|
|
Blood, Vol. 109, Issue 2, 661-669, January 15, 2007
Active MAC-1 (CD11b/CD18) on DCs inhibits full T-cell activation
Blood Varga et al.
109: 661
Supplemental materials for: Varga et al, Vol 109, Issue 2, 661-669
Files in this Data Supplement:
- Figure S1. Phenotype of DC from CD18–/– mice (PDF, 78.2 KB) -
Mature DCs were cultured for 7 days in the presence of GM-CSF and IL-4, labeled with the indicated antibodies, and analyzed by flow cytometry.
- Figure S2. Binding activity of β2 integrins on DCs (PDF, 13.5 KB) -
Day 7, 8, and 9 DCs have been left untreated or were pretreated with 5 mM Mg2+. Cells were then placed into an ICAM-1 binding assay. Human IgG–coated wells served as a specificity control. One representative of 3 experiments is shown.
- Figure S3. DC viability after Mg2+ treatment (PDF, 11.2 KB) -
DCs from CD18–/– mice were either left untreated or activated using 100 ng/mL LPS. Cells were cultured for 72 hours with or without Mg2+ as indicated. Viability was assessed by trypan blue exclusion. Data are mean values (± SEM) of three independent experiments.
- Figure S4. DC–T-cell contact duration after Mg2+ activation (PDF, 30.2 KB) -
Bone marrow DCs (5 × 104) of either CD18+/+ or CD18–/– mice were treated with Mg2+ or not and incubated with 5 × 105 CD18+/+ T cells in a collagen gel. Contacts of T cells and bone marrow DCs were recorded by video microscopy, and individual DC–T-cell pairs were analyzed for their contact times.
- Figure S5. Effect of Mg2+ on isolated T cells (PDF, 24.2 KB) -
T cells from CD18+/+ and CD18–/– mice were plated in triplicate (1 × 105/well) in 96-well plates that had been coated with 10 µg/mL anti/CD3 antibody. Soluble anti-CD28 antibody (5 µg/mL) was added to the culture. T cells were either left untreated or incubated with 10 µg/mL anti-CD18 antibody (GAME46), and/or 5 mM Mg2+, as indicated. After 5 days, 1 µCi (0.037 MBq) 3H-thymidine was added to the wells for the last 18 hours of incubation, cells were harvested, and proliferation was determined by  -counting. One representative experiment of 3 is shown.
- Figure S6. Cellular distribution of Mac-1 (CD11b/CD18) on DCs (PDF, 20 KB) -
DCs (5 × 105) were left untreated or incubated with 5 mM Mg2+ for 18 hours. Then cells were stained on the surface and intracellularly with anti-CD11b antibody, or with an isotype control antibody (shaded graph). Cells were analyzed by flow cytometry.
|
|
