BACKGROUND: Autoimmune uveitis is a leading cause of visual impairment in developed countries in patients of working age. Animal models of experimental autoimmune uveitis (EAU) have been established to serve as a useful template for novel therapeutic approaches. METHODS: Experimental autoimmune uveitis is induced in C57BL/6 mice by subcutaneous application of interphotoreceptor retinoid binding protein in complete Freund's adjuvant and pertussis toxin. Clinical and histological grading is used to assess the inflammation intensity of EAU. RESULTS: The protocol of induction of EAU in mice hides several important aspects, which are crucial for developing the disease. These details have to be addressed to ensure reproducible disease induction. We describe our experience in establishing the model by pointing out the critical steps in EAU protocol which we found important. CONCLUSION: The mouse model of EAU has practical value for preclinical studies, is robust and well established. However, the induction of inflammation of the eye can be quite challenging when important details of the protocol are not recognized and adhered to.

• Klíčová slova
• C57BL/6 mouse strain, chronic posterior autoimmune uveitis, experimental autoimmune uveitis, induction protocol, mouse model,
• MeSH
• adjuvancia imunologická MeSH
• autoimunitní nemoci chemicky indukované   MeSH
• chronická nemoc MeSH
• dráždivé látky MeSH
• Freundovo adjuvans MeSH
• modely nemocí na zvířatech * MeSH
• myši inbrední C57BL MeSH
• oční proteiny * MeSH
• pertusový toxin MeSH
• proteiny vázající retinol * MeSH
• uveitida chemicky indukované   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adjuvancia imunologická MeSH
• dráždivé látky MeSH
• Freundovo adjuvans MeSH
• interstitial retinol-binding protein MeSH Prohlížeč
• oční proteiny * MeSH
• pertusový toxin MeSH
• proteiny vázající retinol * MeSH

AIM: to investigate the dynamics of the T-cell immune response in rabbits with experimental autoimmune uveitis (EAU) of varying severity. MATERIALS AND METHODS: The experiment involved two groups of Chinchilla rabbits (15 rabbits in each group). The model of EAU was created. The clinical picture of intraocular inflammation of varying severity was assessed. The determination of the level of white blood cells (WBC), lymphocytes (Lymphs), CD3+, CD4+, CD8+, and CD16+ in the blood of rabbits was conducted. RESULTS: Group I - moderate and severe uveitis, Group II - uveitis of mild severity. WBC, Lymphs, CD3+, CD4+, CD16+ were elevated and statistically significant in both groups of animals compared to control parameters on all days of the experiment (3, 7, 10, 14, 21 days) (p.

• Klíčová slova
• T-cells response, autoimmune uveitis, experiment,
• MeSH
• autoimunitní nemoci * imunologie   krev   MeSH
• králíci MeSH
• modely nemocí na zvířatech MeSH
• T-lymfocyty imunologie   MeSH
• uveitida * imunologie   krev   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Non-infectious uveitis is considered an autoimmune disease responsible for a significant burden of blindness in developed countries and recent studies have linked its pathogenesis to dysregulation of the gut microbiota. We tested the immunomodulatory properties of two probiotics, Escherichia coli Nissle 1917 (EcN) and E. coli O83:K24:H31 (EcO), in a model of experimental autoimmune uveitis (EAU). To determine the importance of bacterial viability and treatment timing, mice were orally treated with live or autoclaved bacteria in both preventive and therapeutic schedules. Disease severity was assessed by ophthalmoscopy and histology, immune phenotypes in mesenteric and cervical lymph nodes were analyzed by flow cytometry and the gut immune environment was analyzed by RT-PCR and/or gut tissue culture. EcN, but not EcO, protected against EAU but only as a live organism and only when administered before or at the time of disease induction. Successful prevention of EAU was accompanied by a decrease in IRBP-specific T cell response in the lymph nodes draining the site of immunization as early as 7 days after the immunization and eye-draining cervical lymph nodes when the eye inflammation became apparent. Furthermore, EcN promoted an anti-inflammatory response in Peyer's patches, increased gut antimicrobial peptide expression and decreased production of inducible nitric oxide synthase in macrophages. In summary, we show here that EcN controls inflammation in EAU and suggest that probiotics may have a role in regulating the gut-eye axis.

• Klíčová slova
• Escherichia coli Nissle 1917, experimental autoimmune uveitis, macrophages, mucosal immune system, probiotics,
• MeSH
• autoimunitní nemoci terapie   MeSH
• Escherichia coli * MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• probiotika * aplikace a dávkování   farmakologie   MeSH
• střevní sliznice patologie   MeSH
• uveitida terapie   MeSH
• zánět terapie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Autoimmune uveitis is a serious sightthreatening disease that in many cases fails to respond to conventional immunosuppressive or biological therapy. Experimental models used in research allow more detailed study of pathogenesis of the autoimmune process and testing new therapeutic strategies. Recent results show that infection can trigger autoimmune diseases, and some commensal microorganisms are essential in causing disease activity. The aim of this work was to assess the effect of broadspectrum antibiotics - combination of metronidazole and ciprofloxacin or metronidazole alone - on the intensity of intraocular inflammation in experimental autoimmune uveitis (EAU). EAU was induced in mouse strain C57BL/6J by interphotoreceptor retinoid- binding protein in complete Freund's adjuvant and pertussis toxin. The grade of uveitis was assessed clinically and histologically in haematoxylin and eosin- stained tissues. Lymphocytes and macrophages were detected in cryosections using the immunoperoxidase method with antibodies. The therapy was commenced one week before EAU induction and continued throughout the experiment. In addition, metronidazole treatment was also started two weeks before EAU induction. Antibiotics significantly reduced the intensity of uveitis compared to the control group (P < 0.05). The effects of combination of ciprofloxacin and metronidazole and of metronidazole alone were similar when the therapy started one week before EAU induction (P < 0.05). Metronidazole commenced two weeks before EAU induction and throughout the experiment suppressed the intensity of EAU with even higher statistical significance (P < 0.0001). It can be assumed that the high protective effect of metronidazole on EAU intensity may be due not only to its antimicrobial effect, but also to its immunomodulatory activity.

• MeSH
• ciprofloxacin farmakologie   terapeutické užití   MeSH
• metronidazol farmakologie   terapeutické užití   MeSH
• myši inbrední C57BL MeSH
• stupeň závažnosti nemoci MeSH
• uveitida komplikace   farmakoterapie   patologie   MeSH
• zánět komplikace   farmakoterapie   patologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ciprofloxacin MeSH
• metronidazol MeSH

INTRODUCTION: Autoimmune uveitis is a sight threatening disease which in many cases fails to respond to conventional immunosuppressive or biological therapy. The research in experimental models of autoimmune uveitis helps to find new therapeutical strategies. The aim of this study is to present the clinical and histological signs of experimental autoimmune uveitis (EAU) in mice. METHODS: EAU was induced in C57BL/6 mice by subcutaneous application of IRBP (interphotoreceptor retinoid binding protein) in complete Freunds adjuvant and intraperitoneal application of pertussis toxin. Clinical evaluation of uveitis was performed in vivo using special imaging system with otoscope. Histological evaluation of uveitis was performed at day 35 post induction of EAU on hematoxylin and eosin stained frozen sections. Clinical and histological grading was used to assess the inflammation intensity of EAU. RESULTS: The intensity of inflammation is depicted on representative fundus images and histological images of retina at day 35 post induction. CONCLUSION: The model of EAU is robust and reproducible and allows us to study the immunopathological mechanisms of inflammation and its regulation. The inflammatory signs in our model are similar to findings of posterior uveitis of autoimmune etiology in humans, thus we may apply our experimental results in human medicine.

• MeSH
• autoimunitní nemoci chemicky indukované   diagnóza   MeSH
• fundus oculi MeSH
• imunosupresiva MeSH
• modely nemocí na zvířatech * MeSH
• myši inbrední C57BL MeSH
• oční proteiny toxicita   MeSH
• proteiny vázající retinol toxicita   MeSH
• retina patologie   MeSH
• uveitida chemicky indukované   diagnóza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• imunosupresiva MeSH
• interstitial retinol-binding protein MeSH Prohlížeč
• oční proteiny MeSH
• proteiny vázající retinol MeSH

In human, autoimmune uveitis is a leading cause of visual disability and ranks with diabetic retinopathy as a major source of blind registrations in developed countries. Since most cases of non-infectious uveitis are considered to be autoimmune or at least immune-mediated, the management of such patients has rested on appropriate immunosuppression. Some patients, however, despite maximal immunotherapy, fail to respond or are seriously intolerant of the drug therapies. Since its establishment 20 years ago, the model of experimental autoimmune uveoretinitis has served as a useful template for novel therapeutic approaches. The aim of our study was to compare the efficacy of mycophenolate mofetil and cyclophosphamide and golimumab treatment in the mouse model of experimental autoimmune uveitis. The intensity of intraocular inflammation was evaluated histologically in the treatment and control groups. Experimental autoimmune uveitis has been induced in mouse strain C57BL/6 by subcutaneous application of interphotoreceptor retinoid binding protein in complete Freund's adjuvant and pertussis toxin. The treatment was commenced on the day of uveitis induction. Cyclophosphamide was applied intraperitoneally in a single dose (100 mg/kg), mycophenolate mofetil intraperitoneally daily (30 mg/kg or 50 mg/kg), golimumab subcutaneously weekly (70 mg/kg). Sham intraperitoneal injection of a placebo (aqua pro injectione) and untreated mice with experimental autoimmune uveitis served as controls. The results show statistically significant suppression of experimental uveitis both with mycophenolate mofetil and with cyclophosphamide, and thus support its use in human medicine.

• MeSH
• antiflogistika nesteroidní terapeutické užití   MeSH
• cyklofosfamid terapeutické užití   MeSH
• kyselina mykofenolová analogy a deriváty   terapeutické užití   MeSH
• modely nemocí na zvířatech MeSH
• monoklonální protilátky terapeutické užití   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• uveitida farmakoterapie   MeSH
• zánět farmakoterapie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• antiflogistika nesteroidní MeSH
• cyklofosfamid MeSH
• golimumab MeSH Prohlížeč
• kyselina mykofenolová MeSH
• monoklonální protilátky MeSH

• MeSH
• autoimunitní nemoci farmakoterapie   MeSH
• časové faktory MeSH
• imunizace MeSH
• imunosupresiva terapeutické užití   MeSH
• kombinovaná farmakoterapie MeSH
• kožní testy MeSH
• králíci MeSH
• methotrexát terapeutické užití   MeSH
• preklinické hodnocení léčiv MeSH
• triamcinolonacetonid terapeutické užití   MeSH
• uveitida farmakoterapie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• imunosupresiva MeSH
• methotrexát MeSH
• triamcinolonacetonid MeSH

AIMS: To investigate the therapeutic potential of visual stimulation (VS) and BDNF in murine experimental autoimmune uveoretinitis (EAU). MAIN METHODS: Mice were immunized by subcutaneous injection of interphotoreceptor retinoid-binding protein in Freund's complete adjuvant and intravenous injection of pertussis toxin, and were then exposed to high-contrast VS 12 h/day (days 1-14 post-immunization). EAU severity was assessed by examining clinical score, visual acuity, inflammatory markers, and immune cells in the retina. The transcriptome of activated retinal cells was determined by RNA-seq using RNA immunoprecipitated in complex with phosphorylated ribosomal protein S6. The retinal levels of protein products of relevant upregulated genes were quantified. The effect of BDNF on EAU was tested in unstimulated mice by its daily topical ocular administration (days 8-14 post-immunization). KEY FINDINGS: VS attenuated EAU development and decreased the expression of pro-inflammatory cytokines/chemokines and numbers of immune cells in the retina (n = 10-20 eyes/group for each analysis). In activated retinal cells of control mice (n = 30 eyes/group), VS upregulated genes encoding immunomodulatory neuropeptides, of which BDNF and vasoactive intestinal peptide (VIP) also showed increased mRNA and protein levels in the retina of VS-treated EAU mice (n = 6-10 eyes/group for each analysis). In unstimulated EAU mice, BDNF treatment mimicked the protective effects of VS by modulating the inflammatory and stem cell properties of Müller cells (n = 5 eyes/group for each analysis). SIGNIFICANCE: VS effectively suppresses EAU, at least through enhancing retinal levels of anti-inflammatory and neuroprotective factors, VIP and BDNF. Our findings also suggest BDNF as a promising therapeutic agent for uveitis treatment.

• Klíčová slova
• Brain-derived neurotrophic factor (BDNF), Experimental autoimmune uveoretinitis (EAU), Müller cells, Retina, Visual stimulation,
• MeSH
• autoimunitní nemoci * imunologie   metabolismus   MeSH
• cytokiny metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• mozkový neurotrofický faktor * metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• retina metabolismus   účinky léků   MeSH
• retinitida * farmakoterapie   prevence a kontrola   imunologie   MeSH
• uveitida * metabolismus   farmakoterapie   imunologie   MeSH
• vazoaktivní intestinální peptid farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Bdnf protein, mouse MeSH Prohlížeč
• cytokiny MeSH
• mozkový neurotrofický faktor * MeSH
• vazoaktivní intestinální peptid MeSH

The microbiota is a crucial modulator of the immune system. Here, we evaluated how its absence or reduction modifies the inflammatory response in the murine model of experimental autoimmune uveoretinitis (EAU). We induced EAU in germ-free (GF) or conventionally housed (CV) mice and in CV mice treated with a combination of broad-spectrum antibiotics either from the day of EAU induction or from one week prior to induction of disease. The severity of the inflammation was assessed by fundus biomicroscopy or by histology, including immunohistology. The immunophenotyping of T cells in local and distant lymph nodes was performed by flow cytometry. We found that GF mice and mice where the microbiota was reduced one week before EAU induction were protected from severe autoimmune inflammation. GF mice had lower numbers of infiltrating macrophages and significantly less T cell infiltration in the retina than CV mice with EAU. GF mice also had reduced numbers of IFN-γ and IL-17-producing T cells and increased numbers of regulatory T cells in the eye-draining lymph nodes. These data suggest that the presence of microbiota during autoantigen recognition regulates the inflammatory response by influencing the adaptive immune response.

• MeSH
• adaptivní imunita MeSH
• aktivace lymfocytů MeSH
• antibakteriální látky farmakologie   MeSH
• autoantigeny imunologie   MeSH
• autoimunitní nemoci chemicky indukované   imunologie   mikrobiologie   MeSH
• bakteriální nálož účinky léků   MeSH
• gnotobiologické modely MeSH
• interferon gama biosyntéza   MeSH
• interleukin-17 biosyntéza   MeSH
• makrofágy imunologie   MeSH
• mikrobiota * imunologie   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• oči imunologie   patologie   MeSH
• oční proteiny imunologie   MeSH
• proteiny vázající retinol imunologie   MeSH
• průtoková cytometrie MeSH
• regulační T-lymfocyty imunologie   MeSH
• retina imunologie   MeSH
• retinitida chemicky indukované   etiologie   imunologie   mikrobiologie   MeSH
• uveitida chemicky indukované   imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• antibakteriální látky MeSH
• autoantigeny MeSH
• interferon gama MeSH
• interleukin-17 MeSH
• interstitial retinol-binding protein MeSH Prohlížeč
• oční proteiny MeSH
• proteiny vázající retinol MeSH

The gateway reflex is a mechanism by which neural inputs regulate chemokine expression at endothelial cell barriers, thereby establishing gateways for the invasion of autoreactive T cells into barrier-protected tissues. In this study, we hypothesized that rod photoreceptor dysfunction causes remodeling of retinal neural activity, which influences the blood-retinal barrier and the development of retinal inflammation. We evaluated this hypothesis using Gnat1rd17 mice, a model of night blindness with late-onset rod-cone dystrophy, and experimental autoimmune uveoretinitis (EAU). Retinal remodeling and its effect on EAU development were investigated by transcriptome profiling, target identification, and functional validation. We showed that Gnat1rd17 mice primarily underwent alterations in their retinal dopaminergic system, triggering the development of an exacerbated EAU, which was counteracted by dopamine replacement with L-DOPA administered either systemically or locally. Remarkably, dopamine acted on retinal endothelial cells to inhibit NF-κB and STAT3 activity and the expression of downstream target genes such as chemokines involved in T cell recruitment. These results suggest that rod-mediated dopamine release functions in a gateway reflex manner in the homeostatic control of immune cell entry into the retina, and the loss of retinal dopaminergic activity in conditions associated with rod dysfunction increases the susceptibility to autoimmune uveitis.

• Klíčová slova
• Gnat1, NF-κB, STAT3, blood–retinal barrier, dopamine, endothelial cells, experimental autoimmune uveoretinitis, gateway reflex, night blindness, rod-cone dystrophy,
• MeSH
• buněčné linie MeSH
• dopamin metabolismus   MeSH
• endoteliální buňky metabolismus   patologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• NF-kappa B metabolismus   MeSH
• retina metabolismus   patologie   MeSH
• transkripční faktor STAT3 metabolismus   MeSH
• uveitida metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dopamin MeSH
• NF-kappa B MeSH
• Stat3 protein, mouse MeSH Prohlížeč
• transkripční faktor STAT3 MeSH