interleukin-8 and Keratitis--Herpetic

To identify whether histopathologic and immunoassay biomarkers of inflammation are predictive for allograft rejection after penetrating keratoplasty (PKP) for herpes simplex virus (HSV) keratitis.. Retrospective, interventional case series with prospective component of pathologic evaluation of frozen tissue.. Sixty-two consecutive patients with HSV keratitis who underwent PKP.. A chart review and histopathologic examination of the excised host corneal button was performed to identify associations between clinical data and histopathologic presence of inflammation. Enzyme-linked immunosorbent assay for interleukin (IL)-8 and monocyte chemotactic protein-1 (MCP-1) chemokines and immunohistochemical staining for human leukocyte antigen (HLA)-DR and intercellular adhesion molecule-1 (ICAM-1) antigens was also performed in inflamed and noninflamed specimens.. To determine whether the presence of subclinical inflammation at the time of PKP predicts allograft rejection.. Although 81% of patients had clinically quiescent disease, histopathology revealed that 74% had active corneal inflammation, a finding that was associated with the presence of clinical neovascularization (P = 0.01). Allograft rejections were experienced by 34% of the patients in this cohort. The histopathologic presence of inflammation was a risk factor for allograft rejection (P = 0.02). Corneal specimens demonstrating inflammation had significantly increased IL-8 (P = 0.0005) and MCP-1 (P = 0.003) levels, and greater immunoreactivity for HLA-DR and ICAM-1 when compared with specimens without inflammation. Treatment with IL-10 ex vivo significantly inhibited IL-8 (P = 0.006), and MCP-1 (P = 0.01) chemokines, and qualitatively substantially reduced HLA-DR, but not ICAM-1, expression.. Histopathologic inflammation is a risk factor for corneal allograft rejection.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Chemokine CCL2; Chemokines; Child; Child, Preschool; Cornea; Enzyme-Linked Immunosorbent Assay; Female; Graft Rejection; HLA-DR Antigens; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-8; Keratitis, Herpetic; Keratoplasty, Penetrating; Male; Middle Aged; Retrospective Studies; Risk Factors; Transplantation, Homologous; Young Adult

To identify histopathologic features predictive for adverse allograft outcomes following penetrating keratoplasty for herpes simplex virus (HSV) keratitis.. Retrospective, interventional case series of 62 consecutive patients with HSV keratitis who underwent penetrating keratoplasty at the Kellogg Eye Center from 1990 through 2000. A detailed chart review and review of the histopathology of the excised corneal button were performed to identify associations between clinical data (disease activity, vascularity, adverse allograft outcomes) and histopathologic data (inflammation, neovascularization, biomarkers). The main outcome measure was to find histopathologic features that may predict HSV recurrence, allograft rejection, or failure.. Although 81% of patients had clinically quiescent disease, histopathology revealed that 74% had active corneal inflammation, a finding that was associated with the presence of clinical neovascularization (P = .01). Histopathologic inflammation was a risk factor for allograft rejection (P = .02) but not failure (P = .98) or HSV recurrence (P = .45). Histopathologic neovascularization did not predict rejection (P = .19) but did predict failure (P = .002) and HSV recurrence (P = .05). Biomarkers, including HLA-DR, ICAM-1, and IL-8(CXC) and MCP-1 (CC) chemokines, were all significantly increased in fresh corneal specimens demonstrating moderate to severe inflammation. IL-10 treatment ex vivo significantly inhibited HLA-DR, IL-8 (P = .006), and MCP-1 (P = .01) but did not reduce ICAM-1 expression.. Histopathologic inflammation, neovascularization, and the presence of specific biomarkers are risk factors for corneal allograft morbidity.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Chemokine CCL2; Child; Child, Preschool; Cornea; Enzyme-Linked Immunosorbent Assay; Female; Graft Rejection; HLA-DR Antigens; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Keratitis, Herpetic; Keratoplasty, Penetrating; Male; Middle Aged; Recurrence; Retrospective Studies; Risk Factors; Transplantation, Homologous; Young Adult

Granulocyte macrophage colony-stimulating factor (GM-CSF) is thought to play a key role in chronic inflammatory diseases by governing the survival and function of infiltrating neutrophils. The objective of this study was to determine the putative role of GM-CSF in the pathogenesis of human herpetic stromal keratitis (HSK).. Primary human corneal fibroblast (HCF) cultures and a telomerase-immortalized human corneal epithelial (HCE) cell line representative of native HCE were stimulated with the known HSK-inducing cytokines interferon (IFN)-gamma, interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha. Alternatively, the T-cell cytokine IL-17 was added solely or simultaneously. Human neutrophils were incubated with conditioned medium (CM) of the HCF and HCE stimulated with the aforementioned cytokines, or recombinant GM-CSF, and their viability or activation status was determined by flow cytometry. GM-CSF and IL-8 secretion levels in the CM were determined by ELISA. The antibody-dependent cellular cytotoxicity (ADCC) of neutrophils toward herpes simplex virus (HSV)-infected HCFs was determined by flow cytometry. The expression of GM-CSF was determined in HSK and control corneal buttons by real-time RT-PCR and immunohistology.. Compared with IFN-gamma, CM of either cell type stimulated with IL-1beta, or in the case of HCE cells, stimulated with TNF-alpha or IL-17, delayed neutrophil apoptosis significantly. Only in HCFs did IL-17 exhibit a synergistic effect with TNF-alpha. The antiapoptotic activity was attributable in part to the GM-CSF secreted by the activated HCFs and HCE cells. GM-CSF stimulation of neutrophils induced their activation and the secretion of IL-8. GM-CSF did not increase significantly the ADCC reaction of neutrophils toward HSV-infected HCFs. Finally, GM-CSF was expressed in corneas of the patients with HSK but not in control subjects.. The data suggest that GM-CSF, expressed by cornea-resident cells such as HCFs and HCE cells, may play a role in the immunopathogenesis of HSK by prolonging the survival and modulating the effector function of corneal infiltrating neutrophils.

Topics: Antibody-Dependent Cell Cytotoxicity; Cells, Cultured; Corneal Stroma; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Epithelium, Corneal; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Herpesvirus 1, Human; Humans; Immunoenzyme Techniques; Interferon-gamma; Interleukin-1beta; Interleukin-8; Keratitis, Herpetic; Neutrophil Activation; Neutrophils; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Tumor Necrosis Factor-alpha

Herpetic stromal keratitis (HSK) is an immunopathologic disease triggered by infection of the cornea with HSV. Key events in HSK involve the interaction between cornea-infiltrating inflammatory cells and resident cells. This interaction, in which macrophages, producing IL-1 and TNF-alpha, and IFN-gamma-producing Th1 cells play a crucial role, results in the local secretion of immune-modulatory factors and a major influx of neutrophils causing corneal lesions and blindness. The Th1-derived cytokine IL-17 has been shown to play an important role in several inflammatory diseases characterized by a massive infiltration of neutrophils into inflamed tissue. Here we show that IL-17 is expressed in corneas from patients with HSK and that the IL-17R is constitutively expressed by human corneal fibroblasts (HCF). IL-17 exhibited a strong synergistic effect with TNF-alpha on the induction of IL-6 and IL-8 secretion by cultured HCF. Secreted IL-8 in these cultures had a strong chemotactic effect on neutrophils. IL-17 also enhanced TNF-alpha- and IFN-gamma-induced secretion of macrophage-inflammatory proteins 1alpha and 3alpha, while inhibiting the induced secretion of RANTES. Furthermore, considerable levels of IFN-gamma-inducible protein 10 and matrix metalloproteinase 1 were measured in stimulated HCF cultures, while the constitutive secretion of monocyte chemotactic protein 1 remained unaffected. The data presented suggest that IL-17 may play an important role in the induction and/or perpetuation of the immunopathologic processes in human HSK by modulating the secretion of proinflammatory and neutrophil chemotactic factors by corneal resident fibroblasts.

Topics: Adjuvants, Immunologic; Cell Line; Cells, Cultured; Chemokines; Cornea; Drug Synergism; Fibroblasts; Humans; Interleukin-17; Interleukin-6; Interleukin-8; Keratitis, Herpetic; Matrix Metalloproteinase 1; Neutrophil Infiltration; Receptors, Interleukin; Receptors, Interleukin-17; Recombinant Proteins; Stromal Cells; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

Neutrophil invasion is a primary event in the development of herpetic keratitis. It has been reported that HSV-1 infection of keratocytes induces the synthesis of IL-8, a potent neutrophil chemoattractant, while corneal epithelium does not. Nevertheless, little is known about the correlation between neutrophil migration and the production of chemotactic factors by HSV-1-infected corneal cells, especially in epithelial cells which form an initial barrier of the ocular surface. We examined whether human corneal epithelial cells as well as keratocytes could induce neutrophil chemotaxis in response to HSV-1 infection.. Human corneal epithelial cells immortalized with SV40 (HCE) and human keratocytes were infected with HSV-1. The culture fluids collected at 4, 12, 24 h after infection were assayed for human neutrophil chemotaxis using a modified Boyden chamber method. IL-8 levels in these supernatants were measured using enzyme-linked immunosorbent assay (ELISA).. The chemotactic activity induced by HCE and keratocytes after MP strain of HSV-1 infection peaked as early as 4 h postinfection, then declined. Chemotactic activity induced by HSV-1-infected HCE and IL-8 levels on these supernatants paralleled with the infectious virus titer. It was inhibited by monoclonal anti-IL-8 antibody. UV-inactivation of MP strain abrogated neither the induction of chemotactic activity nor IL-8 secretion of infected HCE.. At the early phase of HSV-1 infection, corneal epithelial cells play an important role in inducing neutrophil chemotaxis, which was mediated by IL-8.

Topics: Cell Line; Chemotaxis, Leukocyte; Cornea; Epithelial Cells; Herpesvirus 1, Human; Humans; Interleukin-8; Keratitis, Herpetic; Neutrophils