Pseudomonas corneal ulcer associated with rigid, gas-permeable, daily-wear lenses in a patient infected with human immunodeficiency virus.
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Adult↗
Biomedical subjects
A B Nesburn
Publications and source records attributed to A B Nesburn.
Baculovirus-expressed glycoprotein E (gE) of herpes simplex virus type-1 (HSV-1) protects mice against lethal intraperitoneal and lethal ocular HSV-1 challenge.
We have constructed a recombinant baculovirus expressing high levels of the herpes simplex virus type 1 (HSV-1) glycoprotein E (gE) in Sf9 cells. The expressed gE migrated on gels as a double band with apparent molecular weights of 68 and 70 kDa. The recombinant gE was glycosylated based on its susceptibility to tunicamycin treatment and was transported to the membrane of Sf9 cells based on indirect immunofluorescence. Mice vaccinated with gE developed high serum titers of HSV-1-neutralizing antibodies based on plaque reduction assays. gE vaccination also induced a strong delayed type hypersensitivity (DTH) response to HSV-1. In addition, mice vaccinated with the recombinant gE were protected from both intraperitoneal and ocular lethal HSV-1 challenge. To our knowledge, this is the first report in which vaccination with gE was shown to induce high neutralizing antibody titers, a DTH response, or protection against lethal HSV-1 challenge.
Animals↗
Baculovirus-expressed glycoprotein G of herpes simplex virus type 1 partially protects vaccinated mice against lethal HSV-1 challenge.
The DNA sequence encoding the complete HSV-1 glycoprotein G (gG) was inserted into a baculovirus transfer vector and recombinant viruses expressing gG were isolated. Three gG-related recombinant baculovirus expressed peptides of 37, 42, and 44 kDa were detected by Western blotting using monoclonal antibody to gG. The 42- and 44-kDa species were susceptible to tunicamycin, Endoglycosidase H (Endo-H), and N-glycosidase F (PNGase F) treatments, suggesting that they were glycosylated. Although only very low levels (approximately 1:10) of HSV-1-neutralizing antibody were produced in mice vaccinated with the baculovirus gG, these mice were partially protected from lethal challenge with HSV-1 (75-78% survival) and this level of protection was highly significant (P = 0.002). This is the first report to show that vaccination with HSV-1 gG can provide mice with any level of protection against lethal HSV-1 challenge.
Animals↗
Baculovirus expressed herpes simplex virus type 1 glycoprotein C protects mice from lethal HSV-1 infection.
A recombinant baculovirus (vAc-gC1) was constructed that expresses the glycoprotein C (gC) gene of herpes simplex virus type 1 (HSV-1). When Sf9 cells were infected with this recombinant, a protein that was smaller in size than authentic HSV-1 gC was detected by Western blotting using anti-gC polyclonal antibody. The recombinant gC was susceptible to tunicamycin, partially resistant to Endo-H, and was found on the membrane of Sf9 cells. Antibodies raised in mice to recombinant gC reacted with gC from HSV-1 infected cells and neutralized the infectivity of HSV-1 in vitro. Immunized mice were protected from lethal challenge with HSV-1.
Animals↗
Expression of herpes simplex virus type 1 glycoprotein B in insect cells. Initial analysis of its biochemical and immunological properties.
A recombinant baculovirus (vAc-gB1) was constructed which expresses the glycoprotein B (gB) gene of herpes simplex virus type 1 (HSV-1). When Sf9 cells were infected with these recombinant viruses, a protein that was close in size to authentic HSV-1 gB was detected by gB polyclonal antibody. The recombinant gB was found on the membrane of Sf9 cells and was susceptible to tunicamycin, glycosidase F (PNGase F) and partially susceptible to Endo-H. Antibodies raised in mice to this recombinant recognized viral gB and neutralized the infectivity of HSV-1 in vitro. Mice inoculated with the recombinant gB were protected from lethal challenge with HSV-1.
Animals↗
Baculovirus-expressed glycoprotein H of herpes simplex virus type 1 (HSV-1) induces neutralizing antibody and delayed type hypersensitivity responses, but does not protect immunized mice against lethal HSV-1 challenge.
We have shown previously that herpes simplex virus type 1 (HSV-1) glycoprotein H (gH) expressed by a baculovirus recombinant is transported to the cell surface in the absence of other HSV-1 gene products, and that the expressed gH has an apparent Mr similar to that of authentic HSV-1 gH. We report here that antibodies raised in mice to this baculovirus-expressed gH neutralize the infectivity of HSV-1 in vitro; this neutralizing activity was not complement-dependent. Mice vaccinated with gH also developed delayed type hypersensitivity (DTH) to HSV-1. This is the first report of expressed HSV-1 gH inducing neutralizing antibody or DTH responses in vaccinated animals. In contrast to the gH expressed in mammalian systems, the ability of this baculovirus-expressed gH to induce a neutralizing antibody response may be due to the inability of the mammalian expression system to transport gH to the cell surface. Despite inducing anti-HSV-1 neutralizing antibody and DTH responses, vaccination of mice with gH did not protect the mice against lethal intraperitoneal challenge with HSV-1.
Antibody Formation↗
Expression of herpes simplex virus type 1 glycoprotein I in baculovirus: preliminary biochemical characterization and protection studies.
We have constructed a recombinant baculovirus expressing the herpes simplex virus type 1 (HSV-1) glycoprotein I (gI). Sf9 cells infected with this recombinant virus synthesized gI-related polypeptides with apparent molecular sizes of 52 and 56 kDa. The recombinant gI appeared to be glycosylated, since it was susceptible to both tunicamycin and endoglycosidase H, and the expressed gI was transported to the surface of infected cells as judged by indirect immunofluorescence. Antibodies to the recombinant gI raised in mice neutralized HSV-1 infectivity. Finally, we show here for the first time that vaccination with gI can protect mice against HSV-1 challenge.
Animals↗
A retrospective study of disposable extended wear lenses in 100 patients: year 2.
We present the results of the second year of follow-up of 100 patients wearing disposable contact lenses on an extended wear basis. Thirty-seven males and 63 females participated in the study (mean age: 41.1 years +/- 13.88 (mean +/- 1 standard deviation); and the mean patient follow-up time was 26 months +/- 4.9). A total of 13 patients were lost to follow-up. Seventeen patients discontinued extended wear yet continued to wear soft lenses on a daily wear basis--the reasons for this finding are discussed. A significant decrease in the number of events defined as complications (32 events) was noted when compared to the first year of follow-up (136 events). Seven corneal infiltrates occurred during the second year, compared to 6 such events during the first year of the study. Additional complications included: giant papillary conjunctivitis (6 cases), corneal staining (5 cases), and corneal abrasions (4 cases). Disposable extended wear soft contact lenses offer a reasonable means of vision correction in the properly selected patient. At the same time, caution should be exercised in the fitting and wear of these lenses.
Adolescent↗
The use of the SoftPerm lens for the correction of irregular astigmatism.
We evaluated the performance of the SoftPerm combined rigid gas permeable (RGP) and hydrogel lens for the visual correction of 49 eyes (30 patients) with irregular astigmatism. Follow-up was up to 15 months. An attempt was also made to compare the SoftPerm lens to its predecessor, the Saturn II lens. The most common etiology for irregular astigmatism was keratoconus. Others etiologies were penetrating keratoplasty and refractive surgery. Visual acuity improved significantly. Only two eyes were best corrected to 20/20 before fitting whereas, 13 eyes saw 20/20 after fitting. The rate of lens replacement was 0.22 lenses per eye per month. A variety of complications occurred, including corneal staining, abrasions, and edema. Complications also reflected the relative difficulty of handling of the lens, yet no vision threatening complications occurred. Overall, the SoftPerm lens is a vast improvement over the Saturn II and is a reasonable means of correction of irregular astigmatism.
Adolescent↗
Immunoselection of recombinant baculoviruses expressing high levels of biologically active herpes simplex virus type 1 glycoprotein D.
The DNA sequence encoding the complete herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) was inserted into a baculovirus transfer vector under control of the polyhedrin gene promoter of the baculovirus Autographa california nuclear polyhedrosis virus (AcNPV). After co-transfection of Spodoptera frugiperda (Sf9) insect cells with wild-type AcNPV DNA and the recombinant transfer vector DNA, polyhedrin-negative recombinants that expressed high levels of HSV-1 gD were isolated using immunoaffinity selection with antibody coated magnetic particles followed by plaque purification. These recombinant baculoviruses expressed a protein that was slightly smaller than virion HSV-1 gD made in Vero cells. This recombinant protein was expressed at high levels. The expressed protein was glycosylated, was found on the membrane of Sf9 cells, and reacted with gD specific antibodies. Antibodies raised in mice to the recombinant gD neutralized HSV-1 as measured by plaque reduction assays. Mice inoculated with the recombinant baculovirus were completely protected from lethal challenge with HSV-1.
Animals↗
Identification of a major regulatory sequence in the latency associated transcript (LAT) promoter of herpes simplex virus type 1 (HSV-1).
The latency associated transcript (LAT) gene is the only viral genomic region that is abundantly transcribed during herpes simplex virus type 1 (HSV-1) neuronal latency. As such, it may play an important role in HSV-1 latency and/or reactivation. The regulation of the LAT gene is complex and appears to include a combination of positive and negative functional elements in and near the LAT promoter. In this study, transient CAT assays were used to map the minimal promoter necessary for constitutive activity in neuronal and nonneuronal cells to between nucleotide positions -161 and -2 (relative to the start of LAT transcription). The region from -283 to -161 was able to slightly increase promoter activity of the minimal promoter and appeared to have a larger effect in neuronal derived cells. Gel-shift experiments using nuclear extracts from neuronal and nonneuronal derived cells detected a major factor that bound specifically to the -161 to -2 probe. We designated this factor LAT promoter binding factor (LPBF). Two additional minor factors also bound specifically to the minimal promoter. DNase I footprint analysis and gel-shift competition experiments demonstrated that LPBF bound to a region that includes the palindromic sequence CCACGTGG located at nucleotides -72 to -65. Deletion of this palindrome resulted in a loss of binding of LPBF from the minimal promoter region and an 8- to 30-fold reduction in promoter activity in both neuronal and nonneuronal cells. Thus, LPBF appears to play a major role in LAT promoter regulation.
Animals↗
Cell surface expression of herpes simplex virus type 1 glycoprotein H in recombinant baculovirus-infected cells.
DNA encoding the complete sequence for the herpes simplex virus type 1 (HSV-1) glycoprotein H (gH) was inserted into a baculovirus transfer vector under control of the baculovirus polyhedrin gene promoter. After co-transfection with wild-type baculovirus DNA, recombinants expressing gH were isolated by plaque purification. The baculovirus-expressed HSV-1 gH represented a significant portion of total cellular protein and was several hundred fold more abundant than gH in HSV-1-infected Vero cells. The expressed gH appeared to be glycosylated, since it was similar in size to wild-type HSV-1 gH, was susceptible to both tunicamycin and endoglycosidase-H treatment, and was labeled by [3H]mannose. In contrast to previous reports of gH expressed in mammalian cells, the baculovirus recombinant-expressed gH was abundant on the cell surface as judged by indirect immunofluorescence. To our knowledge, this is the first report of expressed HSV-1 gH being transported to the cell surface in the absence of other HSV-1 gene products and the first report of expressed gH with an apparent molecular weight similar to authentic HSV-1 gH.
Animals↗
A retrospective study of disposable extended-wear lenses in 100 patients.
A retrospective evaluation of 100 patients who wore disposable extended-wear contact lenses is presented. The patients were selected at random and were fitted with one of three lens brands approved for extended wear. There were 37 males and 63 females in the study. The mean age was 40.1 years (range, 14 to 76 years). Sixty-seven of the patients wore nondisposable extended-wear soft contact lenses before being fitted with disposable lenses. Complications were noted and included two instances of infectious ulcerative keratitis and six cases of peripheral corneal infiltrates. It is believed that the rate of complications is due to the ongoing hypoxic effect of prolonged use of extended-wear or other contact lenses and may be improved by providing a greater variety of lens fitting parameters. Disposable extended-wear lenses may be a reasonable means of vision correction in the appropriately selected patient and with proper follow-up. At the same time, caution should be exercised in the fitting and wear of disposable extended-wear lenses.
Adolescent↗
The use of Saturn II lenses in keratoconus.
We studied Saturn II contact lenses in keratoconus patients who were intolerant to other available contact lenses. A total of 24 patients, who were followed for up to 15 months, participated in the study. Visual acuities improved significantly following fitting with the Saturn II contact lens; whereas only one eye in the series was correctable to 20/20 before fitting, eight eyes were corrected to 20/20 following fitting. A great number of lenses had to be replaced during the study because of deposit formation and tearing at the interface (rate of replacement: 0.52 lenses per eye per month). A variety of complications occurred during lens wear, resulting in only six eyes wearing the Saturn II lens at the close of the study (out of 46 eyes that had started wearing the lens). Discomfort due to tightness of the peripheral segment was the most common cause of lens discontinuation. We conclude that the Saturn II lens was inadequate for fitting patients with advanced keratoconus.
Adolescent↗
Activity of herpes simplex virus type 1 latency-associated transcript (LAT) promoter in neuron-derived cells: evidence for neuron specificity and for a large LAT transcript.
By using chloramphenicol acetyltransferase (CAT) assays in neuron-derived cell lines, we show here that promoter activity associated with the herpes simplex virus type 1 latency-associated transcript (LAT) had neuronal specificity. Promoter activity in these transient CAT assays coincided with a DNA region containing excellent RNA polymerase II promoter consensus sequences. Primer extension analysis in a LAT promoter-CAT plasmid construct placed the start of transcription about 28 nucleotides from the first T in the consensus TATA box sequence. Neuronal specificity of this promoter was suggested by examining the effect of sequences upstream of the promoter on CAT activity in neuronal versus nonneuronal cells. In nonneuronal cells, promoter activity was decreased 3- to 12-fold with the addition of upstream sequences. In contrast, in neuron-derived cells, the addition of upstream sequences did not decrease promoter activity. The LAT promoter predicted by our transient CAT assays was located over 660 nucleotides upstream from the 5' end of the previously mapped 2-kilobase (kb) LAT. This unusual location was explained by in situ and Northern (RNA) blot hybridization analyses that suggested that LAT transcription began near the promoter detected in our CAT assays, rather than near the 5' end of the 2-kb LAT. In situ hybridization with neurons from latently infected rabbits detected small amounts of LAT RNA within 30 nucleotides of the consensus TATA box sequence. This suggested that LAT transcription began near this TATA box. Northern blot hybridization of RNA from ganglia of latently infected rabbits revealed a faint 8.3-kb band of the same sense as LAT. We conclude that (i) the LAT promoter has neuronal specificity, (ii) the LAT promoter is located over 660 nucleotides upstream of the 5' end of the previously characterized stable 2-kb LAT, (iii) LAT transcription begins about 28 nucleotides from the first T of the consensus TATA box sequence and extends to near the first available polyadenylation site approximately 8.3 kb away, and (iv) this 8.3-kb RNA may be an unstable precursor of the more stable 2- and 1.3-kb LATs.
Animals↗
Ocular safety and efficacy of an HSV-1 gD vaccine during primary and latent infection.
One potential complication of systemic herpes simplex virus (HSV) vaccination is that subsequent ocular infection may lead to increased immunogenic corneal scarring. Therefore, V52, a genetically engineered vaccinia virus that expresses the HSV-1 glycoprotein gD, was tested for ocular safety and for protection against ocular challenge with a stromal-disease-producing strain (McKrae) of HSV-1. To maximize immune response, rabbits were vaccinated by a series of inoculations. V52-vaccinated rabbits developed significant HSV-1 neutralizing antibody titers; however, they were not as high as those induced by vaccination with live HSV-1 McKrae. One month after the final vaccination, all rabbits were challenged ocularly. Eyes were monitored for 35 days for epithelial keratitis, stromal keratitis, and iritis. In no case was epithelial keratitis, stromal keratitis, or iritis significantly exacerbated by vaccination. The gD V52 recombinant vaccine provided protection against HSV-1 induced epithelial keratitis (P = 0.02) and long-term stromal scarring (P = 0.04). There was no significant reduction in the incidence of trigeminal ganglionic latency in the vaccinated rabbits (P greater than 0.05). Thus, our results indicate that V52, a gD recombinant vaccine probably is safe with regard to corneal scarring, and may provide a small amount of protection against ocular HSV-1 infection. The amount of protection provided was less than that reported in mice and guinea pigs. This suggests that to provide high levels of ocular protection in rabbits (and probably in humans), HSV-1 vaccines may have to elicit a more vigorous immune response than that produced by normal HSV-1 infection.
Animals↗
Sequence of the latency-related gene of herpes simplex virus type 1.
During herpes simplex virus type 1 latency in neurons, RNA is transcribed from one small region of the viral genome. This latency-related RNA (LR RNA) is antisense to the mRNA of the immediate early gene ICPO, partially overlaps the ICPO mRNA, and is suspected of playing some role in helping maintain latency. We report here the sequence of the latency-related gene (LR gene) and its flanking sequences from strain F, and compare it to the available partial sequences of this region from strains 17 syn+ and KOS. Two potential open reading frames (ORFs) were common to all three sequences. Sequence analysis suggested the possibility of promoters in two different regions of the LR gene.
Base Sequence↗
Excimer laser ablation of the human lens at 308 nm with a fiber delivery system.
A 308 nm excimer laser has been used with a fiber delivery system to perform ablation of the human lens. Preliminary results demonstrate the system's ability to ablate lens nucleus and cortex selectively, preserving the anterior and posterior capsules. The total delivered energy necessary to ablate a human lens ranged from 35 to 63 joules. Laser tissue interaction and ablation rates were computed for the different components of the human lens (capsule, cortex, nucleus) for the operatively useful energy densities (fluences). Operative experience suggests that cortex and nucleus can be ablated while preserving the capsule if an adequate irrigation and aspiration system is developed. These results also suggest that this modality may be adequate for performing endocapsular cataract extraction. Laser tissue interactions were also computed at variable distances between the fiber tip and tissue. As this distance increased, the spread of the beam increased and a significant increase in energy was necessary to induce tissue ablation. This was due to the decrease in fluence with increasing distance to the target tissue and/or the absorption and scattering of the delivered energy within a short distance from the fiber tip by the ablated material. Evidence of a sonic effect was also present.
Aged↗