Fig. 5: LncRNA derived MHC class I peptides as cancer vaccines in a colon26 tumour model.
From: Long non-coding RNA-derived peptides are immunogenic and drive a potent anti-tumour response
A Groups of 10 BALB/c mice were vaccinated with ChAdOx1-PepLnc adenoviral vectors expressing a poly-antigen cassette containing 20 lncRNA-derived peptides, or a control ChAdOx1-GFP adenoviral vector. At 9 days post vaccination, half the mice were sacrificed and their splenocytes collected for ELISpot assay (a). The other half received a booster vaccination 4 weeks later with MVA-PepLnc or MVA-GFP as indicated, for a further 9 days (b). Splenocytes were stimulated with the indicated peptide, or a pool of peptides contained within the poly-antigen cassette, and activity was measured in interferon-γ-based ELISpot. DMSO and PHA were used as negative and positive controls respectively; n = 2 independent experiments. B (a) BALB/c mice were vaccinated with ChAdOx1-PepLnc or ChAdOx1-GFP adenoviral vectors. At day 9, post-vaccination mice were challenged with CT26 cells; n = 5 ChAdOx1-GFP, n = 6 ChAdOx1-PepLnc. (b) Absolute tumour volume is presented as mean ± SEM, n = 5 ChAdOx1-GFP, n = 6 ChAdOx1-PepLnc; (c) Absolute tumour volume of individual mice at day 17 (two-tailed Student’s t test; ** p  =  0.0067), n = 5 ChAdOx1-GFP, n = 6 ChAdOx1-PepLnc; box and whiskers are defined as minimum, first quartile, median, third quartile, and maximum of data. (d) Relative body weight of BALB/c mice presented as a mean value, n = 5 ChAdOx1-GFP, n = 6 ChAdOx1-PepLnc. C (a) Colon26-bearing BALB/c mice were vaccinated with DCs pulsed with pools of 15 lncRNA derived peptides at Day 0. As a control, unpulsed dendritic cells were used; n = 6 mice per treatment. (b) Absolute tumour volume in BALB/c mice presented as mean ± SEM, n = 6 mice per treatment; (c) Absolute tumour volume of individual mice at day 12 (two-tailed Student’s t test; *** p  = 0.0004), n = 6 mice per treatment; box and whiskers are defined as minimum, first quartile, median, third quartile, and maximum of data. (d) Relative body weight of BALB/c mice presented as a mean value ± SEM, n = 6 mice per treatment. D Immunohistochemical staining of anti-CD8 (a), anti-CD4 (b), or anti-CD163 (c) in colon26 tumours at 14 days post vaccination with pulsed DCs (see Fig. 5C). Original magnification, 20x, scale bar, 50 μm; and 63x; scale bar, 16 μm. n = 4 independent experiments; d) Optical density is presented as mean ± SD; two-tailed Student’s t test; n = 4 independent experiments (each performed on two separate slides); ** adjusted P value < 0.01.
