Figure 1.

Therapeutic efficacy of haematopoietic stem cell [HSC] gene therapy in Il10rb^-/- IBD mice correlates with gene marking in lamina propria macrophages. [A] A third-generation self-inactivating [SIN] lentiviral vector was used for gene transfer of a codon optimised [co] murine Il10rb cDNA coupled to the mVenus fluorescent reporter driven by the phosphoglycerate kinase [PGK] promoter. [B] Phosphorylation of STAT3 after stimulation with IL10 analysed by flow cytometry in macrophages derived from wild type [WT] or Il10rb^-/- lin^- bone marrow cells. Il10rb^-/- lin^- cells were transduced with a PGK-eGFP [control] or PGK-IL10rb therapeutic vector. [C] Control transduced Il10rb^-/- lin^- bone marrow cells, corrected Il10rb^-/- lin^- or WT lin^- cells were transplanted into irradiated Il10rb^-/- IBD mice and body weights were analysed for 35 days. Values are given as % of initial body weight (n = 4–5 per group, mean ± range [range depicted as shaded area]). [D] Quantification after histological scoring [n = 4–5 per group, individual values with mean ± SD]. [E] Representative light microscopy of H/E stained colon paraffin sections. [F] Correlation of the frequency of gene-marked lymphocytes, granulocytes, and macrophages in the lamina propria with the histological score [mean ± 95% CI]. LTR, long terminal repeat; wPRE, woodchuck hepatitis virus post-transcriptional regulatory element, significances were calculated by two-way ANOVA with Bonferroni’s multiple comparison test, *p <0.05, **p <0.01, ***p <0.001, ****<0.0001, ns not significant. IBD, inflammatory bowel disease; SD, standard deviation; H/E, haematoxylin and eosin; CI, confidence interval; ANOVA, analysis of variance.