Conjunctiva Of Patients Research Articles

A comparison of the effects of propofol and thiopental on tear production in dogs

The tear film plays an important role in maintaining the integrity of the ocular suface. During general anesthesia, tear production is considerably reduced, which requires care to prevent adverse effects that result in diseases of these structures. Studies comparing the effects of induction of anaesthesia with thiopental and propofol on tear production have not been carried out yet. Because these drugs are used in veterinary medicine, we decided to evaluate the tear production in 30 dogs undergoing experimental surgery as well as routine procedures at the veterinary hospital of Federal University of Viçosa. Patients were divided into two groups of equal number. All animals were sedated with clorpromazine and maintained with isoflurane in diluted oxygen. Group 1 was induced with thiopental whereas group 2 with propofol. Schirmer tear test 1 was performed before sedation (T0), 15 minutes after sedation (T1) and 10 minutes after induction of anesthesia (T2) with the drug chosen for one of the groups. There was a significant decrease in tear production for both drugs, but no significant statistical differences were found between them. Thus, considering the results and the way in which the study was conducted, we suggest protecting the cornea and conjunctiva of patients during anesthesia using any of the drugs here evaluated.

Expression of double strand DNA breaks repair genes in pterygium

Purpose: The alteration in the expression of some genes and proteins responsible for chosen DNA repair pathways in pterygium pathogenesis were studied. This study was focused on the examination of the expression of genes and RAD50 protein taking part in homologous recombination.Methods: Peripheral blood lymphocytes, samples of pterygium tissue, samples of conjunctiva of patients suffering from pterygium as well as peripheral blood lymphocytes and conjunctiva of patients from the control group were examined. In order to identify genes products from RNA, Ribonuclease Protection Assai method was applied. LIM15, RAD50, RAD54, RAD52, MRE11, XRCC2, XRCC3, RAD51, RAD51B, RAD51C, RAD51D genes transcripts were detected. Expression of RAD50 protein was analyzed immunohistochemically.Results: Peripheral blood lymphocytes analyses revealed lower level of RAD50 gene expression in the pterygium patients compared to the control group and the increased expression of XRCC2, XRCC3 and RAD51 genes in patients with pterygium, who declared the recurrence of the lesion in comparison to the patients with primary pterygium. Lower expression of the RAD54 gene in pterygium tissue comparing to conjunctiva from the eyes with pterygium was found. An expression of RAD50 gene in the conjunctiva originating from eyes with pterygium in comparison to the conjunctiva of control group was shown to be considerably higher. Expression of RAD50 protein in pterygium squamous epithelial cells was significantly higher than in conjunctiva from control group.Conclusion: There may exist a relationship between pterygium pathogenesis and damages of double strand DNA, however, the elucidation of its exact nature needs further study.

T Cell-Related Costimulatory Molecules in the Conjunctiva of Patients with Severe Allergic Conjunctivitis

We showed previously that costimulatory molecules such as OX40 and 4-1BB participate in the development of murine allergic conjunctivitis (AC). We investigated whether these molecules are expressed in the conjunctiva of patients with severe AC. Giant papillae were collected from 4 patients with severe AC. As a control, bulbar conjunctivas from 4 normal volunteers were used. Frozen sections of the conjunctivas were stained with anti-CD4, anti-CD8, anti-OX40, anti-4-1BB, or control antibodies (Abs) and then incubated with the biotinylated secondary Ab. Alternatively, staining was performed with a biotinylated anti inducible costimulator (ICOS) Ab. After using an avidin-biotin complex kit, the staining levels were graded from 0 to 3. For immunofluorescence analysis, conjunctival sections were stained with primary Abs that had been fluorescently labeled. The conjunctival stroma of all 4 patients with severe AC were infiltrated with CD4+ and CD8+ cells, whereas only 1 control sample contained stromal CD4+ and CD8+ cells. Conjunctivas from patients but none of the control conjunctivas expressed OX40, 4-1BB, and ICOS. The expression levels of these molecules were significantly higher in patients than the control volunteers. Immunofluorescence analysis demonstrated that the OX40 and 4-1BB, but not the ICOS, molecules were expressed by the infiltrating CD4+ and/or CD8+ cells. Together with the data in murine AC, OX40 and 4-1BB are expressed in the conjunctiva of patients with severe AC but not in the control conjunctivas, which suggests that OX40 and 4-1BB participate, at least in part, in the development of severe AC.

Corneal and Conjunctival Sensitivity in Patients With Dry Eye: The Effect of Topical Cyclosporine Therapy

: To evaluate changes in mechanical sensitivity of cornea and conjunctiva in patients with dry eye disease unresponsive to artificial tears therapy and to investigate the effect of topical cyclosporine therapy on sensitivity of the ocular surface. : Thirty-seven patients with dry eye disease and 35 healthy control subjects were enrolled to this prospective study. All patients included in the study completed a 3-month run-in period of using nonpreserved artificial tear supplements, but all failed to achieve adequate subjective or objective improvement. Patients were then instructed to use topical cyclosporine A 0.05% twice a day. Pre- and posttreatment (1, 3, and 6 months) evaluations included, corneal and conjunctival sensitivity testing with the Cochet-Bonnet esthesiometer, subjective symptom scoring, fluorescein and lissamine green staining, Schirmer test, and tear breakup time (BUT). : Conjunctival and corneal sensitivities were significantly lower in patients with dry eye compared with controls (P < 0.0001). In patients with dry eye, corneal sensitivity correlated positively with conjunctival sensitivity and negatively with ocular surface staining scores. Conjunctival sensitivity correlated negatively with the duration of dry eye disease, the total symptom severity score, and the severity of dryness symptom and positively with Schirmer test and tear BUT. Corneal and conjunctival sensitivities did not change significantly after artificial tear therapy (P > 0.05). After topical cyclosporine, statistically significant improvements from baseline were observed in corneal sensitivity at 3- and 6-month visits (P < 0.001). For conjunctival sensitivity, statistically significant improvements from baseline were seen at all follow-up visits (P < 0.0001). Topical cyclosporine treatment also led to significant improvements in symptom scores, Schirmer test, tear BUT, and ocular surface staining scores (P < 0.0001). : The mechanical sensitivity of cornea and conjunctiva to tactile stimulus is reduced in patients with dry eye. Our findings suggest that topical cyclosporine may be effective in improving this reduced mechanical sensitivity of the ocular surface.

Liquid Nitrogen Cryotherapy of Conjunctival Lymphangiectasia: A Case Series

Purpose: To report a case series of conjunctival lymphangiectasia treated with liquid nitrogen cryotherapy. Methods: A 1.5-mm Brymill cryoprobe was applied in a double freeze-thaw method after an incisional biopsy of a portion of the conjunctiva in patients with conjunctival lymphangiectasia. Freeze times were 1 to 2 seconds with thawing of 5 to 10 seconds between treatments. Patients were reexamined at 1 day, 2 weeks, 3 months, 6 months, and yearly following cryotherapy. Results: Five eyes of 4 patients (3 male and 1 female) with biopsy-proven conjunctival lymphangiectasia underwent liquid nitrogen cryotherapy. The average patient age was 53 years. Ocular examination revealed large lymphatic vessels that were translucent and without conjunctival injection. Subjective symptoms included epiphora, ocular irritation, eye redness, and occasional blurred vision. After treatment with liquid nitrogen cryotherapy, the patients’ symptoms and signs resolved within 2 weeks. Lymphangiectasia recurred twice in one patient, at 1 and 3 years postoperatively. In another patient, lymphangiectasia recurred at 6 months. The average time to recurrence in these 3 eyes was 18 months. Average length of follow-up was 24.5 months for all subjects. Conclusion: Liquid nitrogen cryotherapy may be an effective surgical alternative in the treatment of conjunctival lymphangiectasia. Cryotherapy may need to be repeated in some instances. Trans Am Ophthalmol Soc 2009;107:229-233

Immune diagnosis of pure ocular mucous membrane pemphigoid: indirect immunofluorescence versus immunoblot*

Diagnosis of pure ocular mucous membrane pemphigoid (OMMP) is based on the demonstration of linear depositions of IgG, IgA, C3 or combinations thereof alongside the epithelial basal membrane zone with direct immunofluorescence (DIF) or immunoperoxidase (DIP) analysis of a freeze biopsy of the conjunctiva in patients with cicatrizing kerato-conjunctivitis. The aims of this study were: 1) to investigate whether the direct immunoperoxidase method is more sensitive than direct immunofluorescence; 2) to analyse whether circulating antibodies against basement membrane autoantigens can be detected in DIF positive and DIF negative patients and which method is the most sensitive for this. In 11 patients with exclusively cicatrizing keratoconjunctivitis, biopsies were evaluated with DIF and DIP. Serum was investigated with indirect immunofluorescence and immunoblot. Immunofluorescence analysis revealed linear IgG and/or IgA depositions in five out of 11 conjunctival biopsies. These depositions were less intense with immunoperoxidase analysis. Serum immunofluorescence analysis was positive in only one case (20%); immunoblot analysis for circulating antibodies was positive in 4 of 5 patients (80%) with pure OMMP. With the methods used in this study: a) the immunoperoxidase analysis was no more sensitive than direct immunofluorescence, and b) the immunoblot was more sensitive in demonstrating circulating antibodies than indirect immunofluorescence.

Immunohistochemical Expression of HLA-DR in the Conjunctiva of Patients Under Topical Prostaglandin Analogs Treatment

Subclinical inflammation may be observed in patients using topical antiglaucomatous drugs. The objective of this study was to investigate inflammation in conjunctiva of glaucoma patients using prostaglandin analogs, by the detection of an immunogenetic marker (HLA-DR) and compare the effect of 3 different drugs: latanoprost, bimatoprost, and travoprost in the induction of this inflammation. Thirty-three patients with primary open-angle glaucoma were evaluated without and with prostaglandin analogs topical therapy. Imprints of conjunctival cells were obtained, fixed on glass slides, and prepared for immunohistochemical analysis. Before the use of prostaglandin analogs, 4 of the 33 patients evaluated presented expression of HLA-DR in the conjunctiva (mild). After 1 month on prostaglandin analog treatment, all but 1 patient presented HLA-DR staining. HLA-DR expression of these 32 patients was scored as mild (19 patients), medium (11 patients), or intense (2 patients). The differences were statistically significant both when the presence and the increased expression of HLA-DR were considered (P<0.001). When the 3 different groups were analyzed (latanoprost, bimatoprost, and travoprost) no statistically significant difference was found (P=0.27). The use of prostaglandin analogs eye drops provokes a subclinical inflammatory reaction, observed by HLA-DR expression, even after a short period of treatment, independently of the class of the prostaglandin analogs used.

Glycogene Expression in Conjunctiva of Patients with Dry Eye: Downregulation of Notch Signaling

Glycoconjugates regulate a variety of biological events in mucosal surfaces, such as differentiation of postmitotic epithelial cells and maintenance of the wet-surfaced phenotype. This study aimed to identify the repertoire of genes (glycogenes) involved in biosynthesis of glycoconjugates in conjunctiva of normal subjects and patients with dry eye. RNA from conjunctival impression cytology samples was amplified and hybridized to a custom-designed glycogene microarray. Intensity data were converted to expression values and analyzed by ANOVA. Microarray data for selected Notch glycogenes were confirmed by quantitative real-time PCR. Notch receptors and ligands were immunolocalized on conjunctival biopsies by confocal microscopy. By microarray, 424 glycogenes were identified in normal conjunctival epithelium; galectins, glycosyltransferases, mucins, Notch signaling molecules, and proteoglycans were among the most highly expressed. In dry eye, 46 glycogenes were significantly downregulated, including five members of the Notch signaling pathway (Notch1, Notch 2, Notch 3, Jagged1, Delta1), four Wnt signaling molecules (Wnt4, -5A, Frizzled6, -7), and three heparan sulfate glycotransferases (HS2ST1, HS3ST6, EXTL2). Only interferon-induced transmembrane protein 1 was upregulated. By real-time PCR, expression ratios of Notch1, Notch 3, and Jagged1 in dry eye were 0.43, 0.56, and 0.50, respectively, compared to controls (P < 0.05). Notch1, Notch3, and Jagged1 were immunolocalized throughout the conjunctival epithelium, whereas Notch2 and Delta1 were distributed apically. This study revealed the differential glycogene expression profiles in normal subjects and patients with dry eye. Downregulation of Notch signaling in dry eye may result in abnormal differentiation of the conjunctival epithelium and have implications in the pathogenesis of the disease.

Infection control audit and potential sources of infection of a Nigerian Eye Unit

The prevalence of post-surgical eye infections in Nigeria has not been documented. However, anecdotal reports suggest that post-operative endophthalmitis is fairly common in Lagos. This study was done to investigate the sources of post-operative eye infections and the preparedness of the hospital to prevent such infections. A bacteriological survey of all eye surgical procedures between September 2004 and June 2005 was performed. Relevant samples were cultured pre- intra and post operatively as required. In the eye clinic, and operation theatre, infection control procedures and practices were audited using a pre-designed questionnaire. Out of 207 procedures performed, there was no case of post -operative eye infection but potential sources of infections identified included nares of staff, surgical packs, theatre bed, hands of surgeons and conjunctivae of patients. Running water and soap were available for hand washing. There was no specific provision for drying hands in the clinic and staff were observed to carry out hand wash considered ineffective. Staff showed good knowledge of disinfectants and antiseptic use. In the theatre, the staff who handle the steam sterilizer were not formally trained on its use. Although sterile packs were appropriately stored they were not dated. Appropriate types of waste bins were not in use and wastes were not segregated appropriately. Ophthalmic equipment were not adequately disinfected in between patients and use of eye drops for dilatation was communal. Potential sources of post-operative eye infections were identified. Areas of deficiencies in infection control practices, which require proper infection control policies, include handwashing facilities and practices, sterilization procedures, disinfection of ophthalmic equipment, linen management and waste handling.

Normal Conjunctival Flora

Objective: To determine the prevalence of normal conjunctival flora and its types. Study Design: An observational study. Place And Duration of Study: At Ophtha-lmological Department, DHQ/Allied Hospital, PMC, Faisalabad, from November 2006 to February 2007. Patients And Methods: This study was performed on 500 patients, aged form 20 years to 70 years with normal looking eyes. The samples of sterile stuart’s swab were made on conjunctiva of patients, without touching eyelashes or lid margins. The swabs were touched with conjunctiva of both eyes and placed in stuart’s transparent media to shift to microbiological laboratory. Results: Out of 500 swabs 412 (82.4%) showed no growth while 78 (15.6%) showed microbial growth. Out of these 78 swabs, there were 44 (56.41%) having normal flora, while 34 (43.59%) had other pathogens. Conclusion: The normal looking eyes harbour different types of the organisms. Any trauma, accidental hurt or elective surgical procedure is not free from risk of conjunctival infection, keratits or endophthalmitis etc.

Pathergy-Like Reaction Following Intravitreal Triamcinolone Acetonide Injection in a Patient with Behçet Disease

Purpose: To describe a patient with Behçet disease who developed a pathergy-like reaction following intravitreal triamcinolone acetonide injection. Methods: Case report. Results: A 30-year-old man with a history of Behçet disease received an intravitreal injection of triamcinolone acetonide for the treatment of cystoid macular edema. The following day, he was noted to have erythematous swelling around the injection site. The lesion disappeared spontaneously within 2 weeks. Conclusion: Pathergy-like reaction to minor trauma may develop not only on the skin but also in the mucous membranes such as conjunctiva in patients with Behçet disease.

Effects of Sequential Artificial Tear and Cyclosporine Emulsion Therapy on Conjunctival Goblet Cell Density and Transforming Growth Factor-β2 Production

To evaluate the effects of sequential treatment with artificial tears and cyclosporine emulsion on conjunctival goblet cell density and production of transforming growth factor (TGF)-beta2 in patients with dry eye disease. Patients with dry eye disease (N = 6) defined by an Ocular Surface Disease Index symptom score >or=25, Schirmer test 1 <10 mm, and corneal fluorescein and conjunctival lissamine green staining scores >or=3 were treated with artificial tears (Refresh Plus; Allergan, Irvine, CA) 4 times a day for 4 weeks, followed by 0.05% cyclosporine emulsion (Restasis; Allergan) twice a day for 12 weeks. Impression cytology was performed on the bulbar conjunctiva of both eyes at baseline, after artificial tear therapy, and after 6 and 12 weeks of cyclosporine therapy. Goblet cells were counted in 5 representative microscopic fields per membrane in those taken from the temporal and inferior bulbar conjunctiva of the worse eye, and membranes taken from the fellow eye were immunostained for TGF-beta2. There were no differences in mean goblet cell density between baseline and 4 weeks of artificial tears in the temporal and inferior bulbar specimens. After 6 weeks of cyclosporine emulsion, goblet cell density was significantly greater than baseline and artificial tears in the inferior bulbar conjunctiva (P < 0.01). After 12 weeks of cyclosporine emulsion, goblet cell density was significantly greater than baseline and artificial tears in both temporal and inferior bulbar sites (P < 0.01). The number of TGF-beta2-positive goblet cells was also noted to increase after 6 and 12 weeks of cyclosporine therapy (P < 0.001). Cyclosporine emulsion, but not artificial tears, increases goblet cell density and production of the immunoregulatory factor TGF-beta2 in the bulbar conjunctiva in patients with dry eye.

Immunohistochemical findings in prosthesis‐associated giant papillary conjunctivitis

To identify functional subsets of inflammatory cells and expression of cytokines in the conjunctiva of patients with ocular prosthesis-associated giant papillary conjunctivitis (P-GPC). Specific immunohistochemical staining using monoclonal antibodies was performed on biopsy specimens obtained from superior tarsal conjunctiva of 18 patients with P-GPC. The prosthetic eyes were taken as the study group, whereas their fellow eyes were used as matched controls. In normal conjunctiva, mast cells (MCs) were located only in the substantia propria (SP), whereas in P-GPC eyes MCs were also notable in the epithelium in five specimens. Tryptase-chymase-positive MCs (MC(TC)) were predominant both in P-GPC (79%) and in fellow (72%) eyes. MC(TC), CD4(+) lymphocyte, CD8(+) lymphocyte and eosinophil numbers were higher in P-GPC specimens compared with the fellow eyes (P = 0.005, 0.074, 0.012 and 0.025, respectively). Eosinophils were detected in 58.8% of P-GPC specimens and 16.7% of control specimens (P = 0.053). The number of inflammatory cells expressing eotaxin and interleukin (IL)-4 was higher in P-GPC group (P = 0.050 and 0.048, respectively). Nine out of 17 giant papillary conjunctivitis specimens (52.9%) showed eotaxin and IL-4 immunoreactivity, which was considerably higher than the fellow eyes (16.7%) (P = 0.064). These findings suggest that P-GPC is an allergic disease of the eye associated with increased numbers of MC, eosinophils and lymphocytes in the conjunctiva and a remarkable expression of IL-4 and eotaxin both by the conjunctival epithelium and by the inflammatory cells in the SP.

Conjunctival Changes in Children with Kawasaki Disease

To examine the cytopathological changes in the conjunctiva of patients with active Kawasaki disease.Case-control prospective study. Bilateral conjunctival swabs were obtained from 3 groups of children: patients with acute KD (11), age-matched controls (7), and patients with inactive KD (9). The ThinPrep prepared and Papanicolaou stained smears were examined blindly by 2 cytopathologists. The cell count differential of cells was performed and recorded quantitatively and comparisons between the 3 patient groups were made.Only neutrophil counts showed a significant difference among the 3 groups. The average scores for the acute KD, control group, and patients with inactive KD were 3.5, 1.6 and 1.3, respectively. Using the Pearson chi2 test, the difference between the acute KD and the inactive group was statistically significant for both eyes (right p = 0.049, left p = 0.004). Samples from acute KD patients were more cellular. Neutrophils surrounding conjunctival epithelial cells, or "neutrophilic rosetting", were seen in 4 (36%) cases of the active disease group but not in the other groups."Neutrophilic conjunctivitis" is characteristic in patients of acute KD that may be of value in the initial evaluation and subsequent follow-up of KD patients.

Isolated Muller's muscle resection for the correction of blepharoptosis

To assess the outcome of isolated Muller's muscle resection with preservation of conjunctiva in patients with blepharoptosis and good to moderate levator function. This study was designed as a prospective, nonrandomised case series. Thirty-four eyes of 27 blepharoptosis patients were operated on, who were phenylephrine test-negative as well as positive. Open-sky Muller's muscle resection was performed with preservation of the conjunctiva. Main outcome measures were increase in margin reflex distance (MRD1), eyelid contour, and symptoms and signs of dry eye. The mean increase in MRD1 was 2.75 mm. All but one patient (96%) had upper lid margins resting at or up to 1 mm below the limbus and obtained symmetry to within 0.5 mm of the fellow eye. No patients had symptoms or signs of dry eye. Isolated Muller's muscle resection is effective for the correction of ptosis in patients with moderate to good levator function. This is irrespective of the lid's response to phenylephrine. Preservation of conjunctival tissue eliminates concerns about dry eye, and also preserves the full height of the fornix.

Estudo da microbiota fúngica da conjuntiva ocular em portadores de hanseníase e seus comunicantes

To study the conjunctival fungal flora of patients with leprosy who live in the colony hospital and of those who deal with "Hansen" patients. Sixty-one eyes (61 patients) with Hansen's disease and twenty-five healthy eyes (25 individuals) of individuals who deal with patients were investigated. Conjunctival fungi growth occurred in 19.67% and 20% of the patients and healthy individuals, respectively. There was no significant difference (p<0.05) regarding fungi percentage of fungi isolated from the conjunctiva between both groups. Candida ssp was the most frequent fungus isolated from the conjunctiva of patients with Hansen's disease, whereas Penicillium spp was the most frequent fungus isolated from the group of people who deal with these patients. The most frequent yeast isolated from conjunctival samples of leprosy patients was Candida, which could be explained by cellular immunodeficiency of these patients, associated with dry eye and the prolonged use of antibiotics.

Conjunctival FOXP3 in Trachoma: T Cells Not Specified

The article by Faal et al. [1] describes differences in the level of FOXP3 mRNA expression in the conjunctiva of patients with Chlamydia trachomatis infection. The authors suggest that their results may indicate a role for regulatory T cells in the resolution of conjunctival immune response, since FOXP3 mRNA remained elevated even if clinical disease signs were present in the absence of infection. But suggestions about the function of regulatory T cells in diseases based only on the quantification of FOXP3 mRNA should be considered with great caution. According to the current state-of-the-art it is well established that FOXP3 does not specify human regulatory T cells since it is expressed at the mRNA and the protein levels to different extents protein at [2–5]. Thus, without quantification of FOXP3 the single cell level in conjunction with reliable markers of human regulatory T cells, e.g., the recently identified lack of CD127 expression on regulatory T cells [6,7], the potential contribution or association of regulatory T cells with a disease cannot be assessed.

Mooren Ulcer

To determine the pattern and distribution of mononuclear cells, adhesion, and co-stimulatory molecules in the conjunctiva of patients with Mooren ulcer. Conjunctival biopsy specimens were obtained from 6 patients with Mooren ulcer and 6 healthy individuals. Immunohistochemistry was performed on frozen sections of the cryopreserved human conjunctivas using monoclonal antibodies directed against CD1alpha, CD3, CD4, CD8, CD20, CD25, CD57, and CD68 cells; the adhesion molecules E-selectin, vascular cell adhesion molecule-1 (VCAM-1), very late activation-4 (VLA-4), ICAM-1, and LFA-1; and the co-stimulatory molecules CD28, B7-1, B7-2, and CTLA-4. Differences in expression on the conjunctival epithelium from patients with Mooren ulcer and normal subjects were noted only for VCAM-1, VLA-4, ICAM-1, and LFA-1. The ratio of CD4+/CD8+ cells in Mooren ulcer specimens was significantly higher (3.5-fold). However, in the substantia propria, Mooren ulcer specimens revealed significantly increased numbers of CD1alpha+, CD3+, CD4+, CD20+, CD28+, B7-1+, B7-2+, and CD68+ cells. The ratios of CD4+/CD8+ cells and B7-2+/antigen-presenting cells in Mooren ulcer specimens were significantly higher (5-fold). All tested adhesion molecules showed significant up-regulation in the patients' conjunctivas. Mooren ulcer vascular endothelial cells prominently expressed E-selectin, VCAM-1, VLA-4, and ICAM-1 compared with normal conjunctiva. The simultaneous presence of multiple types of inflammatory cells, adhesion, and co-stimulatory molecules in Mooren ulcer conjunctiva suggests that their interaction may contribute to a sustained immune activation as at least part of the pathogenic mechanism of this disorder.

Clearing of globotriaosylceramide from endothelial vessels of ocular conjunctiva in patients with Fabry disease treated with agalsidase alfa

Abstract Introduction: Fabry disease is an X‐linked inborn metabolic error caused by a deficiency of lysosomal α‐galactosidase A. Globotriaosylceramide (Gb3) accumulates in lysosomes of various cells. At the moment, enzyme replacement therapy (ERT) is the only available specific treatment for patients with Fabry disease. Enzyme infusions reduce the Gb3 content of plasma, urine sediment and tissue. In a previous study, we reported the development of a method to specifically detect Gb3 deposits by immunofluorescence in conjunctival biopsies. Aim: The aim of this work is to evaluate the clearance of Gb3 in cells from the conjunctiva in patients with Fabry disease treated with agalsidase alfa. Methods: Conjunctival biopsies were obtained from three hemizygous and two heterozygous patients, before and after 6 months of treatment with agalsidase alfa. The specimens were processed for direct immunofluorescence using an anti‐Gb3 monoclonal antibody and light microscopy using a mouse polyclonal antiserum specific for α‐galactosidase A. Results: Positive immunofluorescence was not observed in cells from the blood vessels of patients with Fabry disease after 6 months of treatment, as compared with the specimens before the treatment, which were positive. We detected Gb3 in stromal/macrophage cells that were distal from the blood vessels where the enzyme was circulating. Specific staining for α‐galactosidase A revealed the presence of this protein in the tissue of patients after ERT, but not before. Conclusion: Agalsidase alfa eliminated Gb3 deposits from the cells of blood vessel walls in patients with Fabry disease after 6 months of treatment. The deposits in stromal cells were not removed, probably because the enzyme must diffuse into the tissue to reach these cells and/or because of the short length of therapy. This method may be useful to evaluate the effectiveness of ERT.

Clearing of globotriaosylceramide from endothelial vessels of ocular conjunctiva in patients with Fabry disease treated with agalsidase alfa

Clearing of globotriaosylceramide from endothelial vessels of ocular conjunctiva in patients with Fabry disease treated with agalsidase alfa