Secondary Transfer of Splenocytes From B10.BR (H2k) Mice Transplanted With C57.BL6 Allogeneic BM and Irradiated (7.5 Gy) Splenocytes Into C57.BL6 and B10.BR Recipients
| Recipient Mice . | B10.BR (H2k) . | B10.BR (H2k) . | C57.BL6 (H2b) . | C57.BL6 (H2b) . |
|---|---|---|---|---|
| BM cells injected | B10.BR | B10.BR | C57.BL6 | C57.BL6 |
| Spleen cells injected | Naı̈ve C57.BL6 Spleen Transplant | Chimeric H2b/H2kSpleen Transplant | Naı̈ve B10.BR Spleen Transplant | Chimeric H2b/H2k Spleen Transplant |
| Clinical GVHD | +++ | None | ++ | None |
| Survival post-BMT | day 21, 21 | day >30, >30 | day 18, >30 | day >30, >30 |
| Weight at death | 18.7, 19 g | >26 g | 20 g | >26 g |
| % Donor H2b T cells in | ||||
| Peripheral blood | 23 | 7 | ND | ND |
| Spleen | 12 | 16 | ND | ND |
| BM | 17 | 3 | ND | ND |
| Recipient Mice . | B10.BR (H2k) . | B10.BR (H2k) . | C57.BL6 (H2b) . | C57.BL6 (H2b) . |
|---|---|---|---|---|
| BM cells injected | B10.BR | B10.BR | C57.BL6 | C57.BL6 |
| Spleen cells injected | Naı̈ve C57.BL6 Spleen Transplant | Chimeric H2b/H2kSpleen Transplant | Naı̈ve B10.BR Spleen Transplant | Chimeric H2b/H2k Spleen Transplant |
| Clinical GVHD | +++ | None | ++ | None |
| Survival post-BMT | day 21, 21 | day >30, >30 | day 18, >30 | day >30, >30 |
| Weight at death | 18.7, 19 g | >26 g | 20 g | >26 g |
| % Donor H2b T cells in | ||||
| Peripheral blood | 23 | 7 | ND | ND |
| Spleen | 12 | 16 | ND | ND |
| BM | 17 | 3 | ND | ND |
B10.BR or C57.BL6 mice were exposed to 10 Gy irradiation in 2 equal fractions and transplanted with 1 × 106 syngeneic BM cells and 25 × 106 splenocytes from naı̈ve B10.BR or C57.BL6 donors (as shown) or 25 × 106splenocytes obtained from B10.BR mice with mixed chimerism (H2b/H2k) that were killed 112 days after receiving 0.5 × 106 C57.BL6 BM cells and 120 × 106 irradiated (7.5 Gy) C57.BL6 splenocytes. Three of 4 recipients of naı̈ve allogeneic splenocytes died or were euthanized between day 18 and 21 with clinical evidence of acute GVHD including the appearance of ruffled fur, decreased motor activity, diarrhea, and “hunched over” posture. The remaining mice were killed at day +30. The peripheral blood, BM, and spleen of B10.BR mice surviving to day +30 were analyzed by flow cytometry for the presence of T cells derived from H2b donor splenocytes. Progeny of the irradiated splenocytes (genetically marked with CD45.1) were not detectable after transfer to secondary BMT recipients.
Abbreviation: ND, not detectable.