Effect of N-9 and ι-carrageenan on Ad26 immunogenicity after Ivag priming. Depoprovera-treated mice (n=5/group) were inoculated Ivag with or without N-9 followed 5 hours later with Ad26-E6E7fus (1.0 × 1010 vp) delivered in CMC or ι-carrageenan. Additional groups, IM Ad26-E6E7fus (IM), Ivag Ad26-mock with N-9 and no carrageenan (mock) and Ivag N-9 without carrageenan (sham) were used as control. Cervicovaginal (A–C, E–F) and blood samples (D–G) were analyzed by H2-Db/E749-57 tetramer staining for the presence of HPV16 E7-specific CD8+ T cells three weeks after immunization. Representative FACS plot for H2-Db/E749-57 tetramer staining on cervicovaginal CD8+ T cells (A, row1) and for CD69 and CD103 expression by tetramer negative (A, row2) and tetramer-positive (A, row3). Data are expressed as mean percentage or mean number of cells per organ ± SEM (N/A, not applicable).