Sequence changes in the human adenovirus type 5 DNA polymerase associated with resistance to the broad spectrum antiviral cidofovir.

Although there is currently no FDA approved antiviral treatment for adenovirus (Ad) infections, the broad spectrum antiviral cidofovir (CDV) has demonstrated potent inhibitory activity against many Ad serotypes in vitro and in an in vivo ocular replication model. The clinical potential of CDV prompted the assessment for the emergence of CDV resistance in Ad5. Serial passage of Ad5 in increasing concentrations of CDV resulted in derivation of four different Ad5 variants with increased resistance to CDV.

The antiviral resistance and replication of cidofovir-resistant adenovirus variants in the New Zealand White rabbit ocular model.

Purpose: To determine the antiviral resistance of three cidofovir (CDV)-resistant variants of adenovirus type 5 (Ad5) and their ability to replicate in the New Zealand White rabbit ocular model.

Methods: Rabbits were inoculated topically in both eyes with the CDV-resistant variants R1, R2, and R3, and the Ad5 parental strain. On day 1, rabbits from each virus inoculation were divided into two topical treatment groups: 0.

The effects of topical nonsteroidal anti-inflammatory drugs on adenoviral replication.

Objective: To evaluate the antiviral activity of topical diclofenac sodium (Voltaren Ophthalmic) and ketorolac tromethamine (Acular) (2 nonsteroidal anti-inflammatory drugs [NSAIDs]) on adenoviral replication in vitro and in the adenovirus (Ad) 5 McEwen-New Zealand rabbit ocular model.

Methods: The 50% inhibitory concentration of ketorolac and diclofenac and their respective preservative components were determined for common ocular adenoviral serotypes (Ad8, Ad19, Ad1, and Ad5). In a series of experiments, Ad5 McEwen-inoculated New Zealand rabbit eyes were treated topically 4 times daily for 18 days with either ketorolac, diclofenac, prednisolone acetate (Pred Forte), or control vehicle (Comfort Tears).

Multiple adenoviral serotypes demonstrate host range extension in the New Zealand rabbit ocular model.

Purpose: Although several human adenoviral serotypes demonstrated the genetic capability of replicating in New Zealand rabbit corneas in organ culture, only a single adenovirus (Ad) serotype, Ad5, has been reported to replicate in vivo in New Zealand rabbit eyes. The purpose of this study was to determine whether additional adenoviral serotypes could extend their host range to the New Zealand rabbit ocular model.

Methods: Six rabbits per viral isolate were inoculated in each eye after corneal scarification with 1.

Topical corticosteroids reverse the antiviral effect of topical cidofovir in the Ad5-inoculated New Zealand rabbit ocular model.

Purpose: To determine how the addition of topical corticosteroids would affect the anti-adenoviral inhibitory effect of topical cidofovir (S-HPMPC) in the Ad5 New Zealand (Ad5/NZ) rabbit ocular model.

Methods: In a series of experiments (two-eye design), Ad5-inoculated/NZ rabbits (10(6) pfu/eye) were treated with 1 of 3 treatment regimens. Group 1 was administered 1% cidofovir (CDV) twice a day for 3 days plus comfort tears four times a day for 14 days.

Isolation of human adenovirus type 5 variants resistant to the antiviral cidofovir.

Purpose: Cidofovir (S-HPMPC) is a potent broad-spectrum antiviral drug with potential clinical application against infections caused by human cytomegalovirus, herpes simplex virus, and adenovirus (AD). This study sought to determine whether variants of AD5 could be isolated in vitro that demonstrated increased resistance to this new antiviral drug.

Methods: Homogenous stocks of wild-type AD5 (ATCC strain VR-5) were generated from isolated plaques grown in A549 cells.

The effects of corticosteroids of adenoviral replication.

Objective: To evaluate the effects of Pred Forte (prednisolone acetate; Allergan Pharmaceutical, Irvine, Calif) on the replication of different adenoviral serotypes in vitro and in the adenovirus type 5/New Zealand rabbit ocular model.

Methods: The 50% inhibitory doses of Pred Forte and its components were determined for common ocular serotypes. The effects of continuous topical treatment with Pred Forte for 18 days were evaluated (eg, conjunctivitis, subepithelial immune infiltrates, and serial ocular viral titers) in the adenovirus 5/New Zealand rabbit ocular model.

Topical HPMPC inhibits adenovirus type 5 in the New Zealand rabbit ocular replication model.

Purpose: To evaluate the antiviral inhibitory activity of HPMPC against ocular adenoviral serotypes in vitro and to determine the therapeutic efficacy and ocular toxicity of treatment with topical HPMPC on established adenovirus type 5 (AD5) McEwen infection in the New Zealand (NZ) rabbit ocular replication model.

Methods: The 50% inhibitory dose (ID50) of HPMPC was determined for various clinical isolates of AD5 and AD8 by plaque assay in A549 cells. In vivo inhibitory effects were measured by serial ocular titers and duration of viral shedding in the AD5-NZ rabbit ocular model.

HPMPC, a broad-spectrum topical antiviral agent, inhibits herpes simplex virus type 1 replication and promotes healing of dendritic keratitis in the New Zealand rabbit ocular model.

Previously, we demonstrated that HPMPC, a new, broad-spectrum antiviral agent, inhibited adenovirus type 5 in the New Zealand (NZ) rabbit ocular model (Cornea 1992; 11:529-33). Historically, no antiviral agent has been demonstrated to be effective against both herpes simplex virus type 1 (HSV-1) and adenovirus eye infections in an experimental animal model. In this study, we compared topical 0.

Prolonged recovery of desiccated adenoviral serotypes 5, 8, and 19 from plastic and metal surfaces in vitro.

Background: Prevention of the spread of epidemic keratoconjunctivitis (EKC) at eye care facilities (doctors' offices, clinics, hospitals) has been a major public health goal for ophthalmology for more than 50 years. The authors explored a potentially contributing attribute of the adenovirus serotypes that cause EKC. Specifically, they investigated the capacity of different clinical and laboratory ocular serotypes (AD8, 19, and 5) to survive for extended periods of time in a desiccated state.

A comparison of enzyme immunoassay and polymerase chain reaction with the clinical examination for diagnosing ocular herpetic disease.

Purpose: The results of two laboratory diagnostic herpes simplex virus (HSV) tests, an enzyme immunoassay (improved Herpchek [iHC]) and the polymerase chain reaction (PCR), were compared with the clinical examination in the diagnosis of HSV. We determined when diagnostic laboratory tests provided the initial diagnosis of HSV ocular disease and when they were only confirmatory.

Methods: The sensitivity and specificity of iHC and PCR were determined using 22 HSV culture-positive clinical samples, 10 adenovirus culture-positive clinical samples, 5 samples from normal conjunctivas, 4 bacterial samples, and 1 sample containing Varicella zoster virus.

Pretreatment with topical 0.1% (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine inhibits adenovirus type 5 replication in the New Zealand rabbit ocular model.

Currently there is no clinically effective antiviral agent for the prevention or treatment of ocular adenoviral infections. Using a paired-eye, masked design, we tested the antiviral efficacy of topical 0.1% (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine in the New Zealand rabbit ocular model after topical and intrastromal inoculation with 100 microliters (4 x 10(5) plaque-forming units per eye) of adenovirus type 5 McEwen, a clinical isolate.

An ocular model of adenovirus type 5 infection in the NZ rabbit.

Ocular adenoviral infections occur worldwide, and currently, there is no ocular animal model for evaluating new antivirals or studying pathogenesis. With a paired-eye design, an ocular model was developed in 32 New Zealand rabbits following topical and intrastromal inoculation with a clinical isolate of adenovirus type 5 (Ad5 McEwen). Clinical signs of infection--conjunctivitis, corneal edema, subepithelial infiltrates, and iritis--and seroconversion were evaluated.

Inhibitory effect of (S)-HPMPC, (S)-HPMPA, and 2'-nor-cyclic GMP on clinical ocular adenoviral isolates is serotype-dependent in vitro.

Currently, there is no effective treatment for ocular adenoviral infections that occur in epidemics worldwide, produce significant patient morbidity, and cause substantial economic losses. We tested several new antivirals in vitro, and found that (S)-HPMPC, (S)-HPMPA, and 2'-nor-cyclic GMP demonstrated significant serotype-dependent inhibitory activity by plaque reduction assay (ID50 = 0.017-17.

Replication of ocular isolates of human adenovirus is serotype-dependent in rabbit corneal organ culture.

The goal of the present in vitro study was to determine the ability of unadapted human adenoviral ocular isolates to replicate in the rabbit cornea. Rabbit corneas grown in organ culture (24 well plate) were inoculated topically with 50 microliters (5 x 10(5) pfu) of different ocular adenoviral serotypes (ATCC and clinical isolates). Control wells (no cornea present) were inoculated in a similar fashion.

Prolonged recoverability of desiccated adenovirus type 19 from various surfaces.

Epidemic keratoconjunctivitis is a highly contagious disease whose transmission has been linked to the ophthalmologist's office. The authors studied the ability of adenovirus 19 (ADV 19) to survive on surfaces commonly found in the office setting. An initial in vitro laboratory experiment demonstrated that ADV 19 in a desiccated state could be recovered up to 8 days from paper, and up to 10 days from cloth, metal, and plastic.

Vanadate promotes reactivation and iontophoresis-induced ocular shedding of latent HSV-1 W in different host animals.

Vanadate is a potent inhibitor of calcium stimulated ATPase, Na, K-ATPase, and may have adrenergic activity. Using the iontophoresis method, we compared vanadate to a BSS control and the standard iontophoresis model (6-hydroxydopamine/epinephrine) by measuring induced ocular shedding of latent HSV-1 in different host animals. Latent trigeminal ganglionic infections were established in Balb/c mice and New Zealand rabbits following corneal inoculation with HSV-1 [W] strain, and later confirmed by cocultivation.

Host species and strain differences affect the ability of an HSV-1 ICP0 deletion mutant to establish latency and spontaneously reactivate in vivo.

HSV-1 latency and reactivation were studied in vivo by spontaneous and iontophoresis-induced ocular viral shedding in New Zealand rabbits, Balb/c and A/J mice latently infected with wild-type KOS, and dl x 3.1, a progeny ICP0 deletion mutant. The presence of trigeminal ganglionic latency was demonstrated by the in vitro methods of cocultivation and in situ hybridization.

The effect of alpha blockade on iontophoresis-induced ocular shedding of latent HSV-1 W in different host animals.

Previous studies demonstrated that non-specific beta blockade promoted ocular shedding of latent HSV-1 in the mouse and rabbit iontophoresis models. The present study examined the effect of topical alpha blockers, thymoxamine and corynanthine, on reactivation and induced ocular shedding of latent HSV-1 W in different host animals. Latent trigeminal ganglionic infection was established in Balb/c mice and New Zealand rabbits following corneal inoculation with HSV-1 W strain, and later confirmed by co-cultivation.

A fast, simple reactivation method for the study of HSV-1 latency in the rabbit ocular model.

We developed a simple, fast, economical, and versatile reactivation method for the study of herpes simplex virus type 1 (HSV-1) latency in the rabbit ocular model. Intrastromal injection of sterile, deionized water induced reactivation and ocular shedding of latent HSV-1 in 21 of 27 eyes (78%) in 93% of New Zealand rabbits. Other control groups (eg, anterior chamber injection of sterile, deionized water; topical administration of sterile, deionized water; intrastromal injection of air; and intrastromal needle track), were less efficient in reactivating latent HSV-1.

A herpes simplex type 1 ICPO deletion mutant demonstrates diminished pathogenicity during acute ocular infection in different host animals.

We compared a previously characterized herpes simplex type 1 alpha 0 deletion mutant, dlx3.1, which produced no functional ICP0, with its wild-type parental strain, KOS, during acute ocular infection of different host animals. Acute pathogenicity of the viral strains in NZ rabbits, Balb/c and A/J mice was evaluated by keratitis scores, ocular and trigeminal ganglionic viral titers, and host survival.

RNA complementary to herpes simplex virus type 1 ICP0 gene demonstrated in neurons of human trigeminal ganglia.

Recent studies with mice have demonstrated abundant RNA transcripts which are complementary (antisense) to the herpes alpha gene ICP0 in latently infected ganglia. We investigated the situation in unselected human trigeminal ganglia. Strand-specific 2.

The antiviral effects of rose bengal and fluorescein.

We evaluated the antiviral effects of rose bengal and fluorescein sodium. The direct antiviral activity was determined by an in vitro direct neutralization assay. The 50% inhibitory dose was 16 micrograms/mL for rose bengal and 460 micrograms/mL for fluorescein.