interleukin-8 and Vitreoretinopathy--Proliferative

Several eye diseases are accompanied by inflammatory processes. The authors examined the expression of the proinflammatory chemokine CXCL8 and the corresponding receptors in healthy human retinas, in cellular membranes from patients with proliferative vitreoretinopathy (PVR) or human glial cell cultures and in an animal model of PVR in rabbit eyes.. The authors used immunohistochemical methods, Western blotting, RT-PCR, and real time RT-PCR to characterize the expression of CXCL8, CXCR1, and CXCR2 in human and rabbit retinas. Functionality of the receptors in cultured glial cells was tested by Ca(2+) imaging.. Immunohistochemical examinations of normal human and rabbit retinas revealed a distinct expression of CXCR1 and CXCR2 in several neuronal cell types. CXCL8 mRNA was demonstrated only by RT-PCR in normal retinas, and receptor expression was confirmed by Western blotting and RT-PCR. The presence of CXCR1 and CXCR2, but not CXCL8, was detected by immunostaining in glial fibrillary acidic protein-positive glial cells of cellular PVR membranes. Immunoreactivity for CXCL8, CXCR1, and CXCR2 was observed in virtually all cultured glial cells and in the human Müller cell line MIO-M1. Müller cells responded to the application of CXCL8 with increased cytosolic Ca(2+) concentrations. In PVR rabbit retinas, CXCR1 expression is increased in Müller cells, and CXCL8 and CXCR2 are strongly expressed in microglial cells.. Expression of CXCL8 and CXCL8 receptors in glial cells of human PVR membranes and rabbit PVR retinas suggests an involvement in glial reactivity. Furthermore, the prominent expression of CXCR1 and CXCR2 in neurons of the healthy human and rabbit retina suggests additional physiological functions.

Topics: Animals; Blotting, Western; Cell Culture Techniques; Cell Membrane; Disease Models, Animal; Fluorescent Antibody Technique, Indirect; Humans; Interleukin-8; Neuroglia; Neurons; Rabbits; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Retina; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Vitreoretinopathy, Proliferative

We examined the levels of vitreous chemokines and Sho (Zheng in Chinese) of Chinese-Korean-Japanese medicine in diabetic patients. Patients undergoing vitrectomy were classified into Group 1 (no diabetic retinopathy), Group 2 (diabetic retinopathy with no or a few new vessels), and Group 3 (diabetic retinopathy with many new vessels). The levels of IL-8, MCP-1, MIP-1alpha, MIP-1beta, and RANTES in the vitreous fluid were measured using cytometric bead array method. Sho was determined by the standard diagnostic method of Chinese-Korean-Japanese medicine. Vitreous levels of IL-8 and MCP-1 in Groups 2 and 3 were higher than those in Group 1. MIP-1alpha, MIP-1beta, and RANTES levels in Groups 2 and 3 were almost the same as those in Group 1. The percentage of patients with Keishibukuryo-gan (Guizhifuling-wan in Chinese) sho in Group 3 was higher than that in Group 1. In conclusion, vitreous levels of IL-8 and MCP-1 were high in patients with diabetic vitreoretinopathy. Keishibukuryo-gan sho may be associated with diabetic vitreoretinopathy.

Topics: Aged; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokines; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Female; Humans; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Prospective Studies; Vitrectomy; Vitreoretinopathy, Proliferative; Vitreous Body

Formation of epiretinal membranes (ERMs) after proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR) results in progressive deterioration of vision, but its pathogenic mechanisms are still unknown. This study was conducted to examine the role of nuclear factor kappa B (NF-kappaB) in the formation of ERMs after PDR and PVR.. ERM samples were obtained by vitrectomy from 10 patients with PDR (aged 53+/-12 years with 14+/-5 years of diabetes), 20 patients with PVR, and 17 patients with idiopathic ERMs. Ten PVR and 17 idiopathic ERM samples were processed for reverse transcription-polymerase chain reaction (RT-PCR) analysis. In addition, 10 PDR and 10 PVR membranes were processed for immunohistochemical analysis.. NF-kappaB mRNA expression levels were significantly higher (10 of 10 versus 9 of 17 subjects in idiopathic ERM, p=0.0119) in PVR subjects. Immunohistochemical analysis showed NF-kappaB protein expression in 8 of the 10 PDR samples as well as all 10 PVR samples, and NF-kappaB positive cells were partially double labeled with glial cell markers. Interestingly, NF-kappaB protein was also overlapped with angiogenic factor interleukin-8 (IL-8) in glial cells as well as vascular endothelial cells.. These results suggest that NF-kappaB is involved in the formation of both glial and vascular endothelial cell components, and that these two cell types might have functional interactions that lead to the enlargement of intraocular proliferative membranes.

Topics: Adult; Aged; Diabetic Retinopathy; Endothelium, Vascular; Epiretinal Membrane; Fluorescent Antibody Technique, Indirect; Humans; Interleukin-8; Middle Aged; Neuroglia; NF-kappa B; NF-kappa B p50 Subunit; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-ret; Receptor Protein-Tyrosine Kinases; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Vitrectomy; Vitreoretinopathy, Proliferative

To examine the contribution of infiltrating cells in the local production of cytokines within the vitreous of patients with proliferative vitreoretinopathy (PVR).. The presence of mRNA coding for IL-6, IL-8, IL-1beta, IL-1alpha, TNFalpha, IFNgamma, IL-12, and HPRT was investigated in 25 vitreous samples from patients with PVR, 11 vitreous samples from patients with retinal detachment (RD) not complicated by PVR, and 10 vitreous samples from patients with macular hole (MH). A quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using an internal competitor was used to investigate these samples. From these samples, 15 PVR, 8 RD, and 8 MH were analysed for the protein levels of the same cytokines using enzyme linked immunosorbent assay (ELISA). Spearman correlation was used to test any association between mRNA and cytokine protein levels, as an indicator of the contribution these cells make to the intravitreal cytokine milieu.. A strong correlation was found between mRNA and their respective cytokine levels (protein products) for IL-6, IL-8, IL-1beta, IL-1alpha, TNFalpha, IFNgamma (Spearman r = 0.83, 0.73, 0.67, 0.91, 0.73, and 0.73 respectively), but not for IL-12. The median levels of IL-6, IL-8, IL-1beta, and IFNgamma mRNA and their respective cytokines were significantly higher (p <0.05) in patients with PVR than in those with macular hole. There was no statistically significant difference in the median levels of IL-1alpha mRNA between PVR and MH but the cytokine IL-1alpha was detected at a significantly higher level in PVR compared with MH patients. Between PVR and RD patients, there was no statistically significant difference in mRNA levels for all the investigated cytokines (p >0.05) except for IL-6 where there was a statistical significance (p= 0.038). In contrast, the median levels of IL-6, IL-8, and IL-1beta cytokines were significantly higher (p <0.05) in patients with PVR than in those with RD, whereas for IL-1alpha and IFNgamma no significant statistical difference was detected between PVR and RD patients (p >0.05). When results of RD and MH patients were compared, a statistical difference was only detected in mRNA levels of INFgamma (p = 0.008). However, no difference was detected for INFgamma (protein product) or for any of the other cytokines between RD and MH patients.. Levels of both protein and mRNA encoding IL-6, IL-8, IL-1beta, and IFNgamma is significantly increased in vitreous samples from patients with PVR. The strong correlation between ELISA detectable cytokines (protein products) and their respective mRNA levels suggest that intravitreal, invasive cells are the major source of these cytokines, with the exception of IL-12. Cells invading the vitreous do not appear to locally produce IL-12 mRNA. This would appear to implicate cells peripheral to the vitreal mass as the major source of this cytokine.

Topics: Adult; Aged; Aged, 80 and over; Case-Control Studies; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hypoxanthine Phosphoribosyltransferase; Interferon-gamma; Interleukin-1; Interleukin-12; Interleukin-6; Interleukin-8; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Statistics, Nonparametric; Tumor Necrosis Factor-alpha; Vitreoretinopathy, Proliferative; Vitreous Body

To determine interleukin 8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) expression in response to mechanical injury in human retinal pigment epithelial (HRPE) cells.. Enzyme linked immunosorbent assay (ELISA) was performed to determine IL-8 and MCP-1 secretion by HRPE cells after mechanical denudation. IL-8 and MCP-1 mRNA expression by HRPE cells was assessed using semiquantitative RT-PCR. The effects of immunosuppressive drugs, dexamethasone (DEX) and cyclosporin A (CSA), as well as immunosuppressive cytokines, interleukin 4 (IL-4), interleukin 10 (IL-10), and interleukin 13 (IL-13), on chemokine expression in HRPE cells after denuding injury were analysed.. Mechanical injury induced HRPE IL-8 mRNA and IL-8 secretion. Although MCP-1 mRNA was enhanced slightly after denuding injury, MCP-1 secretion was not increased. DEX and CSA inhibited HRPE chemokine expression after injury. IL-4 and IL-13 enhanced IL-8 and MCP-1 production by HRPE cells after injury while IL-10 had no effect.. These results suggest that IL-8 may be involved in retinal inflammatory responses to injury and that DEX and/or CSA treatment may help control the inflammatory components of retinal diseases such as proliferative vitreoretinopathy.

Topics: Adult; Analysis of Variance; Chemokine CCL2; Cyclosporine; Dexamethasone; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Humans; Immunohistochemistry; Immunosuppressive Agents; Interleukin-10; Interleukin-13; Interleukin-4; Interleukin-8; Middle Aged; Pigment Epithelium of Eye; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Vitreoretinopathy, Proliferative

To determine whether the infiltrating cells in the vitreous and subretinal fluid of patients with proliferative vitreoretinopathy (PVR) express messenger RNA for various cytokines found in this condition.. The presence of mRNA coding for HPRT, IL-6, IL-1beta, IL-8, and TNFalpha was investigated in 20 vitreous and subretinal fluid (SRF) samples from patients with PVR by reverse transcriptase polymerase chain reaction (RT-PCR). 16 samples from patients with retinal detachment and macular holes were used as controls.. HPRT was detected in all samples of PVR and in 11 (69%) control cases. Patients with PVR demonstrated mRNA for the cytokines tested more often than controls. The difference was statistically significant.. The presence of mRNA encoding for IL-6, IL-1beta, IL-8, and TNFalpha is significantly detected by RT-PCR in vitreous and SRF samples of patients with PVR, indicating local production of these cytokines by vitreous and SRF cells.

Topics: Cytokines; Humans; Hypoxanthine Phosphoribosyltransferase; Interleukin-1; Interleukin-6; Interleukin-8; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha; Vitreoretinopathy, Proliferative

To observe the levels of cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8) and IL-6 in the early inflammatory stages of a rabbit proliferative vitreoretinopathy (PVR) model induced by macrophages.. The vitreous and venous blood of rabbit eyes of the PVR model were obtained and tested with ELISA kits of TNF-alpha, IL-8 and IL-6 and analyzed statistically.. The TNF-alpha level increased at the 7th day after macrophage injection and reached its peak (309 pg/ml) on day 21. IL-8 and IL-6 maintained their higher levels during 7 to 21 days after macrophage injection and the peak levels were 1 325 pg/ml and 998 pg/ml on day 14 respectively. Those showed significant differences compared to control (P

Topics: Animals; Female; Interleukin-6; Interleukin-8; Macrophages; Male; Rabbits; Tumor Necrosis Factor-alpha; Vitreoretinopathy, Proliferative

To investigate whether the chemokines monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) are involved in the pathogenesis of proliferative vitreoretinal disorders and to study their possible interaction with IL-6.. In a prospective study of 125 consecutive patients (125 eyes), vitreous and paired serum samples were obtained and were assayed for MCP-1 and IL-8. Levels of IL-6 were determined by proliferation of the IL-6-dependent hybridoma cell line 7TD1.. Monocyte chemotactic protein-1 was detected in 13 (48%) of 27 vitreous samples from patients with retinal detachment, in five (63%) of eight samples from patients with macular pucker, in 31 (72%) of 43 samples from patients with proliferative vitreoretinopathy, and in 32 (76%) of 42 samples from patients with proliferative diabetic retinopathy, but not in samples from five patients with idiopathic epiretinal membrane. There was a significant (P = .049) correlation between the incidence of MCP-1 detection in retinal detachment, macular pucker, and proliferative vitreoretinopathy groups and the severity of proliferation. Interleukin-8 was detected in two vitreous samples from eyes with retinal detachment, in two samples from eyes with proliferative vitreoretinopathy, and in three samples from eyes with proliferative diabetic retinopathy. Monocyte chemotactic protein-1 levels in the vitreous samples were positively correlated with IL-6 levels (r = .31, P = .01). Interleukin-6 levels were significantly (P = .0097) greater in vitreous samples with than without detectable levels of MCP-1.. Monocyte chemotactic protein-1 is present in a substantial percent of vitreous samples from eyes with proliferative vitreoretinal disorders and may help in stimulating the infiltration of monocytes and macrophages into eyes with these disorders.

Topics: Antibodies, Monoclonal; Chemokine CCL2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-6; Interleukin-8; Prospective Studies; Retinal Detachment; Vitreoretinopathy, Proliferative; Vitreous Body

We investigated the role of interleukin 8 (IL-8) in the pathogenesis of proliferative vitreoretinopathy (PVR). The IL-8 level in the vitreous and the blood of 20 patients with PVR was measured by a specific enzyme-linked immunoassay method. Vitreous from cadaver eyes (n = 20) was used as the control. The IL-8 level in the vitreous was between 31.3 and 65 pg/ml in 40% of eyes with PVR. IL-8 was detected neither in the blood of the patients with PVR nor in the vitreous from cadaver eyes. IL-8 levels in the vitreous did not correlate with either the grade of PVR or the duration of symptoms. These results suggest that IL-8 may play a role in the pathogenesis of PVR.

Topics: Adult; Cadaver; Female; Humans; Interleukin-8; Male; Middle Aged; Time Factors; Vitreoretinopathy, Proliferative; Vitreous Body

To measure the concentration changes of IL-8 during the development of experimental proliferative vitreoretiopathy (PVR) induecd by macrophages and explore the effect on the inflammatory cells at the early stage of the model.. Rabbit PVR model was induced by homogenous macrophages. The vitreous, agueous and vein blood were taken at different periods and IL-8 levels in them were measured with an ELISA kit.. The IL-8 level reached its peak of 1,325 pg/ml at 14th day in the model which was 14 times as the level of controls. The level then decreased to 48 pg/ml at 28th day after macrophage injection.. The changes of IL-8 in the model were parallel to the inflammatory reaction during the development of PVR, suggesting that IL-8 may play an important role the inflammatory cells' infiltrating and migrating to the vitreous.

Topics: Animals; Interleukin-8; Macrophages; Rabbits; Time Factors; Vitreoretinopathy, Proliferative; Vitreous Body

We determined whether interleukin-8, monocyte chemotactic protein-1, and macrophage-colony stimulating factor are present in the vitreous of patients with proliferative diabetic retinopathy (PDR) or proliferative vitreoretinopathy (PVR). The levels of these cytokines were measured by specific enzyme-linked immunoassays in vitreous from 30 patients with PDR, 13 patients with PVR, and 26 control individuals, including 10 cadaver eyes and 16 patients with idiopathic macular holes, idiopathic macular puckers, vitreous hemorrhages, or uncomplicated retinal detachments. Detectable levels of interleukin-8 were found in 90% of vitreous samples of patients with PDR, 85% with PVR, and 58% of control samples. IL-8 was significantly increased in PDR (mean +/- SEM; 25.0 +/- 5.3 ng/ml; p = 0.01), but not in PVR (11.9 +/- 3.9 ng/ml; p = 0.50) compared to control human vitreous (8.5 +/- 2.5 2.5 ng/ml). MCP-1 was detected in 90% of vitreous samples of patients with PDR, 92% with PVR, and 81% of control samples. MCP-1 was significantly increased in PDR (6.2 +/- 0.9 ng/ml, p = 0.001) and PVR (7.7 +/- 2.5 ng/ml, p = 0.001) over the levels in control vitreous (1.2 +/- 0.2 ng/ml). M-CSF was detected in 94% of vitreous samples of patients with PDR, 88% with PVR, and 92% from control vitreous. M-CSF was significantly elevated in PDR (32.3 +/- 8.3 ng/ml, p = 0.03), but not in PVR (23.6 +/- 12.8 ng/ml, p = 0.4) compared to control (10.7 +/- 3.5 ng/ml). Our results suggest that IL-8, MCP-1, and M-CSF participate in the pathogenesis of PDR and PVR.

Topics: Chemokine CCL2; Cytokines; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Macrophage Colony-Stimulating Factor; Vitreoretinopathy, Proliferative; Vitreous Body