Extended Data Figure 6. LTBR ligands and expression of LTBR via mRNA or with deletion and point mutants.

a) IL2 secretion after 24 hour stimulation with CD3/CD28 antibodies. Where indicated, recombinant soluble LTA (1 ng/mL) or LIGHT (10 ng/mL) were added together with CD3/CD28 antibodies. CD4+ T cells from one donor were tested in triplicate. b,c) CD4+ and CD8+ T cells from two donors were co-incubated for 24 hours with CD3/CD28 antibodies or recombinant soluble LTA or LIGHT and then IL2 (b) and IFNγ (c) were measured. (n = 3 biological replicates). d,e) Differentiation phenotype (d) or proliferation (e) after restimulation of tNGFR and LTBR transduced T cells (n = 3 biological replicates) incubated either with IL2 alone or with LTA (1 ng/mL) or LIGHT (10 ng/mL) for the duration of culture. CM: Central memory. EM: Effector memory. Unpaired two-sided t-test p values are shown. f-i) Transient LTBR or tNGFR expression via mRNA nucleofection (f). T-cells were either nucleofected with LTBR or tNGFR mRNA (n = 3 biological replicates), and the surface expression of LTBR (g), tNGFR (h) or four genes upregulated in LTBR cells (i) was monitored over 21 days. At each timepoint the expression of target genes was normalised to matched tNGFR control. j) Schematic representation of FLAG-tagged LTBR mutants. k) LTBR and FLAG expression in T-cells transduced with LTBR mutants.

Error bars indicate SEM.