Analysis of multiple cytokines in aqueous humor of patients with idiopathic macular hole

Abstract Background: Idiopathic macular holes are common ophthalmic manifestations with unknown pathogenesis. Thus far, there has been minimal research regarding the causes of idiopathic macular holes, especially with respect to the underlying immune mechanism. To provide clarity regarding the treatment and prognosis of idiopathic macular holes, specifically regarding the levels of cytokines in affected patients, this study examined and analyzed multiple cytokine levels in aqueous humor from patients with idiopathic macular holes.Methods: This comparative cross-sectional study included 38 patients in two groups: a cataract control group (n=17) and an idiopathic macular hole group (n=21). The levels of 48 cytokines in aqueous humor were detected by multiplex analysis with antibody-coupled magnetic beads. The Kolmogorov–Smirnov test was used to check whether the data were normally distributed; Student’s t-test and the Mann–Whitney U test were used to assess differences in cytokine levels between the two groups. Spearman correlation analysis was used to assess relationships among cytokine levels in the experimental group. Signaling pathways containing cytokines with significantly different expression in the experimental group were identified.Results: There were significant differences in aqueous humor cytokine levels between patients with idiopathic macular holes and patients in the cataract control group. Notably, hepatocyte growth factor (p=0.0001), GM-CSF (p=0.0111), and IFN-γ (p=0.0120) were significantly upregulated in the experimental group, while TNF-α (p=0.0032), GRO-α (p<0.0001), and MIF (p<0.0001) were significantly downregulated in the experimental group. Furthermore, the GM-CSF level showed significant positive correlations with levels of IL-1 (r=0.67904, p<0.001), IL-4 (r=0.76017, p<0.001), and IFN-γ (r=0.59922, p=0.004097) in the experimental group. Moreover, the levels of nerve growth factor and hepatocyte growth factor showed a significant positive correlation (r=0.64951, p=0.001441) in the experimental group.Conclusions: Patients with idiopathic macular holes showed significant variation in aqueous humor immune response after the onset of hole formation, including the recruitment of immune cells and regulation of cytokine expression. Our findings also suggest that it is not appropriate to use patients with macular holes as the control group in studies of aqueous humor cytokine levels in ophthalmic diseases.

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Analysis of multiple cytokines in aqueous humor of patients with idiopathic macular hole

10.21203/rs.3.rs-25621/v2 ◽

2020 ◽

Author(s):

Heping Wang ◽

Yuqi Li ◽

Song Han ◽

Tongtong Niu

Keyword(s):

Growth Factor ◽

Aqueous Humor ◽

Control Group ◽

Gm Csf ◽

Macular Holes ◽

Cytokine Levels ◽

Idiopathic Macular Holes ◽

Ifn Γ ◽

Hepatocyte Growth ◽

Experimental Group

Abstract Background: Idiopathic macular holes are common ophthalmic manifestations with unknown pathogenesis. Thus far, there has been minimal research regarding the causes of idiopathic macular holes, especially with respect to the underlying immune mechanism. To provide clarity regarding the treatment and prognosis of idiopathic macular holes, specifically regarding the levels of cytokines in affected patients, this study examined and analyzed multiple cytokine levels in aqueous humor from patients with idiopathic macular holes. Methods: This comparative cross-sectional study included 38 patients in two groups: a cataract control group (n=17) and an idiopathic macular hole group (n=21). The levels of 48 cytokines in aqueous humor were detected by multiplex analysis with antibody-coupled magnetic beads. The Kolmogorov–Smirnov test was used to check whether the data were normally distributed; Student’s t-test and the Mann–Whitney U test were used to assess differences in cytokine levels between the two groups. Spearman correlation analysis was used to assess relationships among cytokine levels in the experimental group. Signaling pathways containing cytokines with significantly different expression in the experimental group were identified. Results: There were significant differences in aqueous humor cytokine levels between patients with idiopathic macular holes and patients in the cataract control group. Notably, hepatocyte growth factor (p=0.0001), GM-CSF (p=0.0111), and IFN-γ (p=0.0120) were significantly upregulated in the experimental group , while TNF-α (p=0.0032), GRO-α (p<0.0001), and MIF (p<0.0001) were significantly downregulated in the experimental group. Furthermore, the GM-CSF level showed significant positive correlations with levels of IL-1 (r=0.67904, p<0.001), IL-4 (r=0.76017, p<0.001), and IFN-γ (r=0.59922, p=0.004097) in the experimental group. Moreover, the levels of nerve growth factor and hepatocyte growth factor showed a significant positive correlation (r=0.64951, p=0.001441) in the experimental group. Conclusions: Patients with idiopathic macular holes showed significant variation in aqueous humor immune response after the onset of hole formation, including the recruitment of immune cells and regulation of cytokine expression. Our findings also suggest that it is not appropriate to use patients with macular holes as the control group in studies of aqueous humor cytokine levels in ophthalmic diseases.

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Analysis of multiple cytokines in aqueous humor of patients with idiopathic macular hole

10.21203/rs.3.rs-25621/v3 ◽

2020 ◽

Author(s):

Heping Wang ◽

Yuqi Li ◽

Song Han ◽

Tongtong Niu

Keyword(s):

Growth Factor ◽

Aqueous Humor ◽

Control Group ◽

Gm Csf ◽

Macular Holes ◽

Cytokine Levels ◽

Idiopathic Macular Holes ◽

Ifn Γ ◽

Hepatocyte Growth ◽

Experimental Group

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Analysis of multiple cytokines in aqueous humor of patients with idiopathic macular hole

10.21203/rs.3.rs-25621/v1 ◽

2020 ◽

Author(s):

Heping Wang ◽

Yuqi Li ◽

Song Han ◽

Tongtong Niu

Keyword(s):

Macular Hole ◽

Aqueous Humor ◽

Significant Positive Correlation ◽

Control Group ◽

Idiopathic Macular Hole ◽

Positive Correlation ◽

Gm Csf ◽

Macular Holes ◽

Idiopathic Macular Holes ◽

Ifn Γ

Abstract Background Idiopathic macular hole is a common type of ophthalmic disease, and its pathogenesis is unknown. At present, little research about the pathogenesis, especially the immune mechanism, of idiopathic macular hole has been reported. To provide scientific guidance for the treatment and prognosis of idiopathic macular holes, this study examined and analyzed multiple cytokines levels in aqueous humor from patients with idiopathic macular hole. Methods Thirty-eight subjects were divided into cataract control group (n = 17) and idiopathic macular hole group (n = 21). The levels of 48 cytokines in aqueous humor were detected by multiple magnetic beads immunization. Results There were significant differences in cytokine levels in aqueous humor from idiopathic macular holes patients compared with the cataract control group. For example, HGF (p = 0.0001), GM-CSF (p = 0.0111), and IFN-γ (p = 0.0120) were significantly up-regulated, while TNF-α (p = 0.0032), GRO-α (p < 0.0001), and MIF (p < 0.0001) were significantly down-regulated. GM-CSF showed a significant positive correlation with IL-1 (r = 0.67904, p < 0.001), IL-4 (r = 0.76017, p < 0.001), and IFN-γ (r = 0.59922, p = 0.004097). Moreover, NGF and HGF also showed a significant positive correlation (r = 0.64951, p = 0.001441). Conclusion Patients with idiopathic macular hole could exhibit significant variation of the immune reaction in aqueous humor after the disease, including the recruitment of immune cells and the regulation of cytokine expression. This study also implied that it is not rigorous to use macular hole as the control in current studies on aqueous humor cytokines in ophthalmic diseases.

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IMMMUNE MEDIATORS IN IDIOPATHIC UVEITIS

10.1101/2021.08.04.455093 ◽

2021 ◽

Author(s):

Marie-Helene Errera ◽

Ana Pratas ◽

Sylvain Fisson ◽

Thomas Manicom ◽

Marouane Boubaya ◽

...

Keyword(s):

Aqueous Humor ◽

Unsupervised Classification ◽

Control Group ◽

Serum Samples ◽

Gm Csf ◽

Cytokines And Chemokines ◽

Age Related ◽

Order Of Magnitude ◽

Ifn Γ ◽

Endothelial Growth Factor

To investigate which cytokines, chemokines and growth factors are involved in the immunopathogenesis of idiopathic uveitis, and whether cytokine proﬁles are associated with. Serum and aqueous humor (AH) samples of 75 patients with idiopathic uveitis were analyzed by multiplex immunoassay. Infectious controls consisted of 16 patients with ocular toxoplasmosis all conﬁrmed by intraocular ﬂuid analyses. Noninfectious controls consisted of 7 patients with Behçet disease related uveitis and 15 patients with sarcoidosis related uveitis. The control group consisted of AH and serum samples from 47 noninﬂammatory control patients with age-related cataract. In each sample, 27 immune mediators ± IL-21 and IL-23 were measured. In idiopathic uveitis, 13 of the 29 mediators, including most proinﬂammatory and vascular mediators such as IL-6, IL-8, IL-12, G-CSF, GM-CSF, MCP-1, IP-10, TNF-α and VEGF, were signiﬁcantly elevated in the aqueous humor when compared to all controls. Moreover, IL-17, IP-10, and IL-21, were signiﬁcantly elevated in the serum when compared to all controls. We clustered 4 subgroups of idiopathic uveitis using a statistical analysis of hierarchical unsupervised classification, characterized by the order of magnitude of concentrations of intraocular cytokines. The pathogenesis of idiopathic uveitis is characterized by the presence of predominantly proinﬂammatory cytokines and chemokines and vascular endothelial growth factor with high expression levels as compared to other causes of uveitis. There are indications for obvious Th-1/ IL21-Th17 pathways but also IL9-Th9 and increased IFN-γ-inducing cytokine (IL12) and IFN-γ-inducible CXC chemokine (IP-10). The combined data suggest that immune mediator expression is different among idiopathic uveitis. This study suggests various clusters among the idiopathic uveitis group rather than one speciﬁc uveitis entity.

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Elevated Cytokine Levels in Aqueous Humor Are Associated with Peripheral Anterior Synechiae after Penetrating Keratoplasty

International Journal of Molecular Sciences ◽

10.3390/ijms222212268 ◽

2021 ◽

Vol 22 (22) ◽

pp. 12268

Author(s):

Yuki Kusano ◽

Takefumi Yamaguchi ◽

Sota Nishisako ◽

Takehiro Matsumura ◽

Masaki Fukui ◽

...

Keyword(s):

Aqueous Humor ◽

Penetrating Keratoplasty ◽

Corneal Transplantation ◽

Gm Csf ◽

Experimental Glaucoma ◽

Cytokine Levels ◽

Permanent Loss ◽

Ifn Γ ◽

Macrophage Colony ◽

Factor Α

Peripheral anterior synechiae (PAS) after corneal transplantation leads to refractory glaucoma and permanent loss of vision. However, the exact mechanism remains elusive. This study aimed to evaluate the association between cytokine levels in the aqueous humor (AqH) and the progression of PAS after penetrating keratoplasty (PKP). We measured 20 cytokine levels in AqH and assessed the correlation with PAS progression after PKP in 85 consecutive patients who underwent PKP. We also evaluated age-dependent alterations in PAS and cytokine levels in DBA2J mice. PAS developed in 38 (44.7%) of 85 eyes after PKP. The incidence of intraocular pressure increase after PKP was significantly greater in eyes with PAS (26.3%) than in those without PAS (2%, p = 0.0009). The PAS area at 12 months after PKP was significantly positively correlated with the preoperative levels of interleukin (IL)-6, interferon (IFN)-γ and monocyte chemotactic protein (MCP)-1 (p ≤ 0.049). In the DBA2J mice, an experimental glaucoma model that developed PAS at 50 weeks, the AqH levels of IL-2, IL-6, IL-10, IFN-γ, tumor necrosis factor-α, MCP-1 and granulocyte-macrophage colony-stimulating factor (GM-CSF) significantly increased at 50 weeks compared to 8 weeks (p ≤ 0.021). In conclusion, inflammatory alterations in the AqH microenvironment, such as high preoperative specific cytokine levels, can lead to PAS formation and glaucoma.

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IL-10-592 A/C Gene Polymorphism and Cytokine Levels are Associated with Susceptibility to Drug Resistance in Tuberculosis

Turkish Journal of Immunology ◽

10.25002/tji.2020.1235 ◽

2020 ◽

Vol 8 (3) ◽

pp. 103-112

Author(s):

Atefeh SADEGHI SHERMEH ◽

Majid KHOSHMIRSAFA ◽

Ali-Akbar DELBANDI ◽

Payam TABARSI ◽

Esmaeil MORTAZ ◽

...

Keyword(s):

Serum Levels ◽

World Health ◽

Control Group ◽

Drug Resistant ◽

Nucleotide Polymorphisms ◽

Genotype Frequencies ◽

Cytokine Levels ◽

Ifn Γ ◽

Health Organization ◽

And Function

Introduction: Tuberculosis (TB) and especially resistant forms of it have a substantial economic burden on the community health system for diagnosis and treatment each year. Thus, investigation of this field is a priority for the world health organization (WHO). Cytokines play important roles in the relationship between the immune system and tuberculosis. Genetic variations especially single nucleotide polymorphisms (SNPs) impact cytokine levels and function against TB. Material and Methods: In this research SNPs in IFN-γ (+874 T/A) and IL-10 (-592 A/C) genes, and the effects of these SNPs on cytokine levels in a total of 87 tuberculosis patients and 100 healthy controls (HCs) were studied. TB patients divided into two groups: 1) 67 drug-sensitive (DS-TB) and 2) 20 drug-resistant (DR-TB) according to drug sensitivity test using polymerase chain reaction (PCR). For the genotyping of two SNPs, the PCR-based method was used and IFN-γ and IL-10 levels were measured by ELISA in pulmonary tuberculosis (PTB) and control group. Results: In -592A/C SNP, only two genotypes (AA, AC) were observed and both genotypes showed statistically significant differences between DR-TB and HCs (p=0.011). IL-10 serum levels in PTB patients were higher than HCs (p=0.02). The serum levels of IFN-γ were significantly higher in DS-TB patients than that of the other two groups (p<0.001); however, no significant differences were observed for allele and genotype frequencies in IFN-γ +874. Conclusions: Our results suggest that the SNP at -592 position of IL-10 gene may be associated with the susceptibility to DR-TB. However, further investigation is necessary. Keywords: Polymorphism, IFN-γ, IL-10, tuberculosis, drug-resistant tuberculosis

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Aqueous humor cytokine levels through microarray analysis and a sub-analysis based on optical coherence tomography in wet age-related macular degeneration patients

BMC Ophthalmology ◽

10.1186/s12886-021-02152-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jin-Ho Joo ◽

Hyejee Kim ◽

Jae-Ho Shin ◽

Sang Woong Moon

Keyword(s):

Optical Coherence Tomography ◽

Growth Factor ◽

Inflammatory Cytokines ◽

Age Related Macular Degeneration ◽

Gm Csf ◽

Age Related ◽

Ifn Γ ◽

Low Levels ◽

Wet Amd

Abstract Background To identify disease-specific cytokine and growth factor profile differences in the aqueous humor between wet age-related macular degeneration (AMD) patients and age-matched controls and to correlate their levels with the optical coherence tomography (OCT) findings. Methods Aqueous humors were obtained from 13 wet AMD eyes and 10 control eyes. Twenty cytokines and growth factors were measured using a RayBio antibody microarray technology in wet AMD and control eyes. Results The samples obtained from wet AMD patients exhibited a significantly increased expression of MCP-1, MIP-1α, MIP-1β, and vascular endothelial growth factor (VEGF). Subretinal fluid (SRF) patients showed significantly lower levels of proinflammatory cytokines, such as IL-1α and GM-CSF, than those without SRF. Pigment epithelial detachments (PED) patients showed lower levels of inflammatory cytokines, such as GM-CSF, IFN-γ, and TNF-α, than those without PED. Subretinal tissue (SRT) patients showed a higher level of IFN-γ than those without SRT. Compared with the controls, type 1 macular neovascularization (MNV) patients showed increased levels of MCP-1, MIP-1α, and MIP-1β, but not VEGF (p = 0.083). However, type 2 MNV patients showed increased levels of MCP-1 and VEGF (p = 0.040 and p = 0.040). Conclusion Inflammatory cytokines varied according to the type of AMD- and OCT-based parameters. Our observation of low levels of VEGF in patients with type 1 MNV implies that the inhibition of VEGF alone appears to be insufficient treatment for these patients and that cytokines such as MCP-1, MIP-1α, and MIP-1β should be modulated. And the presence of SRF in MNV may be associated with a positive prognosis because we found relatively low levels of proinflammatory cytokines.

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Hepatocyte Growth Factor Is Constitutively Produced by Human Bone Marrow Stromal Cells and Indirectly Promotes Hematopoiesis

Blood ◽

10.1182/blood.v89.5.1560.1560_1560_1565 ◽

1997 ◽

Vol 89 (5) ◽

pp. 1560-1565 ◽

Cited By ~ 2

Author(s):

Kenji Takai ◽

Junichi Hara ◽

Kunio Matsumoto ◽

Gaku Hosoi ◽

Yuko Osugi ◽

...

Keyword(s):

Bone Marrow ◽

Growth Factor ◽

Hepatocyte Growth Factor ◽

Stromal Cells ◽

Mononuclear Cells ◽

Gm Csf ◽

Tumor Growth Factor ◽

Cd34 Cells ◽

Factor Α ◽

Hepatocyte Growth

Bone marrow (BM) stromal cells are required for normal hematopoiesis. A number of soluble factors secreted by these cells that mediate hematopoiesis have been characterized. However, the mechanism of hematopoiesis cannot be explained solely by these known factors, and the existence of other, still unknown stromal factors has been postulated. We showed that hepatocyte growth factor (HGF ) is one such cytokine produced by human BM stromal cells. BM stromal cells were shown to constitutively produce HGF and also to express the c-MET/HGF receptor. The production of HGF was enhanced by addition of heparin and phorbol ester. Dexamethasone and tumor growth factor-β (TGF-β) inhibited the production of HGF. Interleukin-1α (IL-1α) tumor necrosis factor-α (TNF-α), and N6,2′-o-dibutyryl-adenosine-3′:5′-cyclic monophosphate (dbc-AMP) showed no obvious influence on HGF production. Western blot analysis of HGF derived from BM stromal cells showed two bands at 85 and 28 kD corresponding to native and variant HGF, respectively. Addition of recombinant HGF significantly promoted the formation of burst-forming unit-erythroid (BFU-E) and colony-forming unit-granulocyte erythroid macrophage (CFU-GEM) by BM mononuclear cells in the presence of erythropoietin and granulocyte-macrophage colony-stimulating factor (GM-CSF ), but the formation of CFU-GM was not modified. However, HGF had no effects on colony formation by purified CD34+ cells. Within BM mononuclear cells, c-MET was expressed on a proportion of cells (CD34−, CD33+, CD13+, CD14+, and CD15+), but was not found on CD34+ cells. We conclude that HGF is constitutively produced by BM stromal cells and that it enhances hematopoiesis. In addition, expression of c-MET on the stromal cells suggests the presence of an autocrine mechanism, operating through HGF, among stromal cells.

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Proteomic analysis of tears and serum for dry eye disease using ovariectomized cynomolgus monkey

10.21203/rs.2.21889/v1 ◽

2020 ◽

Author(s):

xiaolong yang ◽

Yue Xu ◽

Yun Wang ◽

Chang Li ◽

Xiaofeng Zhang

Keyword(s):

Gene Ontology ◽

Growth Factor ◽

Dry Eye ◽

Cynomolgus Monkey ◽

Enrichment Analysis ◽

Differentially Expressed ◽

Control Group ◽

Differentially Expressed Proteins ◽

Dry Eyes ◽

Experimental Group

Abstract Background Ovariectomized cynomolgus monkey 30 months after surgery was selected as the research object to identify protein changes in tears and serum to provide a reference for the diagnosis and pathogenesis of dry eye in menopausal women. Methods Six cynomolgus monkey were randomly divided into an experimental group and a control group (3 in each group). The experimental group underwent bilateral ovariectomy, while the control group underwent sham surgery with their ovaries reserved. Proteomic analysis was performed by LC-MS/MS on tears and serum collected from two groups. Differentially expressed proteins were identified and were performed cluster analysis, which included gene ontology, the Kyoto Encyclopedia of Genes and Genomes pathway and protein-protein interaction. Results 33 differentially expressed proteins have been identified in tears and17 differentially expressed proteins have been identified in serum. Kyoto Encyclopedia of Genes and Genomes enrichment analysis in tears has discovered Glucagon signaling pathway and neurotrophin signaling pathway may play an important role in the pathogenesis of dry eye. Gene ontology enrichment analysis in serum has discovered insulin-like growth factor binding and growth factor binding in molecular function probably make effort in pathogenesis of dry eye. KEGG analysis in serum has discovered salivary secretion may be the key pathway in pathogenesis of dry eye. Conclusions Protein G7PCH4, Q2PG17 and G7PT55 in tears may be the key protein in pathogenesis of dry eyes. Protein G7P1T1, G7PUN9 and G8F302 in serum may play an important role in pathogenesis of dry eyes.

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Clinical Significance of Hepatocyte Growth Factor and Transforming Growth Factor-Beta-1 Levels in Assessing Disease Activity in Inflammatory Bowel Disease

Canadian Journal of Gastroenterology and Hepatology ◽

10.1155/2020/2104314 ◽

2020 ◽

Vol 2020 ◽

pp. 1-5

Author(s):

Rania Naguib ◽

Wafaa Mohamed El-Shikh

Keyword(s):

Growth Factor ◽

Hepatocyte Growth Factor ◽

Disease Activity ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Inflammatory Responses ◽

Control Group ◽

Inflammatory Bowel ◽

Growth Factor Beta ◽

Hepatocyte Growth

Background. Transforming growth factor-beta (TGF-β) and hepatocyte growth factor (HGF) are inflammatory cytokines which function as key regulators of immunological homeostasis and inflammatory responses. They have been linked to inflammatory bowel diseases (IBD). In this study, we aim to assess the levels of TGF-β and HGF and other inflammatory markers in patients with IBD and correlate them with the disease activity. Study Design. A cross-sectional study involving 100 patients with ulcerative colitis (UC) and 100 patients with Crohn’s disease (CD) and 50 control subjects. TGF-β and HGF levels were measured and correlated with disease activity. Results and Conclusion. Serum levels of TGF-β and HGF were significantly higher in IBD patients compared with the control group. In the UC group, the levels of HGF and TGF-β were significantly higher than in the CD group. Levels of TGF-β and HGF correlate with the activity of IBD.

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