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Inhibitory role of transforming growth factor β2 in experimental autoimmune anterior uveitis

  • Bharati Matta
  • Puran S. Bora
  • Adam J. Neuhouser
  • Nalini S. BoraEmail author
Open Access
Basic Science
  • 54 Downloads

Abstract

Purpose

Experimental autoimmune anterior uveitis (EAAU) is a clinically relevant animal model for human idiopathic anterior uveitis (IAU). The role of the immunomodulator transforming growth factor β2 (TGF-β2) in EAAU pathology is unknown. In this study, we investigated the regulatory role of TGF-β2 in EAAU.

Methods

EAAU was induced in male Lewis rats by footpad injection of melanin-associated antigen (MAA). TGF-β2 was administered intravenously (iv) in MAA-sensitized rats during the induction of EAAU, or after the clinical onset of uveitis. MAA-sensitized rats injected similarly with an equal volume of PBS served as control. Animals were examined daily between days 7 and 30 post-injection for the clinical signs of uveitis using slit lamp biomicroscopy. Animals were sacrificed at various time points and eyes were harvested for histological analysis to assess the course and severity of inflammation. For histopathological analysis, paraffin sections of harvested eyes were stained with hematoxylin and eosin. Popliteal lymph nodes (LNs) were used for CD4+CD25+FoxP3+ T regulatory (Tregs) population analysis and for CD4+ T cell proliferation assay.

Results

Administration of recombinant TGF-β2 during the early stages of EAAU prevented the induction of uveitis. Compared to PBS, the presence of TGF-β2 in the cell culture significantly (p < 0.05) inhibited the proliferation of CD4+ T cells in response to MAA. In MAA-sensitized Lewis rats, iv treatment with recombinant TGF-β2 resulted in significantly (p < 0.05) increased percentage of Tregs compared to animals treated similarly with PBS. Thus, TGF-β2 inhibited the induction of EAAU by inhibiting CD4+ T cell proliferation and increasing the number of Tregs. Injection of TGF-β2 in rats with active EAAU resulted in diminished disease activity. Unfortunately, this treatment did not lead to the early resolution of EAAU.

Conclusions

TGF-β2 plays a critical role in regulation of intraocular inflammation in EAAU. Findings reported in this study improve our understanding of immunopathology of IAU and suggest that recombinant TGF-β2 may be a promising therapeutic agent for human IAU.

Keywords

Anterior uveitis Inflammation Transforming growth factor β2 T regulatory cells CD4+ T cells Experimental autoimmune anterior uveitis 

Introduction

Idiopathic anterior uveitis (IAU) is the most prevalent intraocular inflammatory disease in humans and is characterized by the inflammation of the iris and/or the CB. Visual complications associated with recurrent and/or untreated IAU lead to the permanent loss of vision [1, 2, 3]. Human IAU has been considered an autoimmune disease where an immune response affects only the eye [1, 2, 3]. Evidence from animal models and limited human studies support a crucial role of T cells in the development of uveitis [4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19].

Experimental autoimmune anterior uveitis (EAAU) is an autoimmune ocular inflammatory disease and it serves as a clinically relevant animal model for idiopathic human anterior uveitis. This animal model has been extensively used by us and other investigators to understand the etiopathogenesis of IAU [6, 7, 8, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21]. EAAU is a CD4+ T cell-mediated autoimmune disease that can be induced in Lewis rats by immunization with melanin-associated antigen (MAA) in the hind footpad. The cardinal features of EAAU include lymphocytic infiltration in the iris, the CB, and the anterior chamber of the eye [14, 15, 16, 17, 18, 19, 20, 21].

We have published that the induction of EAAU can be prevented by inducing immune tolerance against MAA [18, 19]. We further reported that the levels of transforming growth factor β2 (TGF-β2) increased in the popliteal lymph nodes (LNs) of tolerized animals [18, 19]. However, the specific role of TGF-β2 in the pathogenesis of EAAU remains unclear. In this study, we investigated the ability of TGF-β2 to modulate inflammatory responses in EAAU.

Materials and methods

Animals

Pathogen-free male Lewis rats (5–6 weeks old) were obtained from Harlan Sprague-Dawley (Indianapolis, IN). This study was approved by the Institutional Animal Care and Use Committee, University of Arkansas for Medical Sciences, Little Rock, AR.

Induction and evaluation of EAAU

MAA was isolated from bovine iris and CB as previously described by us [14, 17]. Male Lewis rats were immunized with 100 μl of stable emulsion containing 75 μg of MAA emulsified (1:1) in complete Freund’s adjuvant (Sigma-Aldrich, St. Louis, MO) using a single-dose induction protocol in the hind footpad as previously described by us [14, 16, 17, 18, 19]. Pertussis toxin was used as an additional adjuvant. Animals were examined daily between days 7 and 30 post-injection for the clinical signs of uveitis using slit lamp biomicroscopy. EAAU was scored by an observer unaware of the experimental design. Intensity of uveitis was scored in a masked fashion on the arbitrary scale of 0 to 4, as follows: 0, normal; 1, dilated iris vessels and thickened iris and ciliary body; exudates in the anterior chamber with protein, a few scattered inflammatory cells, or both; 2, moderate infiltration of inflammatory cells in the iris, ciliary body, or both; moderate number of inflammatory cells within the anterior chamber; 3, heavy infiltration of inflammatory cells within the iris and ciliary body and within the anterior chamber; 4, heavy exudation of cells with dense protein aggregation in the anterior chamber; inflammatory cell deposits on the corneal endothelium. Eyes were also harvested at various time points for histological analysis to assess the course and severity of inflammation using the criteria previously reported [14, 16, 17, 18, 19].

TGF-β2 administration

Recombinant human TGF-β2 (PeproTech, Rocky Hill, NJ) was used for in vivo injections. Human TGF-β2 has high degree of homology (> 90%) with rat TGF-β2 at the amino acid level and it cross-reacts with its rat counterpart (PeproTech). TGF-β2 was administered by two different routes: intravenous (iv) and intraperitoneal (ip) in pilot experiments. Since anterior chamber (AC) injections are not feasible after the onset of EAAU because eyes are inflamed, TGF-β2 was not delivered into the AC of the eye in the current study. In our pilot experiments, a therapeutic effect on EAAU was observed when TGF-β2 was administered intravenously and not intraperitoneally. Therefore, administration of TGF-β2 via iv route was used in the following experiments.

MAA-sensitized Lewis rats received a single iv injection of recombinant TGF-β2 1 day before immunization (day − 1), on the day of immunization (day 0), and 1 day after immunization (day + 1). Control animals were treated similarly with equal volume of sterile phosphate buffered saline (PBS). To perform TGF-β2 dose titration, two different doses—500 ng/injection and 1 μg/injection of recombinant TGF-β2—were used separately in our initial experiments. Since iv injection of 500 ng and 1 μg of TGF-β2 gave same results, 500 ng of TGF-β2 was used in subsequent experiments.

In another set of experiments, Lewis rats immunized with MAA received a single iv injection of recombinant TGF-β2 after the onset of EAAU on days 12 and 14 post-immunization. MAA-sensitized rats treated similarly with equal volume of sterile PBS served as control. Clinical and histopathological examination (described above) was used to determine the onset, severity, and duration of EAAU.

Histology

For histology [18, 19], freshly enucleated rat eyes were fixed in 10% neutral buffered formalin (Sigma-Aldrich) for 24 h at room temperature, dehydrated in ethanol through ascending series of ethanol concentrations and embedded in paraffin. Four-micron sections were stained with hematoxylin and eosin (H&E) purchased from Fisher (Fair Lawn, NJ). Sections were examined using a light microscope (Carl Zeiss Meditec, Inc., Thornwood, NY).

Cell preparation

Popliteal LNs were harvested on day 5 for Treg population analysis and on day 12 for CD4+ T cell proliferation assay. A single-cell suspension was prepared as previously described [18, 19]. Briefly, the tissue was mashed between the frosted slides and cells were filtered through cell strainer and washed once with Dulbecco’s phosphate buffered saline (DPBS). Total lymphocytes were purified using Histopaque gradient (Sigma Aldrich) according to manufacturer’s protocol. The cells were suspended in complete RPMI 1640 culture medium containing l-glutamine (Mediatech, Herndon, VA), 1% (v/v) MEM essential vitamin mixture (Life Technologies, Rockville, MD), NEAA (BioWhittekar, Allendale, NJ), mixture of antibiotics (penicillin 100 U/ml, streptomycin 100 U/ml and amphotericin B 0.25 μg/ml) purchased from BioWhittekar, Allendale, NJ, and 10% (v/v) FCS (Mediatech, Herndon, VA).

Staining for Tregs

The Fc receptor was blocked by incubating the cells with anti-rat CD32 antibody (BD Biosciences, San Jose, CA) for 30 min. The cells were then surface stained using anti-rat CD4-APC and anti-rat CD25-PE antibodies from Biolegend (San Diego, CA). After treating with antibodies for 40 min, cells were washed and stained with FITC labeled FoxP3 antibody (eBiosciences, San Diego, CA) using FoxP3 staining kit according to manufacturer’s instructions (ebiosciences). The stained cells were analyzed using FACS Calibur (BD Biosciences). The percentage of CD4+CD25+FoxP3+ Tregs was calculated using WinMDI [18, 19].

In vitro cell proliferation assay

In vitro T cell proliferation was performed as previously described by the authors [18, 19]. Total lymphocytes were harvested from lymph nodes of MAA-sensitized animals on day 12 post-immunization. The cells were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) using Cell Trace CFSE Cell Proliferation Kit according to manufacturer’s protocol (Molecular Probes, Invitrogen, Carlsbad, CA) and were cultured with MAA (20 μg/ml) for 5 days in the absence and presence of TGF-β2 (10 ng/ml). On day 6, non-adherent cells were collected and labeled with anti-rat CD4-APC antibody. The stained cells were then analyzed using FACS Calibur. The percentage of proliferating cells was calculated using WinMDI.

Statistical analysis

Data were analyzed and compared using Student’s t test or Mann-Whitney U test using Statistica Program (Statsoft, Inc., Tulsa, OK) and differences were considered statistically significant with p < 0.05.

Results

Effect of recombinant TGF-β2 on the induction of EAAU

To study the effect of TGF-β2 on the induction of EAAU, MAA-sensitized Lewis rats received a total of three iv injections (one injection/time point) of recombinant TGF-β2 (500 ng/injection) at the following time points: 1 day before immunization (day − 1), on the day of immunization (day 0), and 1 day after immunization (day + 1). Another set of MAA-sensitized animals treated similarly with an equal volume of PBS served as control. Treatment with TGF-β2 resulted in complete inhibition of induction of EAAU as determined by clinical (Fig. 1a) and histological (Fig. 1c) examination. In these rats, no infiltration of the inflammatory cells was noted in the iris, the CB, and the anterior chamber of the eye. Conversely, similar treatment with PBS did not alter the course or the severity of EAAU (Fig. 1a, b). The iris, the CB, and the anterior chamber of the PBS-treated rat exhibited heavy exudation of inflammatory cells (Fig. 1b).
Fig. 1

Transforming growth factor β2 (TGF-β2) inhibits the induction of experimental autoimmune anterior uveitis (EAAU). Recombinant TGF-β2 or PBS was administered intravenously 1 day before immunization, at the time of immunization, and 1 day post-immunization with melanin-associated antigen (MAA). This treatment with recombinant TGF-β2 prevented the induction of uveitis clinically (a). MAA-sensitized rats treated similarly with phosphate buffered saline (PBS) developed the normal clinical course of EAAU (a). Lewis rats injected with PBS (control) or TGF-β2 as described above were sacrificed at day 18 post-immunization. Representative H&E stained sections of harvested eyes demonstrated heavy infiltration of the inflammatory cells within the iris (I), the ciliary body (CB), and the anterior chamber (AC) of PBS-injected rats (b). In contrast, no inflammatory cells could be detected in the eye of Lewis rats injected iv with TGF-β2 (c) at this time point. The data are expressed as the mean + SD, representative of three independent experiments. n = 15 rats/group. Objective magnification × 20

Mechanism of TGF-β2-mediated prevention of induction of EAAU

We next investigated the underlying mechanisms responsible for inhibition of EAAU induction by TGF-β2 treatment. To address this, we investigated the proliferation of CD4+ T cells in the presence of TGF-β2 (in vitro) as well as determined the effect of TGF-β2 on generation of Tregs (in vivo) using popliteal LNs that drain the site of MAA injection.

Proliferation of CD4+ cells (in vitro)

Since EAAU is a CD4+ T cell-mediated autoimmune disease [16, 17, 18, 19], experiments were carried out to assess the effects of TGF-β2 on the proliferation of CD4+ T cells in response to MAA. Proliferation of CD4+ T cells in the presence and absence of recombinant TGF-β2 was compared using an in vitro cells proliferation assay. Total lymphocytes purified from popliteal LNs of MAA-sensitized animals sacrificed at day 12 post-immunization were labeled with CFSE and were cultured with MAA (20 μg/ml) for 5 days in the presence of TGF-β2 (10 ng/ml) or equal volume of PBS. On day 6, these in vitro cultured CFSE-labeled lymphocytes were labeled with anti-rat CD4-APC and analyzed using flow cytometry to investigate the effect of TGF-β2 on proliferation of CD4+ T cells. Our data shows that compared to PBS (Fig. 2a, c), the presence of TGF-β2 in the cell culture significantly inhibited the proliferation (in vitro) of CD4+ T cells in response to MAA (Fig. 2b, c).
Fig. 2

TGF-β2 inhibits the proliferation (in vitro) of CD4+ T cells isolated from lymph nodes (LNs) harvested at day 12 post-immunization in response to MAA. Density plots (a and b) show representative flow cytometric data and panel c represents cumulative data from three separate experiments. Presence of 10 ng/ml of recombinant TGF-β2 (b and c) in culture inhibited the proliferation of CD4+ T cells in response to MAA compared to the treatment with same volume of PBS (a, c). The data are expressed as the mean + SD, representative of three independent experiments. n = 9 rats/group *p < 0.05

Generation of CD4+CD25+FoxP3+ Tregs (in vivo)

We have reported that LN-derived CD4+CD25+FoxP3+ Tregs cells play a crucial role in the induction of immune tolerance against MAA in EAAU [18, 19]. Therefore, we next determined and compared the percentage of Tregs in popliteal LNs of MAA-sensitized Lewis rats after iv injections of TGF-β2 or PBS. Lewis rats were injected intravenously with recombinant TGF-β2 (500 ng/injection) on day − 1, day 0, and day + 1 post-immunization as described above. Animals were sacrificed on day 5 post-immunization. Total lymphocytes harvested from popliteal LNs were stained for CD4, CD25, and FoxP3 and percentage of CD4+CD25+FoxP3+ Tregs was determined using flow cytometry as described under “Materials and Methods.” Our results (Fig. 3a–c) demonstrate that in MAA-sensitized Lewis rats, iv treatment with recombinant TGF-β2 resulted in increased percentage of Tregs (13.25 + 0.69%) compared with MAA-sensitized animals treated similarly with PBS (8.4 + 0.44%). These differences were statistically significant (p < 0.05).
Fig. 3

TGF-β2 induces T regulatory (Treg) cells in the popliteal LNs. CD4+CD25+FoxP3+ Tregs in the popliteal LNs of MAA-sensitized Lewis rats after TGF-β2 administration on day − 1, day 0, and day + 1 post-immunization via iv route (ac). Lymph nodes were collected on day 5 post-immunization and lymphocytes were stained for CD4, CD25, and FoxP3. Representative density plots show the percentage of Tregs found in animals treated with PBS (a) or recombinant TGF-β2 (b). Cumulative data from three independent experiments is shown in panel c. The data are expressed as the mean + SD, representative of three independent experiments. n = 9 rats/group. *p < 0.05

Effect of recombinant TGF-β2 on EAAU after the onset of uveitis

Experiments were carried out to investigate if active EAAU can be inhibited by administration of recombinant TGF-β2. MAA-sensitized Lewis rats received iv injection of TGF-β2 or PBS separately on two occasions after the onset of the clinical disease (on days 12 and 14 post-immunization) and the disease progression was monitored. A sharp decline in the clinical disease activity was observed in all animals at days 13–18 that received TGF-β2 via iv route (Fig. 4). However, from day 19 post-immunization onward EAAU followed the normal course of the disease in these animals. The duration of EAAU in MAA-sensitized rats that were treated with recombinant TGF-β2 after the onset of uveitis was similar to PBS-treated control rats (Fig. 4).
Fig. 4

TGF-β2 suppresses active EAAU. To investigate the effect on ongoing uveitis, MAA-sensitized Lewis rats received recombinant TGF-β2 or PBS via iv route at days 12 and 14 post-immunization (after the onset of clinical disease). This treatment with TGF-β2 reduced the clinical severity of EAAU until day 18 post-immunization. However, from day 19 post-immunization, EAAU followed the normal course of the disease, similar to that observed in PBS-injected rats. On days 15, 16, 17, and 18 post-immunization, the clinical severity of EAAU in rats treated with TGF-β2 was significantly (p < 0.05) reduced compared to Lewis rats treated with same volume of PBS. The data are expressed as the mean + SD, representative of three independent experiments. n = 9 rats/group

Discussion

The management of patients with IAU remains a significant challenge because no specific treatment is available for this disease. IAU is treated symptomatically only using non-specific therapies such as corticosteroid and immunosuppressive agents [1, 2, 3]. Unfortunately, these treatment modalities do not address the underlying mechanisms and are associated with adverse side effects. Prolonged immunosuppression results in increased morbidity. Due to the lack of safe and effective therapeutic options available to the patients with IAU, further research is required so that novel therapeutic targets can be discovered for this sight-threatening disease.

EAAU is a convenient and reliable animal model of human IAU [6, 7, 8, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21]. Previous reports by us, and in the literature, have shown that CD4+ T cell infiltration into the iris and the CB with inflammatory cell infiltration in the anterior chamber of the eye is the hallmark of EAAU pathology [6, 7, 8, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21]. We have previously reported that TGF-β2 plays an important role in the development of immune tolerance against MAA [18, 19]. In the current study, the immunomodulatory role for TGF-β2 in the induction and progression of EAAU was explored. TGF-β is a cytokine with multiple effects in T cell development, homeostasis, and tolerance [22, 23, 24]. Three isoforms of TGF-β (TGF-β1, TGF-β2, and TGF-β3) are present in mammals [25, 26]. TGF-β2 is the main isoform of TGF-β in the eye and it is produced locally [25]. Importantly, TGF-β2 has been reported to play an important role in the pathogenesis of several vision-threatening ocular diseases [25, 26, 27, 28, 29].

Two sets of in vivo experiments were performed to investigate the protective role for TGF-β2 in EAAU, one at the time of induction and another after the onset of uveitis. In the first set of experiments, recombinant TGF-β2 completely blocked the induction of EAAU when given at the time of induction of disease (days − 1, 0, and + 1 post-immunization). To determine the mechanism by which TGF-β2 inhibited the induction of EAAU, we used a combination of in vitro and in vivo experiments. We specifically analyzed the effect of TGF-β2 on the proliferation of CD4+ T cells and on the generation of Tregs. In our study, the proliferative response of CD4+ T cells to MAA was negligible when cultured in the presence of TGF-β2. We further observed that in MAA-sensitized Lewis rats, treatment with recombinant TGF-β2 resulted in increased percentage of Tregs in the draining popliteal LNs at day 5 post-immunization. It has been reported that TGF-β can induce CD4+ T cells in the periphery to become Tregs [30]. This mechanism may be responsible for the relatively quick development of Tregs in the popliteal LNs of rats treated with TGF-β2 noted in the current study. TGF-β2 has also been implicated to suppress CD4+ T cells [31, 32]. Thus, early increase in number of Tregs in the draining lymph nodes observed in the current study suggests that TGF-β2-induced Tregs suppressed the further activation of immune cells and significantly contributed to the inhibition of EAAU induction.

We previously reported that EAAU is induced in Lewis rats by a CD4+ T cell response to MAA [16, 17, 18, 19], the number of CD4+CD25+FoxP3+ Tregs increased in Lewis rats tolerized against MAA and these Tregs were crucial for the induction of immune tolerance in this animal model [18, 19]. CD4+CD25+FoxP3+ Tregs represent the best characterized subset of suppressive T cell population and play an important role in prevention and/or suppression of autoimmune diseases [33, 34, 35]. It is well recognized that TGF-β2, an immunosuppressive cytokine, triggers the expression of Foxp3 in CD4+ T cells and subsequently converts them to FoxP3+ T regulatory cells [22, 26, 30, 31, 32]. Together, our results suggest that the treatment with TGF-β2 during the early stages of EAAU completely suppressed the immune response by inhibiting CD4+ T cell proliferation and by generating Tregs.

In the second set of in vivo experiments, we tried to mimic the scenario when the patients report to the clinic with active IAU. Keeping that in mind, treatment with TGF-β2 or PBS was withheld until the clinical onset of EAAU and was given on days 12 and 14 post-immunization (after the clinical onset of EAAU). Rats which received PBS developed a prototypical course of EAAU. Interestingly, animals treated with TGF-β2 developed a diminished intraocular inflammation and less severe EAAU. However, this treatment did not result in early resolution of EAAU. Failure of complete suppression of active EAAU by TGF-β2 treatment can be attributable to the fact that in these animals the immune cells are already triggered and primed to perform destined functions.

In conclusion, here, we have reported several novel observations: (i) EAAU did not develop in Lewis rats injected with TGF-β2 on the day before, on the day of, and on the day after MAA immunization; (ii) TGF-β2 mediates inhibition of EAAU induction by diminishing CD4+ T cell proliferation and increasing Tregs; (iii) Lewis rats which received TGF-β2 after the onset of EAAU developed less severe anterior uveitis. Taken together, our results demonstrate that TGF-β2 plays a pivotal role in regulation of intraocular inflammation in EAAU. Disease-modifying effects of TGF-β2 observed in the present study help us in progressing our understanding of the pathogenesis of anterior uveitis and raise the possibility that recombinant TGF-β2 could be a successful therapeutic agent for treatment of human IAU. Since the effect of TGF-β2 was transient when administered systemically, continued intraocular (local) administration of the agent may induce a lasting form of immune suppression.

Notes

Acknowledgements

This work was supported by Pat & Willard Walker Eye Research Center, Jones Eye Institute, University of Arkansas for Medical Sciences, Little Rock, AR.

Compliance with ethical standards

This study was approved by the Institutional Animal Care and Use Committee, University of Arkansas for Medical Sciences, Little Rock, AR.

Conflict of interest

The authors declare that they have no conflict of interest.

Commercial relationships

None.

Ethical statement

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.

References

  1. 1.
    Gritz DC, Wong IG (2004) Incidence and prevalence of uveitis in Northern California. Ophthalmology 111:491–500.  https://doi.org/10.1016/j.ophtha.2003.06.014 CrossRefGoogle Scholar
  2. 2.
    Bora NS, Kaplan HJ (2007) Intraocular diseases - anterior uveitis. Chem Immunol Allergy 92:213–220.  https://doi.org/10.1159/000099272 CrossRefGoogle Scholar
  3. 3.
    Read RW (2006) Uveitis: advances in understanding of pathogenesis and treatment. Curr Rheumatol Rep 8:260–266.  https://doi.org/10.1007/s11926-006-0006-6 CrossRefGoogle Scholar
  4. 4.
    Tang J, Zhu W, Silver PB, Su SB, Chan CC, Caspi RR (2007) Autoimmune uveitis elicited with antigen-pulsed dendritic cells has a distinct clinical signature and is driven by unique effector mechanisms: initial encounter with autoantigen defines disease phenotype. J Immunol 178:5578–5587.  https://doi.org/10.4049/jimmunol.178.9.5578 CrossRefGoogle Scholar
  5. 5.
    Forrester JV, Klaska IP, Yu T, Kuffova L (2013) Uveitis in mouse and man. Int Rev Immunol 32:76–96.  https://doi.org/10.3109/08830185.2012.747524 CrossRefGoogle Scholar
  6. 6.
    Chan CC, Hikita N, Dastgheib K, Whitcup SM, Gery I, Nussenblatt RB (1994) Experimental melanin-protein-induced uveitis in the Lewis rat: immunopathologic processes. Ophthalmology 101:1275–1280.  https://doi.org/10.1016/S0161-6420(94)31199-7 CrossRefGoogle Scholar
  7. 7.
    Smith JR, Rosenbaum JT, Williams KA (2008) Experimental melanin-induced uveitis: experimental model of human acute anterior uveitis. Ophthalmic Res 40:136–140.  https://doi.org/10.1159/000119864 CrossRefGoogle Scholar
  8. 8.
    Smith JR, O'Rourke LM, Becker MD, Cao M, Williams KA, Planck SR, Rosenbaum JT (2000) Anti-rat ICAM-1 antibody does not influence the course of experimental melanin-induced uveitis. Curr Eye Res 21:906–912.  https://doi.org/10.1076/ceyr.21.5.906.5537 CrossRefGoogle Scholar
  9. 9.
    Atalla L, Linker-Israeli M, Steinman L, Rao NA (1990) Inhibition of autoimmune uveitis by anti-CD4 antibody. Invest Ophthalmol Vis Sci 31:1264–1270Google Scholar
  10. 10.
    Becker MD, Adamus G, Davey MP, Rosenbaum JT (2000) The role of T cells in autoimmune uveitis. Ocul Immunol Inflamm 8:93–100.  https://doi.org/10.1076/0927-3948(200006)821-0FT093 CrossRefGoogle Scholar
  11. 11.
    Broekhuyse RM, Winkens HJ, Kuhlmann ED (1996) Intraperitoneally injected melanin is highly uveitogenic. Exp Eye Res 62:199–200.  https://doi.org/10.1006/exer.1996.0024 CrossRefGoogle Scholar
  12. 12.
    Fang IM, Lin CP, Yang CM, Chen MS, Yang CH (2005) Expression of CX3C chemokine fractalkine, and its receptor CX3CR1 in experimental autoimmune anterior uveitis. Mol Vis 11:443–451Google Scholar
  13. 13.
    Yeh PT, Lin FA, Lin CP, Yang CM, Chen MS, Yang CH (2010) Expression of lymphotactin and its receptor, XCX, in Lewis rats with experimental autoimmune anterior uveitis. Graefes Arch Clin Exp Ophthalmol 248:1737–1747.  https://doi.org/10.1007/s00417-010-1435-5 CrossRefGoogle Scholar
  14. 14.
    Bora NS, Kim MC, Kabeer NH, Simpson SC, Tandhasetti MT, Cirrito TP, Kaplan AD, Kaplan HJ (1995) Experimental autoimmune anterior uveitis. Induction with melanin-associated antigen from the iris and ciliary body. Invest Ophthalmol Vis Sci 36:1056–1066Google Scholar
  15. 15.
    Fang IM, Yang CH, Lin CP, Yang CM, Chen MS (2004) Expression of chemokine and receptors in Lewis rats with experimental autoimmune anterior uveitis. Exp Eye Res 78:1043–1055.  https://doi.org/10.1016/j.exer.2004.02.006 CrossRefGoogle Scholar
  16. 16.
    Bora NS, Woon MD, Tandhasetti MT, Cirrito TP, Kaplan HJ (1997) Induction of experimental autoimmune anterior uveitis by a self-antigen: melanin complex without adjuvant. Invest Ophthalmol Vis Sci 38:2171–2175Google Scholar
  17. 17.
    Bora NS, Sohn JH, Kang SG, Cruz JM, Nishihori H, Suk HJ, Wang Y, Kaplan HJ, Bora PS (2004) Type I collagen is the autoantigen in experimental autoimmune anterior uveitis. J Immunol 172:7086–7094.  https://doi.org/10.4049/jimmunol.172.11.7086 CrossRefGoogle Scholar
  18. 18.
    Matta B, Jha P, Bora PS, Bora NS (2008) Tolerance to melanin-associated antigen in autoimmune uveitis is mediated by CD4+CD25+ T-regulatory cells. Am J Pathol 173:1440–1454.  https://doi.org/10.2353/ajpath.2008.080150 CrossRefGoogle Scholar
  19. 19.
    Matta B, Jha P, Bora PS, Bora NS (2010) Antigen specific tolerance inhibits autoimmune uveitis in pre-sensitized animals by deletion and CD4+CD25+ T- regulatory cells. Immunol Cell Biol 88:187–196.  https://doi.org/10.1038/icb.2009.83 CrossRefGoogle Scholar
  20. 20.
    Fang I-M, Yang C-H, Yang C-M (2014) Chitosan oligosaccharides attenuate ocular inflammation in rats with experimental autoimmune anterior uveitis. Mediat Inflamm 2014:1–15.  https://doi.org/10.1155/2014/827847 Google Scholar
  21. 21.
    Hsu Y-R, Chang S-W, Lin Y-C, Yang C-H (2017) MicroRNA-146a alleviates experimental autoimmune anterior uveitis in the eyes of Lewis rats. Mediat Inflamm 2017:1–12.  https://doi.org/10.1155/2017/9601349 Google Scholar
  22. 22.
    Huber S, Schramm C (2006) TGF-beta and CD4+CD25+ regulatory T cells. Front Biosci 11:1014–1023CrossRefGoogle Scholar
  23. 23.
    Yoshimura A, Muto G (2011) TGF-β function in immune suppression. Curr Top Microbiol Immunol 350:127–147.  https://doi.org/10.1007/82_2010_87 Google Scholar
  24. 24.
    Kingsley DM (1994) The TGF-beta superfamily: new members, new receptors, and new genetic tests of function in different organisms. Genes Dev 8:133–146.  https://doi.org/10.1101/gad.8.2.133 CrossRefGoogle Scholar
  25. 25.
    Saika S (2006) TGFbeta pathobiology in the eye. Lab Investig 86:106–115.  https://doi.org/10.1038/labinvest.3700375 CrossRefGoogle Scholar
  26. 26.
    Wan YY, Flavell RA (2007) “Ying-Yang” functions of TGF-β and Tregs in immune regulation. Immunol Rev 220:199–213.  https://doi.org/10.1111/j.1600-065X.2007.00565.x CrossRefGoogle Scholar
  27. 27.
    Yu AL, Moriniere J, Welge-Lussen U (2013) Vitamin E reduces TGF-beta2-induced changes in human trabecular meshwork cells. Curr Eye Res 38:952–958.  https://doi.org/10.3109/02713683.2013.793360 CrossRefGoogle Scholar
  28. 28.
    Swaminathan SS, Oh DJ, Kang MH, Shepard AR, Pang IH, Rhee DJ (2014) TGF-β2-mediated ocular hypertension is attenuated in SPARC-null mice. Invest Ophthalmol Vis Sci 55:4084–4097.  https://doi.org/10.1167/iovs.13-12463 CrossRefGoogle Scholar
  29. 29.
    Pfeiffer N, Voykov B, Renieri G, Bell K, Richter P, Weigel M, Lorenz K, Feindor M et al (2017) First-in-human phase I study of ISTH0036, an antisense oligonucleotide selectively targeting transforming growth factor beta 2 (TGF-β2), in subjects with open-angle glaucoma undergoing glaucoma filtration surgery. PLoS One 12:1–18.  https://doi.org/10.1371/journal.pone.0188899 Google Scholar
  30. 30.
    Chen W, Jin W, Hardegen N, Lei KJ, Marinos N, McGrady G, Wahl SM (2003) Conversion of peripheral CD4+CD25- naive T cells to CD4+CD25+ regulatory T cells by TGF-β induction of transcription factor Foxp3. J Exp Med 198:1875–1886.  https://doi.org/10.1084/jem.20030152 CrossRefGoogle Scholar
  31. 31.
    Fu S, Zang N, Yopp AC, Chen D, Mao M, Chen D, Zang H, Ding Y, Bromberg JS (2004) TGF-beta induces Foxp3 + T-regulatory cells from CD4 + CD25 – precursors. Am J Transplant 4:1614–1627.  https://doi.org/10.1111/j.1600-6143.2004.00566.x CrossRefGoogle Scholar
  32. 32.
    Chen W, Konkel JE (2010) TGF- β and “adaptive” Foxp3+ regulatory T cell. J Mol Cell Biol 2:30–36.  https://doi.org/10.1093/jmcb/mjp004 CrossRefGoogle Scholar
  33. 33.
    Paust S, Cantor H (2005) Regulatory T cells and autoimmune disease. Immunol Rev 204:195–207.  https://doi.org/10.1111/j.0105-2896.2005.00247.x CrossRefGoogle Scholar
  34. 34.
    André S, Tough DF, Lacroix-Desmazes S, Kaveri SV, Bayry J (2009) Surveillance of antigen-presenting cells by CD4+ CD25+ regulatory T cells in autoimmunity: immunopathogenesis and therapeutic implications. Am J Pathol 174:1575–1587.  https://doi.org/10.2353/ajpath.2009.080987 CrossRefGoogle Scholar
  35. 35.
    Selvaraj RK, Geiger TL (2008) Mitigation of experimental allergic encephalomyelitis by TGF-β induced Foxp3+ regulatory T lymphocytes through the induction of anergy and infectious tolerance. J Immunol 180:2830–2838.  https://doi.org/10.4049/jimmunol.180.5.2830 CrossRefGoogle Scholar

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© The Author(s) 2019

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Bharati Matta
    • 1
    • 2
  • Puran S. Bora
    • 1
  • Adam J. Neuhouser
    • 1
  • Nalini S. Bora
    • 1
    Email author
  1. 1.Department of Ophthalmology, Jones Eye Institute, Pat and Willard Walker Eye Research Center, College of MedicineUniversity of Arkansas for Medical SciencesLittle RockUSA
  2. 2.The Feinstein Institute for Medical Research, Northwell HealthManhassetUSA

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