Rapid-Release Griffithsin Fibers for Dual Prevention of HSV-2 and HIV-1 Infections

Abstract

The biologic griffithsin (GRFT) has recently emerged as a candidate to safely prevent sexually transmitted infections (STIs), including human immunodeficiency virus type 1 (HIV-1) and herpes simplex virus 2 (HSV-2). However, to date, there are few delivery platforms that are available to effectively deliver biologics to the female reproductive tract (FRT). The goal of this work was to evaluate rapid-release polyethylene oxide (PEO), polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP) fibers that incorporate GRFT in in vitro (HIV-1 and HSV-2) and in vivo (HSV-2) infection models. GRFT loading was determined via enzyme-linked immunosorbent assay (ELISA), and the bioactivity of GRFT fibers was assessed using in vitro HIV-1 pseudovirus and HSV-2 plaque assays. Afterwards, the efficacy of GRFT fibers was assessed in a murine model of lethal HSV-2 infection. Finally, murine reproductive tracts and vaginal lavage samples were evaluated for histology and cytokine expression, 24 and 72 h after fiber administration, to determine safety. All rapid-release formulations achieved high levels of GRFT incorporation and were completely efficacious against in vitro HIV-1 and HSV-2 infections. Importantly, all rapid-release GRFT fibers provided potent protection in a murine model of HSV-2 infection. Moreover, histology and cytokine levels, evaluated from collected murine reproductive tissues and vaginal lavage samples treated with blank fibers, showed no increased cytokine production or histological aberrations, demonstrating the preliminary safety of rapid-release GRFT fibers in vaginal tissue.

Antiviral lectin Q-Griffithsin suppresses fungal infection in murine models of vaginal candidiasis

“…Additionally, assessing the neutrophilic and monocytes phagocytic killing activity in the presence of Q-GRFT will provide a better understanding of Q-GRFT’s role in VVC. Q-GRFT has shown promise in preventing viral sexually transmitted infections, including HSV-2 and HIV-1 ( Nixon et al., 2013 ; Derby et al., 2018 ; Tyo et al., 2020 ; Tyo et al., 2020 ). Our data demonstrate additional potential for utility of Q-GRFT vaginal dosage forms in both preventing and treating candidiasis, and further support incorporation of Q-GRFT in multipurpose STI prevention modalities.…”

“…Native GRFT is prone to oxidation ( Kramzer et al., 2021 ), and our group has developed an engineered form, Griffithsin-M78Q (Q-GRFT), with improved stability, and similar antiviral activity to GRFT. We and others have demonstrated that GRFT and Q-GRFT are both safe and efficacious in preclinical models of HIV-1 and HSV-2 infection ( O'Keefe et al., 2009 ; Kouokam et al., 2011 ; Férir et al., 2011 ; Nixon et al., 2013 ; Kouokam et al., 2016 ; Derby et al., 2018 ; Girard et al., 2018 ; Günaydın et al., 2019 ; Yang et al., 2019 ; Tyo et al., 2020 ; Tyo et al., 2020 ; Kramzer et al., 2021 ), as well as in early-stage human clinical studies ( Teleshova et al., 2022 ). Recently, we reported a novel antifungal activity of Q-GRFT, with potent growth inhibition of Candida species of human importance including C. albicans , C. parapsilosis , C. krusei , C. glabrata and against strains of the pan-resistant C. auris ( Nabeta et al., 2021 ).…”

Recent advances in electrospun nanofiber vaginal formulations for women's sexual and reproductive health

“…Such combination strategies are referred to as 'multipurpose prevention technology' products. The first article, reporting the fabrication of griffithsin-loaded nanofibers for immediate release, describes the use of the hydrophilic, mucoadhesive and biocompatible polymers PEO, PVA and PVP, and evaluation in vitro for HIV-1 and HSV-2 and also in vivo for HSV-2 (Tyo et al, 2020a). The fibers provided complete and dual-purpose protection against in vitro infections with both viruses, and griffithsin was capable of reducing HSV-2 transmission in mice.…”

Antiviral Electrospun Polymer Composites: Recent Advances and Opportunities for Tackling COVID-19

“…Recently, researchers developed a new type of electrospun fibers that incorporated the GRFT to provide dualpurpose protection against HIV-1 and HSV-2 infections. These fibers were composed of polyethylene oxide (PEO), polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP) (Tyo et al, 2020). Furthermore, Baskakova et al developed electrospun fibers containing acyclovir, ciprofloxacin, and cyanocobalamin to treat cytomegalovirus infections for patients with AIDS (Baskakova et al, 2016).…”