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Blood, Vol. 110, Issue 13, 4319-4330, December 15, 2007
Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells
Blood Duluc et al.
110: 4319
Supplemental materials for: Duluc et al
Files in this Data Supplement:
- Figure S1. M2d suppress T-cell proliferation (JPG, 27.7 KB)
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M , LIF-M, IL-6-M or DC were stimulated for 48 h with 200 ng/ml LPS, washed, and cultured in triplicate at 4 × 103 cells with 105 purified allogenic CD4+ T cells stimulated with 10 µg/ml anti-CD3 mAb (clone OKT3; ATCC) and 20 U/ml IL-2 (Immunotools). After 4 d, cells were pulsed for 16 h with 3H-thymidine (0.25 µCi/well; Amersham). 3H-thymidine incorporation was measured by standard liquid-scintillation counting. Results (mean ± SD, n = 4) are expressed in variation of T cell proliferation defined as follows: % = (A -B) / B × 100, where A proliferation with APC; B, proliferation without APC. * means p<0.05.
- Figure S2. Increased expression of IDO in M2d (JPG, 47.2 KB)
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Monocytes were cultured 5 days in CM in the absence (M) or presence of 200 ng/ml of LIF (LIF-M). After cell lysis, protein extracts were analyzed for IDO expression by western-blotting as previously described (Hill M, The FASEB Journal. 2005;19:1957-1968). GAPDH expression was used for control of protein loading. The result is representative of one out of three experiments.
- Figure S3. M2d suppress T-cell proliferation (JPG, 120 KB)
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48 h LPS-activated M, LIF-M, IL-6-M, TA-M or TAM were cultured at 2 × 104 cells with 105 purified allogenic CD4+ T cells, previously stained with 0.5 µM fluorescent dye CFDA-SE (Molecular Probes), activated with 10 µg/ml anti-CD3 mAb plus 20 U/ml IL-2 in the absence or presence of 200 µM 1-D-methyl-tryptophan (1-MT, Sigma). At d 5, cells were stained with 7-AAD and APC-labelled annexin-V (BD Biosciences). CD4+ T cell proliferation was measured by CFDA-SE dilution on living cells.
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