---|---|
| BM alone | NA (NA) | 0.35 ± 0.06 (1.67 ± 0.31) | NA (NA) | 0.62 ± 0.11 (2.97 ± 0.42) | NA (NA) | 13.88 ± 0.56 (67.33 ± 2.23) |
| Control | 2.11 ± 0.65 (14.04 ± 1.56) | 0.03 ± 0.01 (0.20 ± 0.03) | 2.21 ± 0.45 (15.15 ± 1.75) | 0.56 ± 0.20 (3.80 ± 1.01) | 0.67 ± 0.17 (4.54 ± 0.72) | 1.45 ± 0.27 (10.01 ± 1.25) |
| Rapamycin | 2.83 ± 0.51 (6.97 ± 1.22*) | 0.11 ± 0.03* (0.27 ± 0.02*) | 1.76 ± 0.45 (4.25 ± 0.55*) | 0.79 ± 0.18 (1.91 ± 0.14*) | 1.20 ± 0.19* (2.98 ± 0.54*) | 3.90 ± 0.25* (9.69 ± 1.36) |
| αCD28 | 1.49 ± 0.38 (9.53 ± 1.74*) | 0.05 ± 0.03 (0.29 ± 0.04*) | 2.10 ± 0.87 (12.68 ± 1.90) | 0.69 ± 0.39 (3.94 ± 0.63) | 0.72 ± 0.30 (4.31 ± 0.58) | 1.87 ± 1.19 (10.10 ± 2.13) |
| αCD28 + rapamycin | 0.91 ± 0.23*† (2.91 ± 0.59*†‡) | 0.14 ± 0.00*‡ (0.45 ± 0.03*†‡) | 1.29 ± 0.70 (4.05 ± 2.02*‡) | 0.68 ± 0.11 (2.20 ± 0.27*) | 0.96 ± 0.16 (3.10 ± 0.34*) | 5.65 ± 0.36*†‡ (18.28 ± 0.38*†‡) |
. | Spleen-derived CD4+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived CD4+Ly5.1-, total cells per mouse × 106 (%) . | Spleen-derived CD8+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived CD8+Ly5.1-, total cells per mouse × 106 (%) . | Spleen-derived B220+Ly5.1+, total cells per mouse × 106 (%) . | BM-derived B220+Ly5.1-, total cells per mouse × 106 (%) . |
|---|---|---|---|---|---|---|
| BM alone | NA (NA) | 0.35 ± 0.06 (1.67 ± 0.31) | NA (NA) | 0.62 ± 0.11 (2.97 ± 0.42) | NA (NA) | 13.88 ± 0.56 (67.33 ± 2.23) |
| Control | 2.11 ± 0.65 (14.04 ± 1.56) | 0.03 ± 0.01 (0.20 ± 0.03) | 2.21 ± 0.45 (15.15 ± 1.75) | 0.56 ± 0.20 (3.80 ± 1.01) | 0.67 ± 0.17 (4.54 ± 0.72) | 1.45 ± 0.27 (10.01 ± 1.25) |
| Rapamycin | 2.83 ± 0.51 (6.97 ± 1.22*) | 0.11 ± 0.03* (0.27 ± 0.02*) | 1.76 ± 0.45 (4.25 ± 0.55*) | 0.79 ± 0.18 (1.91 ± 0.14*) | 1.20 ± 0.19* (2.98 ± 0.54*) | 3.90 ± 0.25* (9.69 ± 1.36) |
| αCD28 | 1.49 ± 0.38 (9.53 ± 1.74*) | 0.05 ± 0.03 (0.29 ± 0.04*) | 2.10 ± 0.87 (12.68 ± 1.90) | 0.69 ± 0.39 (3.94 ± 0.63) | 0.72 ± 0.30 (4.31 ± 0.58) | 1.87 ± 1.19 (10.10 ± 2.13) |
| αCD28 + rapamycin | 0.91 ± 0.23*† (2.91 ± 0.59*†‡) | 0.14 ± 0.00*‡ (0.45 ± 0.03*†‡) | 1.29 ± 0.70 (4.05 ± 2.02*‡) | 0.68 ± 0.11 (2.20 ± 0.27*) | 0.96 ± 0.16 (3.10 ± 0.34*) | 5.65 ± 0.36*†‡ (18.28 ± 0.38*†‡) |
Lethally irradiated (1000 cGy) B10.BR recipients were given transplants of 15 × 106 T-cell-depleted marrow cells from B6 donors plus 15 × 106 splenocytes from B6.Ly5.1 donors or with bone marrow alone (BM alone). Recipients of marrow and splenocytes were treated intraperitoneally with either control solvent, anti-CD28 mAb at 100 μg × 1 day (αCD28), rapamycin at 1.5 mg/kg × 14 days (rapamycin), or the combination of both (αCD28 + rapamycin). On day 14 after transplantation, splenocytes were stained for CD4, CD8, B220, H2-Kb, and Ly5.1. Groups contained 3 to 4 recipients. Data shown are mean ± 1 SD. Results are representative of 2 replicate experiments. NA indicates not applicable.
P < .05 compared with BM plus splenocytes
P < .05 compared with rapamycin
P < .05 compared with αCD28