Fig. 7

Blocking effect of AAV-13A7-Fc administration on CD27 and CD62L shedding during DSS administration. A Blood samples were collected three weeks after AAV-13A7-Fc or AAV-14D5-dimHLE i.m. injection (before DSS) or 13 days later (i.e., after DSS 2.5% for 5 days followed by 8 days of regular water). CD62L and CD27 shedding was evaluated by flow cytometry of the indicated gated T cell subsets after incubation, or not, with 150 µM ATP. The top panels illustrate the flow cytometry gating strategy and the bottom panels the percentage of CD27⁺CD62L^high in each indicated T cell subset. B Plasma samples were collected before the start of DSS treatment (three weeks after AAV injection) and after 2.5% DSS + regular water treatment (day 13). Concentrations of unbound nanobody-based biologics were titrated by flow cytometry using a P2X7-expressing cell line and standard curves obtained with the same recombinant biologics produced in vitro. One representative experiment out of three is shown, with n = 5–6 mice per group in each experiment. Statistical analysis was performed using one-way ANOVA with *p < 0.05, **p < 0.01 and ***p < 0.001