Fig. 4: Antisense RNA silencing ameliorates TNBS-induced experimental colitis in humanized NOD/scid/gamma mice.

a NOD/scid/gamma (NSG) female recipients were injected with antisense⁺ CD4 cells, obtained from one healthy blood donor. After 3 weeks, mice were bled and checked for human chimerism. Mice showing >10% human chimerism were sensitized to TNBS. One week after sensitization, mice were administered a single enema of TNBS and given a single injection of FANA-CD39-AS oligonucleotides (or vehicle) intraperitoneally. After 72 h, mice were sacrificed. b Cropped gel of RT-PCR showing positivity for CD39-AS RNA in healthy blood donor CD4 T lymphocytes (representative of three independent experiments with similar results). Full scan gel is provided in Source data file. c Mean ± SEM disease activity index at the time of harvest in TNBS mice treated with vehicle (n = 5) or FANA-CD39-AS oligonucleotides (n = 6; *P = 0.035 using two-sided unpaired t test). d Mean ± SEM colon length (cm) at the harvest in vehicle (n = 5) and FANA-CD39-AS-treated (n = 6) mice (*P = 0.04 using two-sided Mann–Whitney test). e Hematoxylin and eosin staining of colon sections (original magnification, ×10, scale bar: 200 μM); arrows indicate the area magnified in the insets (×20, scale bar: 200 μM); mean ± SEM histology score at the time of harvesting is also shown (**P = 0.002 using two-sided unpaired t test). f Representative dot plots of CD4 and CD39 fluorescence of peripheral blood mononuclear cells isolated from one vehicle and one FANA-CD39-AS oligonucleotide-treated mouse at the time of harvest. Frequency of CD4⁺CD39⁺ cells is indicated in the upper right quadrant. g Mean ± SEM frequency of CD4⁺CD39⁺, CD4⁺FOXP3⁺, CD4⁺IL17⁺, and CD4⁺IL10⁺ cells in the peripheral blood, spleen, mesenteric lymph node (MLN), intra-epithelial lymphocytes (IEL), and lamina propria (LP) lymphocytes at the time of harvest in vehicle (n = 5) or FANA-CD39-AS oligonucleotide (n = 6) treated animals (*P = 0.048 for CD4⁺CD39⁺, *P = 0.049 for CD4⁺IL-17⁺ using two-sided unpaired t test).