title: Effects of Female Sex Hormones on Susceptibility to HSV-2 in Vaginal Cells Grown in Air-Liquid Interface.
abstract: The lower female reproductive tract (FRT) is comprised of the cervix and vagina, surfaces that are continuously exposed to a variety of commensal and pathogenic organisms. Sexually transmitted viruses, such as herpes simplex virus type 2 (HSV-2), have to traverse the mucosal epithelial lining of the FRT to establish infection. The majority of current culture systems that model the host-pathogen interactions in the mucosal epithelium have limitations in simulating physiological conditions as they employ a liquid-liquid interface (LLI), in which both apical and basolateral surfaces are submerged in growth medium. We designed the current study to simulate in vivo conditions by growing an immortalized vaginal epithelial cell line (Vk2/E6E7) in culture with an air-liquid interface (ALI) and examined the effects of female sex hormones on their growth, differentiation, and susceptibility to HSV-2 under these conditions, in comparison to LLI cultures. ALI conditions induced Vk2/E6E7 cells to grow into multi-layered cultures compared to the monolayers present in LLI conditions. Vk2 cells in ALI showed higher production of cytokeratin in the presence of estradiol (E2), compared to cells grown in progesterone (P4). Cells grown under ALI conditions were exposed to HSV-2-green fluorescent protein (GFP) and the highest infection and replication was observed in the presence of P4. Altogether, this study suggests that ALI cultures more closely simulate the in vivo conditions of the FRT compared to the conventional LLI cultures. Furthermore, under these conditions P4 was found to confer higher susceptibility to HSV-2 infection in vaginal cells. The vaginal ALI culture system offers a better alternative to study host-pathogen interactions.
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. leey27@mcmaster.ca.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. leey27@mcmaster.ca.
authors: Yung
authors: Y
authors: Lee
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. dizzels@mcmaster.ca.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. dizzels@mcmaster.ca.
authors: Sara E
authors: SE
authors: Dizzell
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. vwleung@hsph.harvard.edu.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. vwleung@hsph.harvard.edu.
authors: Vivian
authors: V
authors: Leung
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. nazliaisha@gmail.com.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. nazliaisha@gmail.com.
authors: Aisha
authors: A
authors: Nazli
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. zahoorma@mcmaster.ca.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. zahoorma@mcmaster.ca.
authors: Muhammad A
authors: MA
authors: Zahoor
authors: Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Boston, MA 02115, USA. rfichorova@rics.bwh.harvard.edu.|Harvard Medical School, Harvard University, Boston, MA 02115, USA. rfichorova@rics.bwh.harvard.edu.
authors: Raina N
authors: RN
authors: Fichorova
authors: McMaster Immunology Research Centre, Michael G. DeGroote Centre for Learning and Discovery, McMaster University, Hamilton, ON L8S 4K1, Canada. kaushic@mcmaster.ca.|Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. kaushic@mcmaster.ca.
authors: Charu
authors: C
authors: Kaushic
mesh_terms: D018929:Cell Culture Techniques; D004847:Epithelial Cells / Q000187:drug effects* / Q000821:virology*; D005260:Female; D012739:Gonadal Steroid Hormones / Q000378:metabolism*; D006558:Herpes Genitalis / Q000821:virology; D018258:Herpesvirus 2, Human / Q000254:growth & development*; D054884:Host-Pathogen Interactions; D006801:Humans; D008297:Male; D008954:Models, Biological; D014621:Vagina / Q000821:virology; D014776:Virus Cultivation
chemical_list: D012739:Gonadal Steroid Hormones
keywords: HSV-2; Vk2/E6E7 cells; air-liquid interface; estradiol; female genital tract; female sex hormones; herpes simplex virus type 2; medroxyprogesterone acetate; progesterone; vaginal cells