Figure 1.

Biological impact of CD28 alterations. (A) Schematic representation of the different CD28 RNA fusions and PCR design for their screening. Arrows indicate the approximate positions of oligonucleotide primers on the indicated RNA fusions. (B) NF-κB luciferase reporter assay in Jurkat cells monitoring the activity of a CD28 T195P activating mutant and the ICOS(ex1)_CD28(ex2) fusion, compared with CD28 WT. Data are represented as mean+/−SEM from four independent experiments. (C) Spider plot representation of gene sets differentially enriched in patients with (red) (n=14) or without (blue) (n=84) alterations (mutations or fusions) in CD28. A total of 304 signatures, including 23 of 50 hallmark signatures and nine signatures from the curated and immunogenic signature collection of the Molecular Signatures Database (MSigDB), all signatures of interest in lymphoid biology (Signatures database, Staudt’s lab), and two manually annotated sequences linked to TCR signalling were tested. The list of signatures that reached statistical significance is provided in Online Supplementary Table S2. WT: wild-type; Mut/Fus: mutations or fusions.