title: Griffithsin and Carrageenan Combination To Target Herpes Simplex Virus 2 and Human Papillomavirus.
abstract: Extensive preclinical evaluation of griffithsin (GRFT) has identified this lectin to be a promising broad-spectrum microbicide. We set out to explore the antiviral properties of a GRFT and carrageenan (CG) combination product against herpes simplex virus 2 (HSV-2) and human papillomavirus (HPV) as well as determine the mechanism of action (MOA) of GRFT against both viruses. We performed the experiments in different cell lines, using time-of-addition and temperature dependence experiments to differentiate inhibition of viral attachment from entry and viral receptor internalization. Surface plasmon resonance (SPR) was used to assess GRFT binding to viral glycoproteins, and immunoprecipitation and immunohistochemistry were used to identify the specific glycoprotein involved. We determined the antiviral activity of GRFT against HSV-2 to be a 50% effective concentration (EC50) of 230 nM and provide the first evidence that GRFT has moderate anti-HPV activity (EC50 = 0.429 to 1.39 μM). GRFT blocks the entry of HSV-2 and HPV into target cells but not the adsorption of HSV-2 and HPV onto target cells. The results of the SPR, immunoprecipitation, and immunohistochemistry analyses of HSV-2 combined suggest that GRFT may block viral entry by binding to HSV-2 glycoprotein D. Cell-based assays suggest anti-HPV activity through α6 integrin internalization. The GRFT-CG combination product but not GRFT or CG alone reduced HSV-2 vaginal infection in mice when given an hour before challenge (P = 0.0352). While GRFT significantly protected mice against vaginal HPV infection when dosed during and after HPV16 pseudovirus challenge (P < 0.026), greater CG-mediated protection was afforded by the GRFT-CG combination for up to 8 h (P < 0.0022). These findings support the development of the GRFT-CG combination as a broad-spectrum microbicide.
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Keith
authors.initials: K
authors.lastname: Levendosky
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Mizenina
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Martinelli
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Ninochka
authors.initials: N
authors.lastname: Jean-Pierre
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Larisa
authors.initials: L
authors.lastname: Kizima
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Aixa
authors.initials: A
authors.lastname: Rodriguez
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Kleinbeck
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Thierry
authors.initials: T
authors.lastname: Bonnaire
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Robbiani
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Zydowsky
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, NCI at Frederick, Frederick, Maryland, USA.
authors.forename: Barry R
authors.initials: BR
authors.lastname: O'Keefe
authors.affiliation: Center for Biomedical Research, Population Council, New York, New York, USA jromero@popcouncil.org.
authors.forename: José A
authors.initials: JA
authors.lastname: Fernández-Romero
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002351:Carrageenan / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D004338:Drug Combinations; D004357:Drug Synergism; D005260:Female; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology; D006367:HeLa Cells; D006558:Herpes Genitalis / Q000188:drug therapy* / Q000821:virology; D018258:Herpesvirus 2, Human / Q000187:drug effects* / Q000502:physiology; D052162:Human papillomavirus 16 / Q000187:drug effects / Q000502:physiology; D052161:Human papillomavirus 18 / Q000187:drug effects / Q000502:physiology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D030361:Papillomavirus Infections / Q000188:drug therapy* / Q000821:virology; D037121:Plant Lectins / Q000494:pharmacology*; D014709:Vero Cells; D053585:Virus Attachment / Q000187:drug effects; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D004338:Drug Combinations; D037121:Plant Lectins; D002351:Carrageenan; C499654:griffithsin protein, Griffithsia