Quantitative Analysis of Interleukin-6 in Vitreous from Patients with Proliferative Vitreoretinal Diseases

doi:10.1016/S0021-5155(00)00290-2

Japanese Journal of Ophthalmology

Volume 45, Issue 1, January–February 2001, Pages 40-45

https://doi.org/10.1016/S0021-5155(00)00290-2 Get rights and content

Abstract

Purpose: To explore immunological mechanisms in the pathogenesis of proliferative vitreoretinal diseases, we measured the concentration of interleukin-6 in the vitreous body and serum from patients with proliferative diabetic retinopathy (PDR), proliferative vitreoretinopathy (PVR), and premacular fibrosis. To evaluate immunological etiology, interleukin-6 levels in each disease were compared with disease severity.

Methods: Clinical samples were obtained at the beginning of pars plana vitrectomy from 30 eyes of 26 patients with PDR, 12 eyes of 12 patients with PVR, and 10 eyes of 10 patients with premacular fibrosis. Interleukin-6 was quantitated with an enzyme-linked immunosorbent assay.

Results: The levels of detectable interleukin-6 in the vitreous specimens ranged from 22.8 to 666.4 pg/mL in the PDR patients and from 28.2 to 416.3 pg/mL in the PVR patients. No interleukin-6 was detected in the vitreous specimens from patients with premacular fibrosis or in any serum samples from patients. Interleukin-6 levels of vitreous specimens from PDR patients were higher than those from PVR patients (P < .02, Mann–Whitney U-test). There was no correlation between clinical severity and interleukin-6 levels in vitreous specimens from either PDR or PVR patients.

Conclusion: Our results indicated that cell-mediated immunity is involved in the pathogenesis of proliferative vitreoretinal diseases.

Introduction

Preretinal membranes in patients with proliferative vitreoretinal diseases, such as proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR), are thought to be caused by migration and proliferation of various types of cells, including pigment epithelial cells, fibroblasts, glial cells, vascular endothelial cells, and macrophages.1, 2, 3, 4, 5, 6 It has been reported that these phenomena are modified by various cytokines and growth factors7, 8, 9, 10 that are released from both normal and damaged retina. When retinal ischemia or retinal detachment occurs, these damaged areas may release cytokines. Many studies have been performed to resolve the pathogenesis of proliferative retinal diseases. Autoimmune mechanisms have been thought to play a role in the pathogenesis of proliferative retinal diseases.11, 12, 13 Rahi and Addison¹⁴ have suggested a relationship between immunological mechanisms and the development of microangiopathy. Baudouin et al15, 16, 17, 18, 19, 20, 21 found ocular deposits of immunogloblin, complement, and human leukocyte antigen in patients with PDR and PVR in a series of studies using an immunohistochemical method.

Interleukin-6 is known to be a potent cytokine in cell-mediated immunity that plays an important role in the regulation of immune response, acute phase reactions, and hematopoiesis. Abnormal production of interleukin-6 has been implicated in many kinds of diseases, including autoimmune diseases, chronic inflammation, and lymphoid malignancies. To explore immunological mechanisms in the pathogenesis of proliferative vitreoretinal diseases, we measured the concentration of interleukin-6 in the vitreous body and serum from patients with PDR, PVR, and premacular fibrosis and compared it with the severity of each disease.

Section snippets

Patients

Clinical samples were obtained at the beginning of pars plana vitrectomy from 30 eyes of 26 patients with PDR (Table 1), 12 eyes of 12 patients with PVR (Table 2), and 10 eyes of 10 patients with premacular fibrosis (Table 3). The PDR patients included 17 men and 9 women, ranging in age from 28 to 67 years (average: 52 years). The duration of diabetes in these patients ranged from 2 to 30 years. Of 26 PDR patients, 25 patients had non-insulin-dependent diabetes (type II). Only 1 patient was

Results

Interleukin-6 levels could be measured in 28 of 30 vitreous specimens (93%) from patients with PDR and in 7 of 12 specimens (58%) from patients with PVR Table 1, Table 2. In the vitreous specimens from patients with premacular fibrosis, no interleukin-6 was detected (Figure 1). The levels of detectable interleukin-6 in the vitreous specimens ranged from 22.8 to 666.4 pg/mL (136.1 ± 26.1 pg/mL; mean ± SEM) in the PDR patients and from 28.2 to 416.3 pg/mL (69.9 ± 33.7 pg/mL; mean ± SEM) in the

Discussion

Preretinal membranes of PDR patients contain newly formed vessels and extracellular matrix that were affected by liberated angiogenic factors produced by the ischemic retina, so-called nonperfusion areas. The characteristics of these angiogenic factors involved in retinal angiogenesis have not yet been clarified.

The main complication of rhegmatogenous retinal detachment is PVR. This complication results from migration and proliferation of various types of cells, including those derived from

References (29)

R. Machemer et al.
Pigment epithelial proliferation in human retinal detachment with massive periretinal proliferation
Am J Ophthalmol
(1978)
S.J. Ryan
The pathophysiology of proliferative vitreoretinopathy in its management
Am J Ophthalmol
(1985)
P. Wiedemann
Growth factors in retinal diseasesproliferative vitreoretinopathy, proliferative diabetic retinopathy, and retinal degeneration
Surv Ophthalmol
(1992)
C.J. Brinkman et al.
Cell-mediated immunity after retinal detachment as determined by lymphocyte stimulation
Am J Ophthalmol
(1978)
C. Baudouin et al.
Immunohistopathologic findings in proliferative diabetic retinopathy
Am J Ophthalmol
(1988)
C. Baudouin et al.
Immunohistologic study of proliferative vitreoretinopathy
Am J Ophthalmol
(1989)
C. Baudouin et al.
Class II antigen expression in diabetic preretinal membranes
Am J Ophthalmol
(1990)
A. Waage et al.
Interleukin-6 in synovial fluid from patients with arthritis
Clin Immunol Immunopathol
(1989)
G.F. Bottazzo et al.
Role of aberrant HLA-DR expression and antigen presentation in induction of endocrine autoimmunity
Lancet
(1983)
H.D. Van et al.
Glial cell proliferation in human retinal detachment with massive periretinal proliferation
Am J Ophthalmol
(1977)

D.A. Newsome et al.

Human massive periretinal proliferation. In vitro characteristics of cellular components

Arch Ophthalmol

(1981)

P.S. Hiscott et al.

Retinal pigment epithelial cells in epiretinal membranesan immunohistochemical study

Br J Ophthalmol

(1984)

M. Weller et al.

Demonstration of mononuclear phagocytes in a human epiretinal membrane using a monoclonal anti-human macrophage antibody

Graefes Arch Clin Exp Ophthalmol

(1988)

B. Kirchhof et al.

Pathogenesis of proliferative vitreoretinopathy. Modulation of retinal pigment epithelial cell functions by vitreous and macrophages

Dev Ophthalmol

(1989)

Cited by (28)

Cathepsin-D, Adiponectin, TNF-α, IL-6 and hsCRP plasma levels in subjects with diabetic foot and possible correlation with clinical variables: A multicentric study
2012, Foot
Citation Excerpt :
The association between the low levels of adiponectin with low levels of HDL-cholesterol might represent an independent cardiovascular risk factor, whereas high levels of adiponectin are associated with high levels of HDL-cholesterol indicating a protective risk profile [18]. TNF-α may act as local intensification signals in pathological processes associated with chronic inflammation [19,20]. These cytokines may mediate the synthesis of acute phase proteins which are able to initiate and support inflammatory process in the vascular wall.
Diabetic foot, characterized by a pronounced inflammatory reaction, decreased collagen content and biosynthesis and accelerated degradation are crucial in wound healing. Cathepsin D, an aspartic endopeptidases implicated in cell growth, apoptosis, and its inhibitor has been reported to reverse the inhibition of collagen biosynthesis in wounded rat skin with diabetes. To date, the increased proteolytic activity of Cathepsin D in diabetic foot has not been evaluated and the pathogenic significance of the inflammation has received little attention. Of the patients [with ulcer (n = 211) and without ulcer (n = 208)], 89.73% had type 2 diabetes. Subjects with diabetic foot ulcer showed higher median plasma level of Cathepsin D [556.3 (312.6–587.3) RFC/ml vs 306.3 (92.6–337.3) RFC/ml], TNF-α [96.6 (79.9–121.5) ng/ml vs 8.4 (7.1–9.20) ng/ml], IL-6 [32.2 (8.52–48.4) ng/ml vs 4.9 (4.5–5.6) ng/ml], hsCRP [12.6 (11.2–14.1) mg/ml vs 3.90 (3.50–4.60) mg/ml] and lower median plasma levels of adiponectin [8.50 (7.10–9.5) ng/ml vs 13.3 (12.1–14.2) ng/ml]. A positive correlation was found between grades of ulcer, BMI, A1c and retinopathy for Cathepsin D, for adiponectin, between grades of ulcer, BMI, retinopathy, nephropathy & smoking, for IL-6, between grades of ulcer, BMI, nephropathy, CAD & smoking, for hsCRP, grades of ulcer, BMI, LDL-C, nephropathy & smoking, while total cholesterol, nephropathy, PAD, smoking and neuropathy for TNF-α.
Intraocular expression of serum amyloid A and interleukin-6 in proliferative diabetic retinopathy
2011, American Journal of Ophthalmology
Because serum amyloid A can regulate angiogenesis, we searched for an association between serum amyloid A and interleukin-6 (IL-6), as proinflammatory factors, and proliferative diabetic retinopathy (PDR).
Retrospective, comparative study.
Seventy-six patients (76 eyes) with PDR and 31 patients (31 eyes) with nondiabetic ocular disease (control group), including idiopathic epiretinal membranes (8 eyes) and idiopathic macular holes (23 eyes), were enrolled. Enzyme-linked immunosorbent assay, dual-color immunofluorescence staining, and semiquantitative reverse-transcription polymerase chain reaction were used to examine the serum amyloid A and IL-6 levels in vitreous and plasma, expression of protein and mRNA of serum amyloid A in the excised membranes, respectively.
Vitreous serum amyloid A and IL-6 levels in the study group were significantly higher than those in the control group (both P < .001), whereas the plasma concentrations of serum amyloid A and IL-6 did not vary significantly between the groups (P = .555 and P = .621, respectively). A significant correlation was observed between the vitreous and plasma levels of serum amyloid A in subjects with PDR (r = 0.525; P < .001). In fibrovascular membranes of the study group, colocalization of endothelial marker CD31 with serum amyloid A and colocalization of fibrillar structure markers fibronectin with serum amyloid A were observed. Expression of serum amyloid A mRNA was significantly higher in fibrovascular membranes with PDR than in idiopathic epiretinal membranes (P = .004).
Serum amyloid A and IL-6 may be involved with the inflammatory process in the development of PDR. Local expression of serum amyloid A may exist in PDR.
TNF-α is an independent serum marker for proliferative retinopathy in type 1 diabetic patients
2008, Journal of Diabetes and its Complications
Citation Excerpt :
IL-6 acts in synergy with IL-1β and TNF-α (Song & Kellum, 2005). Several studies demonstrated a correlation between IL-6 and diabetic retinopathy (Abu el Asrar et al., 1992; Funatsu et al., 2002; Kauffmann et al., 1994; Kojima, Yamada, & Tamai, 2001; Shimizu, Funatsu, Yamashita, Yamashita, & Hori, 2002). In the present study, we did find trends toward a positive correlation between IL-6 and PDR, but the number of samples exceeding the detection limit for IL-6 was too few to allow any further statistical analyses.
This study aimed to determine if there are any associations between serum levels of inflammatory markers and proliferative retinopathy (PDR) in type 1 diabetic patients.
A cross-sectional design was utilized for this study.
One hundred twenty-eight type 1 diabetic patients underwent stereo fundus photography according to the Early Treatment Diabetic Retinopathy Study and were divided into two retinopathy groups: no or nonproliferative retinopathy (NDR/NPDR; n=62) and PDR (n=66). Serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, soluble vascular cellular adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1), P-selectin, and high-sensitivity C-reactive protein (hsCRP) were analyzed. Statistical analysis was performed using nonparametric Mann–Whitney U test and multivariate logistic regression analysis.
Patients with PDR had higher levels of TNF-α [7.0 pg/ml (<4–17) vs. 6.0 pg/ml (<4–25); P=.009], sVCAM-1 [860 ng/ml (360–2120) vs. 700 ng/ml (310–1820); P<.001], and P-selectin [180 ng/ml (39–400) vs. 150 ng/ml (42–440); P=.017; figures are expressed as median (range)]. There were no differences in serum levels of sICAM-1 or hsCRP. IL-1β was not detectable in any patient, and IL-6 was detectable in only 22.7% of the patients. In multivariate logistic regression analysis, TNF-α was the single, persistent, independent determinant inflammatory marker for PDR.
The association between TNF-α and PDR in type 1 diabetic patients suggests that inflammation might play a role in the pathogenesis of proliferative diabetic retinopathy.
Elevated intravitreal interleukin-6 levels in patients with proliferative diabetic retinopathy
2006, Canadian Journal of Ophthalmology
Background: We conducted this study to elucidate the possible role of interleukin-6 (IL-6) in the pathogenesis of proliferative diabetic retinopathy (PDR).
Methods: Together with pertinent clinical and laboratory data, intravitreal and serum concentrations of IL-6 were determined in 8 patients with PDR by means of enzyme-linked immunosorbent assay (ELISA). The results were compared with data from 8 nondiabetic control subjects undergoing vitrectomy.
Results: Significantly higher intravitreal IL-6 concentrations were found in patients with PDR (mean [SD], 755 [177] pg/mL) compared with control subjects (93 [151] pg/mL) (p = 0.001). The serum IL-6 levels in the PDR group were lower than the measurable threshold of the ELISA kit (<0.16 pg/mL). Diabetic patients with macular edema had a higher mean (SD) level of intravitreal IL-6 (896 [73] pg/mL) compared with patients without macular edema (613 [119] pg/mL) (Mann-Whitney U test, p = 0.03). Correlation analysis did not reveal any significant association between intravitreal IL-6 levels and patient age, duration of either diabetes mellitus or vitreous hemorrhage, panretinal photocoagulation, type of current medical therapy, hyperglycemia, or the biochemical indicators of renal function.
Interpretation: IL-6, a proinflammatory cytokine, may have a role in PDR. Intraocular production of IL-6 appears to be responsible for the elevated intravitreal levels observed.
Contexte: Cette étude avait pour objet d’élucider le rôle que pourrait jouer l’interleukine-6 (IL-6) dans la pathogénèse de la rétinopathie diabétique proliférante (RDP).
Méthodes: En joignant des résultats cliniques et de laboratoire, nous avons établi les concentrations intravitréennes et sériques d’IL-6 chez 8 patients atteints de RDP, au moyen du dosage par la méthode E.L.I.S.A. Les résultats ont été comparés à ceux de 8 de témoins non diabétiques subissant une vitrectomie.
Résultats: Des concentrations d’IL-6 significativement plus élevées ont été trouvées chez les patients atteints de RDP (moyenne [ÉT], 755 [177] pg/mL), comparativement aux témoins (93 [151] pg/mL) (p = 0,001). Les taux sériques d’IL-6 chez le groupe atteint de RDP étaient inférieurs au seuil mesurable par la méthode E.L.I.S.A. (<0,16 pg/mL). Les patients diabétiques ayant un œdème maculaire avaient un taux intravitréen moyen [ÉT] plus élevé d’IL-6 (896 [73] pg/mL), par rapport à ceux ayant un œdème maculaire (613 [19] pg/mL) (Mann-Whitney U test, p = 0,03). L’analyse corrélative n’a pas révélé d’association significative entre les niveaux intravitréens d’IL-6 et l’âge des patients, la durée du diabète sucré ou de l’hémorragie du vitré, la photocoagulation panrétinienne, le type de thérapie médicale en cours, l’hyperglycémie ou les indicateurs biochimiques de la fonction rénale.
Interprétation: L’IL-6, cytokine proinflammatoire, peut jouer un rôle dans la RDP. La production intraoculaire d’IL-6 semble être responsable des taux intravitréniens élevés observés.
The role of cytokines and trophic factors in epiretinal membranes: Involvement of signal transduction in glial cells
2006, Progress in Retinal and Eye Research
Idiopathic epiretinal membranes (ERMs) in the macular region can cause a reduction in vision and sometimes recurs after surgical removal, but its pathogenic mechanisms are still unknown. On the other hand, the presence of secondary ERMs has been associated with various clinical conditions including proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). Recent studies have shown a significant association between clinical grades of PDR or PVR, and the expression levels of specific cytokines and/or growth factors in the vitreous fluid. Expression of these factors and their receptors are also observed in secondary ERMs. ERMs are composed of many cell types such as retinal pigment epithelial cells and vascular endothelial cells, however the role of glial cells is yet unclear. Interestingly, glial cells in ERMs express some trophic factor receptors and transcription factors, such as NF-κB, suggesting an involvement of glial signal transduction in the pathogenesis of ERMs. In this review, we summarize recent progress regarding the clinical and laboratory findings of ERMs.
Vitreous Fluid Biomarkers
2006, Advances in Clinical Chemistry
Citation Excerpt :
PVR is associated with increased expression and action of various cytokines and growth factors. These factors include IL‐6 [183–186], IL‐8 [187], ICAM‐1 [188, 189], MCP‐1 [190, 191], PDGF [192], FGF [186, 192], HGF [193], PEDF [194], TGF‐β [195], TNF [196], and others. In one study, Kon et al. [197] evaluated the vitreous fluid concentration of TGF‐β2, bFGF, IL‐1β, IL‐6, and protein in patients with retinal detachment to determine the value of these mediators in predicting the future development of PVR.
Interactions between cells and the network of secreted proteins are associated with the ocular disease. In most cases, clinical appearcance is sufficiently diagnostic. However, in cases of nonspecific or atypical clinical presentation, diagnostic sampling of vitreous fluid can aid diagnosis and treatment for ocular disease. Progresses in the basic sciences, particularly molecular biology, and advances in surgical instrumentation have greatly enhanced the diagnostic armamentarium. These developments also have led to a better understanding of the pathophysiological processes involved in ocular diseases and have prompted evolution of new therapeutic modalities. In this chapter, we review techniques for vitreous fluid sampling and biomarker quantitation thereof. The molecular biology of bioactive vitreous fluid factors is also discussed with respect to their clinical involvement in the development of ocular disease.

View all citing articles on Scopus

View full text

Quantitative Analysis of Interleukin-6 in Vitreous from Patients with Proliferative Vitreoretinal Diseases

Abstract

Introduction

Section snippets

Patients

Results

Discussion

Am J Ophthalmol

Am J Ophthalmol

Surv Ophthalmol

Am J Ophthalmol

Am J Ophthalmol

Am J Ophthalmol

Am J Ophthalmol

Clin Immunol Immunopathol

Lancet

Glial cell proliferation in human retinal detachment with massive periretinal proliferation

Am J Ophthalmol

Human massive periretinal proliferation. In vitro characteristics of cellular components

Arch Ophthalmol

Retinal pigment epithelial cells in epiretinal membranesan immunohistochemical study

Br J Ophthalmol

Demonstration of mononuclear phagocytes in a human epiretinal membrane using a monoclonal anti-human macrophage antibody

Graefes Arch Clin Exp Ophthalmol

Pathogenesis of proliferative vitreoretinopathy. Modulation of retinal pigment epithelial cell functions by vitreous and macrophages

Dev Ophthalmol