FIGURES AND TABLES
FROM:
TGF
is responsible for skin tumour infiltration by macrophages enabling the tumours to escape immune destruction
Scott N Byrne, Matthew C Knox and Gary M Halliday
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Figure 1.
TGF
-transfected tumour cells (PGWTGF) secrete elevated levels of TGF and undergo progressive tumour growth in vivo. (a) TGF production was assessed in supernatant collected from in vitro-cultured tumour cells using an enzyme-linked immunosorbent assay kit, mean+s.e.m. from four replicates shown. (b) Pooled tumour growth (mean diameter
s.e.m.) and (c) tumour incidence measurements from five independent experiments, PGW; n=16 mice and PGWTGF; n=48 mice (where s.e.m. is not obvious it is too small to be observed in this scale). Fully regressed tumours were included as zero for the calculation of mean diameter. *
P<0.05. TGF, transforming growth factor-1.
Figure 2.
Progressor skin tumours are infiltrated by a significantly greater percentage of macrophages while regressor tumours are populated by dendritic cells. (a) The first column of dot plots show the level of isotype staining on whole tumour cell preparations compared to the level of CD45 and CD11c staining (second column). Four-colour flow cytometry identified CD11c+CD45+ DC (R1) and CD11c-CD45+ non-DC inflammatory cell infiltrates (R2). The third column of dot plots show the level of isotype staining for CD11b and MHC II on CD45+ cells compared to the expression of CD11b and MHC II on DC (R1; fourth column) and non-DC inflammatory cells (R2; last column of dot plots). TAM were identified as CD11c-CD45+CD11b+ (last column in (a)). Dot plots are representative of four separate experiments. (b) The percentage of CD11c+CD45+ CD11b+ DC or CD11c-CD45+CD11b+ TAM from four separate experiments were pooled. Mean+s.e.m. shown. * P<0.05. DC, dendritic cells; TAM, tumour-associated macrophages.
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Figure 3.
TGF alters the APC ratio and enhances macrophage phagocytosis in vitro. (a) A total of 2 
106 whole spleen single-cell suspensions were incubated at various TGF concentrations for 24 h. CD11c+CD11b+ DC (
) and CD11c-CD11b+ Macs (
) were identified by flow cytometry and the percentage of cells at the various TGF concentrations was determined. (b) Fluorescently labelled latex beads were added for the final 3 h of incubation and the level of bead uptake was determined by flow cytometry. The numbers shown in the histograms refer to the number of beads that had been phagocytosed by each cell. The total number of phagocytic DC () and Macs () was analysed by flow cytometry and the change induced by TGF is shown in the right panel. APC, antigen-presenting cell; DC, dendritic cells; TGF, transforming growth factor-1.
Figure 4.
Macrophages are the dominant phagocytic cells infiltrating progressing skin tumours. (a) Injected intravenously 18 h prior to tumour removal and analysis by four-colour flow cytometry. The phenotype of the cells that had taken up beads were confirmed as either CD11c+CD11b+MHC IIhigh DC (bold histogram) or CD11c-CD11b+MHC II+ TAM (shaded histogram). Progressor tumour dot plots only are displayed as regressor dot plots looked similar. (b) Significantly more beads were taken up by any type of cell (percentage of total) within regressor (solid bar) compared to progressor (open bar) tumours (n=7). (c) Analysis of the percentage of each APC subset (DC in solid bars statistically compared to TAM in the open bar) that had taken up beads (n=7). In all cases mean+s.e.m. shown. * P<0.05. APC, antigen-presenting cell; DC, dendritic cells; TAM, tumour-associated macrophages.
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