Figure 8: LTα3 induces PNAd expression on tumour vasculature by signalling through TNFRs.
Infiltration of naive OT-I cells into i.p. (a) or s.c. (b) B16-OVA tumours in the indicated mice was determined. n=7 (a) or 4 (b) per group. PNAd expression (c,d) and Ccl21 expression in i.p. (c,e) or s.c. (d) tumours were quantified. n=4 per group. (f) Expression of Lta and Tnfa mRNA in sorted CD3+CD8+ T cells from i.p. (left panel) and s.c. (right panel) tumours was determined by quantitative reverse transcriptase–PCR. Expression levels are displayed relative to Hprt. (g) Endogenous CD8 T-cell accumulation in day 14 i.p. B16-OVA tumours growing in WT, Rag1−/− and Rag1−/− mice repleted with either TNFα−/− or LTα−/− CD8 T-cells (n=3 per group) was determined by immunofluorescence. (h) PNAd expression in i.p. tumours from the indicated mice was determined. n=4 per group. (i) Expression of TNFR1 and TNFR2 on CD31+ endothelial cells from i.p. (dashed lines) and s.c. (solid lines) B16-OVA tumours was determined by flow cytometry. Shaded region represents background staining in FMO controls. (j) CD31 and PNAd expression in i.p. tumours from WT→TNFR1/2−/− or TNFR1/2−/−→WT bone marrow chimeras. Data (mean+s.e.m.) are pooled from two experiments (a,b) or representative of two (c–j) independent experiments. ns: P>0.05, *P<0.05, **P<0.01, ***P<0.001 by unpaired t-test (d,e) or one-way analysis of variance with Tukey’s post test (a–c,h). Scale bars=100 μm.
