Recipients given day 15 treatment showed similar proteinuria and survival curve to the control recipients. Decrease Donor T Cell Percentages or Yields After HCT Lethally irradiated BALB/c recipients were transplanted with spleen cells (75106) and BM cells (2.5 106) from DBA/2 donors and injected i.v. with either rat IgG or anti-CD20 mAb (40 mg/Kg) the following day after HCT. Recipients were sacrificed 7, 15 and 25 days after HCT. Percentage and yield as well as a representative pattern of splenic CD5. 1+ TCR+ T cells is shown from 4 mice each group per time point. NIHMS592754-supplement-02.tif (1.0M) GUID:?7E818828-1765-4D7D-A719-7C10E42E47E2 03: Supp Fig 3 C Low-dose C57BL/6 CD8+ T Cells Induced Severe cGVHD in Recipients Given WT BM But Induced Little Signs of cGVHD in Recipients Given Ig?/? BM Lethally irradiated BALB/c recipients were given a low dose of donor C57BL/6 CD8+ T cells (0.5106) and either WT donor BM or Ig?/? donor BM (2.5106). Recipients were monitored for clinical GVHD, including (A) body weight loss, (B) clinical cutaneous cGVHD score and (D) survival. A representative photograph taken at day 60 is shown (n=4). NIHMS592754-supplement-03.tif (11M) GUID:?64F087A2-7954-4D5C-B19B-A58E178C2BE3 04: Supp Fig 4 C Administration of Anti-CD20 mAb Does Not Prevent Acute GVHD Lethally irradiated BALB/c recipients were injected with 5106 whole spleen cells and 2.5 106 TBCD-BM cells from C57BL/6 donors and injected i.v. with either rat IgG or anti-CD20 mAb (40 mg/Kg) the following day after HCT. Recipients given TBCD-BM alone were used as controls. Recipients were monitored for clinical GVHD, including body weight change, clinical GVHD score, and survival (n=8 from two replicate experiments). NIHMS592754-supplement-04.tif (354K) GUID:?F37BD853-8F0F-43E8-B45A-C8540E0DA3E0 05: Supp Fig 5 C Treatment With Anti-CD20 mAb After GVHD Onset Does Not Ameliorate GVHD Lethally irradiated BALB/c recipients were injected with 1.25106 whole spleen cells and 2.5 106 TBCD-BM cells from C57BL/6 donors and injected i.v. Pdpn with either rat IgG or anti-CD20 mAb (40 mg/Kg) starting on day 45 after disease onset, with follow-up injections on day 50 and 55. Recipients were monitored for clinical GVHD, including body weight change, clinical cutaneous GVHD, and survival (n=4 from two replicate experiments). NIHMS592754-supplement-05.tif (3.0M) GUID:?D046C46B-33AA-4F10-A592-9E1C76F90451 Abstract Chronic graft-versus-host disease (cGVHD) is an autoimmune-like syndrome, and donor B cells play important roles in augmenting its pathogenesis. B cell-depleting anti-CD20 mAb has been administered before or after cGVHD onset for preventing or treating cGVHD in clinic. Stevioside Hydrate Although administration before onset appeared to Stevioside Hydrate be more effective, the effect is variable and sometimes minimal. Here, we used two mouse cGVHD models to evaluate the preventive and therapeutic effect of anti-CD20 mAb. With the model of DBA/2 donor to MHC-matched BALB/c recipient, one intravenous injection of anti-CD20 mAb (40 mg/kg) the following day or on day 7 after HCT when serum autoantibodies were undetectable effectively prevented induction of cGVHD and preserved strong graft-versus-leukemia (GVL) effect. The separation of GVL effect from GVHD was associated with a significant reduction of donor CD4+ T cell proliferation and expansion, and protection of host thymic medullary epithelial cells. Anti-CD20 mAb administration also Stevioside Hydrate prevented expansion of donor T cells and induction of cGVHD in another mouse model of C57BL/6 donor to MHC-mismatched BALB/c recipients. In contrast, administration of anti-CD20 mAb after GVHD onset was not able to effectively deplete donor B cells or ameliorate cGVHD in either model. These results indicate that administration of anti-CD20 mAb prior to signs of cGVHD can prevent induction of autoimmune-like cGVHD while preserving GVL effect; there is little effect if administered after cGVHD onset. This provides new insights into clinical prevention and therapy of cGVHD with B cell-depleting reagents. Introduction Allogeneic hematopoietic cell transplantation (HCT) is a curative therapy for hematological malignancies such as leukemia and lymphoma . While donor T cells including CD4+ and CD8+ in transplants play a critical role in mediating graft-versus-leukemia/lymphoma (GVL) effects and preventing tumor relapse, alloreactive T cells also mediate a severe side effect called graft-versus-host disease (GVHD), a major obstacle for widespread application of allogeneic HCT [2C6]. While both CD4+ and CD8+ T cells can induce GVHD, CD8+ T cells are more potent than CD4+ T cells in mediating GVL effect [7C15]. GVHD is initiated by alloreactive T cell infiltration of GVHD target tissues (i.e. gut, skin, liver, lung, and thymus) in recipients conditioned with total body irradiation (TBI) or high dose chemotherapy ..