Expression of cyclo-oxygenase-2 and downstream enzymes in diabetic fibrovascular epiretinal membranes.

BACKGROUND/AIMS: The inducible enzyme cyclo-oxygense-2 (COX-2) and its metabolic products are important mediators for angiogenesis. We investigated the expression of COX-2 and its downstream enzymes microsomal prostaglandin-E synthase (mPGES)-1, cytosolic PGES (cPGES) and thromboxane synthase (TXS), and correlated it with vascular endothelial growth factor (VEGF) expression and level of vascularisation in proliferative diabetic retinopathy (PDR) epiretinal membranes. METHODS: Membranes from five patients with active PDR and nine patients with inactive PDR were studied by immunohistochemistry. RESULTS: Vascular endothelial cells expressed COX-2, mPGES-1 and VEGF in 11, nine and six membranes, respectively. TXS was expressed in stromal cells in 12 membranes. There was no immunoreactivity for cPGES. There were significant correlations between number of blood vessels expressing CD34 and number of blood vessels expressing COX-2 (r(s) = 0.858; p<0.001), mPGES-1 (r(s) = 0.743; p = 0.002) and VEGF (r(s) = 0.845; p = 0.001) and number of cells expressing TXS (r(s) = 0.74; p = 0.002). The number of blood vessels expressing CD34 (p = 0.007), COX-2 (p = 0.027) and VEGF (p = 0.008) and stromal cells expressing mPGES-1 (p = 0.003), TXS (p = 0.04) and VEGF (p = 0.017) were significantly higher in active membranes than in inactive membranes. CONCLUSION: COX-2 and its metabolic products might contribute to PDR angiogenesis.

Expression of antiapoptotic and proapoptotic molecules in diabetic retinas.

PURPOSE: To investigate the expression of the antiapoptotic and proapoptotic markers in diabetic retinas. METHODS: In total, 12 donor eyes from six subjects with diabetes mellitus, and 10 eyes from five nondiabetic subjects without known ocular disease serving as control subjects were examined. Immunohistochemical techniques were used with antibodies directed against cyclooxygenase-2 (Cox-2), Akt (protein kinase B), Mcl-1, Bad, cytochrome c, apoptosis-inducing factor (AIF), tumour necrosis factor receptor-1-associated death domain protein (TRADD), and Fas-associated death domain protein (FADD). RESULTS: In retinas from all subjects without diabetes, cytoplasmic immunoreactivity for the antiapoptotic molecules Cox-2, Akt, and Mcl-1 was noted in ganglion cells. Cytoplasmic immunostaining for Cox-2 was also noted in the retinal pigment epithelial cells. Weak immunoreactivity for the mitochondrial apoptogenic proteins cytochrome c, and AIF was noted in the inner segments of photoreceptors, in the inner one-third of the outer plexiform layer, in cells in the inner nuclear layer, in the inner plexiform layer, and in ganglion cells. There was no immunoreactivity for the other antibodies tested. All diabetic retinas showed de novocytoplasmic immunoreactivity for Bad in ganglion cells, and in occasional cells in the inner nuclear layer. Upregulation of cytochrome cand AIF immunoreactivity was noted. Cox-2, Akt, and Mcl-1 immunoreactivity was not altered in the diabetic retinas. There was no immunoreactivity for TRADD, and FADD. CONCLUSIONS: Ganglion cells in diabetic and nondiabetic retinas express the antiapoptotic molecules Cox-2, Akt, and Mcl-1. Retinal ganglion cells express the proapoptotic molecule Bad in response to diabetes-induced neuronal injury. Diabetic retinas show upregulation of the mitochondrial proteins cytochrome c, and AIF.

Expression of growth factors in the conjunctiva from patients with active trachoma.

PURPOSE: The blinding complications of trachoma are associated with progressive conjunctival fibrosis due to excessive accumulation of extracellular matrix (ECM) components. We studied the processes involved in the regulation of fibrosis in trachoma by investigating the expression of the fibrogenic and angiogenic connective tissue growth factor (CTGF) and basic fibroblast growth factor (bFGF), the angiogenic vascular endothelial growth factor (VEGF), the angiogenesis-associated endothelial cell marker CD105 (endoglin), and the ECM protein tenascin in the conjunctiva. METHODS: Conjunctival biopsy specimens from six patients with active trachoma, and six control subjects were studied by immunohistochemical techniques using monoclonal and polyclonal antibodies directed against CTGF, bFGF, VEGF, CD105, and tenascin. RESULTS: In the normal conjunctiva, weak immunoreactivity for VEGF was observed in epithelial cells. There was no immunoreactivity for the other antibodies. In all trachoma specimens, immunoreactivity for CTGF and bFGF was localized in monocytes/macrophages, positive for the CD68 marker. Strong immunoreactivity for VEGF was observed in epithelial cells and on vascular endothelial cells. CD105 immunoreactivity was observed on vascular endothelial cells. Immunoreactivity for tenascin was noted in the upper substantia propria. CONCLUSIONS: These findings suggest that macrophages play an active role in conjunctival scarring, upregulated local production of CTGF, bFGF, and VEGF contributes to both fibrous tissue growth and angiogenesis, vascular endothelial cells are activated and are undergoing active angiogenesis, and deposition of tenascin reflect remodelling of the conjunctiva in trachomatous conjunctivitis.

Immunopathogenesis of conjunctival remodelling in vernal keratoconjunctivitis.

PURPOSE: To study the processes involved in mediating conjunctival remodelling in vernal keratoconjunctivitis (VKC) by investigating the expression of integrin receptors, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), transforming growth factor-beta(TGF-beta), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and Ki67 antigen, which is a marker for cell proliferation. METHODS: Conjunctival biopsy specimens from 16 patients with active VKC and nine control subjects were studied by immunohistochemical techniques using monoclonal and polyclonal antibodies directed against the integrin alpha3 and alpha6 subunits, EGFR, VEGF, TGF-beta, bFGF, PDGF, and Ki67 antigen. The phenotype of inflammatory cells expressing growth factors was examined by double immunohistochemistry. RESULTS: In the normal conjunctiva, very weak immunoreactivity was observed for EGFR and VEGF in epithelial cells, and for alpha3 and alpha6 integrin subunits on basal epithelial cells, and on vascular endothelial cells in the upper substantia propria. There was no immunoreactivity for the other antibodies. In VKC specimens, strong staining for alpha3 and alpha6 integrin subunits was observed on the membranes of basal and suprabasal epithelial cells, and all vascular endothelial cells. Immunoreactivity for Ki67 antigen was observed in the nuclei of the basal and suprabasal epithelial cells. Strong immunoreactivity was observed for EGFR in the deeper layers of the epithelium, and for VEGF in all epithelial cells. Inflammatory cells expressing EGFR, VEGF, TGF-beta, bFGF, and PDGF were noted in 8, 9, 11, 10, and 10 specimens, respectively. The majority of inflammatory cells expressing growth factors were eosinophils (45+/-4%) and monocytes/macrophages (35+/-4%). CONCLUSIONS: Chronic conjunctival inflammation in VKC is associated with increased staining of alpha3, and alpha6 integrin subunits, EGFR, VEGF, TGF-beta, bFGF, and PDGF that might mediate conjunctival remodelling.

Inducible nitric oxide synthase and vascular endothelial growth factor are colocalized in the retinas of human subjects with diabetes.

PURPOSE: Nitric oxide (NO) mediates vascular endothelial growth factor (VEGF)-induced angiogenesis and vascular hyperpermeability. This study was undertaken to study the cellular distribution of inducible nitric oxide synthase (iNOS) and VEGF in the retinas from human subjects with diabetes mellitus. In addition, glial reactivity and peroxynitrite generation were detected by immunolocalization of glial fibrillary acidic protein (GFAP) and nitrotyrosine, respectively. METHODS: Eight post-mortem eyes from four consecutive subjects with diabetes mellitus and eight eyes from four subjects without diabetes and without known ocular disease were prospectively collected and examined. We used immunohistochemical techniques and antibodies directed against iNOS, VEGF, GFAP, and nitrotyrosine. RESULTS: In retinas from all subjects without diabetes, weak GFAP immunoreactivity was confined to nerve fibre and ganglion cell layers. There was no immunoreactivity for iNOS, nitrotyrosine, and VEGF. All diabetic retinas showed GFAP induction in Müller cells and GFAP upregulation in nerve fibre and ganglion cell layers. All diabetic retinas showed cytoplasmic immunoreactivity for iNOS, and VEGF in ganglion cells, cells in the inner nuclear layer, and glial cells. In serial sections, ganglion cells and cells in the inner nuclear layer expressing VEGF were localized in the same area of iNOS-expressing ganglion cells and cells in the inner nuclear layer. Six retinas from three subjects with diabetes showed immunoreactivity for nitrotyrosine in vascular endothelial cells in inner retinal layer. CONCLUSIONS: iNOS and VEGF are colocalized in diabetic retinas. Increased GFAP immunoreactivity is a pathological event in the retina during diabetes.

Immuno-histochemical evaluation of conjunctival remodelling in vernal keratoconjunctivitis.

PURPOSE: To study the expression of the extracellular matrix (ECM) proteins, tenascin, laminin, and fibronectin in the conjunctiva of patients with active vernal keratoconjunctivitis (VKC). METHODS: Conjunctival biopsy specimens were obtained from nine patients with active VKC and 6 normal control subjects. The presence and distribution of tenascin, laminin, and fibronectin were assessed microscopically with immunohistochemical techniques and a panel of monoclonal and polyclonal antibodies directed against tenascin, laminin, and fibronectin. RESULTS: In normal conjunctiva, weak immunoreactivity for tenascin was localized to the walls of blood vessels in the upper substantia propria. Weak immunoreactivity for laminin was located at the epithelial-stromal junction and in the walls of blood vessels. Staining for fibronectin was absent. In VKC specimens, intense immunoreactivity for tenascin was noted in the substantia propria associated with the inflammatory infiltrate and in the perivascular stroma. Intense immunoreactivity for laminin around all stromal vessels and fibrillar immunoreactivity among basal epithelial cells were noted. There was no immunoreactivity for fibronectin. CONCLUSION: Our data indicate increased deposition of tenascin and laminin in the conjunctiva from patients with active VKC. Our findings suggest that tenascin and laminin might play distinct roles in chronic inflammation seen in VKC.

[Headaches in ophthamology]. / Les céphalées en ophtalmologie.

PURPOSE: To determine the frequency of ocular abnormalities in patients with headache seen for an ocular examination. METHODS: All 944 consecutive new patients with headache examined during a 3-year period (1984-1986) at the Department of Ophthalmology, University of Kinshasa, Democratic Republic of the Congo, were enrolled in this retrospective study. All patients received an ocular examination which included measurement of visual acuity, refraction, slit lamp examination, and fundoscopy. Other examinations such as measurement of intraocular pressure, neuro-ophthalmological examination, visual field and fluoangiography were performed when needed. RESULTS: The frequency of patients with headache examined during this period was 15.6% (944 out of 6,066 patients) or one patient out of 16 new patients. Their average age was 30+/-13.5 years. Females (60%) complained more frequently of headache than males (40%) (p<0.05). Migraine was found in 3.9% of patients. Ocular abnormalities were found in 505 (64%) patients and included refractive disorders (44%), lesions of the posterior segment (17%), abnormalities of the conjunctiva (12%), abnormalities of the anterior segment (12%), presbyopia (11%), and ocular motor palsies, heterophoria, convergence insufficiency and other ocular abnormalities (4%). Of 224 patients with ametropia, 154 (69%) were myopic with or without astigmatism. When specified, headache was most often fronto-occipital. Headache was most frequently associated with decreased vision, eye pain, epiphora, foreign body sensation, itching, and photophobia. CONCLUSION: Ocular examination could be necessary in cases of patients who complain of headache.

Expression of T lymphocyte chemoattractants and activation markers in vernal keratoconjunctivitis.

BACKGROUND/AIMS: T lymphocytes are present in increased numbers in the conjunctiva of patients with vernal keratoconjunctivitis (VKC) and their activation has a central role in the pathogenesis of the chronic allergic inflammatory reactions seen in VKC. The aims of this study were to examine the expression of three recently described potent T lymphocyte chemoattractants, PARC (pulmonary and activation regulated chemokine), macrophage derived chemokine (MDC), and I-309, the MDC receptor CCR4, and T lymphocyte activation markers, CD25, CD26, CD62L, CD71, and CD30, and to correlate them with the counts of CD3(+) T lymphocytes in the conjunctiva of patients with VKC. METHOD: Conjunctival biopsy specimens from 11 patients with active VKC, and eight control subjects were studied by immunohistochemical techniques using a panel of monoclonal and polyclonal antibodies directed against PARC, MDC, I-309, CCR4, CD25, CD26, CD62L, CD71, and CD30. The numbers of positively stained cells were counted. The phenotype of inflammatory cells expressing chemokines was examined by double immunohistochemistry. RESULTS: In the normal conjunctiva, vascular endothelial cells in the upper substantia propria showed weak immunoreactivity for CD26. There was no immunoreactivity for the other antibodies. VKC specimens showed inflammatory cells expressing PARC, MDC, and I-309. The numbers of PARC(+) inflammatory cells were higher than the numbers of MDC(+) and I-309(+) inflammatory cells and the mean values of the three groups differed significantly (17.0 (SD 10.1); 9.5 (9.9), and 4.3 (7.9), respectively, p = 0.0117, ANOVA). The numbers of PARC(+) inflammatory cells had the strongest correlation with the numbers of CD3(+) T lymphocytes. Few CCR4(+) inflammatory cells were observed in only three specimens. Double immunohistochemistry revealed that all inflammatory cells expressing chemokines were CD68(+) monocytes/macrophages. The numbers of CD25(+) T lymphocytes were higher than the numbers of CD26(+), CD62L(+), CD71(+), and CD30(+) T lymphocytes and the mean values of the five groups differed significantly (46.2 (27.9), 30.7 (16.0), 20.1 (8.6), 7.8 (7.7), and 6.5 (4.0), respectively, p <0.001, ANOVA). The numbers of CD25(+) T lymphocytes had the strongest correlation with the numbers of CD3(+) T lymphocytes. CONCLUSION: These results suggest a potential role for PARC, MDC, and I-309 in attracting T lymphocytes into conjunctiva in VKC. T lymphocytes in VKC are activated and express several activation markers which might contribute to the pathogenesis of VKC.

Ocular emergencies in Kinshasa (Democratic Republic of Congo).

PURPOSE: To determine frequencies of ocular emergencies and identify their nature. DESIGN: Observational case series. METHODS: In a retrospective study, the records of all 118 consecutive patients seen in emergency room during an eleven-month period were reviewed. RESULTS: Ocular emergencies represented 4% of the 2917 new patients visiting the department of Ophthalmology during this time. There was a 2.1/1 male to female preponderance and a peak age of presentation between 11 and 30 years. The mean age was 26 years +/- 17. Ocular trauma (68 patients) accounted for over two-fourths (57.6%) of the total cases. Only 16% of patients presented within 48 hours. Fifty-one percent of injuries occurred to the left eye, 38% to the right, and 10% bilaterally. The commonest ocular injury problems were eyelid laceration (13 patients, 19.1%), post-traumatic iritis (12 patients, 17.6%), and corneal laceration and penetration (10 patients, 14.7%), accounting for 51% (35 patients) of the total. Home- and work-related ocular injuries accounted for 54% of all ocular injuries. Thirty-three percent of all ocular injuries were caused by assault and fight, and 15% were related to motor vehicle accident. For the non-traumatic ocular emergencies, the main aetiological factor was inflammation (18%). CONCLUSION: Our study showed that males account for the majority of eye injuries and this class is more prone to assault-related injuries. In our country prevention strategies must take account of these.

Color vision in 42 Congolese patients with tuberculosis receiving ethambutol treatment.

PURPOSE: To study color vision in Congolese patients with tuberculosis receiving ethambutol therapy. METHODS: A prospective, descriptive study of color vision test in patients with systemic tuberculosis receiving ethambutol was performed between April 1995 and January 1998 at the Department of Ophthalmology, University of Kinshasa. Color vision tests were assessed with pseudoisochromatic plates (the Ishihara Pseudo-isochromatic Plates), the AO-HRR (American Optical Handy Rand Rittler), the Bölle and Kastel anomaloscope, Farnsworth-Munsell test (the D-15 and the FM-100). RESULTS: There were 42 patients with a mean age of 33 years (range, 14 to 75 years). The color vision of all the patients was found to be normal as measured by the Ishihara pseudoisochromatic plates. One (2%) patient showed color vision defect (anarchic axis] with the OA-HRR test. Three (7%) of 42 patients displayed blue-yellow color axis or anarchic axis color vision test on the D-15 test. Fifteen (36%) of 42 patients had high total error scores at the Farnsworth-Munsell 100 test. The color axis was as follows: anarchic axis (13.1%), red-green-color and blue-yellow-color combined axis (13.1%), blue-yellow color axis (7.5%). Results of the Bölle and Kastel anomaloscope were normal in all patients. CONCLUSION: Our results confirm the importance of color vision examinations in the detection of the complications of ethambutol treatment.

Presenting signs of retinoblastoma in Congolese patients.

OBJECTIVE: To report the relative frequency of the signs of presentation in Congolese children with retinoblastoma. METHODS: A retrospective study was undertaken of all (29) patients with retinoblastoma examined between January 1995 and December 2000. RESULTS: There were 19 (66%) boys and 10 (34%) girls. For all cases, mean age at diagnosis was 2.94 years +/- 1.6 (range, four months to six years). For bilateral cases, it was 1.12 year +/- 1.4 (range, two months to three years) whereas for unilateral cases it was 3.23 years +/- 1.5 (range, four months to six years) (P = 0.016). More than seven distinct signs were identified. Leukocoria was the most common presenting sign in 49% of diagnosed cases followed by proptosis (28%). Other signs were strabismus, red eye, anterior scleral staphyloma, hyphema and buphthalmia. CONCLUSION: Strabismus seemed to be uncommon whereas proptosis is important in our small series when compared to signs reported in the developed world.

Langerhans' cells in vernal keratoconjunctivitis express the costimulatory molecule B7-2 (CD86), but not B7-1 (CD80).

PURPOSE: Vernal keratoconjunctivitis (VKC) is associated with T-helper 2 (TH2)-like cell response and increased immunoglobulin (Ig) E production. Recent studies have suggested that interactions between costimulatory molecules B7 on antigen-presenting cells and CD28 on T cells are critical for successful antigen presentation and the development of the TH2 immune response. The objective of this study was to examine the expression of costimulatory molecules CD28, B7-1 (CD80) and B7-2 (CD86) in conjunctival biopsies from patients with active VKC and normal controls. METHODS: Conjunctival biopsy specimens from 15 subjects with active VKC, and 8 control subjects, were studied by immunohistochemical techniques using a panel of monoclonal and polyclonal antibodies directed against CD28, B7-1 and B7-2 molecules. The phenotype of inflammatory cells expressing costimulatory molecules was examined by sequential double immunohistochemistry. RESULTS: In the normal conjunctiva, B7-2 was expressed on a few mononuclear cells in the epithelium and substantia propria in 5 of 8 specimens. There was no immunoreactivity for CD28 or B7-1. In VKC specimens, few B7-1+ mononuclear cells were noted in the substantia propria in 7 of 15 specimens. B7-2 was expressed on mononuclear cells in the epithelium and substantia propria in all specimens. Compared with normal controls, VKC specimens showed significantly more mononuclear cells expressing B7-2 (30.5+/-14.1 vs 1.88+/-2.5; p < 0.001). In VKC specimens, the numbers of mononuclear cells expressing B7-2 were significantly higher than the numbers of mononuclear cells expressing B7-1 (30.5+/-14.1 vs 2.3+/-3.1; p < 0.001). CD28 was expressed on mononuclear cells in the epithelium and substantia propria in 14 specimens. Colocalisation studies revealed that the majority of mononuclear cells expressing B7-2 were CD1a+ Langerhans' cells, and that the mononuclear cells expressing CD28 were CD3+ T lymphocytes. CONCLUSIONS: B7-2 is more widely and prominently expressed by Langerhans' cells compared with B7-1. The interaction of B7-2 with CD28 may mediate the development of the TH2 immune response in VKC. Thus the manipulation of this pathway could be an important target for the development of future therapies in VKC.

Expression of the inducible isoform of nitric oxide synthase in the retinas of human subjects with diabetes mellitus.

PURPOSE: Inducible nitric oxide synthase has been implicated in the pathogenesis of cerebral ischemic damage, in the angiogenic process and in diabetic vascular damage. This study was undertaken to determine whether inducible nitric oxide synthase is present in the retinas from human subjects with diabetes mellitus. METHODS: This was an experimental immunohistochemical prospective study. Ten postmortem eyes from five subjects with diabetes mellitus, 10 eyes from five subjects without diabetes and without known ocular disease, and two eyes from one subject with unilateral ocular ischemic syndrome secondary to severe carotid artery obstruction were examined. We used immunohistochemical techniques and antibodies directed against inducible nitric oxide synthase, glial fibrillary acidic protein, and vimentin. The main outcome measure was immunoreactivity for these antibodies. RESULTS: Immunoreactivity for inducible nitric oxide synthase was not observed in retinas from all subjects without diabetes and without ocular disease. Six retinas from three subjects with diabetes and nonproliferative retinopathy, and the retina from the eye with ocular ischemic syndrome showed immunoreactivity for inducible nitric oxide synthase in cells with elongated processes. Based on morphology and on glial fibrillary acidic protein and vimentin immunoreactivity, this inducible nitric oxide synthase immunoreactivity appeared to localize to retinal Müller glial cells. CONCLUSIONS: These observations suggest that Müller cells may be involved in the microvascular remodeling of the diseased retina and that high concentrations of nitric oxide produced by inducible nitric oxide synthase could contribute to neurotoxicity and angiogenesis that occur in diabetic retinopathy.

Expression of chemokine receptors in vernal keratoconjunctivitis.

BACKGROUND/AIMS: Chemokines are small peptides which are potent activators and chemoattractants for leucocyte subpopulations. Their action is mediated by a family of seven transmembrane spanning G-protein coupled receptors. The aims of this study were to examine the expression of the chemokine receptors CCR1, CCR3, CCR5, CXCR3, and CXCR4 in the conjunctiva of patients with vernal keratoconjunctivitis (VKC) and to investigate the phenotype of inflammatory cells expressing these chemokine receptors. METHODS: Conjunctival biopsy specimens from 16 patients with active VKC, and eight control subjects were studied by immunohistochemical techniques using a panel of monoclonal antibodies directed against human CCR1, CCR3, CCR5, CXCR3, and CXCR4. The phenotype of inflammatory cells expressing chemokine receptors was examined by double immunohistochemistry. RESULTS: In the normal conjunctiva, few inflammatory cells expressed CXCR3 in five of eight specimens. There was no immunoreactivity for CCR1, CCR3, CCR5, and CXCR4. In VKC specimens, membranous immunoreactivity for CXCR3 was noted on inflammatory cells in all specimens. Compared with control specimens, VKC specimens showed significantly more inflammatory cells expressing CXCR3 (54.3 (SD 34.3) v 3.3 (5.0); p<0.001). Few CCR1+, CCR3+, CCR5+, and CXCR4+ inflammatory cells were observed in only three of 16 specimens. Double immunohistochemistry revealed that all CXCR3 positive inflammatory cells were CD3 positive T lymphocytes and that 61.7% (3.7%) of the infiltrating T lymphocytes were reactive for CXCR3. CONCLUSIONS: CXCR3 is the predominant chemokine receptor and is expressed abundantly on T lymphocytes in the conjunctiva of patients with active VKC. These data suggest a potential role for CXCR3 receptors in the regulation of lymphocyte recruitment within conjunctiva of VKC patients. New therapeutic strategies that block CXCR3 may inhibit T lymphocyte recruitment and suppress adverse inflammatory reactions.

Gelatinase B in vernal keratoconjunctivitis.

OBJECTIVES: To investigate the expression of gelatinase B in the conjunctiva of patients with vernal keratoconjunctivitis (VKC) and the cellular source of this enzyme. METHODS: Conjunctival biopsy specimens from 12 patients with active VKC and 12 control subjects were studied using immunohistochemical techniques and a monoclonal antibody against gelatinase B. The phenotype of gelatinase B(+) inflammatory cells was examined using double immunohistochemical analysis and monoclonal antibodies against eosinophil peroxidase or macrophage CD68. Quantitative zymography was used to compare the activity of gelatinase B in conjunctival biopsy specimens from 10 patients with active VKC and 7 control subjects. RESULTS: Gelatinase B was detected in a few polymorphonuclear cells in 8 control specimens. All VKC specimens showed gelatinase B immunoreactivity in the epithelial and stromal inflammatory infiltrate. Compared with control specimens, VKC specimens showed significantly more gelatinase B-positive cells (mean +/- SD, 40.8 +/- 29.9 vs 10.3 +/- 2.4; P<.02). Most gelatinase B-positive cells were eosinophils (90.2% +/- 3.6%). Zymography revealed that gelatinase B levels in VKC specimens were significantly higher than the levels found in normal conjunctiva (3780.3 +/- 3541.0 vs 610.1 +/- 397.1 scanning units; P<.03). CONCLUSIONS: These findings suggest overexpression of gelatinase B by eosinophils in VKC specimens and participation of gelatinase B in the pathologic changes in VKC. CLINICAL RELEVANCE: Control of the release and/or activation of gelatinase B in eosinophils may provide a new therapeutic strategy for treating VKC.

Immunology of trachomatous conjunctivitis.

Trachoma, a chronic follicular conjunctivitis caused by infection with Chlamydia trachomatis, is a leading cause of preventable blindness. The blinding complications are associated with progressive conjunctival scarring. Our immunohistochemical studies of conjunctival biopsies from children with active trachoma demonstrated the presence of both humoral and cell-mediated immune responses. Antichlamydial antibodies can neutralize Chlamydiae, block attachment and internalization of the organism, and can produce partial immunity. Our observations suggest a role for T-lymphocytes and cell mediated immunity in the genesis of conjunctival scarring. Conjunctival epithelial cells expressed major histocompatibility complex (MHC) class II antigens which might allow conjunctival epithelial cells to present Chlamydial antigens to T-cells enhancing the immune response. The epithelial cells expressing MHC class II antigens might present autoantigens to T-cells leading to induction of an autoimmune reaction. We have demonstrated that the conjunctival epithelial cells from patients with trachoma expressed interleukin (IL)-1 alpha and IL-1 beta. In addition, we have detected cytoplasmic expression of IL-1 alpha, IL-1 beta, tumor necrosis factor alpha and platelet-derived growth factor by macrophages. These cytokines have the potential to influence the remodeling and fibrosis observed in trachoma. Alterations of extracellular matrix components and collagen metabolism occur in the conjunctival tissue from patients with trachoma. New collagen type V formation was noted in active trachoma and scarred trachoma. The conjunctival tissue from patients with active trachoma contained increased amounts of collagen types I, III and IV. Scarred trachoma is characterized by marked increase in basement membrane--collagen IV and marked decrease in collagen types I and III. In addition, we demonstrated increased activity of gelatinase B and numbers of inflammatory cells containing gelatinase B in trachoma patients suggesting that this enzyme might be involved in matrix degradation and promotion of conjunctival scarring in trachoma.

Expression of CD23/CD21 and CD40/CD40 ligand in vernal keratoconjunctivitis.

PURPOSE: The overproduction of immunoglobulin E (IgE) antibodies is associated with vernal keratoconjunctivitis (VKC). CD23/CD21 and CD40/CD40 ligand (CD40L) interactions have been proposed to be involved in the regulation of IgE synthesis. The purpose of the present study was to investigate the presence and distribution of CD23, CD21, CD40 and CD40L in the conjunctiva from patients with active VKC. METHODS: Conjunctival biopsy specimens from 8 subjects with active VKC and 6 control subjects were studied. We used immunohistochemical techniques and a panel of monoclonal antibodies (mAbs) directed against CD23, CD21, CD40 and CD40L. In addition, a panel of mAbs were used to characterise the composition of the inflammatory infiltrate. RESULTS: In the normal conjunctiva, basal epithelial cells and vascular endothelial cells in the upper substantia propria showed a constitutive very weak immunoreactivity for CD40. The immunoreactivity for CD23, CD21 and CD40L was absent. In VKC specimens, the stromal inflammatory infiltrate was orgnanised as a diffuse infiltrate and as small lymphoid follicles consisting of CD20+ B lymphocytes intermingled with smaller numbers of CD3+ T lymphocytes, and CD68+ monocytes/macrophages. Lymphocytes in the centre of the lymphoid follicles showed CD23 and CD21 immunoreactivity. CD40 immunoreactivity in epithelial cells and vascular endothelial cells was stronger in VKC specimens than in control eyes. Furthermore, the majority of mononuclear cells, including T and B lymphocytes, showed immunoreactivity for CD40. CD40L immunoreactivity was not detected. CONCLUSION: B lymphocytes in the lymphoid follicles expressing CD23, CD21 and CD40 are activated and might be precursors of IgE-producing B cells. These results suggest that the conjunctiva might contribute to IgE synthesis.

Risk factors for open-angle glaucoma: a case-control study.

The objective of this study was to identify possible risk factors associated with open-angle glaucoma (OAG). A case-control study included patients seen at an ophthalmologic clinic. Cases were all consecutive new patients with either unilateral or bilateral OAG detected during the study period. Controls were a random sample of all other patients aged 30 or more, seen in the same department in the same period. Data on demographic, anthropometric and diet habits as well as medical characteristics were collected from 144 patients by medical examination and interview. The study took place at the University Department of Ophthalmology and general private practice of ophthalmology, both in the city of Kinshasa. Forty consecutive patients with OAG and 104 controls were chosen randomly between all consecutive non OAG patients. Odds ratio (OR) are presented for the relation between OAG and age, sex, ethnicity, family history of glaucoma, the length of stay in Kinshasa, body mass index, hypertension, diabetes mellitus, cigarette smoking, alcohol, diet habits. Adjusted odds ratio resulting from stepwise logistic regression was employed. Results indicate: family history of glaucoma (OR, 18; 95% CI, 5.80-59.00; P < 0.001), age (OR, 1.05; 95% CI, 1.01-1.09; P = 0.025), body mass index (OR, 1.09; 95% CI, 1.01-1.18; P = 0.05), hyperopia (OR, 2.9; 95% CI, 1.05-7.08; P = 0.03), Mongo ethnic group (OR, 3.5; 95% CI, 1.11-12.20; P = 0.03) and consumption of rice (OR, 4.6; 95% CI, 1.65-12.20; P = 0.004) conferred a significantly greater risk of OAG. This study seems confirm that Mongo ethnic group is associated with an increased risk of OAG.

Topical 0.1% indomethacin solution versus topical 0.1% dexamethasone solution in the prevention of inflammation after cataract surgery. The Study Group.

PURPOSE: To compare the efficacy and safety of topical 0.1% indomethacin with 0.1% dexamethasone after cataract surgery. METHODS: 145 patients (indo = 71, dexa = 74) were enrolled in a randomised, double-masked study and received one drop 4 times a day of indomethacin or dexamethasone for 1 month. RESULTS: Proteinic flare and cellular Tyndall decreased with time in both groups, with a difference in favour of indomethacin for cellular Tyndall on day 30 (p = 0.046). Conjunctival hyperaemia was less pronounced in the dexamethasone group on day 30 (p = 0.03). Tolerance of both drugs was good. CONCLUSION: 0.1% Indomethacin solution appears to be as safe and efficient as 0.1% dexamethasone eyedrops in the management of post-operative inflammation and could be a good alternative to the use of steroids.

Chemokines in the limbal form of vernal keratoconjunctivitis.

BACKGROUND/AIMS: Chemokines are a family of low molecular weight cytokines that attract and activate leucocytes. The CC chemokines act on eosinophils, basophils, monocytes, and lymphocytes, suggesting that they play an important part in allergic diseases. The aims of this study were to investigate the expression of the CC chemokines, RANTES, eotaxin, monocyte chemotactic protein (MCP) 1, MCP-2, and MCP-3 in the conjunctiva of patients with vernal keratoconjunctivitis (VKC) and to determine the cellular source of these chemokines. METHODS: Conjunctival biopsy specimens from nine subjects with active VKC, and six control subjects were studied by immunohistochemical techniques using a panel of monoclonal and polyclonal antibodies directed against RANTES, eotaxin, MCP-1, MCP-2, and MCP-3. The phenotype of inflammatory cells expressing chemokines was examined by sequential double immunohistochemistry. RESULTS: In the normal conjunctiva, superficial epithelial cells showed a constitutive, weak cytoplasmic expression of eotaxin. Few inflammatory cells in the perivascular areas expressed RANTES, MCP-1, MCP-2, and MCP-3. In VKC specimens, the epithelium showed intense cytoplasmic eotaxin staining in all cells, and cytoplasmic RANTES staining mainly in the superficial layers. Furthermore, RANTES and eotaxin were expressed on the vascular endothelium mainly in the upper substantia propria. Compared with normal controls, VKC specimens showed significantly more inflammatory cells expressing RANTES, eotaxin, MCP-1, and MCP-3 (p<0.001, 0.0028, 0.0092, and <0. 001, respectively). In VKC specimens, the numbers of inflammatory cells expressing RANTES were significantly higher than the numbers of inflammatory cells expressing eotaxin, MCP-1, and MCP-2 (all p values <0.001). Colocalisation studies revealed that the majority of inflammatory cells expressing chemokines were CD68 positive monocytes/macrophages. CONCLUSIONS: These results demonstrate an increase in the expression of RANTES, eotaxin, MCP-1, and MCP-3 in the conjunctiva of patients with VKC compared with control subjects. These data suggest a potential role for these chemokines in the pathogenesis of VKC. Antagonists of chemokine receptors may provide new therapeutic modalities in VKC.