RESUMO
BACKGROUND: Ultraviolet B irradiation confers strong resistance against experimental autoimmune encephalomyelitis, a model of multiple sclerosis. This protection by ultraviolet B is independent of vitamin D production but causes isomerization of urocanic acid, a naturally occurring immunosuppressant. METHODS: To determine whether UCA isomerization from trans to cis is responsible for the protection against experimental autoimmune encephalomyelitis afforded by ultraviolet B, trans- or cis-urocanic acid was administered to animals and their disease progression was monitored. RESULTS: Disease incidence was reduced by 74% in animals exposed to ultraviolet B, and skin cis-urocanic acid levels increased greater than 30%. However, increasing skin cis-urocanic acid levels independent of ultraviolet B was unable to alter disease onset or progression. CONCLUSIONS: It is unlikely that urocanic acid isomerization is responsible for the ultraviolet B-mediated suppression of experimental autoimmune encephalomyelitis. Additional work is needed to investigate alternative mechanisms by which UVB suppresses disease.
Assuntos
Encefalomielite Autoimune Experimental/metabolismo , Esclerose Múltipla/metabolismo , Raios Ultravioleta , Ácido Urocânico/química , Ácido Urocânico/efeitos da radiação , Animais , Progressão da Doença , Encefalomielite Autoimune Experimental/terapia , Feminino , Isomerismo , Camundongos , Camundongos Endogâmicos C57BL , Esclerose Múltipla/terapia , Terapia Ultravioleta , Ácido Urocânico/administração & dosagemRESUMO
Urocanic acid is a major ultraviolet (UV)-absorbing chromophore. Chitins are highly crystalline structures that are found predominantly in crustacean shells. Alpha-chitin consists of microfibers that contain nanofibrils embedded in a protein matrix. Acid hydrolysis is a common method used to prepare chitin nanofibrils (NFs). We typically obtain NFs by hydrolyzing chitin with acetic acid. However, in the present study, we used urocanic acid to prepare urocanic acid chitin NFs (UNFs) and examined its protective effect against UVB radiation. Hos: HR-1 mice coated with UNFs were UVB irradiated (302 nm, 150 mJ/cm²), and these mice showed markedly lower UVB radiation-induced cutaneous erythema than the control. Additionally, sunburn cells were rarely detected in the epidermis of UNFs-coated mice after UVB irradiation. Although the difference was not as significant as UNFs, the number of sunburn cells in mice treated with acetic acid chitin nanofibrils (ANFs) tended to be lower than in control mice. These results demonstrate that ANFs have a protective effect against UVB and suggest that the anti-inflammatory and antioxidant effects of NFs influence the protective effect of ANFs against UVB radiation. The combination of NFs with other substances that possess UV-protective effects, such as urocanic acid, may provide an enhanced protective effect against UVB radiation.
Assuntos
Quitina/química , Nanofibras , Raios Ultravioleta/efeitos adversos , Ácido Urocânico/farmacologia , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Eritema/prevenção & controle , Masculino , Camundongos , Camundongos Pelados , Pele/efeitos dos fármacos , Pele/patologia , Pele/efeitos da radiação , Queimadura Solar/prevenção & controle , Ácido Urocânico/administração & dosagem , Ácido Urocânico/químicaRESUMO
New treatment modalities are needed in atopic dermatitis. We evaluated the pharmacokinetics, safety, tolerability, and efficacy of topical cis-urocanic acid (cis-UCA) cream in randomised vehicle-controlled double-blinded clinical trials. The subjects received 5% cis-UCA emulsion cream and control vehicle on volar forearms after right-left randomisation. Study 1: 16 healthy subjects received one dose on the skin and, a week later, on DMSO-irritated skin. Study 2: 16 healthy subjects received 2 daily doses for 10 days. Study 3: 13 patients with mild to moderate disease were treated on selected skin lesions twice daily for 28 days. Study treatments were well tolerated. cis-UCA remained close to endogenous levels in plasma and urine. cis-UCA reduced transepidermal water loss (TEWL) both in healthy subjects and in the patients. Eczema area severity index and physician's global assessment improved from baseline with both treatments. cis-UCA cream improved skin barrier function and suppressed inflammation in the human skin.
Assuntos
Dermatite Atópica/tratamento farmacológico , Fármacos Dermatológicos/administração & dosagem , Pele/efeitos dos fármacos , Ácido Urocânico/administração & dosagem , Administração Cutânea , Dermatite Atópica/diagnóstico , Dermatite Atópica/imunologia , Fármacos Dermatológicos/efeitos adversos , Fármacos Dermatológicos/farmacocinética , Método Duplo-Cego , Esquema de Medicação , Emulsões , Finlândia , Humanos , Estudos Prospectivos , Índice de Gravidade de Doença , Pele/imunologia , Pele/patologia , Fatores de Tempo , Resultado do Tratamento , Ácido Urocânico/efeitos adversos , Ácido Urocânico/farmacocinéticaRESUMO
BACKGROUND: cis-Urocanic acid (cis-UCA) is an endogenous immunosuppressive molecule of the epidermis. OBJECTIVES: We investigated the effects of topical cis-UCA creams (2·5% and 5%) in acute and subacute mouse models of skin inflammation. METHODS: Acute skin irritation was induced by applying dimethyl sulphoxide (DMSO) on the earlobe of CD-1 mice. Topical cis-UCA, hydrocortisone (1%) or tacrolimus (0·1%) were applied 10 min later. In another model, subacute inflammation was provoked and maintained by three applications of 12-O-tetradecanoylphorbol-13-acetate (TPA) on the ears of NMRI mice on days 1, 2 and 4. The test products were applied topically twice a day during 6 days. RESULTS: In the acute DMSO model, cis-UCA creams suppressed ear swelling at 1 h significantly more efficiently than hydrocortisone (P < 0·01) and tacrolimus (P < 0·001). Ear swelling was significantly inhibited by cis-UCA (P < 0·001) in the subacute TPA model as well. The 5% cream also decreased erythema, whereas tacrolimus enhanced skin reddening. Treatments with cis-UCA did not affect TPA-induced infiltration of neutrophils to the skin. In contrast to hydrocortisone, cis-UCA did not reduce epidermal thickness. CONCLUSIONS: The results suggest that cis-UCA - unlike hydrocortisone and tacrolimus - is efficient in both acute and subacute skin inflammation, attenuating skin oedema and erythema. Topical drug therapy with cis-UCA may provide a safe and effective drug treatment modality in inflammatory skin disorders.
Assuntos
Fármacos Dermatológicos/farmacologia , Toxidermias/tratamento farmacológico , Edema/tratamento farmacológico , Eritema/tratamento farmacológico , Ácido Urocânico/farmacologia , Doença Aguda , Administração Cutânea , Animais , Fármacos Dermatológicos/administração & dosagem , Dimetil Sulfóxido/toxicidade , Toxidermias/etiologia , Irritantes/toxicidade , Masculino , Camundongos , Infiltração de Neutrófilos/efeitos dos fármacos , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/toxicidade , Ácido Urocânico/administração & dosagemRESUMO
Exposure to UVB radiation before antigen delivery at an unirradiated site inhibits functional immunological responses. Mice treated dorsally with suberythemal low-dose UVB and immunized with ova in abdominal skin generated ova-specific CD8 T cells with a significantly decreased activation, expansion, and cytotoxic activity compared with unirradiated mice. UVB also impaired the delayed-type hypersensitivity (DTH) reaction to ova. Transfer of CD4âºCD25âºcells from UVB-exposed mice did not suppress the ova-specific CD8 T-cell response or DTH reaction in unexposed mice, confirming that systemic low-dose UVB does not induce long-lived functional regulatory CD4âºCD25⺠T cells. Repairing cyclobutane pyrimidine dimer-type DNA damage and blocking aryl hydrocarbon receptor signaling also did not reverse the immunosuppressive effect of UVB on ova-specific CD8 T cells and DTH, suggesting that cyclobutane pyrimidine dimers and the aryl hydrocarbon receptor are not required in systemic low-dose UVB-induced immunosuppression. The known UVB chromophore, cis-urocanic acid, and reactive oxygen species triggered the inhibition of DTH caused by UVB, but they were not involved in the modulation of CD8 T cells. These findings indicate that systemic low-dose UVB impedes the primary response of antigen-specific CD8 T cells by a novel mechanism that is independent of pathways known to be involved in systemic suppression of DTH.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos da radiação , Inflamação/patologia , Pele/imunologia , Pele/patologia , Raios Ultravioleta , Administração Tópica , Animais , Antioxidantes/farmacologia , Antígenos CD4/metabolismo , Linfócitos T CD8-Positivos/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Reparo do DNA/efeitos da radiação , Relação Dose-Resposta à Radiação , Feminino , Hipersensibilidade Tardia/complicações , Hipersensibilidade Tardia/imunologia , Hipersensibilidade Tardia/patologia , Inflamação/complicações , Inflamação/imunologia , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia , Dímeros de Pirimidina/metabolismo , Receptores de Hidrocarboneto Arílico/antagonistas & inibidores , Pele/efeitos dos fármacos , Pele/efeitos da radiação , Baço/efeitos dos fármacos , Baço/patologia , Baço/efeitos da radiação , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/efeitos da radiação , Ácido Urocânico/administração & dosagem , Ácido Urocânico/farmacologiaRESUMO
BACKGROUND/PURPOSE: Trans-urocanic acid is isomerized to cis-urocanic acid (C-UCA) by ultraviolet radiation. C-UCA suppresses immunity in vitro and in vivo in animals; its effect on human skin is unknown. We sought to determine whether its topical application to normal skin suppresses induction of immunity to dinitrochlorobenzene (DNCB). METHODS: Forty subjects applied C-UCA (0%, 0.02%, 0.2%, or 2%) for 17 days. A 40-mcg dose of DNCB was then applied to induce immunity. Subjects were challenged for immunity at 6-week follow-up by occluding doses of DNCB (0, 3.125, 6.25, or 12.5 mcg) on untreated normal skin. Induced immunity was measured by area of erythema and induration 2 and 4 days postchallenge. RESULTS: No significant differences were found in incidence of sensitization by C-UCA concentration (P=.59). DNCB sensitization developed in all 10 subjects induced through 0% C-UCA (placebo); only 23 of 30 patients were sensitized through skin treated with C-UCA. Mean areas of erythema and induration induced through C-UCA-treated skin were less than those in controls (P < 0.05). The number of Langerhans cells in C-UCA-treated skin was unaffected. Laboratory tests of immune function and lymphocyte numbers were unchanged. CONCLUSION: Topically applied C-UCA blunts normal induction responses to a cutaneous sensitizer.
Assuntos
Imunossupressores/efeitos adversos , Pele/imunologia , Raios Ultravioleta/efeitos adversos , Ácido Urocânico/efeitos adversos , Adulto , Dinitroclorobenzeno/administração & dosagem , Dinitroclorobenzeno/imunologia , Eritema/imunologia , Feminino , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/imunologia , Células de Langerhans/imunologia , Masculino , Pessoa de Meia-Idade , Estereoisomerismo , Fatores de Tempo , Ácido Urocânico/administração & dosagem , Ácido Urocânico/imunologiaRESUMO
OBJECTIVE: To present a novel treatment approach for urinary bladder cancer, protodynamic therapy, which comprises inhibition of cancer cell proliferation by intracellular acidification; cis-urocanic acid (cis-UCA) was investigated as a protodynamic drug in bladder cancer cell cultures and compared with conventional chemotherapeutic agents. MATERIALS AND METHODS: The moderately differentiated cell line 5637 and the poorly differentiated T24 cell line were exposed to cis-UCA for 0.25-2 h, and to epirubicin, doxorubicin, cisplatin and paclitaxel for 2 h, to simulate drug exposure on intravesical instillation. The combination of cis-UCA and chemotherapeutic agents was also studied. Cell viability was measured with a colorimetric assay. RESULTS: cis-UCA inhibited proliferation and suppressed the survival of cells at an extracellular pH
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma de Células de Transição/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Fotoquimioterapia/métodos , Neoplasias da Bexiga Urinária/tratamento farmacológico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/administração & dosagem , Relação Dose-Resposta a Droga , Doxorrubicina/administração & dosagem , Sinergismo Farmacológico , Epirubicina/administração & dosagem , Humanos , Concentração de Íons de Hidrogênio , Paclitaxel/administração & dosagem , Ácido Urocânico/administração & dosagemRESUMO
OBJECTIVE: To evaluate the effects of cis-urocanic acid (cis-UCA) on mammary gland (MG) inflammation and injury associated with Escherichia coli-induced mastitis in dairy cows. ANIMALS: 12 lactating dairy cows (36 MGs). PROCEDURES: At 7-week intervals, a different MG in each cow was experimentally inoculated with E coli. At 6-hour intervals from 6 to 36 hours after inoculation, the inoculated MG in each cow was infused with 40 mL of saline (0.9% NaCl) solution, 12.5mM cis-UCA, or 25mM cis-UCA (4 cows/group); ultimately, each cow received each treatment. Immediately prior to and at various time points after inoculation and treatment, milk samples were collected. Bacterial CFUs, somatic cell counts (SCCs), N-acetyl-beta-D-glucosaminidase (NAGase) and lactate dehydrogenase (LDH) activities, and concentrations of bovine serum albumin, tumor necrosis factor-alpha, and cis-UCA were quantified in each milk sample. Results-Compared with findings in saline solution-treated MGs, NAGase and LDH activities in milk samples from cis-UCA-treated MGs were lower. Cis-UCA had no effect on milk SCCs and milk concentrations of bovine serum albumin and tumor necrosis factor-alpha. Furthermore, cis-UCA had no adverse effect on bacterial clearance; CFUs of E coli in MGs treated with saline solution or cis-UCA were equivalent. CONCLUSIONS AND CLINICAL RELEVANCE: In cows, milk NAGase and LDH activities were both lower in E coli-infected MGs infused with cis-UCA than in those infused with saline solution, which suggests that cis-UCA reduced mastitis-associated tissue damage. Furthermore, these data indicated that therapeutic concentrations of cis-UCA in milk can be achieved via intramammary infusion.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Inflamação/tratamento farmacológico , Mastite Bovina/tratamento farmacológico , Ácido Urocânico/farmacologia , Animais , Bovinos , Relação Dose-Resposta a Droga , Escherichia coli/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/veterinária , Feminino , Fatores de Tempo , Ácido Urocânico/administração & dosagemRESUMO
The p53 tumor suppressor gene is the most frequently mutated gene identified in many tumors, including hepatocellular carcinoma (HCC). Gene therapy using the p53 gene has been proposed and performed with inactivation of p53 function. However, there have been few reports of nonviral vector-mediated p53 gene delivery in HCC. In this study, the wild-type p53 (wt-p53) gene was transfected into human hepatocellular carcinoma cell line HepG(2) using the urocanic acid-modified chitosan (UAC) as a nonviral vector, and transfection efficiency was determined by FACS analysis. UAC-mediated p53 transfection in HepG(2) cells resulted in high expression levels of wt-p53 mRNA and protein and significant cellular growth inhibition. DAPI staining and Annexin V/PI double-staining assay revealed apoptosis occurrence in HepG(2) cells after treatment with UAC/pEGFP-p53 complexes. In in vivo studies, intratumoral injection of UAC/pEGFP-p53 complexes into BALB/c nude mice bearing HepG(2) cells clearly suppressed tumor growth, and significantly induced apoptosis. These results demonstrated that UAC-mediated efficient p53 gene transfer could induce apoptosis thereby significantly inhibiting the growth of HepG(2) cells in vitro and in vivo, and suggested that UAC-mediated p53 gene delivery might be a promising approach for HCC gene therapy.
Assuntos
Carcinoma Hepatocelular/terapia , Quitosana/administração & dosagem , Terapia Genética/métodos , Neoplasias Hepáticas/terapia , Proteína Supressora de Tumor p53/genética , Ácido Urocânico/administração & dosagem , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Transfecção/métodosRESUMO
A new nonviral gene vector--urocanic acid-coupled chitosan (UAC) was prepared by the reaction of the activated urocanic acid (UA) with the amine group on the chitosan (CTS). The structure of UAC was confirmed with FT-IR, 1H NMR and element analysis. The influencing factors of substitution values were studied by orthogonal test, and the substitution values of UAC increased with the prolongation of activating time of UA and the increasing ratio of UA to CTS. The condensation ability and the resistance to DNase I of UAC/pDNA were evaluated by agarose gel electrophoresis, and UAC showed good condensation ability with pDNA, well protecting pDNA from the degradation by DNase I. The particle size and zeta potential were evaluated by zetasizer, and the results showed that the UAC/pDNA complex was well stable and could easily enter into cells. The transfection studies were performed with HepG2 cells in vitro. It showed that the in vitro transfection of UAC/pDNA was efficient in HepG2 cells and could express more green fluorescent proteins than that of CTS. So the UAC is easy to prepare and a promising non-viral gene vector.
Assuntos
Quitosana/administração & dosagem , Terapia Genética/métodos , Vetores Genéticos , Ácido Urocânico/administração & dosagem , Quitosana/síntese química , Quitosana/metabolismo , DNA/genética , DNA/metabolismo , Desoxirribonuclease I/metabolismo , Sistemas de Liberação de Medicamentos , Células Hep G2 , Humanos , Tamanho da Partícula , Plasmídeos , Transfecção , Ácido Urocânico/síntese química , Ácido Urocânico/metabolismoRESUMO
The low efficiency of conventional therapies in achieving long-term survival of patients with lung cancer calls for development of novel treatment options. Although several genes have been investigated for their antitumor activities through gene delivery, problems surrounding the methods used, such as efficiency, specificity, and toxicity, hinder application of such therapies in clinical settings. Aerosol gene delivery as nonviral and noninvasive method for gene therapy may provide an alternative for a safer and more effective treatment for lung cancer. In this study, imidazole ring-containing urocanic acid-modified chitosan (UAC) designed in previous study was used as a gene carrier. The efficiency of UAC carrier in lungs was confirmed, and the potential effects of the programmed cell death protein 4 (PDCD4) tumor suppressor gene on three major pathways (apoptosis, cell cycle, and angiogenesis) were evaluated. Aerosol containing UAC/PDCD4 complexes was delivered into K-ras null lung cancer model mice through the nose-only inhalation system developed by our group. Delivered UAC/PDCD4 complex facilitated apoptosis, inhibited pathways important for cell proliferation, and efficiently suppressed pathways important for tumor angiogenesis. In summary, results obtained by Western blot analysis, immunohistochemistry, and terminal deoxynucleotidyl transferase-mediated nick end labeling assay suggest that our aerosol gene delivery technique is compatible with in vivo gene delivery and can be applied as a noninvasive gene therapy.
Assuntos
Proteínas Reguladoras de Apoptose/administração & dosagem , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Quitosana/administração & dosagem , Genes ras , Neoplasias Pulmonares/tratamento farmacológico , Neovascularização Patológica/prevenção & controle , Proteínas de Ligação a RNA/administração & dosagem , Ácido Urocânico/administração & dosagem , Administração por Inalação , Aerossóis , Animais , Proteínas Reguladoras de Apoptose/uso terapêutico , Sobrevivência Celular/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout , Proteínas de Ligação a RNA/uso terapêuticoRESUMO
Cutaneous exposure to the pyrethroid insecticide permethrin significantly suppresses contact hypersensitivity (CH) response to oxazolone in C57BL/6N mice. Additionally, cis-urocanic acid (cUCA), an endogenous cutaneous chromophore isomerized to its active form following exposure to ultraviolet radiation, modulates cell-mediated cutaneous immune responses. This study describes cutaneous immune alterations following combined topical permethrin and intradermal cUCA exposure. Female C57BL/6N mice were administered 5, 50 or 100 microg cUCA daily for 5 consecutive days. CH was then evaluated by the mouse ear swelling test (MEST) response to oxazolone. Decreased responses of 52.3%, 76.3% and 76.3%, respectively, as compared to controls were observed. Then, mice were co-exposed to 5 microg cUCA daily for 5 days and 1.5, 5, 15, or 25 microL permethrin, on either day 1, 3 or 5 of the cUCA treatment to evaluate combined immunomodulatory effects of the two chemicals, or cUCA daily for 5 days followed by permethrin on day 3, 5, or 7 after the last cUCA injection to demonstrate prolonged immunosuppressive effects. Two days after final treatment, mice were sensitized with oxazolone and MEST was performed. Mice receiving five cUCA injections and permethrin topically on cUCA injection day 1 showed up to 93.3% suppression of MEST compared to vehicle control. CH was suppressed by 87.5%, 86.6% and 74.2% in mice treated with 25 muL permethrin on days 3, 5 and 7 after cUCA, respectively, compared to vehicle control. Taken together, these data indicate co-exposure to cUCA and permethrin profoundly suppresses cell-mediated cutaneous immunity.
Assuntos
Dermatite de Contato/prevenção & controle , Derme/efeitos dos fármacos , Derme/patologia , Permetrina/farmacologia , Ácido Urocânico/farmacologia , Animais , Dermatite de Contato/tratamento farmacológico , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Permetrina/administração & dosagem , Ácido Urocânico/administração & dosagemRESUMO
Nonviral delivery systems for gene therapy have been increasingly proposed as safer alternatives to viral vectors. Chitosan is considered to be a good candidate for the gene delivery system since it is already known as a biocompatible, biodegradable, and low toxic material with high cationic charge potential. However, the use of chitosan for gene delivery is limited due to low transfection efficiency. To enhance the transfection efficiency, water-soluble chitosan (WSC) was coupled with urocanic acid (UA) bearing imidazole ring which can play the crucial role in endosomal rupture through proton sponge mechanism. The urocanic acid-modified chitosan (UAC) was complexed with DNA, and UAC/DNA complexes were characterized. The sizes of UAC/DNA complexes under physiological condition (109-342 nm) were almost same as those of chitosan-DNA complexes. UAC also showed good DNA binding ability, high protection of DNA from nuclease attack, and low cytotoxicity. The transfection efficiency of chitosan into 293T cells was much enhanced after coupling with UA and increased with an increase of UA contents in the UAC.
Assuntos
Quitina/análogos & derivados , Quitina/administração & dosagem , DNA/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Terapia Genética/métodos , Ácido Urocânico/administração & dosagem , Linhagem Celular , Quitina/química , Quitosana , DNA/genética , Relação Dose-Resposta a Droga , Humanos , Ácido Urocânico/químicaRESUMO
BACKGROUND/PURPOSE: Cutaneous cis-urocanic acid (cUCA) or ultraviolet B exposure has been shown to cause diminished cutaneous contact hypersensitivity (CH) and to induce systemic tolerance (increased regulatory T lymphocytes) in mice. Permethrin is also a known CH inhibitor, but the molecular mechanisms are currently poorly understood. In this study, CH was evaluated in four strains of mice: an immunosensitive strain (C57BL/6N), an immunoresistant strain (SvImJ), a strain developed from C57BL/6N mice but genetically altered at both the tumor necrosis factor-alpha receptors (TNFalphap55R and p75R), and a strain developed from C57BL/6N but genetically deleted at the interferon-gamma (IFNgamma) locus. METHODS: CH was evaluated in each group via oxazolone challenge following a 5-day exposure to intradermal (ID) cUCA or a single exposure to topical permethrin, or co-exposure to both chemicals in 5-week-old female C57BL/6N, SvImJ, and C57BL/6N mice genetically altered at the TNFalpha or IFNgamma locus. RESULTS: A 5-day exposure to ID cUCA or a single exposure to topical permethrin resulted in diminished CH response in C57BL/6N mice, and this effect was exacerbated with concurrent exposure to both chemicals. CH in SvImJ was both cUCA- and permethrin-resistant relative to C57BL/6N mice, as 5-day cUCA or a single exposure to permethrin did not diminish CH, nor did concurrent exposure to cUCA and permethrin. Mice deleted at both TNFalphaR loci displayed similar but somewhat blunted diminished CH responses to cUCA or permethrin. This trend became significant with combined chemical exposure. IFNgamma knockout mice displayed similar diminished CH responses to cUCA or permethrin alone. Unlike C57BL/6N mice, the IFNgamma knockout mice did not show a further reduction in CH with combined chemical exposure. CONCLUSIONS: These results suggest the following: (1)Mouse strains show variable susceptibility to permethrin- and cUCA-induced immunomodulation. (2)TNFalpha may be involved in the immunomodulatory effects of cUCA and permethrin. (3)IFNgamma may be required for the more than additive depression of CH caused by cUCA+permethrin.
Assuntos
Adjuvantes Imunológicos/farmacologia , Dermatite de Contato/imunologia , Pele/imunologia , Raios Ultravioleta , Ácido Urocânico/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Administração Cutânea , Animais , Dermatite de Contato/patologia , Feminino , Injeções Intradérmicas , Interferon-alfa/efeitos dos fármacos , Interferon-alfa/efeitos da radiação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Permetrina/administração & dosagem , Permetrina/farmacologia , Pele/efeitos dos fármacos , Pele/efeitos da radiação , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/efeitos da radiação , Ácido Urocânico/administração & dosagemRESUMO
UV radiation induces skin cancer primarily by its DNA-damaging properties, but also by its capacity to suppress the immune system. The photoisomer of urocanic acid (UCA), cis-UCA, is an important mediator of UV-induced immunosuppression and is involved in the inhibition of tumor immunity. The immunomodulatory cytokine IL-12 is known to counteract many of the immunosuppressive effects of UV radiation, including UV-induced immune tolerance. In this study, we addressed whether IL-12 also reverts the immunosuppressive activities of cis-UCA. Cis-UCA inhibits the ability of Langerhans cells to present tumor Ags for primary and secondary tumor immune responses. IL-12 treatment completely prevented the suppression by cis-UCA. IL-12 also protected mice from cis-UCA-induced suppression of contact hypersensitivity responses. To study the effects of cis-UCA on Ag-processing and Ag-presenting function in vitro, Langerhans cells were treated with UCA isomers and incubated with OVA or OVA peptide(323-339) before exposure to OVA-specific transgenic T cells. Cis-, but not trans-UCA suppressed Ag presentation, which was completely reversed upon addition of IL-12. Since these findings suggest that cis-UCA may play an important role in photocarcinogenesis by inhibiting a tumor immune response, mice were chronically UVB irradiated to induce skin cancer. Whereas all mice in the control groups developed tumors, mice treated with a mAb with specificity for cis-UCA showed a significantly reduced tumor incidence. These data strongly indicate the importance of cis-UCA during photocarcinogenesis and support the concept of counteracting cis-UCA as an alternative strategy to prevent UV-induced skin cancer, possibly via the application of IL-12.
Assuntos
Apresentação de Antígeno/imunologia , Antígenos de Neoplasias/metabolismo , Imunossupressores/antagonistas & inibidores , Interleucina-12/fisiologia , Células de Langerhans/imunologia , Neoplasias Induzidas por Radiação/imunologia , Raios Ultravioleta , Ácido Urocânico/antagonistas & inibidores , Animais , Apresentação de Antígeno/efeitos dos fármacos , Antígenos de Neoplasias/imunologia , Dermatite de Contato/imunologia , Dermatite de Contato/prevenção & controle , Epitopos de Linfócito T/imunologia , Feminino , Soros Imunes/administração & dosagem , Imunização Passiva , Imunossupressores/farmacologia , Injeções Intraperitoneais , Células de Langerhans/efeitos dos fármacos , Células de Langerhans/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos A , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/prevenção & controle , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Células Tumorais Cultivadas , Ácido Urocânico/administração & dosagem , Ácido Urocânico/imunologia , Ácido Urocânico/farmacologiaRESUMO
Urocanic acid (UCA) has been shown to mediate the UVB radiation-induced immunosuppression initiated in the skin by UV-induced isomerization from the trans to the cis isomer. However, the mechanism by which cis-UCA acts is still unclear. Therefore, the present study was undertaken to determine the effect of trans- and cis-UCA on cyclic adenosine 3',5'-monophosphate (cAMP) synthesis in human dermal fibroblasts, Golden Syrian hamster hepatocytes and in the human adenocarcinoma cell line, HT29. Neither trans- nor cis-UCA was able to stimulate cAMP synthesis directly in any of the models tested. In human dermal fibroblasts, cis-UCA, in contrast to trans-UCA, specifically inhibited cAMP synthesis induced by either prostaglandin (PG) E1 or PGE2 with a maximum inhibitory effect of 25-30% at cis-UCA concentrations greater than 1 microM and half-maximum inhibitory effect (EC50) observed at 35 nM. The effect of cis-UCA was not to stimulate phosphodiesterase and cAMP breakdown. The inhibitory effect of cis-UCA (an imidazole derivative) was not mediated through stimulation of the alpha 2-adrenergic receptor. The inhibitory effect of cis-UCA on stimulated cAMP synthesis was a function of the cell density and was only significant when the fibroblasts were confluent or postconfluent. In contrast to the studies with human dermal fibroblasts, an inhibitory effect of cis-UCA was not observed in either isolated hamster hepatocytes or HT29 cells, in which cAMP synthesis was stimulated by glucagon and vasoactive intestinal peptide, respectively. These results point to a possible regulation of cAMP synthesis in fibroblasts as one mechanism by which cis-UCA exerts its biological effect in the skin.
Assuntos
AMP Cíclico/biossíntese , Ácido Urocânico/farmacologia , Agonistas alfa-Adrenérgicos/farmacologia , Alprostadil/farmacologia , Animais , Tartarato de Brimonidina , Células Cultivadas , Colo/efeitos dos fármacos , Cricetinae , Dinoprostona/farmacologia , Epinefrina/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Células HT29 , Histamina/farmacologia , Humanos , Fígado/efeitos dos fármacos , Mesocricetus , Quinoxalinas/farmacologia , Ácido Urocânico/administração & dosagemRESUMO
Trans-urocanic acid (trans-UCA) accumulates in the upper layers of the epidermis and can be isomerized to cis-UCA by UV light irradiation. Cis-urocanic acid possesses immunosuppressive properties that have led to its consideration as one of the initiators of UV-induced immunosuppression. High quantities of cis-UCA persist in human skin for prolonged periods in the summer months. In the present study, mice were injected intradermally with trans-UCA and cis-UCA three times a week for 4 weeks in order to ascertain the long-term effects of the presence of these compounds in the skin. The weight of mice and of their spleens were unaffected by the cis- or trans-UCA treatment. A decrease in thymus weight, accompanied by an increase in lymph node weight, was detected in the cis-UCA-treated mice compared with trans-UCA-treated mice and untreated controls. A net accumulation of lymphocytes and dendritic cells (DC) in lymph nodes was evident following cis-UCA treatment but the percentage of both CD4+ and CD8+ lymphocytes as well as Ia+ DC remained constant among the different treatment groups, indicating that there was no specific migration or proliferation of a particular subset of cells. The in vitro lymphoproliferative response of lymph node cells to the mitogen concanavalin A was significantly sup pressed by cis-UCA treatment. The density of Langerhans cells in the epidermis of the ears was not altered by the chronic cis-UCA treatment. However, chronic cis-UCA treatment did suppress the mixed skin lymphocyte reaction response utilizing epidermal cells from the ears (an uninjected area of skin), indicating a systemic suppression. Compared with trans-UCA treatment, chronic cis-UCA treatment did not cause a significant reduction in the contact hypersensitivity response to oxazolone or the delayed hypersensitivity response to herpes simplex virus. Thus, chronic treatment with cis-UCA led to the suppression of some, but not all, of the immune parameters that are affected by UVB irradiation.
Assuntos
Ácido Urocânico/farmacologia , Animais , Contagem de Linfócito CD4 , Linfócitos T CD8-Positivos , Divisão Celular/efeitos dos fármacos , Dermatite de Contato , Feminino , Hipersensibilidade Tardia , Isomerismo , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Teste de Cultura Mista de Linfócitos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Tamanho do Órgão/efeitos dos fármacos , Fenótipo , Timo/efeitos dos fármacos , Ácido Urocânico/administração & dosagemAssuntos
Fármacos Dermatológicos/uso terapêutico , Psoríase/tratamento farmacológico , Ácido Urocânico/uso terapêutico , Administração Cutânea , Administração Tópica , Adulto , Idoso , Antralina/administração & dosagem , Antralina/uso terapêutico , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Fármacos Dermatológicos/administração & dosagem , Epiderme/efeitos dos fármacos , Humanos , Pessoa de Meia-Idade , Curativos Oclusivos , Pomadas , Índice de Gravidade de Doença , Estereoisomerismo , Alcatrões/uso terapêutico , Ácido Urocânico/administração & dosagemAssuntos
Dermatite Alérgica de Contato/tratamento farmacológico , Imunossupressores/uso terapêutico , Ácido Urocânico/uso terapêutico , Administração Tópica , Alérgenos/administração & dosagem , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/química , Testes do Emplastro , Estereoisomerismo , Ácido Urocânico/administração & dosagem , Ácido Urocânico/químicaRESUMO
A single specific epidermal photoreceptor for the immunosuppressive action of UV radiation has not been defined, although separate evidence is accruing in favour of each of two candidates, trans-urocanic acid and DNA. In Monodelphis domestica, specific photoreactivation repair of UV radiation-induced pyrimidine dimers has been shown to abrogate the suppression of contact hypersensitivity (CHS), thus suggesting that DNA is the target for this impairment. However, the both haired and hairless mice, immunosuppressive effects of UV radiation have been reproduced by the exogenous administration of the UV photoproduct of urocanic acid, cis-urocanic acid. We show here that the epidermis of M. domestica contains urocanic acid, that UV irradiation of the shaved dorsal skin has resulted in an increase in epidermal cis-urocanic acid and that the topical application of a cis-urocanic acid-containing lotion significantly depressed the capacity of Monodelphis to respond to contact sensitisers, in a manner analogous to these responses in the hairless mouse. Therefore in Monodelphis, suppression of CHS by UV irradiation appears to involve both urocanic acid photo-isomerisation and epidermal DNA damage.