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Biomedical subjects

D E Willey

Publications and source records attributed to D E Willey.

16 recordsLinked to original sources
PubMed1992

Antiviral effects of herpes simplex virus specific anti-sense nucleic acids.

We have targeted mRNA sequences encompassing the translation initiation codon of the essential herpes simplex virus type 1 (HSV-1) IE3 gene with three kinds of anti-sense molecule. Addition of a 15mer oligodeoxyribonucleoside methylphosphonate to tissue culture cells resulted in suppression of viral replication. HSV-1 replication was also inhibited in cultured cells containing anti-sense vectors expressing transcripts complementary to the IE3 mRNA. We have also constructed a ribozyme which upon base pairing with the target IE3 mRNA induces cleavage at the predicted GUC site. A major obstacle to anti-sense studies in animals is drug delivery of preformed antisense molecules to ganglionic neurons, the site of HSV latency and reactivation. We speculate as to how this may be accomplished through carrier compounds which are taken up by nerve terminals and transported by retrograde axoplasmic flow. By the same route, HSV itself may be used as an anti-sense vector.

Animals↗
PubMed1992

Persistence of herpes simplex virus DNA in rabbit corneal cells.

Corneal cell cultures were established from the corneas of rabbits killed during a period of latency 118 d after ocular infection with the RE strain of herpes simplex virus (HSV). DNA was isolated from frozen cell pellets of 42 cell cultures that did not develop viral cytopathic effects during 44 d in culture. Using the polymerase chain reaction (PCR) to amplify HSV thymidine kinase (TK) gene sequences, HSV-specific DNA was detected in 15 of 42 culture-negative cell cultures. Subsequent reamplification, using nested primers that were complementary to HSV TK sequences internal to the orginal primers, resulted in eight additional culture-negative samples showing positive hybridization for HSV TK DNA. Twenty three of the 42 virus culture-negative corneal cell cultures tested by PCR were found to contain HSV genetic material. Detailed examination of the clinical histories of the eyes from which the corneal cultures were obtained showed no correlation between increased frequency or severity of epithelial disease, stromal disease, or virus shedding and more frequent isolation of virus or detection of HSV-specific DNA. These studies document that HSV DNA residues in the corneas of HSV-infected rabbits up to 118 d post-infection. About 10% of the eyes contained virus that could be reactivated in culture, whereas an additional 55% of the eyes contained DNA sequences homologous to a portion of the HSV TK gene.

Animals↗
PubMed1991

Detection of herpes simplex virus DNA sequences in corneal transplant recipients by polymerase chain reaction assays.

Polymerase chain reaction (PCR) assays were used to amplify herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) sequences in DNA extracted from formalin-fixed, paraffin embedded corneas of patients undergoing corneal transplantation. PCR reamplification with an internal (nested) set of primers was required for detection in 10 of the 12 positive corneas indicating very low abundance of viral sequences. Three of the positive corneal samples were from failed corneal grafts. Overall, TK sequences were detected in 8 of 11 corneas from subjects with a past history of herpes keratitis and in 4 of 11 corneas from subjects with no past history of herpetic eye disease.

Adult↗
PubMed1991

Ocular acyclovir delivery by collagen discs: a mouse model to screen anti-viral agents.

Discs 1.6 mm in diameter trephined from corneal collagen shields were used to deliver acyclovir (ACV) to the cornea of mice inoculated with herpes simplex virus type 1 (HSV-1). In the first minute after application to the cornea, there was a 23% decrease of ACV in the discs. After the first minute, ACV clearance from the discs appeared to be exponential with a half-life of 21 minutes. Treatment given 3 times a day reduced HSV-1 titer in tear film, corneal tissue, and trigeminal ganglia. This animal model should be useful to conserve novel potential anti-viral drugs undergoing initial screening.

Acyclovir↗
PubMed1988

A recombinant vaccinia virus expressing herpes simplex virus type 1 glycoprotein B induces cytotoxic T lymphocytes in mice.

Spleen cells from BALB/c (H-2d) mice vaccinated with vgB11, a recombinant vaccinia virus which expresses glycoprotein B (gB) of herpes simplex virus type 1 (HSV-1), lysed EMT6 (H-2d) target cells infected with vgB11 or with HSV-1 but did not lyse uninfected EMT6 cells or infected L-929 (H-2k) target cells. Unlabelled target cell competition of lysis showed that only syngeneic cells infected with vgB11 or HSV-1 inhibited lysis of radiolabelled HSV-1-infected targets. These results demonstrate that vgB11 induces H-2-restricted anti-HSV-1 cytotoxic T lymphocytes and that gB is the target antigen.

Animals↗
PubMed1987

Expression of herpes simplex virus 1 glycoprotein B by a recombinant vaccinia virus and protection of mice against lethal herpes simplex virus 1 infection.

The herpes simplex virus 1 (HSV-1) strain F gene encoding glycoprotein gB was isolated and modified at the 5' end by in vitro oligonucleotide-directed mutagenesis. The modified gB gene was inserted into the vaccinia virus genome and expressed under the control of a vaccinia virus promoter. The mature gB glycoprotein produced by the vaccinia virus recombinant was glycosylated, was expressed at the cell surface, and was indistinguishable from authentic HSV-1 gB in terms of electrophoretic mobility. Mice immunized intradermally with the recombinant vaccinia virus produced gB-specific neutralizing antibodies and were resistant to a lethal HSV-1 challenge.

Animals↗
PubMed1987

Intentional reactivation of latent ocular herpes infection during BVDU therapy.

Sixteen adult New Zealand white rabbits with previously confirmed herpes simplex virus type 1 (HSV-1) infections were stimulated by iontophoresis of 6-hydroxydopamine into the cornea and were followed-up by topical epinephrine treatment to confirm latency. A total of 224 ocular cultures were obtained, of which 73 were positive for HSV during the seven day cycle. Twenty-seven of the 32 eyes (84%) had at least one positive culture. Animals were randomly divided into two treatment groups. Upon repeat stimulation (Cycle 2), concurrent with oral and topical bromovinyl-deoxyuridine (BVDU) therapy, only 3/104 ocular cultures were HSV positive, while 24/112 ocular cultures from placebo-treated animals were positive. Anti-HSV serum titers were comparable before and after BVDU therapy and the HSV isolates from BVDU treated animals did not develop drug resistance (i.e., ED-50 values were approximately 0.1-0.2 ug/ml both before and after therapy). It was concluded that BVDU had a demonstrable therapeutic effect on the recovery of HSV-1 from ocular cultures during intentional reactivation, but the latent ganglionic infection was not eliminated.

Animals↗
PubMed1985

Sequential analysis of antibody responses in serum, aqueous humor and tear film during latent and induced recurrent HSV infections.

To study HSV specific antibody responses during latent HSV infection and induced HSV ocular shedding, rabbits were infected binocularly with McKrae strain HSV-1. The titer and class of anti-HSV antibody response in serum, aqueous humor and eye washes were determined sequentially during latent HSV infection and following intentional reactivation. In all instances the only antibody class detected was IgG. The highest anti-HSV titers were in the serum and aqueous humor, whereas a significantly lower level of anti-HSV IgG was found in eye washes. Anti-HSV IgG antibody titers were consistently 100-fold higher in serum than in the aqueous humor, which suggests an absence of local ocular antibody synthesis. Neither anti-HSV IgM nor secretory IgA antibodies were detected in any samples.

Animals↗
PubMed1984

Efficacy of BW759 (9-[[2-hydroxy-1(hydroxymethyl)ethoxy]methyl]guanine) against herpes simplex virus type 1 keratitis in rabbits.

A promising new nucleoside analog, 9-[[2-hydroxy-1-(hydroxymethyl)ethoxy] methyl]-guanine (BW759), which is structurally similar to acyclovir, was tested against acute herpetic keratitis in the rabbit ocular model. Topical 1-0.1% BW759 given 3-5x per day gave beneficial results in that corneal epithelial involvement, conjunctivitis, iritis, and corneal clouding were reduced even when chemotherapy was initiated at 3 days postinoculation. Under the same conditions, topical BW759 therapy gave slightly better results than acyclovir, and both were better than idoxuridine therapy. Mortality rate and colonization of the trigeminal ganglia by HSV-1 were unaffected by BW759 therapy. Duration of virus, shed into the tear film was reduced by BW759.

Acyclovir↗
PubMed1984

Reactivation of murine latent HSV infection by epinephrine iontophoresis.

Iontophoresis of epinephrine into the cornea of previously infected mice was used in an attempt to induce reactivation of latent herpes simplex virus (HSV) infection of the trigeminal ganglia. BALB/c mice infected with HSV-1 strain McKrae following corneal scarification developed a latent infection of the trigeminal ganglia within 15 days. At 28 days postinfection, mice were subjected to a 3-day cycle of iontophoresis of epinephrine (0.01%) into the cornea. Ocular shedding of HSV occurred in 16/23 (70%) of stimulated mice; these animals did not shed HSV in the 3-day period prior to iontophoresis. Spontaneous shedding of HSV, however, was noted in 3/97 (3%) mice not subjected to epinephrine iontophoresis. "Infectious" virus was isolated only from the trigeminal ganglia of stimulated mice, whereas "latent" virus was isolated from the trigeminal ganglia of both stimulated and nonstimulated mice. All virus isolates were verified to be HSV by neutralization with a known HSV-1 antiserum. This ocular system thus allows for the study of the full spectrum of latent HSV infections, including latency, ganglionic reactivation, and peripheral virus shedding.

Animals↗
PubMed1984

Antibody response following implantation of xenogenic lenticules.

Rabbits underwent implantation of either lyophilized or fresh porcine lenticules into the central or peripheral cornea. All animals were followed for up to four months by slit-lamp examination and macrophotography to determine implant rejection. Serum antibody levels to soluble porcine cornea extract were determined by an enzyme-linked immunosorbant assay. Only those animals receiving lenticules into the peripheral cornea experienced a rejection and developed an antibody response to the porcine cornea extract. The production of antibody preceded the appearance of vascularization of the implanted lenticules. Thus, the site of lenticule implantation, not the type of tissue preparation, determined the outcome of the graft.

Animals↗
PubMed1983

A specular microscopic viewing system for donor corneas.

A system for routine specular microscopic assessment of donor corneal endothelium including a chamber that allows both storage and viewing of the donor tissue was evaluated. A specular microscope modified to enable noncontact scanning of donor cornea from the endothelial side was used. Forty-five human corneas were examined upon arrival at the eyebank. Storage in the new chamber was compared with that in standard vials for 13 paired corneas. This study demonstrates the feasibility and simplicity of routine specular microscopy on donor corneal tissue, and discusses its advantages.

Adolescent↗
PubMed1983

Effect of intensive acyclovir therapy during artificial reactivation of latent herpes simplex virus.

The chemotherapeutic effect of intensive acyclovir (ACV) treatment was evaluated in New Zealand white rabbits with confirmed latent herpes simplex virus type 1 (HSV-1). Acyclovir was administered orally (1 mg/ml of drinking water), topically (three times per day), and intramuscularly (16.5 mg/kg body wt twice per day) beginning 36 days after ocular inoculation and continued for 4 consecutive weeks, during which time the rabbits underwent concurrent stimulation by bilateral iontophoresis of 0.01% epinephrine into the cornea to induce reactivation of latent virus. None of the eight rabbits receiving intensive ACV shed virus in their tear film sample during 4 weeks of concurrent epinephrine iontophoresis and antiviral therapy. All five rabbits receiving placebo treatment shed HSV in tear film samples. The total number of positive ocular samples was 14 during the first week of dual treatment (5/5 animals positive) and decreased to 2 positive samples (2/5 animals) during the fourth week of dual treatment. Upon sacrifice, HSV was isolated from 8 of 16 trigeminal ganglia of ACV-treated animals and 6 of 10 placebo-treated animals. No increased resistance to ACV was detectable in any of the virus isolated from trigeminal ganglia. Antibody and cellular immune responses were not significantly different between the ACV-treated and placebo-treated groups.

Acyclovir↗