interleukin-8 and Conjunctivitis--Allergic

Extract of Perilla frutescens enriched for rosmarinic acid, a polyphenolic phytochemical, suppresses allergic immunoglobulin responses and inflammation caused by polymorphonuclear leukocytes (PMNL) in mice. However, few placebo-controlled clinical trials have examined the efficacy and safety of polyphenolic phytochemicals for treatment of allergic inflammatory diseases in humans. The present study determined whether oral supplementation with rosmarinic acid is an effective intervention for patients with seasonal allergic rhinoconjunctivitis (SAR). In this 21-day, randomized, double-blind, age-matched, placebo-controlled parallel group study, patients with mild SAR were treated daily with extract of Perilla frutescens enriched for rosmarinic acid (200 mg [n=10] or 50 mg [n=9]) or placebo (n=10). Patients recorded symptoms daily in a diary. Profiles of infiltrating cells and concentrations of eotaxin, IL-1beta, IL-8, and histamine were measured in nasal lavage fluid. Serum IgE concentrations and routine blood tests were also examined. As compared with placebo supplementation, supplementation with extract of Perilla frutescens enriched for rosmarinic acid resulted in a significant increase in responder rates for itchy nose, watery eyes, itchy eyes, and total symptoms (P<0.05). Active treatment significantly decreased the numbers of neutrophils and eosinophils in nasal lavage fluid (P<0.05 vs. placebo). Patients reported no adverse events, and no significant abnormalities were detected in routine blood tests. In conclusion, extract of Perilla frutescens enriched for rosmarinic acid can be an effective intervention for mild SAR at least partly through inhibition of PMNL infiltration into the nostrils. Use of this alternative treatment for SAR might reduce treatment costs for allergic diseases.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Chemokine CCL11; Chemokines, CC; Cinnamates; Conjunctivitis, Allergic; Depsides; Double-Blind Method; Eosinophils; Female; Histamine; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Interleukin-1; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Neutrophils; Perilla frutescens; Phytotherapy; Plant Preparations; Rhinitis, Allergic, Seasonal; Rosmarinic Acid; Treatment Outcome

CCL20, the single chemokine ligand for CCR6, contributes to recruiting CCR6-expressing memory B cells, memory T cells, Th17 cells and dendritic cells, and is involved in regulating immune responses, homeostasis, and inflammation in mucosal tissues.. CCL20 messenger RNA (mRNA) expression was analyzed in the conjunctival epithelium in an in vivo study of patients with vernal keratoconjunctivitis (VKC group) and healthy volunteers (control group) using impression cytology. In vitro analysis of CCL20 mRNA was performed using cultured conjunctival epithelial cells (CECs). Real-time polymerase chain reaction was used to assess IL-8 and eotaxin-2 mRNA expression for comparison with CCL20 mRNA expression.. In the control group, CCL20 mRNA expression was present in all conjunctival locations. However, CCL20 mRNA expression was significantly higher in the upper palpebral conjunctiva in the severe VKC group than in the mild VKC and control groups (p < 0.05, Steel test). In vitro stimulation of CECs with lipopolysaccharide (LPS) significantly increased CCL20 expression in a concentration-dependent manner that was significantly correlated with expression of IL-8 (p < 0.001, Spearman's rank correlation coefficient), but not eotaxin-2.. We conclude that CCL 20 mRNA expression in the conjunctival epithelium plays a crucial role in regulating homeostasis at the ocular surface and in exacerbation of VKC.

Topics: Adolescent; Adult; Cells, Cultured; Chemokine CCL20; Chemokine CCL24; Child; Conjunctiva; Conjunctivitis, Allergic; Epithelial Cells; Female; Gene Expression Regulation; Healthy Volunteers; Humans; Interleukin-8; Male; Real-Time Polymerase Chain Reaction; RNA, Messenger; Young Adult

The purpose of present studies was to determine the involvement of NFκB and STAT6 transcription factors in the production of cytokines by the fibroblasts and epithelial cells in conjunctiva.. An in vitro model of allergic conjunctivitis was developed by sensitizing and challenging rat mast cells with anti-dinitrophenyl (DNP) IgE and DNP-BSA, and then using the conditioned medium to stimulate rat conjunctival fibroblasts. Chemokines (eotaxin-1, IL-8, and RANTES -- Regulated and Normal T cell Expressed and Secreted) released from cells into the medium was determined by ELISA. Human conjunctival fibroblasts and epithelial cells were also directly stimulated with exogenous cytokines tumor necrosis factor (TNF)-α or IL-4. Degradation of IκB-α and phosphorylation of STAT6 were assessed by immunoblotting. For inhibition of NFκB or STAT6 activation, upstream regulators IκB kinase and Janus protein tyrosine kinases (JAK) were inhibited by use of BMS-345541 and JAK inhibitor 1. An in vivo model of conjunctivitis was also produced in rats by intraperitoneal injection of ovalbumin (OA) with aluminum hydroxide and challenge at 21 d with OA eye drops.. Stimulated rat mast cells released TNF-α and IL-4. TNF-α induced NFκB activation in rat and human conjunctival fibroblasts and epithelial cells, and caused production and release of cytokines IL-8 and RANTES. IL-4 activation of STAT6 did not cause release of these cytokines. Only fibroblasts produced the eosinophil-recruiting cytokine, eotaxin-1, after treatment with TNF-α- plus IL-4. As observed in the cultured cells, allergic stimulation in the in vivo model caused degradation of IκB-α in conjunctiva, and infiltration of eosinophils and other inflammatory cells.. Activated NFκB was found to be a major transcription factor for the release of cytokines from conjunctival cells and intensification of the allergic response. Inhibition of the NFκB pathway by therapeutic drugs may be an important objective for the treatment of human allergic conjunctivitis.

Topics: Animals; Cells, Cultured; Chemokine CCL11; Chemokines; Conjunctiva; Conjunctivitis, Allergic; Culture Media, Conditioned; Epithelial Cells; Fibroblasts; Interleukin-4; Interleukin-8; Male; Mast Cells; NF-kappa B; Rats; Rats, Sprague-Dawley; STAT6 Transcription Factor; Tumor Necrosis Factor-alpha

To investigate the role of the epithelium in severe allergic conjunctivitis.. We first investigated the expression of protease-activated receptors (PARs) in cultured human conjunctival epithelial cells and fibroblasts by reverse transcription-polymerase chain reaction. Next, we examined whether mite allergen-stimulated cells release chemokines and whether physiological protease inhibitors such as secretory leukocyte protease inhibitor (SLPI) and α1-antitrypsin can inhibit their production. We also looked at the expression of thymic stromal lymphopoietin (TSLP) in giant papillae of patients with vernal keratoconjunctivitis and examined whether the as Toll-like receptor 3 ligand polyinosinic:polycytidylic acid (poly I:C) can induce expression of TSLP in cultured human conjunctival epithelial cells.. PAR 1, PAR2, and PAR3 were expressed in cultured human conjunctival epithelial cells and fibroblasts at mRNA level. These epithelial cells released interleukin (IL) 6 and IL-8, with an upregulation in their gene expression, in response to the serine protease activity of mite allergens. This response was inhibited by SLPI and α1-antitrypsin. Transforming growth factor β1 decreased the production of SLPI in corneal and conjunctival epithelial cells. TSLP was expressed in giant papillae epithelium in patients with vernal keratoconjunctivitis at mRNA and protein levels. Poly I:C induced expression of TSLP in cultured conjunctival epithelial cells at mRNA level. Costimulation with TSLP and IL-33 had a synergistic effect for IL-13 mRNA expression in cultured human mast cells.. Imbalance between protease of mite allergens and innate protease inhibitors of the epithelium may induce inflammation and disrupt barrier function. Viral infection may induce expression of TSLP via Toll-like receptors and release IL-33 by necrosis. These phenomena promote excessive allergic reactions; hence, the epithelium takes "center stage" in allergic conjunctivitis.

Topics: Allergens; alpha 1-Antitrypsin; Animals; Cells, Cultured; Conjunctiva; Conjunctivitis, Allergic; Epithelial Cells; Fibroblasts; Gene Expression Regulation; Humans; Interleukin-6; Interleukin-8; Mites; Receptors, Cytokine; Receptors, Proteinase-Activated; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Secretory Leukocyte Peptidase Inhibitor; Transforming Growth Factor beta1

To evaluate the variation of chemokines in tears of contact lens wearers.. The subjects were divided into the three groups: a control group consisting of 26 eyes of 26 healthy volunteers without contact lenses a contact lens group (CL group) consisting of 30 eyes of 30 healthy contact lens wearers without ocular surface disorders, and a giant papillary conjunctivitis group (GPC group) consisting of 9 eyes of 9 patients with giant papillary conjunctivitis caused by contact lens wearing. Tear samples were taken by the modified Schirmer I method using a filter paper. Tear samples were eluted and analyzed for chemokines including interleukin-8 (IL-8), eotaxin-2, and pulmonary and activation-regulated CC chemokine (PARC) by the antibody array method. Concentrations of IL-8, eotaxin-2, and PARC in tears were determined quantitatively by enzyme-linked immunosorbent assay (ELISA).. Using the antibody array method, the expression of IL-8, eotaxin-2, and PARC in the GPC group was 4-fold higher or greater than in the control group. In the measurement by ELISA, IL-8 levels in the GPC group (1154.5 +/- 1739.3 pg/ml) (mean +/- SD) were significantly higher (Kruskal-Wallis test, p < 0.01) than in the control (75.2 +/- 88.7 pg/ml) and CL (153.6 +/- 252.8 pg/ml) groups. The eotaxin-2 levels in tears did not show a statistical difference among the three groups. The PARC level in tears of the GPC group (2859.6 +/- 2299.9 pg/ml) was significantly higher than in the control(589.0 +/- 324.8 pg/ml) and CL (671.7 +/- 536.2 pg/ml) groups (Kruskal-Wallis test, p < 0.05).. Wearing a contact lens per se disorders, does not cause chemokine variation in tears. However, an increase of IL-8 which induces neutrophilic invasion and an increase of PARC which induces lymphocyte invasion play an important roles in increasing the risk factor of GPC when wearing contacts.

Topics: Adolescent; Adult; Chemokine CCL24; Chemokines; Chemokines, CC; Conjunctivitis, Allergic; Contact Lenses; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Male; Middle Aged; Tears

To evaluate whether interleukin-8 (IL-8) and RANTES (regulated on activation, normal T-cell expressed and secreted) concentrations in the supernatants of conjunctival epithelial samples from patients with vernal keratoconjunctivitis (VKC) correlate with the number of infiltrating eosinophils or neutrophils and with the severity of corneal lesions.. Thirty-four patients with VKC, 5 patients with seasonal allergic conjunctivitis, and 10 volunteers without allergic diseases were enrolled in this study. Conjunctival epithelial cells were collected by brush cytology and the number of inflammatory cells was counted. The chemokine expression in the cells was investigated by immunocytochemistry and the chemokine concentrations of the cell suspensions were measured by enzyme-linked immunosorbent assay.. The percentages of eosinophils and neutrophils in cell suspensions from VKC patients with corneal erosion or ulcer were higher than those from subjects with clear corneas or superficial punctate keratopathy. IL-8 concentrations in the supernatant of samples correlated significantly with the percentages of neutrophils and eosinophils in paired cell suspensions. No correlation was observed between RANTES and the percentages of eosinophils. Positive staining for IL-8 was observed in the cytosol of conjunctival epithelial cells.. IL-8 in the extracellular space of the conjunctival epithelium may play a role in the recruitment of neutrophils and possibly eosinophils and in the pathogenesis of corneal damage in severe allergic diseases.

Topics: Adolescent; Adult; Aged; Chemokine CCL5; Child; Conjunctiva; Conjunctivitis, Allergic; Corneal Ulcer; Cytological Techniques; Enzyme-Linked Immunosorbent Assay; Eosinophils; Epithelium; Female; Humans; Immunoenzyme Techniques; Interleukin-8; Male; Neutrophil Infiltration

To evaluate the effects of topical ocular drugs with histamine H1-antagonist activity on histamine-stimulated phosphatidylinositol turnover and interleukin (IL) 6 and IL-8 secretion from human conjunctival epithelial cells.. Primary human conjunctival epithelial cell cultures were stimulated with histamine in the presence or absence of test drugs. Phosphatidylinositol turnover was quantified by ion exchange chromatography and cytokine content of supernatants by enzyme-linked immunosorbent assay.. Antazoline hydrochloride, emedastine difumarate, levocabastine hydrochloride, olopatadine hydrochloride, and pheniramine maleate attenuated histamine-stimulated phosphatidylinositol turnover and IL-6 and IL-8 secretion. Emedastine was the most potent in ligand binding, phosphatidylinositol turnover, and IL-6 secretion, with dissociation constant and 50% inhibitory concentrations of 1-3 nmol/L. Olopatadine, antazoline, and pheniramine exhibited similar H1-binding affinities (32-39 nmol/L). However, olopatadine was approximately 10-fold more potent as an inhibitor of cytokine secretion (50% inhibitory concentration, 1.7-5.5 nmol/L) than predicted from binding data, while antazoline and pheniramine were far less potent (20- to 140-fold) in functional assays. Levocabastine (dissociation constant, 52.6 nmol/L) exhibited greater functional activity (50% inhibitory concentration, 8-25 nmol/L) than either antazoline or pheniramine.. Histamine-stimulated phosphatidylinositol turnover and cytokine secretion by human conjunctival epithelial cells are attenuated by compounds with H1-antagonist activity. However, antihistaminic potency alone does not predict anti-inflammatory potential. Olopatadine, emedastine, and levocabastine were notably more potent than pheniramine and antazoline.. Selected topical ocular drugs with antihistaminic activity may offer therapeutic advantages to patients with allergic conjunctivitis by inhibiting proinflammatory cytokine secretion from human conjunctival epithelial cells.

Topics: Cells, Cultured; Chromatography, Ion Exchange; Conjunctiva; Conjunctivitis, Allergic; Dose-Response Relationship, Drug; Epithelial Cells; Histamine; Histamine H1 Antagonists; Humans; Interleukin-6; Interleukin-8; Ophthalmic Solutions; Phosphatidylinositols