Figure 8

Epithelial colonized L. jensenii preserve potent anti-HIV properties. Western blot from 24 h sterile supernatants collected from L. jensenii-colonized vaginal (Vk2/E6E7) and endocervical (End1E6E7) epithelial cells demonstrate consistent preservation of modified Cyanovirin-N (mCV-N) expression in mCV-N producing strains. (Figure 8a) mCV-N producing bioengineered strains (L. jensenii 1153–1666, 2666 and 3666) located in lanes #2, 3 and 4 are contrasted to L. jensenii 1153 WT in lane #1, the β-glucuronidase expressing strain L. jensenii 1153–1646 in lane #5, MALP-2 control in lane #6, and medium control in lane #7. (Figure 8b) A mCV-N standard in lane #1 is compared to the mCV-N producing L. jensenii strains: L. jensenii 1153–1666 and 3666 in lanes #4 and #5 in contrast to the green florescent protein expressing strain L. jensenii 1153-gfp in lane #6, MALP-2 in lane #3 and medium control in lane #2. (Figure 8c) No loss to filtration is observed in western blot analyses of mCV-N before and after spiking one ml of media with one μg mCV-N. (Figure 8d) gp120 binding activity in one representative mCV-N producing L. jensenii 1153–1666 strain detected by a gp120 binding assay in sterile supernatants collected from 24 h L. jensenii colonized vaginal (Vk2/E6E7) epithelial culture. Data are from one representing three independent experiments.