Journal: Viruses

Article Title: Avirulins, a Novel Class of HIV-1 Reverse Transcriptase Inhibitors Effective in the Female Reproductive Tract Mucosa

doi: 10.3390/v11050408

Figure Lengend Snippet: Cytotoxicity and antiviral activity of Avirulins in female reproductive tract epithelium. ( a – c ) Immortalized epithelial cell lines derived from the vaginal wall, ectocervix, and endocervix (VK2, Ect1, and End1, respectively) were treated with Avirulin lead compounds Av-5, Av-14, and Av-26 at a range of concentrations from 0.78–100 µM for 24 h. Cytotoxicity was measured using a commercially available luminescence-based cytotoxicity assay, CytotoxGlo. Cytotoxicity of treatments was compared to DMSO vehicle control, with the detergent Triton-X 100 for complete cell death. ( n = 4, error = SEM). ( d ) Av-5 was incubated with clarified vaginal fluid at 37 °C for 2 h, then diluted with culture media and applied to TZM-bl. Cells were infected with BaL for 24 h, then inhibition was determined using the luciferase reporter assay. Cervico–vaginal fluid from healthy women is inherently antiviral, as shown in the vehicle control. n = 3, error = SEM * = p > 0.05. ( e ) Cytotoxicity of vaginal fluids and 5 µM Av-5 on TZM-bl measured using CytotoxGlo assay. Cervico–vaginal fluid (CVF) dilutions with Av-5 or equivalent DMSO were incubated at 37 °C for 2 h, then applied to TZM-bl. Cell death at 24 h was determined by subtraction of total cell luminescence from dead cell luminescence. n = 3, error = SEM.

Article Snippet: The vaginal, ectocervical, and endocervical epithelium derived cell lines VK2, Ect1, and End1, respectively, were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained with keratinocyte serum free media (KSFM) supplemented with bovine pituitary extract, epidermal growth factor, and calcium chloride in tissue culture treated plates, as per ATCC instructions.

Techniques: Activity Assay, Derivative Assay, Cytotoxicity Assay, Control, Incubation, Infection, Inhibition, Luciferase, Reporter Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Extracellular vesicles from vaginal Gardnerella vaginalis and Mobiluncus mulieris contain distinct proteomic cargo and induce inflammatory pathways

doi: 10.1038/s41522-024-00502-y

Figure Lengend Snippet: bEV preparations from ( a ) NYC culture medium (control), ( b ) L. crispatus , ( c ) G. vaginalis , and ( d ) M. mulieris were labeled with rhodamine B isothiocyanate and observed in the cytoplasm of ectocervical (Ect1), endocervical (End1), and vaginal epithelial (VK2) cells after 1, 4, and 24 h of incubation. All scale bars are 20 μm.

Article Snippet: Ectocervical (Ect/E6E7, ATCC CRL-2614), endocervical (End1/E6E7, ATCC CRL-2615), and vaginal (VK2/E6E7, ATCC CRL-2616) human epithelial cell lines (American Type Culture Collection, Manassas, VA) were cultured in keratinocyte-serum-free media (K-SFM) supplemented with 0.1 ng/mL epidermal growth factor and 50 μg/mL bovine pituitary extract (Gibco, Life Technologies), 100 U/mL penicillin, and 100 μg/mL of streptomycin at 37 °C in a 5% CO 2 humidified incubator.

Techniques: Control, Labeling, Incubation

Journal: NPJ Biofilms and Microbiomes

Article Title: Extracellular vesicles from vaginal Gardnerella vaginalis and Mobiluncus mulieris contain distinct proteomic cargo and induce inflammatory pathways

doi: 10.1038/s41522-024-00502-y

Figure Lengend Snippet: IL-8 expression measured by ELISA increases with increasing doses of bEVs from G. vaginalis and M. mulieris , but not L. crispatus and NYC culture medium (control), relative to nontreated ( a ) ectocervical, ( b ) endocervical, and ( c ) vaginal epithelial cells. d Exposure to bacterial EVs does not induce significant changes in lactate dehydrogenase release. 29-plex Luminex array identified a multi-cytokine response to bEVs from G. vaginalis and M. mulieris , but not L. crispatus , expressed as fold-changes relative to NYC culture medium control-treated ( e ) ectocervical, ( f ) endocervical, and ( g ) vaginal epithelial cells ( n = 3 per treatment). Error bars indicate standard deviation and asterisks indicate adjusted p -values < 0.05 (*), < 0.01 (**), < 0.001 (***), and <0 .0001 (****) via one-way ANOVA with Tukey’s correction for multiple comparisons.

Article Snippet: Ectocervical (Ect/E6E7, ATCC CRL-2614), endocervical (End1/E6E7, ATCC CRL-2615), and vaginal (VK2/E6E7, ATCC CRL-2616) human epithelial cell lines (American Type Culture Collection, Manassas, VA) were cultured in keratinocyte-serum-free media (K-SFM) supplemented with 0.1 ng/mL epidermal growth factor and 50 μg/mL bovine pituitary extract (Gibco, Life Technologies), 100 U/mL penicillin, and 100 μg/mL of streptomycin at 37 °C in a 5% CO 2 humidified incubator.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, Luminex, Standard Deviation