Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts

Peters, Donna M.; Dowd, Nathan; Brandt, Curtis R.; Compton, Teresa

doi:10.1007/bf02723060

Cited by 7 publications

(4 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The isolation procedures for the primary keratocytes were described previously (42). Cells infected with virus were grown in DMEM supplemented with 2% serum.…”

Section: Cells and Virusesmentioning

confidence: 99%

Inhibition of Herpes Simplex Virus Type 1 Infection by Cationic β-Peptides

Akkarawongsa

Potocky²,

English³

et al. 2008

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Previously, it was shown that cationic ␣-peptides derived from the human immunodeficiency virus TAT protein transduction domain blocked herpes simplex virus type 1 (HSV-1) entry. We now show that cationic oligomers of ␤-amino acids ("␤-peptides") inhibit HSV-1 infection. Among three cationic ␤-peptides tested, the most effective inhibition was observed for the one with a strong propensity to adopt a helical conformation in which cationic and hydrophobic residues are segregated from one another ("globally amphiphilic helix"). The antiviral effect was not cell type specific. Inhibition of virus infection by the ␤-peptides occurred at the postattachment penetration step, with a 50% effective concentration of 3 M for the most-effective ␤-peptide. The ␤-peptides did not inactivate virions in solution, nor did they induce resistance to infection when cells were pretreated with the ␤-peptides. The ␤-peptides showed little if any toxicity toward Vero cells. These results raise the possibility that cationic ␤-peptides may be useful antiviral agents for HSV-1 and demonstrate the potential of ␤-peptides as novel antiviral drugs.Herpes simplex virus type 1 (HSV-1) is a significant human pathogen causing mucocutaneous lesions primarily in the oral mucosa (cold sores), as well as other sites. More-severe diseases caused by HSV-1 infection include encephalitis, meningitis, and blinding keratitis (65), and HSV-1 is the leading cause of blindness due to infection in developed countries (5). Following an initial infection, HSV-1 establishes latent infection of neurons in sensory ganglia of the host (29), from where it periodically reactivates and causes recurrent lesions at the site of primary infection. To date, none of the currently approved antivirals can eliminate an established latent infection. Because of the difficulties dealing with latency, preventing HSV-1 from entering the cell is an attractive antiviral strategy.HSV-1 entry is a complex process, involving multiple components on both the cell plasma membrane and the viral envelope. The initial interaction involves the binding of viral glycoprotein C (gC) or gB to cell surface heparan sulfate proteoglycan (58). Four viral glycoproteins, gB, gD, and the gH-gL heterodimer, are essential for the subsequent membrane fusion and entry steps (56,57). Following attachment, gD interacts with any of three cellular receptors: herpes virus entry mediator, nectin-1, and 3-O-sulfated heparan sulfate, leading to a conformational change in gD (12,13,21,22,34,39,54,62). This conformational change in gD is believed to trigger the formation of the fusion complex, which is thought to involve the sequential binding of gB to gD, followed by the binding of gH-gL to the gB-gD complex (10,20,23,24,36,41,50,62).

show abstract

“…The isolation procedures for the primary keratocytes were described previously (42). Cells infected with virus were grown in DMEM supplemented with 2% serum.…”

Section: Cells and Virusesmentioning

confidence: 99%

Inhibition of Herpes Simplex Virus Type 1 Infection by Cationic β-Peptides

Akkarawongsa

Potocky²,

English³

et al. 2008

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…Previous studies have shown that HPV wt-E6/E7 induces morphological mesenchymal type changes. 28 Therefore, the morphological characteristics of clones were also examined. Figure 2D shows that control and E6 Δ /E7 Δ grow in tight colonies, while wt-E6/E7 expression induces a mesenchymal-like change in morphology and cells grow in a non-adherent manner.…”

Section: Resultsmentioning

confidence: 99%

A non-oncogenic HPV 16 E6/E7 vaccine enhances treatment of HPV expressing tumors

et al. 2012

View full text Add to dashboard Cite

Human papillomaviruses (HPVs) are the causative factor for greater than 90% of cervical cancers and 25% of head and neck cancers. The incidence of HPV positive (+) head and neck squamous cell carcinomas (HNSCCs) has greatly increased in the last 30 years. E6 and E7 are the two key viral oncoproteins that induce and propagate cellular transformation. An immune response generated during cisplatin/radiation therapy improves tumor clearance of HPV(+) cancers. Augmenting this induced response during therapy with an adenoviral HPV16 E6/E7 vaccine improves long term survival in preclinical models. Here we describe the generation of an HPV16 E6/E7 construct, which contains mutations that render E6/E7 non-oncogenic, while preserving antigenicity. These mutations do not allow E6/E7 to degrade p53, pRb, PTPN13, or activate telomerase. Non-oncogenic E6/E7 (E6Δ/E7Δ) expressed as a stable integrant, or in the [E1-, E2b-] adenovirus, lacks the ability to transform human cells while retaining the ability to induce an HPV specific immune response. Moreover, E6Δ/E7Δ plus chemotherapy/radiation statistically enhances clearance of established HPV(+) cancer in vivo.

show abstract

“…The derivation and characterization of the human keratocyte cells was described previously. 27 The HTM-1 cells were SV40 T antigen transformed human trabecular meshwork cells and have been described previously. 28 Following incubation overnight at 37°C, 20 L of medium containing the desired concentration (two-fold dilution series) of peptides were added to the cells.…”

Section: Dye-reduction Assaymentioning

confidence: 99%

Corneal Toxicity of Cell-Penetrating Peptides That InhibitHerpes simplexVirus Entry

Akkarawongsa¹,

Cullinan

Zinkel

et al. 2006

Journal of Ocular Pharmacology and Therapeutics

View full text Add to dashboard Cite

Cell-penetrating peptides (CPPs) inhibit Herpes simplex virus entry at low micromolar concentrations and may be useful either as prophylactic or therapeutic agents for herpetic keratitis. The aim of this study was to assess the in vitro and in vivo toxicity of three CPPs-EB, TAT-C, and HOM (penetratin)-for the cornea. Incubation of primary (HK320) or immortalized (THK320) human keratocytes with the EB peptide (up to 100 microM), bHOMd (up to 200 microM), or TAT-C (up to 400 microM) resulted in no evidence of toxicity using a formazan dye-reduction assay. Similar results were obtained with a human trabecular meshwork cell line (TM-1), primary human foreskin fibroblasts (DP-9), Vero, and HeLa cells with EB and TATC. The bHOMd peptide showed some toxicity in Vero and HeLa cells, with CC50 values of 70 and 93 microM, respectively. The EB peptide did not inhibit macromolecular synthesis in Vero cells at concentrations below 150 microM, although cell proliferation was blocked at concentrations of EB above 50 microM. In vivo toxicity was assessed by applying peptides in Dulbecco's modified Eagle's medium to the cornea 4 times daily for 7 d. At concentrations 1000 times the IC50 values, the EB and bHOM peptides showed no toxicity, whereas TAT-C caused some mild eyelid swelling. Some slight epithelial cell sloughing was seen with the bKLA peptide in vivo. These results suggest that these CPPs-and EB in particular-have a favorable toxicity profile, and that further development is warranted.

show abstract

Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts

Cited by 7 publications

References 12 publications

Inhibition of Herpes Simplex Virus Type 1 Infection by Cationic β-Peptides

Inhibition of Herpes Simplex Virus Type 1 Infection by Cationic β-Peptides

A non-oncogenic HPV 16 E6/E7 vaccine enhances treatment of HPV expressing tumors

Corneal Toxicity of Cell-Penetrating Peptides That InhibitHerpes simplexVirus Entry

Contact Info

Product

Resources

About