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James T. Rosenbaum - One of the best experts on this subject based on the ideXlab platform.
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The NLRP3 inflammasome is active but not essential in Endotoxin-Induced Uveitis
Inflammation Research, 2012Co-Authors: Holly L. Rosenzweig, April Woods, Jenna S. Clowers, Stephen R. Planck, James T. RosenbaumAbstract:Objective The inflammasome complex involving caspase-1 and nucleotide-binding domain, leucine-rich repeat containing protein (NLRP)3, also known as NALP3 or cryopyrin is important for host responses to microbial pathogens and several autoinflammatory diseases. We investigated the extent to which NLRP3 and caspase-1 control ocular interleukin (IL)-1β production and severity of Uveitis (intraocular inflammatory disease) in an established, acute inflammatory Uveitis model, Endotoxin-Induced Uveitis (EIU). Methods Expression of NLRP3, its adaptor molecule ASC, also known as PYCARD (PYD and CARD domain containing), and caspase-1 were examined by immunoblotting. IL-1β production was measured by enzyme-linked immunosorbent assay (ELISA). Using knockout mice, roles for caspase-1 and NLRP3 were examined in Uveitis induced by intraocular injection of Escherichia coli lipopolysaccharide (LPS). Results NLRP3, ASC, and caspase-1 proteins are constitutively expressed in eye tissue. During EIU, IL-1β protein production increases; this requires the presence of both caspase-1 and NLRP3. However, severity of EIU is not altered by deficiency in either caspase-1 or NLRP3, as assessed by both intravital microscopy and histology. Conclusions These data identify the importance of the NLRP3 inflammasome for IL-1β production in the eye, yet indicate that its participation in EIU is nonessential.
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The NLRP3 inflammasome is active but not essential in Endotoxin-Induced Uveitis
Inflammation Research, 2011Co-Authors: Holly L. Rosenzweig, April Woods, Jenna S. Clowers, Stephen R. Planck, James T. RosenbaumAbstract:Objective The inflammasome complex involving caspase-1 and nucleotide-binding domain, leucine-rich repeat containing protein (NLRP)3, also known as NALP3 or cryopyrin is important for host responses to microbial pathogens and several autoinflammatory diseases. We investigated the extent to which NLRP3 and caspase-1 control ocular interleukin (IL)-1β production and severity of Uveitis (intraocular inflammatory disease) in an established, acute inflammatory Uveitis model, Endotoxin-Induced Uveitis (EIU).
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Susceptibility to endotoxin induced Uveitis is not reduced in mice deficient in BLT1, the high affinity leukotriene B4 receptor
British Journal of Ophthalmology, 2004Co-Authors: Justine R. Smith, K. Subbarao, D.t. Franc, B. Haribabu, James T. RosenbaumAbstract:Aim: To investigate the role of arachidonic acid derived chemotactic factor, LTB4, in the development of endotoxin induced Uveitis (EIU), using mice deficient in the BLT1 gene which encodes the high affinity LTB4 receptor. Methods: BLT1 gene deficient and wild type BALB/c mice were injected intravitreally with Escherichia coli 055:B5 lipopolysaccharide (250 ng/2 μl). Number of leukocytes invading the anterior chamber 24 hours later were counted on tissue cross sections. Results: In all mice, EIU was characterised by a polymorphonuclear and mononuclear cell infiltrate. Numbers of infiltrating cells did not differ significantly between control and BLT1 gene knockout mice. Conclusion: Chemotactic factors other than LTB4 are primarily responsible for leukocyte migration into the eye during murine EIU.
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Interleukin-6 does not mediate Endotoxin-Induced Uveitis in mice: Studies in gene deletion animals
Investigative Ophthalmology & Visual Science, 1998Co-Authors: James T. Rosenbaum, Paul Kievit, Jong Moon Park, Stephen R. PlanckAbstract:PURPOSE. Interleukin-6 (IL-6) has been strongly implicated in anterior Uveitis based on its presence in aqueous humor from infected eyes and its inflammatory effects when injected intravitreally into rats. We used IL-6-deficient mice to test further the hypothesis that IL-6 contributes to the development of Endotoxin-Induced Uveitis. METHODS. Uveitis was scored by histologic analysis of C3H/HeN mice 24 hours after intravitreal injections of up to 200 ng of recombinant murine IL-6. Uveitis was similarly measured in IL-6-deficient mice and congenic controls 24 hours after intravitreal injection of 250 ng of Esherichia colt endotoxin. Reverse transcription-polymerase chain reaction was used to detect mRNAs for several cytokines at 3 hours postinjection. The IL-6 concentration in aqueous humor samples was determined with a bioassay using the murine B9 plasmacytoma cell line. RESULTS. Direct injection of IL-6 did not induce Uveitis. Mice genetically deficient in IL-6 developed Endotoxin-Induced Uveitis that was comparable or more severe than congenic control mice. Compensatory. changes in the expression of mRNA for other cytokines were not detected in irises from the IL-6-deficient mice. In IL-6-competent mice that received bilateral endotoxin injections, no correlation was found between the number of infiltrating cells in one eye and the IL-6 concentration in the aqueous humor of the contralateral eye. CONCLUSIONS. In marked contrast to previous conclusions with rats, IL-6 was not sufficient for inducing Uveitis in mice. Additionally, IL-6 was not necessary for the development of Uveitis subsequent to intravitreal injection of endotoxin in mice.
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THE EFFECT OF GENETIC DEFICIENCY OF ADHESION MOLECULES ON THE COURSE OF Endotoxin-Induced Uveitis
Current Eye Research, 1998Co-Authors: Stephen R. Planck, Jong Moon Park, Leslie M. O'rourke, Jose Carlos Gutierrez-ramos, James T. RosenbaumAbstract:PURPOSE. Multiple adhesion molecules of the selectin, integrin, and immunoglobulin-like families are involved in the migration of leukocytes out of the bloodstream into inflamed tissues. This study addresses the question of which adhesion molecules are specifically involved in Endotoxin-Induced Uveitis. METHODS. Mice genetically deficient in p-selectin, ICAM-1, s 2 -integrin, or controls received intravitreal injections of endotoxin. Eyes were harvested 24 h later and inflammation was evaluated by histologic and immunohistochemical assays of infiltrating cells. RESULTS. Mice lacking either P-selectin or s 2 -integrin had less inflammation than controls (median cells/section: 64 for P-selectin knockout vs 130 for controls, p = 0.02, n = 17 per group; 244 for s 2 -integrin knockouts, n = 14, vs 355 for controls, n = 17, p = 0.05). Neither gene deletion significantly changed the ratio of infiltrating neutrophils to macrophages. ICAM-1 knockouts tended to have fewer infiltrating cells (median 22 cells/section...
Chi-chao Chan - One of the best experts on this subject based on the ideXlab platform.
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Endotoxin-Induced Uveitis in cyclooxygenase-2-deficient mice.
Investigative Ophthalmology & Visual Science, 2004Co-Authors: Nadine Tuaillon, Defen Shen, Chi-chao ChanAbstract:PURPOSE. Endotoxin-Induced Uveitis (EIU) is a model that mimics human acute anterior Uveitis. Cyclooxygenase (COX)-2 is an enzyme that initiates the conversion of arachidonic acid (AA) into prostaglandins (PGs), whereas 5-lipoxygenase (5-LO) generates leukotrienes (LT). The purpose of this study was to delineate the role of COX-2 in acute ocular inflammation. METHODS. EIU was induced in wild-type (WT), heterozygotic (COX-2 +/- ) and COX-2 null (COX-2 -/- ) mice by injection of lipopolysaccharide (LPS). Other mice were coinjected with LPS and IFNy. Ocular histology, serum cytokines, and AA products determined by ELISA, and relevant ocular messengers determined by RT-PCR were compared among the different groups. RESULTS. Histology showed that the EIU score was significantly enhanced in COX-2 / mice in comparison to WT and COX-2 +/- . PGE 2 was increased in WT and COX-2 +/- EIU but not in COX-2 / EIU. LTB 4 in serum and ocular 5-LO transcripts were increased in COX-2 / EIU mice in comparison with WT and COX-2 +/- EIU mice. IL-6 increased, whereas IFNγ decreased both in serum and ocular transcripts in COX-2 / EIU mice in comparison with WT and COX-2 +/- . Furthermore, EIU was suppressed in mice treated with recombinant IFNγ, as shown by the decreased EIU scores, the presence of serum LTB 4 and IL-6 and ocular 5-LO and IL-6 mRNA, and the increases in serum IFNγ and ocular IFNy, particularly in COX-2 -/- mice. CONCLUSIONS. These data suggest that disturbance of the AA pathway exacerbates EIU in COX-2-deficient mice. IFNγ moderately reverses this exacerbation and protects against EIU.
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MCP-1 Expression in Endotoxin-Induced Uveitis
Investigative Ophthalmology & Visual Science, 2002Co-Authors: Nadine Tuaillon, De Fen Shen, Ravi B. Berger, Bao Lu, Barrett J. Rollins, Chi-chao ChanAbstract:Abstract Monocyte chemoattractant protein (MCP)-1 (CCL-2) is a chemokine with chemoattractant properties for monocytes, memory T cells, natural killer cells, mast cells, and basophils. To delineate the role played by MCP-1 in acute anterior Uveitis, a common ocular inflammation, MCP-1(-/-) mice and wild-type matched control mice were analyzed for the development of Endotoxin-Induced Uveitis (EIU) in response to subcutaneous injection of a sublethal dose of lipopolysaccharide (LPS). EIU was induced in MCP-1(-/-) and wild-type control mice by a single subcutaneous injection of Salmonella typhimurium LPS endotoxin at day 0. Alternatively, MCP-1(-/-) mice were injected subcutaneously with LPS plus recombinant MCP-1 at day 0 and with recombinant MCP-1 6 hours later. Mice were killed at day 1 or 3 after injection. Serum levels of IL-1alpha, IL-1beta, IL-6, IFN-gamma, TNF-alpha, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage inflammatory protein (MIP)-1alpha, MIP-2, regulated on activation normal T-cell expressed and secreted (RANTES), and MCP-1 were determined by ELISA. Eyes were collected and analyzed histologically and by RT-PCR for MCP-1, IFN-gamma, IL-6, TNF-alpha, beta-actin, MCP-5, RANTES, KC, inflammatory protein (IP)-10, and toll-like receptor (TLR)-4. EIU was strongly reduced in MCP-1(-/-) mice compared with wild-type control mice. The number of ocular inflammatory cells was significantly reduced. Moreover, intraocular IFN-gamma transcription was increased. EIU was induced in MCP-1(-/-) mice by co-administration of recombinant rat MCP-1 and LPS. Data indicate that MCP-1 plays a crucial role in the induction of EIU. MCP-1 may be a new therapeutic strategy for acute anterior Uveitis.
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Recurrent intraocular inflammation in Endotoxin-Induced Uveitis.
Investigative Ophthalmology & Visual Science, 2000Co-Authors: Alexander T. Kozhich, Chi-chao Chan, Igal Gery, Scott M. WhitcupAbstract:PURPOSE: Endotoxin-Induced Uveitis (EIU) in rats and mice peaks 24 hours after endotoxin injection and is commonly assumed to be a monophasic disease. This study examined intraocular inflammation at later time points to determine whether endotoxin injection can induce recurrent intraocular inflammation in strains of mice with high or moderate levels of susceptibility to EIU. METHODS: EIU was elicited in two mouse strains with high (C3H/HeN) and moderate (FVB/N) susceptibility, by means of intraperitoneal injections of Salmonella typhimurium endotoxin. Inflammatory cells in the anterior and posterior segments of the eye were counted by a masked observer on histologic sections of eyes from 1 to 17 days after endotoxin injection. RESULTS: A bimodal distribution of inflammatory cell infiltration was noted in eyes from C3H/HeN mice. As previously reported, inflammation peaked at 24 hours after endotoxin injection. However, a second, more pronounced peak of intraocular inflammation occurred approximately 5 days after endotoxin injection. FVB/N mice had a single peak of intraocular inflammation 4 days after injection. CONCLUSIONS: Endotoxin injection in C3H/HeN elicits recurrent intraocular inflammation. The previously unrecognized second peak of inflammation is more severe than the initial inflammatory disease. Studies on this second inflammatory peak may be useful in determining the pathogenesis of recurrent Uveitis in humans.
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Biphasic Ocular Inflammatory Response to Endotoxin-Induced Uveitis in the Mouse
Archives of Ophthalmology, 2000Co-Authors: De Fen Shen, Margaret A. Chang, Dawn M. Matteson, Ronald Buggage, Alexander T. Kozhich, Chi-chao ChanAbstract:Objective To examine the kinetics and mechanisms of Endotoxin-Induced Uveitis in the mouse. Methods C3H/HeN mice were injected subcutaneously with 0.3 mg of Salmonella typhimurium lipopolysaccharide (LPS) in 0.1 mL of phosphate-buffered saline solution or phosphate-buffered saline solution alone in 3 separate experiments; mice were killed after 1, 3, 5, and 7 days. In 2 other separate experiments, mice were killed 1, 3, 6, and 24 hours after LPS injection. All eyes were collected for histological examination, immunohistochemical analyses, aqueous protein level determination, and reverse transcriptase–polymerase chain reaction for ocular interleukin (IL)1α, IL-6, tumor necrosis factor α, and granulocyte-macrophage colony-stimulating factor messenger RNA (mRNA). Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor α and IL-6 levels in aqueous and serum samples. Results Results were consistent for all experiments. Numbers of ocular inflammatory cells and levels of aqueous protein peaked 1 and 5 days after LPS injection. Control mice did not develop inflammation. Serum and aqueous IL-6 and ocular IL-6 mRNA levels peaked at 1 day and subsided at 3 days. However, ocular IL-1α, tumor necrosis factor α, and granulocyte-macrophage colony-stimulating factor mRNA appeared, peaked, and subsided at 3, 5, and 7 days, respectively. Predominant infiltrating cells were neutrophils at 1 day and macrophages at 5 days. Although no ocular inflammatory cells were detected before 24 hours after LPS injection, tumor necrosis factor α mRNA was noticed at 1 hour, peaked at 3 hours, and disappeared at 6 hours and granulocyte-macrophage colony-stimulating factor mRNA was spotted only at 3 hours after LPS injection. Conclusions The ocular inflammatory response to C3H/HeN mouse Endotoxin-Induced Uveitis is biphasic for 7 days. The first wave appears at day 1 and subsides by day 3. A second, higher peak appears at day 5. The 2 inflammatory waves are related to the kinetics of the different cytokines released in the eye. This is in contrast to the rat monophasic Endotoxin-Induced Uveitis model, which has only one peak of intense inflammation associated with cytokine release. Clinical Relevance A biphasic inflammatory response associated with cytokine release lasting several days is observed in C3H/HeN mice with Endotoxin-Induced Uveitis. Because human anterior Uveitis has a tendency to be recurrent in nature, this might be a better experimental model.
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Topical liposome-encapsulated FK506 for the treatment of Endotoxin-Induced Uveitis
Ocular Immunology and Inflammation, 1998Co-Authors: Scott M. Whitcup, Uwe Pleyer, Sabine Lutz, Manabu Mochizuki, Chi-chao ChanAbstract:Purpose: Liposome preparations of FK506 improve the penetration of topically administered drug into the aqueous humor. The purpose of the experiment was to compare topically administered high-dose oil-dissolved FK506 (OD-FK506) and low-dose liposome-bound FK506 (LB-506) for the treatment of Endotoxin-Induced Uveitis (EIU). Methods: Endotoxin-Induced Uveitis was produced in female Lewis rats with Salmonella typhimurium endotoxin. Four hours prior to endotoxin injection, one eye received 20 µl eyedrops every four hours containing either high-dose OD-FK506 at 3 mg/ml (N=20), low-dose LB-FK506 at 0.16 mg/ml (N=19), prednisolone acetate 1% (N=20), or empty liposomes (N=20). Eyes were enucleated 24 hours after endotoxin injection and inflammatory cells were counted on histologic sections by two masked observers. Results: The mean number of infiltrating inflammatory cells per section ± S.E.M. was 127.8 ± 20.1, 76.8 ± 16.7, 75.0 ± 19.1, and 3.6 ± 0.4 for animals treated with empty liposomes, LB-FK506, OD-FK506, a...
Salvatore Musumeci - One of the best experts on this subject based on the ideXlab platform.
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Effect of chitinase inhibitors on Endotoxin-Induced Uveitis (EIU) in rabbits
Pharmacological Research, 2008Co-Authors: Claudio Bucolo, Maria Musumeci, Adriana Maltese, Filippo Drago, Salvatore MusumeciAbstract:The acidic mammalian chitinase (AMCase) is significantly increased in tears of human allergic conjunctivitis. The aim of the study was to investigate the effects of chitinase inhibitors, allosamidin and caffeine versus dexamethasone, in rabbit Endotoxin-Induced Uveitis (EIU). EIU was induced in rabbits by a single intravitreal injection of 100 ng/10 μl lipopolysaccharide (LPS). Drugs at four different concentrations (0.1, 0.01, 0.001 and 0.0001 mM) were topically applied to the rabbit eye five times in 24 h. Tears were collected at 0, 6 and 24 h after LPS to measure the AMCase activity. The effect of treatment was also evaluated at the same time by slit lamp examination. Tear AMCase activity increased 6 and 24 h after LPS injection. The AMCase activity was significantly inhibited in all treated groups with all doses of allosamidin and caffeine except with the lowest concentration. A higher AMCase inhibition at 24 h was found with allosamidin and caffeine compared to dexamethasone. Moreover, topical administration of allosamidin, caffeine and dexamethasone produced a remarkable reduction of inflammatory signs, in the order: dexamethasone > caffeine > allosamidin. AMCase inhibitors showed in this rabbit model of Uveitis a notable control of inflammatory response with a significant reduction of AMCase activity in tears with caffeine and allosamidin. These results support the key role of AMCase in the pathogenesis of human ocular inflammatory diseases and the therapeutic effect of AMCase inhibitors on experimental Uveitis.
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Effect of chitinase inhibitors on Endotoxin-Induced Uveitis (EIU) in rabbits
Pharmacological Research, 2008Co-Authors: Claudio Bucolo, Maria Musumeci, Adriana Maltese, Filippo Drago, Salvatore MusumeciAbstract:The acidic mammalian chitinase (AMCase) is significantly increased in tears of human allergic conjunctivitis. The aim of the study was to investigate the effects of chitinase inhibitors, allosamidin and caffeine versus dexamethasone, in rabbit Endotoxin-Induced Uveitis (EIU). EIU was induced in rabbits by a single intravitreal injection of 100 ng/10 ??l lipopolysaccharide (LPS). Drugs at four different concentrations (0.1, 0.01, 0.001 and 0.0001 mM) were topically applied to the rabbit eye five times in 24 h. Tears were collected at 0, 6 and 24 h after LPS to measure the AMCase activity. The effect of treatment was also evaluated at the same time by slit lamp examination. Tear AMCase activity increased 6 and 24 h after LPS injection. The AMCase activity was significantly inhibited in all treated groups with all doses of allosamidin and caffeine except with the lowest concentration. A higher AMCase inhibition at 24 h was found with allosamidin and caffeine compared to dexamethasone. Moreover, topical administration of allosamidin, caffeine and dexamethasone produced a remarkable reduction of inflammatory signs, in the order: dexamethasone > caffeine > allosamidin. AMCase inhibitors showed in this rabbit model of Uveitis a notable control of inflammatory response with a significant reduction of AMCase activity in tears with caffeine and allosamidin. These results support the key role of AMCase in the pathogenesis of human ocular inflammatory diseases and the therapeutic effect of AMCase inhibitors on experimental Uveitis. ?? 2008 Elsevier Ltd. All rights reserved.
E. Shirasawa - One of the best experts on this subject based on the ideXlab platform.
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The role of platelet-activating factor in cell infiltration in Endotoxin-Induced Uveitis in guinea pigs
Current Eye Research, 1998Co-Authors: F. Tsuji, E. ShirasawaAbstract:PURPOSE. We investigated the role of platelet-activating factor (PAF) in cell infiltration in Endotoxin-Induced Uveitis (EIU) in guinea pigs. METHODS. To elicit EIU, lipopolysaccharide (LPS) was injected into the anterior chamber of the eye. Cell numbers in the aqueous humor after LPS injection were determined by flow cytometry. PAF and prostaglandin E 2 (PGE 2) production after LPS injection were also examined. RESULTS. Intracameral injection of LPS induced cell infiltration into the anterior chamber, and platelet-activating factor (PAF) was detected in the aqueous humor. In addition, topical apafant (PAF antagonist) partially inhibited cell infiltration. Intracameral injection of PAF scarcely induced cell infiltration but the reaction with EIU was accelerated by intracameral injection of a small amount of PAF. CONCLUSIONS. These results suggest that PAF has weak direct activity on cell infiltration in intraocular inflammation but enhances intraocular inflammation.
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The effects of betamethasone derivatives on Endotoxin-Induced Uveitis in guinea pigs
Inflammation Research, 1997Co-Authors: F. Tsuji, K. Sawa, T. Ikuse, E. ShirasawaAbstract:Objective and Design: We reported previously that the betamethasone derivative betamethasone dipropionate behaves as an anti-glucocorticoid in rat Endotoxin-Induced Uveitis (EIU). In the present study, we produced EIU in guinea pigs and investigated the effects of betamethasone dipropionate on the EIU.¶ Material: Male Hartley guinea pigs were used.¶ Treatment: Glucocorticoids were instilled into the eye.¶ Method: To elicit EIU, lipopolysaccharide (LPS) was injected into the anterior chamber of the eye. Cell numbers in the aqueous humor after LPS injection were determined by flow cytometry. Prostaglandin E_2 (PGE_2) production after LPS injection into the anterior chamber was also examined.¶ Results: Intracameral injection of LPS (1 μg/eye) induced cell infiltration into the anterior chamber and PGE_2 production. Betamethasone dipropionate inhibited cell infiltration and PGE_2 production more strongly than betamethasone. These results suggest that betamethasone dipropionate is a potent glucocorticoid in guinea pigs.¶ Conclusions: Structure-activity relationships of glucocorticoids in the guinea pig EIU model may differ from those in the rat EIU model.
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The effects of betamethasone derivatives on Endotoxin-Induced Uveitis in rats
Experimental Eye Research, 1997Co-Authors: F. Tsuji, K. Sawa, Masatomo Kato, Hiroyuki Mibu, E. ShirasawaAbstract:Abstract The anti-inflammatory effects of betamethasone and its derivatives (betamethasone 21-phosphate, betamethasone 21-acetate, betamethasone 17-valerate, clobetasol 17-propionate and betamethasone dipropionate) on Endotoxin-Induced Uveitis (EIU) in rats was investigated. Among the compounds examined, betamethasone, betamethasone 21-phosphate, betamethasone 21-acetate and clobetasol 17-propionate strongly inhibited cell infiltration into the aqueous humor in the EIU by instillation into the eye in a dose-dependent manner (0.01–1.0%) and by systemic administration (1 mg kg −1 ). On the other hand, betamethasone 17-valerate and betamethasone dipropionate showed only weak inhibitory effects or enhancment of cell infiltration by instillation into the eye (0.01–1.0%) and by systemic administration (1 mg kg −1 ). Combined systemic administration of betamethasone dipropionate and betamethasone reduced the inhibitory effect of betamethasone on cell infiltration and gene expression of IL-1β. In an in vitro interleukin-8 (IL-8) release assay, betamethasone showed stronger inhibition of the IL-8 release from rat peritoneal exudate cells (PEC) than betamethasone dipropionate, and simultaneous addition of betamethasone dipropionate with betamethasone reduced the inhibitory effect of the latter. These results suggest that the betamethasone derivative betamethasone dipropionate might behave as an anti-glucocorticoid in rats.
Stephen R. Planck - One of the best experts on this subject based on the ideXlab platform.
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The NLRP3 inflammasome is active but not essential in Endotoxin-Induced Uveitis
Inflammation Research, 2012Co-Authors: Holly L. Rosenzweig, April Woods, Jenna S. Clowers, Stephen R. Planck, James T. RosenbaumAbstract:Objective The inflammasome complex involving caspase-1 and nucleotide-binding domain, leucine-rich repeat containing protein (NLRP)3, also known as NALP3 or cryopyrin is important for host responses to microbial pathogens and several autoinflammatory diseases. We investigated the extent to which NLRP3 and caspase-1 control ocular interleukin (IL)-1β production and severity of Uveitis (intraocular inflammatory disease) in an established, acute inflammatory Uveitis model, Endotoxin-Induced Uveitis (EIU). Methods Expression of NLRP3, its adaptor molecule ASC, also known as PYCARD (PYD and CARD domain containing), and caspase-1 were examined by immunoblotting. IL-1β production was measured by enzyme-linked immunosorbent assay (ELISA). Using knockout mice, roles for caspase-1 and NLRP3 were examined in Uveitis induced by intraocular injection of Escherichia coli lipopolysaccharide (LPS). Results NLRP3, ASC, and caspase-1 proteins are constitutively expressed in eye tissue. During EIU, IL-1β protein production increases; this requires the presence of both caspase-1 and NLRP3. However, severity of EIU is not altered by deficiency in either caspase-1 or NLRP3, as assessed by both intravital microscopy and histology. Conclusions These data identify the importance of the NLRP3 inflammasome for IL-1β production in the eye, yet indicate that its participation in EIU is nonessential.
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The NLRP3 inflammasome is active but not essential in Endotoxin-Induced Uveitis
Inflammation Research, 2011Co-Authors: Holly L. Rosenzweig, April Woods, Jenna S. Clowers, Stephen R. Planck, James T. RosenbaumAbstract:Objective The inflammasome complex involving caspase-1 and nucleotide-binding domain, leucine-rich repeat containing protein (NLRP)3, also known as NALP3 or cryopyrin is important for host responses to microbial pathogens and several autoinflammatory diseases. We investigated the extent to which NLRP3 and caspase-1 control ocular interleukin (IL)-1β production and severity of Uveitis (intraocular inflammatory disease) in an established, acute inflammatory Uveitis model, Endotoxin-Induced Uveitis (EIU).
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Interleukin-6 does not mediate Endotoxin-Induced Uveitis in mice: Studies in gene deletion animals
Investigative Ophthalmology & Visual Science, 1998Co-Authors: James T. Rosenbaum, Paul Kievit, Jong Moon Park, Stephen R. PlanckAbstract:PURPOSE. Interleukin-6 (IL-6) has been strongly implicated in anterior Uveitis based on its presence in aqueous humor from infected eyes and its inflammatory effects when injected intravitreally into rats. We used IL-6-deficient mice to test further the hypothesis that IL-6 contributes to the development of Endotoxin-Induced Uveitis. METHODS. Uveitis was scored by histologic analysis of C3H/HeN mice 24 hours after intravitreal injections of up to 200 ng of recombinant murine IL-6. Uveitis was similarly measured in IL-6-deficient mice and congenic controls 24 hours after intravitreal injection of 250 ng of Esherichia colt endotoxin. Reverse transcription-polymerase chain reaction was used to detect mRNAs for several cytokines at 3 hours postinjection. The IL-6 concentration in aqueous humor samples was determined with a bioassay using the murine B9 plasmacytoma cell line. RESULTS. Direct injection of IL-6 did not induce Uveitis. Mice genetically deficient in IL-6 developed Endotoxin-Induced Uveitis that was comparable or more severe than congenic control mice. Compensatory. changes in the expression of mRNA for other cytokines were not detected in irises from the IL-6-deficient mice. In IL-6-competent mice that received bilateral endotoxin injections, no correlation was found between the number of infiltrating cells in one eye and the IL-6 concentration in the aqueous humor of the contralateral eye. CONCLUSIONS. In marked contrast to previous conclusions with rats, IL-6 was not sufficient for inducing Uveitis in mice. Additionally, IL-6 was not necessary for the development of Uveitis subsequent to intravitreal injection of endotoxin in mice.
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THE EFFECT OF GENETIC DEFICIENCY OF ADHESION MOLECULES ON THE COURSE OF Endotoxin-Induced Uveitis
Current Eye Research, 1998Co-Authors: Stephen R. Planck, Jong Moon Park, Leslie M. O'rourke, Jose Carlos Gutierrez-ramos, James T. RosenbaumAbstract:PURPOSE. Multiple adhesion molecules of the selectin, integrin, and immunoglobulin-like families are involved in the migration of leukocytes out of the bloodstream into inflamed tissues. This study addresses the question of which adhesion molecules are specifically involved in Endotoxin-Induced Uveitis. METHODS. Mice genetically deficient in p-selectin, ICAM-1, s 2 -integrin, or controls received intravitreal injections of endotoxin. Eyes were harvested 24 h later and inflammation was evaluated by histologic and immunohistochemical assays of infiltrating cells. RESULTS. Mice lacking either P-selectin or s 2 -integrin had less inflammation than controls (median cells/section: 64 for P-selectin knockout vs 130 for controls, p = 0.02, n = 17 per group; 244 for s 2 -integrin knockouts, n = 14, vs 355 for controls, n = 17, p = 0.05). Neither gene deletion significantly changed the ratio of infiltrating neutrophils to macrophages. ICAM-1 knockouts tended to have fewer infiltrating cells (median 22 cells/section...