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PubMedDate not supplied

Identification of M.leprae in conjunctiva of leprosy patients using the superior tarsal conjunctiva scrape technique.

The technique of superior tarsal conjunctiva scrape was used for identifying M.leprae in the conjunctiva in 56 leprosy patients (all of them multibacillary, some untreated and others treated with multidrug therapy). The technique of tarsal conjunctiva scrape was shown to be more suitable than conjunctival biopsy for identifying lepra bacilli. This technique is also easier to perform and has shown a statistical relation between bacilloscopical index of skin (BIsk) and bacilloscopical index of tarsal conjunctiva (BIconj) values. Thus, if the bacilli can be identified at tarsal conjunctiva we can assume greater systemic bacillary load in the patients.

Adult↗
PubMed1988

Detection of ocular mucus in normal human conjunctiva and conjunctiva from patients with cicatricial pemphigoid using lectin probes and histochemical techniques.

Conjunctival biopsies from patients with cicatricial pemphigoid affecting the conjunctiva and patients undergoing cataract surgery (normal conjunctiva) were snap-frozen, cryostat sectioned and incubated with fluorescein-conjugated lectins; peanut agglutinin (PNA), Helix pomatia agglutinin (HPA), soybean agglutinin (SBA), wheat germ agglutinin (WGA) and succinylated wheat germ agglutinin (S-WGA). Controls consisted of preincubating the lectins with the appropriate blocking sugars before applying the lectins to the sections. PNA and HPA stained the mucus granules contained in the conjunctival goblet cells but did not stain mucus or glycocalyx at the ocular surface distal to the goblet cells. Native WGA and S-WGA had high affinities for conjunctival goblet cells and the apical epithelial cell layers. Native WGA stained mucus and glycocalyx at the ocular surface. This staining of the ocular surface by WGA was confirmed at the transmission electron microscopic level using WGA conjugated to ferritin. Cicatricial pemphigoid patients in this study had reduced numbers of goblet cells; however, those goblet cells which were observed in cicatricial pemphigoid conjunctiva stained positively with HPA, PNA, WGA, and SWGA as did goblet cells in normal conjunctiva.

Conjunctiva↗
PubMed1978

Malignant melanoma of the conjunctiva and the iris in a case of primary acquired melanosis of the conjunctiva.

A case of primary acquired melanosis of the conjunctiva is reported. During the 18 years of observation the patient developed a malignant melanoma in the iris, and seven years later a malignant melanoma appeared in the lower lid. The iris melanoma may be an extension from the precancerous melanosis, but the most likely explanation is that the condition represents two independent diseases.

Aged↗
PubMed2003

Ag-specific recognition, activation, and effector function of T cells in the conjunctiva with experimental immune-mediated blepharoconjunctivitis.

PURPOSE: To investigate antigen (Ag) specificity, activation, and effector function of the Ag-specific T cells involved in the development of experimental immune-mediated blepharoconjunctivitis (EC), an experimental conjunctivitis. METHODS: EC was induced in Brown Norway rats by injection of ovalbumin (OVA)-specific T cells followed by OVA challenge with eye drops. Eyes, including the conjunctivas, were harvested at different time points after challenge. The dependence of EC onset on the challenging Ag was assessed by challenge with an irrelevant Ag or stimulatory OVA peptides. To show the infiltration of transferred T cells into the conjunctiva, T cells were labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) before transfer. The activation of T cells in the conjunctiva was assessed by measuring phosphorylation of Lck-associated molecules by Western blot analysis. Conjunctivas were also examined by immunohistochemistry and used for reverse transcription-polymerase chain reaction to determine the phenotype of the infiltrating cells and cytokine, chemokine, and chemokine receptor expression. To investigate infiltration of non Ag-specific T cells into the conjunctiva, ragweed (RW)-primed lymphocytes were transferred into OVA-specific T-cell receptor transgenic (DO11.10) mice. The mice were then challenged with RW and the conjunctivas were harvested for immunohistochemistry to detect T cells derived from DO11.10 mice. RESULTS: EC was induced only when challenged with OVA protein or stimulatory OVA peptides, and CFSE-labeled transferred cells were found in the conjunctiva. Phosphorylation of Lck and an 85-kDa Lck-associated molecule were observed in the conjunctiva 6 hours after challenge. Many cytokines and chemokines began to be expressed at 6 hours, and individual expression patterns over time correlated well with the infiltration patterns of different inflammatory cells. In DO11.10 mice that received RW-primed lymphocytes, T cells derived from the recipient mice infiltrated the conjunctiva after RW challenge. CONCLUSIONS: Ag-specific T cells initiate EC by first infiltrating the conjunctiva, where they become activated by the specific Ag in the conjunctiva.

Animals↗
PubMed2005

Interleukin-4-mediated infiltration of eosinophils into the conjunctiva and its suppression by interferon-gamma.

PURPOSE: Interleukin (IL)-4 is a T helper (Th)2 cytokine that plays an important role in the development of allergic reactions. It has been suggested that IL-4 is responsible for the infiltration of eosinophils into the conjunctiva during the development of allergic conjunctivitis. However, it is still unclear whether IL-4 is able to induce this eosinophilic infiltration on its own. We investigated whether subconjunctival injection of IL-4 can induce eosinophils to infiltrate into the conjunctiva. METHODS: Brown Norway rats were subconjunctivally injected with IL-4, eotaxin, or phosphate buffered saline (PBS), and the conjunctivas were harvested for histologic analysis (including immunohistochemistry) 6, 12, 18, and 24 hr later. The harvested conjunctivas were also subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to examine chemokine expression. In a separate experiment, the effect of coadministering interferon-gamma (IFN-gamma) along with IL-4 was examined. RESULTS: The subconjunctival injection of IL-4 induced eosinophil infiltration into the conjunctiva in a dose-dependent manner. IL-4 was as potent as eotaxin. The eosinophilic infiltration started 6 hr after the injection and persisted for up to 24 hr after the injection. Other infiltrating cell phenotypes were noted but were also observed in conjunctivas injected with PBS alone. RT-PCR analysis demonstrated that IL-4 injection did not upregulate chemokine RNA expression in the conjunctiva. Coinjection of IFN-gamma suppressed the infiltration of eosinophils into the conjunctiva induced by IL-4 injection. CONCLUSIONS: The subconjunctival injection of IL-4 specifically induces eosinophils to infiltrate into the conjunctiva. In addition, IFN-gamma in the conjunctiva can counteract this effect of IL-4.

Animals↗
PubMed1999

Detection and identification of Mycoplasma conjunctivae in infectious keratoconjunctivitis by PCR based on the 16S rRNA gene.

A specific PCR assay based on unique sequences of the rrs genes (16S rRNA) of Mycoplasma conjunctivae was developed for direct detection and identification of this pathogen from clinical material. DNA from eye swabs was amplified after a simple lysis step by either a single PCR with the M. conjunctivae specific primer pair McoR1 and McoF1, or by a nested PCR with the Mycoplasma genus specific primer pair MOLIGEN1-L and 16UNI-R in the first step and McoR1 and McoF1 in the second step. The specificity of the primer pair McoR1 and McoF1 was verified with purified DNA from the type strain, from 17 field isolates of M. conjunctivae and from several Mollicutes which are phylogenetically related to M. conjunctivae or which can be isolated from the same host animals. This method identified mycoplasma isolates from goat, sheep, ibex and chamois originating from different countries as M. conjunctivae. No cross amplifications with other mycoplasmas which are related to M. conjunctivae were observed. Eye swab samples containing known numbers of M. conjunctivae cells were analysed after direct lysis of the material. The detection level was estimated to be 20 cells per swab when the nested PCR procedure was used and 2 x 10(5) by the single PCR method. In an experimental infection model of sheep, the nested PCR method for detection of M. conjunctivae gave results which were comparable to mycoplasmal culture. These are the implications for diagnostic purposes: M. conjunctivae isolates can be identified by the one-step PCR method, whereas for detection and identification of M. conjunctivae in clinical material the two-step method should be used (higher sensitivity).

Animals↗
PubMed2001

Sensations evoked by selective mechanical, chemical, and thermal stimulation of the conjunctiva and cornea.

PURPOSE: To study the sensations evoked by selective mechanical, chemical, and thermal stimulation of the conjunctiva and compare them with those elicited by similar stimulation of the cornea. METHODS: Six young subjects participated in the study. Using a gas esthesiometer, selective mechanical (air puffs at flows from 0 to 264 ml/min), chemical (0--80% CO(2) in air), and thermal (air at temperatures from -10 degrees C to +80 degrees C) stimulation was performed on the center of the cornea and on the temporal conjunctiva. The intensity, degree of irritation, stinging and burning pain components, and thermal characteristics of the evoked sensation were evaluated after each stimulus in separate, 10-cm continuous visual analogue scales (VASs). The ability of the subjects to identify the quality of the stimulus applied to the cornea and the conjunctiva was also studied. RESULTS: The subjective intensity and thermal components (cooling or warming) of the sensation reported after mechanical, chemical, and heat stimulation were similar in the conjunctiva and cornea, although lower VAS scores were always reported in the conjunctiva for the irritation and the stinging and burning pain components. In the cornea, stimulation with low temperatures was perceived as a cooling sensation with an irritative component. In the conjunctiva, cooling was perceived as a purely cold sensation. Subjects showed similar discrimination capability in the cornea and the conjunctiva for the various types of stimuli. CONCLUSIONS: Sensations evoked in the cornea by selective mechanical, chemical, and heat and cold stimulation always presented an irritation component. In the conjunctiva, stimuli of the same intensity are always perceived as less irritating than in the cornea. Cold and other non-noxious subqualities of sensation can be evoked in the conjunctiva.

Adult↗
PubMed1975

Orthotopic transplantation of alogeneic canine conjunctiva.

In previous papers it has been shown that simultaneous transplantation of conjunctiva to the corneal-scleral region, and skin to the inside of the pinna will afford good opportunities to follow both the onset and the terminal phase of the allograft reaction. It has also been stated that conjunctiva grafted alone will have a longer survival time (12.1 days), than when grafted simultaneously with skin (7.5 days). In order to find out whether the localization to the corneal-scleral region per se had anything to do with this prolongation of the survival time, conjunctiva was grafted in 8 dogs to the palpebral surface of the nictitating membrane, i.e. more peripherally in the type used as grafts in previous experiments were weighed and examined histologically to compare the "tissue doses" --quantitatively and quanlitatively--transferred to the recipient when transplanting conjunctiva plus skin, or conjunctiva alone. Conjunctiva alone grafted to the nictitating membrane had a mean survival time of 10.5 days which does not significantly differ from the mean survival time of conjunctiva grafted to the cornealscleral region. Various possible reasons for the significantly prolonged survival time of conjunctiva alone, compared with that of conjunctiva when grafted with skin, are discussed in some detail. The author suggests that the most likely explanation of the prolonged survival time of the conjunctival grafts applied alone is that this operation means transfer of an antigen dose so low that the mobilization of the immune defence system is delayed.

Animals↗
PubMed2005

In vivo findings of the bulbar/palpebral conjunctiva and presumed meibomian glands by laser scanning confocal microscopy.

OBJECTIVE: To report in vivo laser scanning confocal microscopic findings of the normal human conjunctiva and presumed meibomian glands and to investigate the potential application of this confocal microscope (HRT II Rostock Cornea Module) as a diagnostic device for normal and abnormal ocular surface structures. METHOD: Four healthy volunteers (51-, 49-, 31-, and 30-year-old men) participated in this study. The temporal bulbar conjunctiva, approximately 5 mm away from the limbus, and the upper tarsus conjunctiva of each patient were examined in vivo by a laser confocal microscope. Also, presumed meibomian glands beneath the tarsal conjunctiva were examined. RESULTS: In all subjects, the in vivo laser confocal microscope obtained similar images. In the superficial epithelial cell layer of the bulbar conjunctiva, numerous bright nuclei were observed. We also observed many Langerhans cells with characteristic dendritic morphology and goblet cells with relatively homogeneous brightness in the bulbar conjunctiva. Most interestingly, the palpebral conjunctival epithelium with invaginations or crypts was observed. In addition, weblike structures, presumably meibomian glands, were observed beneath the palpebral conjunctival epithelium. CONCLUSIONS: These results indicate that in vivo laser confocal microscopy can be used not only to visualize the bulbar conjunctiva but the upper tarsus conjunctiva and possibly subconjunctival meibomian glands. Further investigations of conjunctival and meibomian gland pathologies may further elucidate the potential of this device in clinical ophthalmology.

Adult↗
PubMed2002

Role of enhanced expression of m-CSF in conjunctiva affected by cicatricial pemphigoid.

PURPOSE: Local proliferation of macrophages has been reported to augment the inflammatory response in various human and experimental diseases. Macrophage accumulation in the submucosa is also an important feature in the pathogenesis of ocular cicatricial pemphigoid (OCP). In the present study, the role of local proliferation of macrophages in conjunctiva affected by OCP and the relationship between local proliferation of macrophages and expression of macrophage-colony-stimulating factor (m-CSF) in such conjunctiva were examined. METHODS: Biopsy specimens from the conjunctiva of 10 untreated patients with active OCP and from 5 normal subjects were studied for the expression of m-CSF, macrophages, and proliferating cell nuclear antigen (PCNA), a cell cycle protein, by immunohistochemistry. Dual staining for CD68 (a cell surface marker for macrophages) and PCNA was also performed to identify proliferating macrophages. In addition, fibroblasts isolated from conjunctiva of normal individuals and from patients with OCP were studied for the expression of m-CSF by immunostaining and real-time PCR. To identify the factors that induce m-CSF in conjunctival fibroblasts, the fibroblasts were incubated with different concentrations of interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha, and the levels of m-CSF mRNA were determined by real-time PCR and the amount of m-CSF produced was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Normal conjunctiva showed weak expression of m-CSF in the conjunctival epithelial cells and stroma. Conjunctival expression of m-CSF protein was significantly (P < 0.0001) increased in conjunctival biopsy specimens from patients with OCP. m-CSF was detected in the infiltrating macrophages, stromal cells (presumably fibroblasts), and conjunctival epithelial cells. Compared with normal control conjunctival tissue, a 1.2-fold increase in the expression of mRNA for m-CSF was detected by real-time PCR in the conjunctival tissue obtained from patients with OCP. Increased expression of m-CSF correlated significantly (P < 0.0004) with an increased stromal accumulation of macrophages in conjunctival biopsy specimens of patients with OCP. A number of these accumulated macrophages (CD68-positive) were found to be proliferating (PCNA-positive). In addition, fibroblasts isolated and cultured from conjunctiva of patients with OCP showed significantly increased (1.7-fold) expression of m-CSF compared with normal conjunctival fibroblasts. When conjunctival fibroblasts were treated with IL-1alpha or TNF-alpha, real-time PCR and ELISA detected an increased level of m-CSF. CONCLUSIONS: An increased expression of m-CSF was observed in conjunctiva from patients with active OCP. There was a positive correlation between expression of m-CSF and accumulation of macrophages in conjunctival biopsy sections obtained from patients with OCP. Increased expression of m-CSF, mainly by conjunctival fibroblasts and infiltrating inflammatory cells, may play an important role in the regulation of local proliferation of macrophages in OCP. In the conjunctiva of patients with OCP, this process could augment or enhance the local inflammatory response and tissue injury consequent to it.

Antigens, CD↗
PubMed1996

[Laser Doppler flowmetry of the bulbar conjunctiva as a monitor of the cerebral blood flow].

In order to develop a non-invasive monitor of cerebral blood flow during aortic arch surgery, reliability of the laser Doppler flowmetry of the bulbar conjunctiva was evaluated. The blood flow of the bulbar conjunctiva may represent the cerebral blood flow because the ophthalmic artery originating from the internal carotid artery supplies the blood to the bulbar conjunctiva. A laser Doppler flowmeter (Advance Co., ALF201) and a specially designed probe was prepared to measure the blood flow of the bulbar conjunctiva. The probe was designed to stay in a fixed position on the bulbar conjunctiva when the eye lid is closed, so that it can maintain a stable measurement. The shape of probe is a small box, 7.5 x 4.0 x 2.0 mm connected to a glass fiber cable 2 mm in diameter, as the signal to convey to the flowmeter. Eight mongrel dogs were used for the experimental study. The brachiocephalic vessels were exposed to measure the pressure and the blood flow of the carotid arteries. The blood flow of the cerebral cortex and the bulbar conjunctiva was measured by the laser Doppler flowmetry. When the carotid artery was clamped, the blood flow of the bulbar conjunctiva fell instantaneously in 1.35 +/- 0.85 seconds and reached the plateau in 9.60 +/- 4.15 seconds. The flow returned to the previous value in 7.70 +/- 3.75 seconds after the clamp was removed. A significant positive correlation between the blood flow of the bulbar conjunctiva and that of the cerebral cortex was observed (r = 0.78, p < 0.001, n = 2). When the extracorporeal circulation was used, the waveform of the bulbar conjunctival blood flow was synchronized to the small pulse wave generated by the pump. Although the measurement of the bulbar conjunctival blood flow by laser Doppler flowmetry is an indirect method of monitoring the cerebral blood flow, it can provide non-invasive, continuous, and reliable measurement. We conclude that this is a useful method to monitor the cerebral blood flow during aortic arch surgery.

Animals↗
PubMedDate not supplied

Cyclosporin A inhibits eosinophilic infiltration into the conjunctiva mediated by type IV allergic reactions.

BACKGROUND: Eosinophils are important effector cells in severe allergic conjunctivitis such as vernal keratoconjunctivitis. Infiltration of eosinophils into the conjunctiva is mediated by type I and type IV allergic reactions. Cyclosporin A (CsA) eye drops are administered therapeutically for severe allergic conjunctivitis, but the mechanism by which CsA acts, that is, by inhibiting type I, type IV or both types of allergic reactions, is not known. We investigated whether CsA eye drops inhibit type I, type IV or both types of allergic reactions in the conjunctiva. METHODS: Experimental immune-mediated blepharoconjunctivitis (EC) was induced in BALB/c mice by either active immunization or passive immunization by transfer of ragweed (RW)-primed splenocytes and RW-specific IgE, followed by RW challenge to the conjunctiva. These mice were treated in eye drops with vehicle, 0.1% CsA, 0.5% CsA or 0.1% betamethasone five times (1 and 2 h before RW challenge and 1, 2 and 3 h after RW challenge). Twenty-four hours after the challenge, the conjunctivas were harvested for histological analysis to evaluate eosinophilic infiltration. To evaluate effects of CsA eye drops on systemic immune responses, sera and spleens were collected from actively immunized mice at the time of sacrifice to examine serum IgE levels and cellular immune responses, respectively. RESULTS: CsA eye drops significantly inhibited eosinophilic infiltration into the conjunctiva in actively immunized EC-developing mice compared with vehicle-treated mice. The CsA-induced inhibition was similar to inhibition induced by 0.1% betamethasone. Serum IgE levels and splenocyte responses in CsA-treated mice were equivalent to those in vehicle-treated mice. Betamethasone treatment inhibited eosinophilic infiltration into the conjunctiva induced by both splenocyte transfer and IgE transfer, while CsA treatment inhibited infiltration induced by splenocyte transfer. CONCLUSIONS: CsA eye drops inhibited eosinophilic infiltration into the conjunctiva without affecting systemic immune responses. CsA predominantly inhibits eosinophilic infiltration by interfering with the type IV allergic reaction in the conjunctiva.

Animals↗