RESUMEN
AIM: To use a confocal microscope to characterise the treated and untreated courses of fungal keratitis. METHODS: In the first experiment, Aspergillus fumigatus stromal keratitis was produced in both eyes of seven New Zealand white rabbits. In the second experiment, keratitis was induced in right eyes of 20 rabbits. Group 1 rabbits were treated with topical fluconazole, group 2 rabbits received oral fluconazole, and group 3 rabbits were used as controls. The rabbits were examined with a slit lamp and confocal microscope 2, 6, 10, 14, and 20 days after inoculation. The corneal cultures were taken on days 2, 14, and 20 and biopsies were taken on days 2 and 22. RESULTS: On days 14 and 22 confocal microscopy was more sensitive than culture technique in both treated and untreated animals, since not all cases of fungal keratitis can be cultured. CONCLUSION: This study indicates that confocal microscopy is a rapid and sensitive diagnostic tool for both the early diagnosis and non-invasive follow up of fungal keratitis
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Aspergilosis/patología , Aspergillus fumigatus/aislamiento & purificación , Queratitis/patología , Microscopía Confocal/métodos , Administración Oral , Administración Tópica , Animales , Antifúngicos/administración & dosificación , Aspergilosis/tratamiento farmacológico , Células Cultivadas , Córnea/microbiología , Córnea/patología , Fluconazol/administración & dosificación , Queratitis/tratamiento farmacológico , Queratitis/microbiología , Masculino , ConejosRESUMEN
Keratocytes express MHC class I molecules constitutively, and keratocytes stimulated with IFN-gamma express MHC class II molecules. Unstimulated keratocytes constitutively express B7-1 and ICAM-1, as well as low levels of CD40 and 4-1BBL. These findings indicate that keratocytes may deliver both antigen-specific and costimulatory signals to CD4(+) and CD8(+) T cells. To demonstrate that keratocytes expressing B7-1 provide a costimulatory signal to T cells, CD4(+) or CD8(+) mouse T cells were incubated with anti-CD3 mAb and irradiated keratocytes. Enhanced proliferation of both CD4(+) and CD8(+) T cells occurred, and could be inhibited by anti-B7-1 mAb, indicating T cell costimulatory activity by B7-1 on the keratocytes. To demonstrate that keratocytes can deliver an antigen-specific signal, CD4(+) and CD8(+) T cells from herpes-infected mice were incubated with HSV-1-infected, irradiated keratocytes. The resulting T cell proliferation and production of Th1 cytokines (IL-2, IFN-gamma) indicated T cell activation by antigens presented by the infected keratocytes. These results show that keratocytes in the corneal stroma of the mouse can function as antigen-presenting cells and, thus, may play a role in immune-mediated stromal inflammation such as herpetic stromal keratitis.
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Células Presentadoras de Antígenos/fisiología , Córnea/citología , Animales , Antígenos CD/análisis , Antígeno B7-1/análisis , Antígeno B7-2 , Antígenos CD40/análisis , Células Cultivadas , Citocinas/biosíntesis , Femenino , Antígenos de Histocompatibilidad Clase II/análisis , Queratitis/inmunología , Activación de Linfocitos , Glicoproteínas de Membrana/análisis , Ratones , Ratones Endogámicos BALB C , Células del Estroma/fisiologíaRESUMEN
PURPOSE: To determine the effect of the topical ocular hypotensive drug, isopropyl unoprostone, a docosanoid molecule with very weak prostaglandin activity, on herpes keratitis in the rabbit eye. METHODS: For acute disease, rabbit corneas inoculated with the corticosteroid-sensitive F(MP)E strain of herpes simplex virus type 1 were treated with various combinations of 0.12% isopropyl unoprostone, latanoprost, trifluridine, benzalkonium chloride 0.02%, dexamethasone sodium phosphate, ketorolac tromethamine, or saline solution beginning 1 day after infection. Severity of keratitis was evaluated in a masked manner. For recurrent disease, rabbit corneas infected with McKrae strain herpes simplex virus type 1 were treated with unoprostone or saline solution on postinfection days 25 to 42, and the presence or absence of lesions was recorded. RESULTS: Eyes treated with unoprostone showed significantly less severe disease than saline-treated or latanoprost-treated eyes during acute infection. Unoprostone-treated and saline-treated eyes showed no significant difference in the frequency of recurrent lesions. Eyes treated with latanoprost and/or dexamethasone, separately or in combination, showed increased severity of acute herpes simplex virus keratitis, whereas benzalkonium chloride 0.02%--treated eyes showed no significant difference, compared with saline treatment. Trifluridine resulted in rapid healing. CONCLUSIONS: Unoprostone did not increase the severity or recurrence rate of herpes simplex virus keratitis. Unoprostone requires twice-a-day administration, compared with once-a-day for latanoprost, and unoprostone lowers intraocular pressure less than latanoprost. Nevertheless, unoprostone's superior safety profile may make its use advantageous. Benzalkonium chloride alone did not make the keratitis worse.
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Antihipertensivos/uso terapéutico , Dexametasona/análogos & derivados , Dinoprost/uso terapéutico , Presión Intraocular/efectos de los fármacos , Queratitis Herpética/tratamiento farmacológico , Prostaglandinas F Sintéticas/uso terapéutico , Enfermedad Aguda , Administración Tópica , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/uso terapéutico , Antihipertensivos/administración & dosificación , Dexametasona/administración & dosificación , Dexametasona/uso terapéutico , Dinoprost/administración & dosificación , Dinoprost/análogos & derivados , Quimioterapia Combinada , Femenino , Queratitis Herpética/fisiopatología , Latanoprost , Masculino , Soluciones Oftálmicas , Prostaglandinas F Sintéticas/administración & dosificación , Conejos , Distribución Aleatoria , RecurrenciaRESUMEN
PURPOSE: The2',5'-oligoadenylate-dependent RNase L gene functions in the interferon-inducible RNA decay pathway known as the 2-5A system. The purpose of this study was to determine whether the absence of this gene affects the pathogenesis of herpes simplex virus type 1 (HSV-1) ocular infection in the mouse. METHODS: HSV-1 (strain McKrae) was applied bilaterally to unscarified corneas of RNase L-null mice and congenic controls. To evaluate the severity of herpetic keratitis, slit lamp examinations (SLE) were performed every other day for 14 days. To study corneal histology and apoptosis, HSV-1-inoculated RNase-L-null and congenic control mice, as well as mock-inoculated mice (apoptosis negative control), were killed at 6 and 18 hours postinoculation (PI). Uninoculated mice that underwent corneal scarification (apoptosis positive control) were killed 2 hours after scarification. Eyes were dissected and the corneas processed for light and transmission electron microscopy and the TUNEL assay. RESULTS: In comparison with the congenic control mice, RNase L-null mice showed significantly more severe herpetic keratitis (PI day 8, SLE score, mean +/- SEM: 3.27 +/- 0.10 vs. 2.34 +/- 0.06; P: < 0.001) and significantly higher mortality (PI day 14, 70% vs. 20%; P: < 0.001). Few apoptotic cells were seen in HSV-1-infected RNase L-null mice, although DNA fragmentation consistent with apoptosis was detected in the corneas of congenic control mice 6 and 18 hours after HSV-1 inoculation and in uninfected mice with scarified corneas. Signs of apoptosis were not present in the mock-infected corneas. Electron microscopic evidence of keratocytic apoptosis was detected only in the uninfected scarified corneas and the HSV-1-infected congenic control corneas. CONCLUSIONS: The increased severity of ocular disease and increased mortality in the RNase L-null mice provides evidence, for the first time, that the 2-5A system contributes to protection during ocular herpetic infection. The reduced frequency of apoptosis in these mice suggests that one possible mechanism for this protective effect could be the induction of apoptosis in corneal cells as a means of reducing the spread of infectious virus.
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Córnea/virología , Endorribonucleasas/fisiología , Herpesvirus Humano 1/patogenicidad , Queratitis Herpética/mortalidad , Queratitis Herpética/virología , Animales , Córnea/ultraestructura , Fragmentación del ADN , Etiquetado Corte-Fin in Situ , Queratitis Herpética/enzimología , Queratitis Herpética/patología , Ratones , Ratones Noqueados , Virulencia , Replicación ViralRESUMEN
An understanding of the cellular genes whose expression is altered during HSV reactivation will enable us to better understand host responses and biochemical pathways involved in the process. Furthermore, this knowledge could allow us to develop gene-targeted inhibitors to prevent viral reactivation. Mice latent with HSV-1 strain McKrae and uninfected control mice were subjected to hyperthermic stress (43 degrees C for 10 min) and their trigeminal ganglia (TG) collected 1 h later. Two additional groups included HSV-1 latently infected and uninfected mice not subjected to hyperthermic stress. Poly A+ mRNA was enriched from total mouse TG RNA and reverse transcribed using MMLV RT. Radioactively labeled cDNAs were analyzed by microarray analysis. A stress/toxicology array of 149 mouse genes on a nylon membrane was used. The labeled cDNAs prepared from latently infected, stressed mice demonstrated 3-fold or greater increases in certain mRNA-early response genes (ERGs) compared to cDNAs from uninfected, stressed control mice. The ERG mRNAs that showed increases included two heat shock proteins (HSP60 and HSP40), a basic transcription factor (BTF T62), a DNA repair enzyme, two kinases [MAP kinase and a stress-induced protein kinase (SADK)], an oxidative stress-induced protein, a manganese superoxide dismutase precursor-2 (SOD-2), and cyclooxygenase 2 (COX-2). The gene expression in unstressed, infected TGs was similar to the gene expression in unstressed, uninfected controls. These results suggest that there is a significant difference in the ERG expression profile in latently infected TGs undergoing stress-induced reactivation compared to uninfected TGs.
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Expresión Génica , Herpesvirus Humano 1/genética , Ganglio del Trigémino/metabolismo , Latencia del Virus , Animales , Femenino , Perfilación de la Expresión Génica , Herpesvirus Humano 1/fisiología , Hipertermia Inducida , Ratones , Ratones Endogámicos BALB C , Análisis de Secuencia por Matrices de Oligonucleótidos , Activación ViralRESUMEN
Ocular virus infections remain an important cause of corneal and external disease. Herpes simplex, the most important, is easily treated when it is confined to the epithelium. New studies indicate that herpetic stromal disease and iritis are effectively treated with a combination of corticosteroid and antiviral without additional risk. Recurrences of ocular herpetic disease can be reduced with acyclovir given orally; the benefit seems to be greatest in patients who have had at least one episode of stromal keratitis. Herpes zoster can be treated with either acyclovir or famciclovir, but to be effective, treatment must be initiated within 72 hours of onset. Early treatment reduces the risk of post-herpetic neuralgia and may reduce the risk of ocular complications. Adenovirus infection (epidemic keratoconjunctivitis) is often spread by the ophthalmologist. New medications such as cidofovir appear to be effective against the adenoviruses in non-human systems and may have some effect in man, although previously, drugs that appeared to have an effect in vitro have proven to be ineffective in the clinical setting.
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Enfermedades de la Córnea/virología , Infecciones Virales del Ojo/tratamiento farmacológico , Infecciones por Adenovirus Humanos/tratamiento farmacológico , Animales , Antivirales/uso terapéutico , Ensayos Clínicos como Asunto , Enfermedades de la Córnea/tratamiento farmacológico , Herpes Zóster Oftálmico/tratamiento farmacológico , Humanos , Queratitis Herpética/tratamiento farmacológico , Tracoma/tratamiento farmacológicoRESUMEN
PURPOSE: To investigate the migration of herpes simplex virus type 1 (HSV-1) between latently infected and naive corneal tissues and trigeminal ganglion (TG) in rabbits after penetrating keratoplasty (PKP) and transcorneal epinephrine iontophoresis. METHODS: Two mutants, genetically constructed from HSV-1 strain 17syn+, were used to inoculate rabbit corneas: 17deltaPst, a latency associated transcript (LAT) negative, low-reactivating virus and 17Pr, a high-reactivating, LAT-positive rescuant of 17deltaPst. Latently infected rabbits were given corneal allografts from naive rabbits, and naive rabbits received grafts from latently infected rabbits. Ninety days after PKP, groups of the transplanted rabbits were induced to reactivate by transcorneal epinephrine iontophoresis, but others were not induced. Viral shedding was monitored by tear film cultures. Rabbits were killed 5 days after iontophoresis. Transplanted grafts, recipient corneal rims, and corresponding TG were obtained. Nucleic acids were extracted and amplified for detection of HSV-1 DNA and viral gene transcription. RESULTS: In naive rabbits receiving grafts transplanted from rabbits latently infected with 17Pr (LAT+), 3 of 6 corneal rims contained HSV DNA after induction. In contrast, none of the 5 corneal rims from naive rabbits receiving grafts from rabbits latent with 17deltaPst (LAT-) contained viral DNA. Viral DNA and gene transcripts were detected in 2 of 6 TG from naive rabbits that received grafts from 17Pr (LAT+) latently infected rabbits. In recipient corneal rims and TG of latently infected rabbits receiving grafts from naive rabbits, viral DNA concentration was significantly greater with induced reactivation, compared with the results in noninduced rabbits. The amount of viral DNA in naive grafts transplanted into 17Pr (LAT+) latently infected rabbits was significantly higher with induction than without induction (P = 0.018). More viral DNA and viral gene transcripts were found in tissues from rabbits latently infected with 17Pr (LAT+) than in rabbits latently infected with 17deltaPst (LAT-). CONCLUSIONS: Corneas from latently infected rabbits contain HSV-1 DNA that can replicate after induced reactivation. Viral migration can occur in both anterograde and retrograde directions between the transplanted graft and the recipient corneal rim and TG. The LAT negative HSV-1 construct 17deltaPst has a significantly reduced ability to replicate and migrate.
Asunto(s)
Córnea/virología , Herpesvirus Humano 1/fisiología , Queratitis Herpética/virología , Queratoplastia Penetrante , Latencia del Virus/fisiología , Animales , Córnea/inervación , Cartilla de ADN/química , ADN Viral/análisis , Epinefrina/farmacología , Expresión Génica/genética , Genes Virales/genética , Supervivencia de Injerto , Herpesvirus Humano 1/genética , Iontoforesis , Queratitis Herpética/patología , Reacción en Cadena de la Polimerasa , Conejos , Lágrimas/virología , Ganglio del Trigémino/virología , Activación Viral/efectos de los fármacos , Esparcimiento de Virus/fisiologíaRESUMEN
PURPOSE: To determine if lamellar keratoplasty in rabbits latently infected with herpes simplex virus type 1 (HSV-1) would stimulate graft recipients to shed virus and induce viral-specific corneal lesions. METHODS: Rabbits latently infected with HSV-1 received lamellar allografts in one eye from normal uninfected rabbits and the contralateral eyes served as unoperated controls. Normal rabbits received lamellar grafts from rabbits latently infected with HSV-1. For 1 week after surgery, slit-lamp examination and ocular swab sampling were performed daily to assess viral reactivation. RESULTS: The occurrence of positive swab cultures and corneal epithelial lesions after lamellar keratoplasty was significantly higher in operated eyes of latently infected rabbits when compared to the control eyes. Ocular shedding or recurrent lesions were not observed in the normal rabbits receiving corneal grafts from latently infected donors. CONCLUSIONS: These results indicated that lamellar keratoplasty induces HSV-1 shedding and recurrent epithelial lesions in the eyes of rabbits latently infected with HSV-1, which received lamellar grafts, but not in the eyes of normal rabbits given lamellar grafts from HSV-1 latently infected rabbits. It seems that the site of viral latency is not the anterior corneal stroma or the epithelium.
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Trasplante de Córnea/efectos adversos , Epitelio Corneal/virología , Herpesvirus Humano 1/fisiología , Queratitis Herpética/etiología , Activación Viral , Animales , Epitelio Corneal/patología , Herpesvirus Humano 1/aislamiento & purificación , Queratitis Herpética/patología , Conejos , Recurrencia , Lágrimas/virología , Trasplante Homólogo , Latencia del Virus/fisiología , Esparcimiento de Virus/fisiologíaRESUMEN
A 43-year-old white woman with a history of multiple ocular surgeries, including 4 penetrating keratoplasties, developed a concentric retrocorneal membrane at the graft periphery in the right eye. A white-light, tandem, scanning confocal microscope using a 24x/0.60 contact objective was used to examine the right eye in vivo. At the endothelial layer, confocal microscopic images similar to corneal epithelial cells were detected at the graft periphery. Unlike normal endothelial cells, the imaged cells demonstrated easily recognizable nuclei.
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Enfermedades de la Córnea/patología , Epitelio Corneal/patología , Queratoplastia Penetrante/efectos adversos , Adulto , Enfermedades de la Córnea/etiología , Femenino , Humanos , Microscopía ConfocalRESUMEN
BACKGROUND: The purpose of the study was to assess the appearance of lattice corneal dystrophy by means of white-light confocal microscopy. METHODS: Two consecutive patients with lattice corneal dystrophy were prospectively examined. In vivo white-light tandem-scanning confocal microscopy was performed in the right eye of the first patient. Her left eye had undergone penetrating keratoplasty 4 years earlier. Histologic findings of the corneal button were compared with confocal microscopic findings of the right eye. The other patient was monocular and confocal microscopy was performed only in the non-seeing eye. RESULTS: In both patients, linear and branching structures with changing reflectivity and poorly demarcated margins were visualized in the stroma. The linear structures measured approximately 40-80 microm in width. CONCLUSION: Lattice corneal dystrophy presents characteristic linear images on confocal microscopy and should not be misdiagnosed as fungal hyphae in cases of corneal infection.
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Córnea/patología , Distrofias Hereditarias de la Córnea/patología , Microscopía Confocal , Anciano , Córnea/cirugía , Distrofias Hereditarias de la Córnea/cirugía , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Humanos , Queratoplastia Penetrante , Masculino , Persona de Mediana Edad , Estudios ProspectivosRESUMEN
OBJECTIVE: To compare topical cidofovir with topical trifluridine for the prevention and treatment of herpes simplex type 1 stromal keratitis in rabbits. METHODS: The RE strain of herpes simplex virus 1 was injected into the central stroma of both eyes of New Zealand white rabbits. Two to 3 days after virus inoculation, the rabbits were randomized to treatment groups of 10 each and treated with 1% trifluridine administered 5 or 7 times a day, 1%, 0.5%, or 0.2% cidofovir administered twice a day, fluorometholone administered twice a day, or balanced salt solution (BSS) administered twice a day (control) until day 21 after injection. The treated corneas were examined 3 times a week and the severity of stromal keratitis was graded in a masked fashion. To evaluate the ability of cidofovir to treat established stromal disease, groups of 10 rabbits each were inoculated with herpes simplex virus and treated with 1% cidofovir twice a day, 1% trifluridine 5 times a day, fluorometholone twice a day, or BSS twice a day beginning on day 7 after virus inoculation through day 21. RESULTS: Treatment with 0.2% cidofovir twice a day was not effective in preventing the appearance of stromal disease (P = .89), whereas treatment with 0.5% (P<.001) or 1% (P<.001) cidofovir twice a day or 1% trifluridine 5 times a day (P<.001) or 7 times a day (P = .006) significantly reduced the appearance of stromal keratitis on the 8 evaluation days, compared with BSS treatment (F test analysis of variance). There was no difference between the eyes treated with 0.5% cidofovir twice a day and those treated with 1% trifluridine 5 times a day. Treatment with 1% cidofovir was not effective in treating established stromal disease. CONCLUSIONS: Cidofovir and trifluridine are highly effective in preventing the appearance of herpetic stromal disease. Cidofovir is as effective as, but no more effective than, trifluridine in this model. Neither cidofovir nor trifluridine benefits established stromal disease, however. CLINICAL RELEVANCE: Cidofovir is a new, potent antiviral that seems similar in efficacy to trifluridine and is effective in the prevention of the development of stromal herpes, but is not effective in the treatment of established stromal disease in which hypersensitivity predominates.
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Antivirales/uso terapéutico , Sustancia Propia/efectos de los fármacos , Citosina/análogos & derivados , Herpesvirus Humano 1/efectos de los fármacos , Queratitis Herpética/prevención & control , Organofosfonatos , Compuestos Organofosforados/uso terapéutico , Administración Tópica , Animales , Antivirales/administración & dosificación , Cidofovir , Sustancia Propia/virología , Citosina/administración & dosificación , Citosina/uso terapéutico , Femenino , Herpesvirus Humano 1/fisiología , Queratitis Herpética/tratamiento farmacológico , Queratitis Herpética/virología , Masculino , Soluciones Oftálmicas/administración & dosificación , Soluciones Oftálmicas/uso terapéutico , Compuestos Organofosforados/administración & dosificación , Conejos , Distribución Aleatoria , Trifluridina/administración & dosificación , Trifluridina/uso terapéuticoRESUMEN
AIMS: To report the appearances of cornea guttata and Fuchs' endothelial dystrophy from white light confocal microscopy. METHODS: Seven eyes of four consecutive patients with cornea guttata were prospectively examined. Of the seven eyes, three also had corneal oedema (Fuchs' dystrophy). In vivo white light tandem scanning confocal microscopy was performed in all eyes. Results were compared with non-contact specular microscopy. RESULTS: Specular microscopy was precluded by corneal oedema in one eye. In the remaining six eyes, it demonstrated typical changes including pleomorphism, polymegathism, and the presence of guttae appearing as dark bodies, some with a central bright reflex. In all seven eyes, confocal microscopy revealed the presence of round hyporeflective images with an occasional central highlight at the level of the endothelium. Changes in cell morphology and size were readily appreciated. CONCLUSION: By comparison with specular microscopy, the hyporeflective images with an occasional central highlight seen on confocal microscopy are consistent with the presence of guttae. Confocal microscopy may confirm the diagnosis of cornea guttata and Fuchs' endothelial dystrophy by demonstrating the presence of guttae. This technique is especially valuable in cases of corneal oedema, where specular microscopy may fail to visualise the endothelium. However, specular microscopy should remain the method of choice to evaluate the endothelium, principally because it is easier to use.
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Distrofia Endotelial de Fuchs/diagnóstico , Microscopía Confocal/métodos , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Estudios ProspectivosRESUMEN
PURPOSE: To report the distinguishing characteristics of posterior polymorphous corneal dystrophy (PPMD) using confocal microscopy. MATERIAL AND METHODS: Two consecutive patients with PPMD were prospectively examined using a white-light tandem scanning confocal microscope with a 24x/0.60 contact objective. RESULTS: At the level of Descement's membrane, roundish hyporeflective images were found in 1 patient. In the other patient, hyporeflective bands were detected. In both patients, patchy hyperreflective areas were identified. CONCLUSION: Confocal microscopy may allow diagnosis of PPMD by demonstrating the alterations in Descement's membrane. This technique is especially valuable in cases of endothelial decompensation, where slit-lamp and specular microscopy may fail to demonstrate changes in Descement's membrane.
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Distrofias Hereditarias de la Córnea/patología , Microscopía Confocal , Anciano , Lámina Limitante Posterior/patología , Femenino , Humanos , Persona de Mediana Edad , Estudios Prospectivos , Grabación en VideoRESUMEN
PURPOSE: CTLA4, a high-affinity ligand of B7, can, in soluble form, prevent antigen-driven T-cell activation by blocking CD28-B7 interaction and can thereby prevent immune graft rejection. In this study, we tested the capacity of soluble CTLA4-Ig alone or in combination with UV-B irradiation to suppress corneal allograft rejection in rabbits. METHODS: Corneas from Dutch belted rabbits were incubated in corneal storage medium containing 0, 1, 10, 25, or 250 microg/ml of CTLA4-Ig for 18 h and were then transplanted into the vascularized or nonvascularized corneas of New Zealand White rabbit recipients. A series of donor corneas were exposed to UV-B irradiation alone or a combination of irradiation and CTLA4-Ig to determine if these two treatments would have an additive effect in prolonging graft survival. The fate and clinical condition of the allografts were evaluated by slit-lamp photomicroscopic observation and corneal-thickness measurements. Grafts that were rejected were processed for histopathologic and immunohistochemical analysis to determine the characteristics of cells infiltrating the grafts. RESULTS: Grafts placed in nonvascularized corneas showed no differences in survival times, regardless of treatment. Among the grafts placed in vascularized corneas, those incubated with CTLA4-Ig at a concentration of 250 microg/ml failed within 7-14 days. Histopathologic and immunocytochemical examination revealed a dense accumulation of immune inflammatory cells, especially class II major histocompatibility complex (MHC)-expressing, antigen-presenting cells, in the failed grafts. Grafts incubated with CTLA4-Ig at concentrations of 1 and 10 microg/ml had mean survival times greater than the control, untreated corneal allografts. Some of the grafts in these two treatment groups survived for the 100-day observation period, whereas none of the grafts in the other treatment groups survived to this end point. UV-B irradiated grafts incubated with CTLA4-Ig at a concentration of 1 microg/ml appeared to have longer survival times and fewer rejections compared with control, untreated grafts and grafts treated with UV-B or CTLA4-Ig alone. CONCLUSION: The results show that the CTLA4-Ig coreceptor blocking agent can prolong corneal allograft survival in vascularized graft sites and that UV-B irradiation followed by incubation in CTLA4-Ig may prolong graft survival better than either treatment alone. These results suggest that agents that prevent second-signal interaction between antigen-presenting cells and T lymphocytes may be useful for inhibiting corneal allograft rejection.
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Antígenos de Diferenciación/farmacología , Córnea/efectos de los fármacos , Trasplante de Córnea , Rechazo de Injerto/prevención & control , Inmunoconjugados , Inmunosupresores/farmacología , Proteínas Recombinantes de Fusión/farmacología , Abatacept , Animales , Antígenos CD , Antígeno CTLA-4 , Terapia Combinada , Córnea/efectos de la radiación , Femenino , Supervivencia de Injerto/efectos de los fármacos , Supervivencia de Injerto/efectos de la radiación , Fragmentos Fc de Inmunoglobulinas , Masculino , Conejos , Trasplante Homólogo , Rayos UltravioletaRESUMEN
PURPOSE: To determine whether topically applied latanoprost increases the severity of acute herpes simplex keratitis, the rate of recurrence of herpes keratitis, or both, in the rabbit. METHODS: To determine the effect on severity of acute herpetic keratitis, the corneas of New Zealand white rabbits were infected with either the less-corticosteroid-sensitive McKrae strain or the corticosteroid-sensitive F(MP)E strain of herpes simplex virus type 1. Rabbits were randomly assigned to twice-a-day treatment with latanoprost 0.005%, dexamethasone sodium phosphate 0.1%, or balanced saline solution within 3 days of infection and evaluated daily for up to 13 days after infection. The severity of keratitis was graded in a masked manner. To determine the effect on recurrences of herpetic keratitis, animals infected with McKrae strain herpes simplex virus type 1 that survived to day 32 after infection were randomized to treatment with latanoprost 0.005% or balanced saline solution and evaluated for the presence of corneal lesions from postinfection day 32 to day 47. RESULTS: In the severity studies, treatment of F(MP)E-infected corneas with latanoprost or dexamethasone significantly worsened herpetic keratitis; by postinfection day 5, F(MP)E-infected eyes treated with dexamethasone or latanoprost demonstrated significantly higher severity scores than the eyes treated with balanced saline solution (P = .0001 and .008, respectively). Scores of McKrae-infected corneas treated with latanoprost or dexamethasone were not significantly different from scores of balanced saline solution-treated corneas. In the recurrence study, treatment with latanoprost significantly increased the appearance of clinical recurrences in McKrae-infected eyes, compared with balanced saline solution treatment (P = .0064). CONCLUSION: Latanoprost may worsen acute herpetic keratitis in the rabbit eye and increase the risk of recurrences in latently infected animals.
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Córnea/virología , Herpesvirus Humano 1/crecimiento & desarrollo , Queratitis Herpética/virología , Prostaglandinas F Sintéticas/farmacología , Activación Viral/efectos de los fármacos , Enfermedad Aguda , Administración Tópica , Animales , Córnea/patología , Dexametasona/farmacología , Femenino , Queratitis Herpética/patología , Latanoprost , Masculino , Prostaglandinas F Sintéticas/administración & dosificación , Conejos , RecurrenciaRESUMEN
AIMS: To report the appearances of iridocorneal endothelial (ICE) syndrome from real time, white light confocal microscopy. METHODS: Three consecutive patients, each with ICE syndrome, were examined prospectively. Corneal specular and confocal microscopic examinations were performed in all three patients. In the first patient, a penetrating keratoplasty was performed and the cornea was examined by light and scanning electron microscopy. No surgery was performed in the remaining two patients. RESULTS: In the first patient corneal oedema prevented endothelial specular microscopy. Confocal microscopy performed before penetrating keratoplasty successfully revealed abnormal epithelial-like endothelial cells. Histological examinations of the cornea following penetrating keratoplasty revealed the presence of multilayered endothelial cells with epithelial features (microvilli). In the remaining two patients, specular microscopy showed the presence of ICE cells with typical dark/light reversal. Confocal microscopy demonstrated groups of endothelial cells with epitheloid appearances. In all three patients, the contralateral endothelial appearance was normal by specular and confocal microscopy, except for moderate endothelial polymegathism in one patient. Epithelial-like endothelial cells were characterised by prominent nuclei on confocal microscopy. CONCLUSIONS: The application of confocal microscopy indicates that the ICE syndrome is characterised by epitheloid changes in the endothelium. Confocal microscopy may be used to diagnose the ICE syndrome by demonstrating epithelial-like endothelial cells with hyperreflective nuclei. This technique is especially of value in cases of corneal oedema, since specular microscopy may fail to image the endothelium in such cases.
Asunto(s)
Enfermedades de la Córnea/patología , Glaucoma/patología , Iritis/patología , Microscopía Confocal/métodos , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Síndrome , Agudeza VisualRESUMEN
BACKGROUND: In this study, we determined the binding characteristics of F(ab')2 alloantibody fragments to corneal antigens and assessed the capacity of these antibody fragments to protect corneal allografts from immune attack. METHODS: Goat anti-rabbit alloantibodies were pepsin-digested and labeled with 125I, and the time course of association and dissociation of the F(ab')2 fragments was determined. Corneal allografts were incubated in unlabeled F(ab')2 fragments and transplanted into allogeneic recipients, and the graft survival times were recorded. RESULTS: Binding of radiolabeled F(ab')2 fragments to rabbit cornea cells reached a maximum at 12 hr. At 32 degrees C (rabbit corneal temperature), the radiolabel eluted rapidly from the cornea, reaching baseline at 72 hr. At 4 degrees C (corneal graft storage temperature), significant amounts remained associated with the cornea at 96 hr. Mean survival time for grafts incubated in F(ab')2 anti-rabbit fragments was significantly greater than that of grafts incubated in nonimmune F(ab')2 fragments. Three of the corneal allografts incubated in goat F(ab')2 anti-rabbit fragments survived for 100 days, whereas the longest surviving control allograft incubated in goat F(ab')2 nonimmune fragments was rejected on day 24. Preincubation of corneas in unlabeled, immune F(ab')2 fragments followed by incubation in radiolabeled, immune F(ab')2 fragments suggested that antigen masking was not a factor in the prolongation of graft survival. CONCLUSION: Based on the binding and release kinetics and the graft survival times, it appears that the protective effect of immune F(ab')2 fragments extends well beyond the binding interval of the antibody fragments to corneal cell membranes.
Asunto(s)
Trasplante de Córnea/inmunología , Supervivencia de Injerto , Isoanticuerpos/inmunología , Animales , Unión Competitiva , Córnea/inmunología , Femenino , Cabras , Fragmentos Fab de Inmunoglobulinas/inmunología , Conejos , Trasplante HomólogoAsunto(s)
Enfermedades de la Córnea/patología , Epitelio Corneal/patología , Enfermedades de la Córnea/tratamiento farmacológico , Epitelio Corneal/efectos de los fármacos , Femenino , Humanos , Microscopía Confocal , Persona de Mediana Edad , Soluciones Oftálmicas/uso terapéutico , Nitrato de Plata/uso terapéuticoRESUMEN
PURPOSE: This study aimed to detect corneal conditions presenting with linear images on white light confocal microscopy and to analyze their distinguishing characteristics. METHODS: In 1996 and 1997, 153 eyes of 110 patients with various corneal conditions were examined. In vivo examination of the cornea was performed by using a white-light tandem scanning confocal microscope. Images were captured by using a video camera and stored on S-VHS video tapes. In this retrospective study, patient charts and confocal microscopic video records were reviewed. Conditions with linear images were looked for, and the images were analyzed and compared. RESULTS: The only structures presenting as linear images on confocal microscopy in normal subjects consisted of corneal nerves. The following pathologic conditions also had linear images on confocal microscopy: corneal vascularization, mycotic keratitis, lattice corneal dystrophy, and posterior polymorphous dystrophy. Each condition could be identified based on its reflectivity, delineation, size, branching pattern, and location in the cornea. CONCLUSION: Different corneal conditions present with linear images on confocal microscopy. Correct identification is critical to avoid misdiagnosis.