|
|
Blood, Vol. 109, Issue 9, 4097-4104, May 1, 2007
Selective targeting of the LIGHT-HVEM costimulatory system for the treatment of graft-versus-host disease
Blood Xu et al.
109: 4097
Supplemental materials for: Xu et al, Blood, Vol. 109, Issue 9, 4097-4104
Files in this Data Supplement:
- Figure S1. Biological characteristics of LBH1 (JPG, 99.8 KB)
-
(A, top) 293T cells transfected with either full-length mouse HVEM (filled histogram) or control vector (open histogram) were stained with 1 µg LBH1 (right) or control hamster IgG (left), followed by PE-conjugated anti–hamster IgG Ab. Human 293T transfected with full-length mouse LIGHT (middle) or mouse BTLA (bottom) were stained with 2 µg mouse HVEM-mouse Ig fusion protein (filled histogram) or control mouse Ig protein (open histogram) in the presence of 10 µg/mL LBH1 (right) or control hamster IgG (left). Staining intensity of fusion protein was detected by FITC-conjugated anti–mouse Ig Ab. (B) Agonistic activity of LBH1 was examined in vitro by T cell costimulation assay. Tissue culture plate was coated with graded doses of anti–CD3 mAb (2.5-20 µg/mL) and further with 10 µg/mL LBH1 (black triangles) or control hamster IgG (white squares). Purified B6 T cells (1 × 106 cells/mL) were cultured in the wells for 3 days, and the proliferation activity during the last 15 hours was measured by 3H-thymidine incorporation assay. As positive control, 3 µg/mL anti–CD28 mAb was added in the wells at the beginning of T-cell culture (white circles).
- Figure S2. Preferential reduction of LIGHT-KO donor T cells compared to WT donor T cells in the single BDF1 host (JPG, 62.9 KB)
-
Purified Thy 1.1 WT B6 T cells (7 × 106 cells) were mixed with Thy1.2 LIGHT-KO B6 T cells (7 × 106 cells) and transferred intravenously into BDF1 recipient mice. After 11 days, absolute number of donor T cells in the host spleen was assessed by FACS analysis calculating H-2Kd-negative, Thy1.2, or Thy1.1-positive donor T cells. As control, the mixture of same numbers of Thy1.1 WT B6 T cells and Thy1.2 WT B6 T cells were transferred intravenously into BDF1 mice, followed by analysis of cell number on day 11. Each dot represents the data of individual mouse. *P = .038.
- Figure S3. Minor role of host-derived LIGHT in allo-CTL generation (JPG, 63.7 KB)
-
Either WT (black squares) or LIGHT-KO (white squares) B6 mice were exposed to sublethal irradiation (6 Gy), followed by an intravenous injection of 5 × 107 BALB/c spleen cells on day 0. On day 7, host spleen cells were harvested, and anti-allo CTL activity was assessed by 51Cr-release assay against EL4 (H-2b) and A20 (H-2d) tumors as target cells.
|
|
