Fig. 5 | Nature Communications

Fig. 5

From: Antigen-selective modulation of AAV immunogenicity with tolerogenic rapamycin nanoparticles enables successful vector re-administration

Fig. 5

Control of anti-AAV8 capsid memory T cell responses with SVP[Rapa]. a Protocol outline. Male C57BL/6 mice (n = 4) were treated i.v. with AAV8-luc vector (2.5 × 1012 vg kg−1) together with SVP[Rapa] or with SVP[empty] on day 0 and with a second dose on day 77 with AAV8-luc vector (5 × 1011 vg kg1) together with SVP[Rapa] or SVP[empty] control. Spleens were collected at day 19 and day 80 after priming and boosting, respectively. b IFN-γ ELISpot responses after 7 days of in vitro restimulation with an AAV8 peptide pool performed in splenocytes collected 19 or 80 days following initial vector injection. The symbols represent individual animals and the bars represent mean ± s.d. (n = 4, ***p < 0.05, ns not significant, two-tailed, unpaired t-test). c Protocol outline. Donor mice (n = 15) were left untreated or treated i.v with 4 × 1011 vg kg1 of AAV8-RFP vector. 62 days later, spleens were collected and CD4+ T cells were purified. 1 × 107 of CD4+ T cells were then adoptively transferred into four groups of recipient mice (n = 6). Animals were treated with 4 × 1011 vg kg−1 of AAV8-RFP vector together with SVP[Rapa] or SVP[empty] control on day 1 post adoptive transfer and with 4 × 1011 vg kg−1 of AAV8-SEAP together with SVP[Rapa] or SVP[empty] on 21 post adoptive transfer. The tabular legend represents the summary of the treatment groups and the symbols represent the different cohorts of recipient mice. d, e Analysis of anti-AAV8 IgG antibodies in donor mice (d) and in recipient mice post adoptive transfer (e) measured by ELISA. d, e Data shown as mean ± s.d. (n = 6, *p < 0.05, ****p < 0.0001, refers to significance between cohorts 3 and 4 as determined by two-way ANOVA with Tukey post hoc test). SVP[Rapa] treatment consisted of 50 µg of rapamycin