Figure 1.

Improved rAAV-Mediated Gene Expression with Tacrolimus Treatment

(A–C) β-galactosidase staining after AAV8CMVLacZ injection without tacrolimus treatment (A, 2 weeks; B, 4 weeks; C, 8 weeks). (D–F) β-galactosidase staining after AAV8CMVLacZ injection with tacrolimus treatment (D, 8 weeks; E, 24 weeks; F, 42 weeks). AAV8CMVLacZ expression persisted for 16 weeks after transduction. Following tacrolimus co-treatment, AAV8CMVLacZ expression remained detectable for 42 weeks after transduction, and cell infiltration was observed. (G–I) β-galactosidase staining after AAV9CMVLacZ injection without tacrolimus treatment (G, 2 weeks; H, 8 weeks; I, 16 weeks). (J–L) β-galactosidase staining after AAV9CMVLacZ injection with tacrolimus treatment (J, 8 weeks; K, 24 weeks; L, 42 weeks). (M and N) FLAG staining after AAV8CMVM3FLAG injection without tacrolimus treatment (M, 16 weeks; N, 24 weeks). (O and P) FLAG staining after AAV8CMVM3FLAG injection with tacrolimus treatment (O, 24 weeks; P, 42 weeks). (Q–S) Immunostaining of inflammatory cells in serial section of B 4 weeks after AAV8CMVLacZ injection without tacrolimus treatment (Q, CD68 staining; R, CD4 staining; S, CD8 staining). (T and U) FLAG (T) and hematoxylin and eosin (U) staining 42 weeks after AAV9CMVM3FLAG injection with tacrolimus treatment. AAV9CMVM3FLAG expression remained detectable for 42 weeks after transduction. Cell infiltration was observed but to a lesser degree than that observed with the AAV8 vector. The same muscle fiber is indicated by an asterisk (∗) or star (☆). FLAG staining is marked with arrowheads. Scale bar, 100 μm. AAV, adeno associated virus; CMV, human cytomegalovirus immediate early enhancer and promoter; M3, microdystrophin; CD, cluster of differentiation.