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Blood, Vol. 112, Issue 10, 4080-4089, November 15, 2008
Mononuclear myeloid-derived "suppressor" cells express RAE-1 and activate natural killer cells
Blood Nausch et al.
112: 4080
Supplemental materials for: Nausch et al
Files in this Data Supplement:
- Figure S1. Gr-1+CD11b+F4/80+ cells accumulate in RMA-RAE-1γ and B16BL6 tumor-bearing mice and express RAE-1 (JPG, 54.4 KB)
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(A) C57BL/6 mice were injected s.c. with with 1 × 106 RMA-S lymphoma cells. Cell numbers of PBL/ml blood from untreated (day 0) and tumor-bearing mice at the indicated time points were assessed by cell counting. The percentages of Gr-1+ CD11b+F4/80+ cells among total PBL were determined by flow cytometry and calculated as absolute cell numbers of Gr-1+ CD11b+F4/80+ cells/ml blood. Data represent the mean ± SD of 5 animals per group. (B) C57BL/6 mice were injected s.c. with 1 × 105 RMA-RAE 1γ lymphoma or B16BL6 melanoma cells. Cell numbers of PBL/ml blood from untreated (day 0) and tumor-bearing mice at day 21 or 24 after tumor cell inoculation were assessed by cell counting. The percentages of Gr 1+CD11b+F4/80+ cells among total PBL were determined by flow cytometry and calculated as absolute cell numbers of Gr 1+CD11b+F4/80+ cells/ml blood. Data represent the mean ± SEM of 4 (RMA-RAE 1γ) or 8 (B16BL6) mice per group. (C) PBL of naïve and tumor-bearing mice at day 21 or day 24 after tumor cell inoculation were stained with anti-Gr 1, anti-F4/80 and anti–panRAE-1 mAbs. The expression of RAE 1 (black solid lines) on Gr-1+CD11b+F4/80+ cells was analyzed and compared to a matched-isotype control Ig (grey filled histograms). One representative flow cytometric analysis out of 8 animals per group is shown. Asterisks indicate statistical significance (p<0.01) determined by Student’s t-test. Similar results were obtained in 3 (A) or 2 (B, C) independent experiments.
- Figure S2. Flow cytometric analysis of FACS™-sorted Gr-1+CD11b+F4/80+ and Gr 1+CD11b+F4/80− cells (JPG, 29.7 KB)
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PBL from tumor-bearing mice were prepared as described in Experimental Procedures and Gr 1+CD11b+F4/80+ and Gr 1+CD11b+F4/80− cells were sorted by flow cytometry. Sorted cell were analyzed for the obtained purity by flow cytometry.
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