Fig. 4

Evaluation of AAV-13A7-Fc blocking and of AAV-14D5-dimHLE potentiating vectors on the activity of P2X7 over time and evaluation of the kinetics of production of nanobody-based biologics in vivo after a single i.m. injection of each AAVnano vector. AAV-13A7-Fc or AAV-14D5-dimHLE vectors were injected i.m. at a dose of 10¹¹ vg/mouse and blood cells or sera were collected at the indicated time points. A Blood cells were incubated with 150 µM or 30 µM ATP, as indicated, and the percentages of CD4⁺CD62L^high cells were determined by flow cytometry as an evaluation of P2X7 activity on the surface of the gated CD4⁺ T cells. The first two bars in each graph correspond, respectively, to negative and positive control cells collected from untransduced control mice treated or not with the indicated concentration of ATP. Results represent mean values ± SEM, with n = 2–3 mice per group. B The concentration of unbound 13A7-Fc and 14D5-dimHLE nanobody-based biologics in the sera collected at each indicated time point was titrated by flow cytometry using a P2X7-expressing cell line and standard curves obtained with the same recombinant biologics produced in vitro. Results represent mean values ± SEM, with n = 3 mice per group