RESUMO
BACKGROUND: Increasing evidence suggests photoprotection by oral supplementation with ß-carotene and lycopene. OBJECTIVES: To examine the capacity of lycopene-rich tomato nutrient complex (TNC) and lutein, to protect against ultraviolet (UV)A/B and UVA1 radiation at a molecular level. METHODS: In a placebo-controlled, double-blinded, randomized, crossover study two active treatments containing either TNC or lutein were assessed for their capacity to decrease the expression of UVA1 the radiation-inducible genes HO1, ICAM1 and MMP1. Sixty-five healthy volunteers were allocated to four treatment groups and subjected to a 2-week washout phase, followed by two 12-week treatment phases separated by another 2 weeks of washout. Volunteers started either with active treatment and were then switched to placebo, or vice versa. At the beginning and at the end of each treatment phase skin was irradiated and 24 h later biopsies were taken from untreated, UVA/B- and UVA1-irradiated skin for subsequent reverse transcriptase polymerase chain reaction analysis of gene expression. Moreover, blood samples were taken after the washout and the treatment phases for assessment of carotenoids. RESULTS: TNC completely inhibited UVA1- and UVA/B-induced upregulation of heme-oxygenase 1, intercellular adhesion molecule 1 and matrix metallopeptidase 1 mRNA, no matter the sequence (anova, P < 0·05). In contrast, lutein provided complete protection if it was taken in the first period but showed significantly smaller effects in the second sequence compared with TNC. CONCLUSIONS: Assuming the role of these genes as indicators of oxidative stress, photodermatoses and photoageing, these results might indicate that TNC and lutein could protect against solar radiation-induced health damage.
Assuntos
Carotenoides/administração & dosagem , Luteína/administração & dosagem , Protetores contra Radiação/administração & dosagem , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Administração Oral , Adolescente , Adulto , Análise de Variância , Estudos Cross-Over , Suplementos Nutricionais , Método Duplo-Cego , Feminino , Heme Oxigenase-1/genética , Heme Oxigenase-1/efeitos da radiação , Humanos , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/efeitos da radiação , Licopeno , Solanum lycopersicum , Masculino , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/efeitos da radiação , Pessoa de Meia-Idade , Estresse Oxidativo/efeitos da radiação , Regulação para Cima , Adulto JovemRESUMO
BACKGROUND: The bacterial endotoxin lipopolysaccharide (LPS) represents a prime pathogenic factor of peri-implantitis because of its ability to adhere tenaciously to dental titanium implants. Despite this, the current therapeutic approach to this disease remains based mainly on bacterial decontamination, paying little attention to the neutralization of bioactive bacterial products. The purpose of the present study was to evaluate whether irradiation with low-energy neodymium-doped:yttrium, aluminum, and garnet (Nd:YAG) laser, in addition to the effects on bacterial implant decontamination, was capable of attenuating the LPS-induced inflammatory response. METHODS: RAW 264.7 macrophages or human umbilical vein endothelial cells were cultured on titanium disks coated with Porphyromonas gingivalis LPS, subjected or not to irradiation with the Nd:YAG laser, and examined for the production of inflammatory cytokines and the expression of morphologic and molecular markers of cell activation. RESULTS: Laser irradiation of LPS-coated titanium disks significantly reduced LPS-induced nitric oxide production and cell activation by the macrophages and strongly attenuated intercellular adhesion molecule-1 and vascular cell adhesion molecule expression, as well as interleukin-8 production by the endothelial cells. CONCLUSION: By blunting the LPS-induced inflammatory response, Nd:YAG laser irradiation may be viewed as a promising tool for the therapeutic management of peri-implantitis.
Assuntos
Implantes Dentários/microbiologia , Materiais Dentários , Células Endoteliais/efeitos da radiação , Lasers de Estado Sólido/uso terapêutico , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos da radiação , Porphyromonas gingivalis/fisiologia , Titânio , Animais , Linhagem Celular , Tamanho Celular/efeitos dos fármacos , Tamanho Celular/efeitos da radiação , Células Cultivadas , Citocinas/efeitos dos fármacos , Citocinas/efeitos da radiação , Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/citologia , Imunofluorescência , Células Gigantes/efeitos dos fármacos , Células Gigantes/efeitos da radiação , Humanos , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/efeitos da radiação , Interleucina-8/efeitos dos fármacos , Interleucina-8/efeitos da radiação , Lipopolissacarídeos/efeitos da radiação , Ativação de Macrófagos/efeitos da radiação , Macrófagos/efeitos dos fármacos , Camundongos , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Óxido Nítrico/efeitos da radiação , Doses de Radiação , Veias Umbilicais/citologia , Molécula 1 de Adesão de Célula Vascular/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/efeitos da radiaçãoRESUMO
PURPOSE: In this article, we investigate the role of tumor necrosis factor-alpha (TNF) in the initiation of acute damage to the blood-brain barrier (BBB) and brain tissue following radiotherapy (RT) for CNS tumors. METHODS AND MATERIALS: Intravital microscopy and a closed cranial window technique were used to measure quantitatively BBB permeability to FITC-dextran 4.4-kDa molecules, leukocyte adhesion (Rhodamine-6G) and vessel diameters before and after 20-Gy cranial radiation with and without treatment with anti-TNF. Immunohistochemistry was used to quantify astrogliosis post-RT and immunofluorescence was used to visualize protein expression of TNF and ICAM-1 post-RT. Recombinant TNF (rTNF) was used to elucidate the role of TNF in leukocyte adhesion and vessel diameter. RESULTS: Mice treated with anti-TNF showed significantly lower permeability and leukocyte adhesion at 24 and 48 h post-RT vs. RT-only animals. We observed a significant decrease in arteriole diameters at 48 h post-RT that was inhibited in TNF-treated animals. We also saw a significant increase in activated astrocytes following RT that was significantly lower in the anti-TNF-treated group. In addition, immunofluorescence showed protein expression of TNF and ICAM-1 in the cerebral cortex that was inhibited with anti-TNF treatment. Finally, administration of rTNF induced a decrease in arteriole diameter and a significant increase in leukocyte adhesion in venules and arterioles. CONCLUSIONS: TNF plays a significant role in acute changes in BBB permeability, leukocyte adhesion, arteriole diameter, and astrocyte activation following cranial radiation. Treatment with anti-TNF protects the brain's microvascular network from the acute damage following RT.
Assuntos
Astrócitos/efeitos da radiação , Barreira Hematoencefálica/efeitos da radiação , Irradiação Craniana , Fator de Necrose Tumoral alfa/fisiologia , Animais , Especificidade de Anticorpos , Arteríolas/anatomia & histologia , Arteríolas/efeitos dos fármacos , Arteríolas/efeitos da radiação , Astrócitos/efeitos dos fármacos , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/fisiologia , Encéfalo/irrigação sanguínea , Encéfalo/efeitos da radiação , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Neoplasias do Sistema Nervoso Central/radioterapia , Dextranos/farmacocinética , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/farmacocinética , Imunofluorescência , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Contagem de Leucócitos , Leucócitos/efeitos dos fármacos , Leucócitos/fisiologia , Leucócitos/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Permeabilidade/efeitos dos fármacos , Permeabilidade/efeitos da radiação , Doses de Radiação , Proteínas Recombinantes/farmacologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/farmacologia , Vênulas/anatomia & histologia , Vênulas/efeitos dos fármacos , Vênulas/efeitos da radiaçãoRESUMO
In this study, with the aim of developing a cell-based in vitro photosensitization assay, we examined whether changes of CD86 and CD54 expression on cells of a human monocytic cell line, THP-1, could be used to assess the photosensitizing potential of chemicals. First, we identified suitable conditions of UV-irradiation (irradiation dose; 5.0 J/cm(2), irradiation intensity; 1.7 mW/cm(2)) by investigating the effect of UV-irradiation on CD86 and CD54 expression on untreated or 6-methylcoumarin (a representative photoallergen)-treated THP-1 cells (irradiation method). However, acridine, a representative photo-irritant, augmented CD86 and CD54 expression on THP-1 cells, apparently via induction of reactive oxygen species (ROS). In order to abolish the effect of ROS, we examined CD86 and CD54 expression on THP-1 cells treated with pre-irradiated chemicals (pre-irradiation method). We found that UV-irradiated photoallergens, but not photo-irritants, enhanced CD86 and/or CD54 expression on the THP-1 cells. Finally, based on the results of irradiation, non-irradiation, and pre-irradiation with 18 test chemicals, we built a decision tree, which allows us to distinguish between photoallergens and photo-irritants. We suggest that this system may be useful for in vitro evaluation of the photoallergic potential of chemicals.
Assuntos
Monócitos/efeitos dos fármacos , Fármacos Fotossensibilizantes/toxicidade , Testes de Toxicidade/métodos , Raios Ultravioleta , Acridinas/toxicidade , Alérgenos/toxicidade , Antígeno B7-2/genética , Antígeno B7-2/efeitos da radiação , Linhagem Celular , Cumarínicos/toxicidade , Árvores de Decisões , Relação Dose-Resposta à Radiação , Humanos , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/efeitos da radiação , Irritantes/toxicidade , Monócitos/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismoRESUMO
The expression of adhesion molecules Intercellular adhesion molecule-1 (ICAM-1) and Vascular cell adhesion molecule-1 (VCAM-1) is increased in lesional and in non-lesional skin of psoriatic patients, and play role in pathogenesis of the disease. PUVA and UVB therapy are important treatments of psoriasis vulgaris. It has been demonstrated that UVA and UVB therapies reduce expression of these molecules. In this investigation, phototherapy was used to treat psoriatic patients. The expression of these molecules was examined by immunohistochemical method in lesional and non-lesional skin of 10 patients with psoriasis vulgaris before and after treatment. Results showed increased expression of ICAM-1 molecules in keratinocytes, in perivascular infiltrate--lymphocytes, and in endothelial cells. The expression of VCAM-1 molecules was also increased, although with less intensity then ICAM-1. After therapy, the expression of the adhesion molecules decreased together with a marked improvement of the disease. In conclusion, study demonstrated that phototherapy improves psoriasis vulgaris probably through mechanisms acting on the adhesions molecules. Adverse reactions due to intense or long lasting UVA (PUVA) and UVB therapies are immunosuppression and damage of DNA which can lead to development of non-melanocytic skin tumors like basal cell carcinoma and squamous cell carcinoma, as well as melanoma.
Assuntos
Molécula 1 de Adesão Intercelular/efeitos da radiação , Psoríase/metabolismo , Psoríase/terapia , Terapia Ultravioleta , Molécula 1 de Adesão de Célula Vascular/efeitos da radiação , Células Cultivadas , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Terapia PUVA , Psoríase/patologia , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
The ionizing radiation used in cancer therapy frequently produces damage to normal tissues and induces complex responses, including inflammation. The upregulation of the intercellular adhesion molecule-1 (ICAM-1) in response to numerous inducing factors is associated with inflammation. Therefore, this study examined the molecular mechanisms responsible for ICAM-1 expression induced by gamma-irradiation (gammaIR). ICAM-1 mRNA and cell surface expression were induced in A549 human lung epithelial cells after exposing them to gammaIR. Catalase expression and activity were also increased in gammaIR-treated cells. Treatment of the gammaIR-treated cells with catalase resulted in a significant increase in the ICAM-1 cell surface expression level. The catalase inhibitor 3-amino-1,2,4-triazole (AT) reduced the level of ICAM-1. Electrophoretic mobility shift assay (EMSA) analysis showed that activating protein 1 (AP-1) was activated by gammaIR, whereas NF-kappaB was not. Specific Jun N-terminal kinase (JNK) inhibition attenuated the upregulation of gammaIR stimulated ICAM-1. Western blot analysis revealed a marked elevation in activation of JNK. In addition, pretreatment with AT resulted in a decrease in the level of JNK phosphorylation and AP-1 activation. Overall, data suggest that induction of ICAM-1 expression by gammaIR is associated with catalase. Furthermore, catalase, JNKs, and AP-1 activation induce ICAM-1 upregulation through a sequential process.
Assuntos
Raios gama/efeitos adversos , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Catalase/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Ativação Enzimática , Humanos , Inflamação , MAP Quinase Quinase 4/metabolismo , RNA Mensageiro/análise , Lesões por Radiação/fisiopatologia , Fator de Transcrição AP-1/metabolismo , Regulação para CimaRESUMO
Hapten sensitization through UV-exposed skin induces systemic immune suppression, which is experimentally demonstrated by inhibition of contact hypersensitivity (CHS). Although this UV-induced effect has been shown to be mediated by inhibition of the afferent phase of the CHS, the UV effects on the efferent (elicitation) phase remain unknown. In this study, UV effects on endothelial ICAM-1 expression at elicitation sites were first examined. Mice were sensitized by hapten application onto UV-exposed back skin, and ears were challenged 5 days later. ICAM-1 up-regulation at nonirradiated elicitation sites following hapten challenge was eliminated by UV exposure on sensitization sites distant from elicitation sites. To assess whether loss of the ICAM-1 up-regulation at elicitation sites contributed to UV-induced immunosuppression, we examined CHS responses in UV-exposed ICAM-1-deficient (ICAM-1(-/-)) mice that genetically lacked the ICAM-1 up-regulation. ICAM-1(-/-) mice exhibited reduced CHS responses without UV exposure, but UV exposure did not further reduce CHS responses in ICAM-1(-/-) mice. Furthermore, ICAM-1 deficiency did not affect the afferent limb, because ICAM-1(-/-) mice had normal generation of hapten-specific suppressor and effector T cells. This UV-induced immunosuppression was associated with a lack of TNF-alpha production after Ag challenge at elicitation sites. Local TNF-alpha injection before elicitation abrogated the UV-induced CHS inhibition with increased endothelial ICAM-1 expression. TNF-alpha production at elicitation sites was down-regulated by IL-10, a possible mediator produced by hapten-specific suppressor T cells that are generated by UV exposure. These results indicate that UV exposure inhibits CHS by abrogating up-regulation of endothelial ICAM-1 expression after Ag challenge at elicitation sites.
Assuntos
Dermatite de Contato/metabolismo , Dermatite de Contato/prevenção & controle , Molécula 1 de Adesão Intercelular/biossíntese , Pele/efeitos da radiação , Raios Ultravioleta , Regulação para Cima/efeitos da radiação , Animais , Anticorpos Monoclonais/administração & dosagem , Citotoxicidade Imunológica/genética , Citotoxicidade Imunológica/efeitos da radiação , Dermatite de Contato/imunologia , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Endotélio Vascular/efeitos da radiação , Terapia de Imunossupressão , Injeções Intradérmicas , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/fisiologia , Molécula 1 de Adesão Intercelular/efeitos da radiação , Interleucina-10/imunologia , Luz , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pele/imunologia , Pele/metabolismo , Baço/citologia , Baço/imunologia , Baço/efeitos da radiação , Baço/transplante , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/efeitos da radiação , Fator de Necrose Tumoral alfa/administração & dosagem , Regulação para Cima/genética , Regulação para Cima/imunologiaRESUMO
OBJECTIVE: Alterations in multiple functions of the microvasculature occur in response to gamma irradiation and are thought to contribute to radiation-induced end organ damage by inducing inflammatory responses, particularly leukocyte infiltration into the affected area. Endothelial cell adhesion molecules (ECAMs) mediate leukocyte adhesion and migration. Here, we validate a method to study the effect of Leksell gamma knife stereotactic radiosurgery on the expression of ECAMs on human cerebral endothelium at 0, 24, 48, and 72 hours after irradiation. METHODS: A human brain endothelial cell line (IHEC) was cultured on 12-mm coverslips and subjected to 50 Gy of collimated gamma irradiation with the Leksell gamma knife (Elekta Instruments, Inc., Atlanta, GA). Lactate dehydrogenase release was measured at 24, 48, and 72 hours after irradiation and caspase-3 at 24, 48, 72, 96, and 120 hours. ECAM expression was measured at postirradiation intervals of 0, 24, 48, and 72 hours by cell enzyme-linked immunoabsorbent assay. We used a cell irradiator composed of two chambers. The upper chamber holds the coverslips firmly in place while they are immersed in media. The lower chamber is connected to a peristaltic pump, which pumps water into the chamber and maintains the media in the upper chamber at 37 degrees C through convection. RESULTS: None of the ECAMs tested was significantly elevated compared with the control basally. Twenty-four hours after irradiation, intercellular adhesion molecule 1 was significantly elevated on brain endothelial cells but there was no significant elevation of E-selectin. Vascular cell adhesion molecule 1 was increased slightly but not significantly and decreased at 48 hours. At 72 hours, E-selectin expression was significantly increased; intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 were not altered relative to sham controls. CONCLUSION: Increased ECAM expression and lactate dehydrogenase release support the idea that the cerebral microvasculature undergoes an inflammatory response after Leksell gamma knife stereotactic radiosurgery.
Assuntos
Encéfalo/efeitos da radiação , Selectina E/análise , Selectina E/efeitos da radiação , Endotélio/efeitos da radiação , Raios gama , Molécula 1 de Adesão Intercelular/análise , Molécula 1 de Adesão Intercelular/efeitos da radiação , Radiocirurgia , Apoptose/efeitos da radiação , Caspase 3 , Caspases/análise , Caspases/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Circulação Cerebrovascular/efeitos da radiação , Humanos , Técnicas In Vitro , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/efeitos da radiação , Microcirculação/efeitos da radiação , Fatores de TempoRESUMO
CD8(+) CTL play important roles against malignancy in both active and passive immunotherapy. Nonetheless, the success of antitumor CTL responses may be improved by additional therapeutic modalities. Radiotherapy, which has a long-standing use in treating neoplastic disease, has been found to induce unique biologic alterations in cancer cells affecting Fas gene expression, which, consequently, may influence the overall lytic efficiency of CTL. Here, in a mouse adenocarcinoma cell model, we examined whether exposure of these tumor cells to sublethal doses of irradiation 1) enhances Fas expression, leading to more efficient CTL killing via Fas-dependent mechanisms in vitro; and 2) improves antitumor activity in vivo by adoptive transfer of these Ag-specific CTL. Treatment of carcinoembryonic Ag-expressing MC38 adenocarcinoma cells with irradiation (20 Gy) in vitro enhanced Fas expression at molecular, phenotypic, and functional levels. Furthermore, irradiation sensitized these targets to Ag-specific CTL killing via the Fas/Fas ligand pathway. We examined the effect of localized irradiation of s.c. growing tumors on the efficiency of CTL adoptive immunotherapy. Irradiation caused up-regulation of Fas by these tumor cells in situ, based on immunohistochemistry. Moreover, localized irradiation of the tumor significantly potentiated tumor rejection by these carcinoembryonic Ag-specific CTL. Overall, these results showed for the first time that 1) regulation of the Fas pathway in tumor cells by irradiation plays an important role in their sensitization to Ag-specific CTL; and 2) a combination regimen of tumor-targeted irradiation and CTL promotes more effective antitumor responses in vivo, which may have implications for the combination of immunotherapy and radiation therapy.
Assuntos
Neoplasias do Colo/imunologia , Neoplasias do Colo/radioterapia , Citotoxicidade Imunológica/efeitos da radiação , Imunoterapia Adotiva/métodos , Linfócitos T Citotóxicos/efeitos da radiação , Linfócitos T Citotóxicos/transplante , Regulação para Cima/imunologia , Receptor fas/biossíntese , Animais , Anticorpos Bloqueadores/farmacologia , Anticorpos Monoclonais/farmacologia , Apoptose/imunologia , Apoptose/efeitos da radiação , Divisão Celular/imunologia , Divisão Celular/efeitos da radiação , Neoplasias do Colo/patologia , Relação Dose-Resposta à Radiação , Epitopos de Linfócito T/imunologia , Epitopos de Linfócito T/efeitos da radiação , Proteína Ligante Fas , Feminino , Humanos , Molécula 1 de Adesão Intercelular/biossíntese , Molécula 1 de Adesão Intercelular/efeitos da radiação , Ligantes , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Transplante de Neoplasias , Linfócitos T Citotóxicos/imunologia , Fatores de Tempo , Células Tumorais Cultivadas/imunologia , Células Tumorais Cultivadas/patologia , Células Tumorais Cultivadas/efeitos da radiação , Células Tumorais Cultivadas/transplante , Regulação para Cima/efeitos da radiação , Receptor fas/fisiologia , Receptor fas/efeitos da radiaçãoRESUMO
PURPOSE: To investigate the effects of short-term administration of dexamethasone (DEX) on radiation-induced responses in the mouse lung, focusing on expression of pro-inflammatory cytokine and related genes. METHODS AND MATERIALS: At indicated times after thoracic irradiation and/or drug treatment, mRNA expression levels of cytokines (mTNF-alpha, mIL-1 alpha, mIL-1 beta, mIL-2, mIL-3, mIL-4, mIL-5, mIL-6, mIFN-gamma) and related genes in the lungs of C3H/HeN mice were measured by RNase protection assay. RESULTS: Radiation-induced pro-inflammatory cytokine mRNA expression levels in lung peak at 6 h after thoracic irradiation. DEX (5 mg/kg) suppresses both basal cytokine mRNA levels and this early response when given immediately after irradiation. However, by 24 h, in mice treated with DEX alone or DEX plus radiation, there was a strong rebound effect that lasted up to 3 days. Modification of the early radiation-induced response by DEX did not change the second wave of cytokine gene expression in the lung that occurs at 1 to 2 weeks, suggesting that early cytokine gene induction might not determine subsequent molecular events. A single dose of DEX attenuated, but did not completely suppress, increases in cytokine mRNA levels induced by lipopolysaccharide (2.5 mg/kg) treatment, but, unlike with radiation, no significant rebound effect was seen. Five days of dexamethasone treatment in the pneumonitic phase also inhibited pro-inflammatory cytokine gene expression and, again, there was a rebound effect after withdrawal of the drug. CONCLUSIONS: Our findings suggest that short-term use of dexamethasone can temporarily suppress radiation-induced pro-inflammatory cytokine gene expression, but there may be a rebound after drug withdrawal and the drug does little to change the essence and course of the pneumonitic process.
Assuntos
Anti-Inflamatórios/farmacologia , Dexametasona/farmacologia , Interleucinas/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/efeitos da radiação , Fator de Necrose Tumoral alfa/metabolismo , Animais , Anti-Inflamatórios/administração & dosagem , Dexametasona/administração & dosagem , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Interleucinas/efeitos da radiação , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C3H , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , RNA Mensageiro/efeitos da radiação , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/efeitos da radiaçãoRESUMO
BACKGROUND: Recent studies have demonstrated that a tanning lamp emitting predominantly ultraviolet (UV) A induces significant yields of the type of potentially mutagenic DNA damage that are associated with the onset of skin cancer (i.e. cyclobutane pyrimidine dimers). UV-induced immunosuppression is also an important event leading to skin cancer. OBJECTIVES: To the modulation of key immunological molecules following exposure to a broad-spectrum UVB lamp and a predominantly UVA-emitting tanning lamp using model in vitro systems. METHODS: We compared secretion and mRNA expression of interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha in normal human epidermal keratinocytes, and interferon (IFN)-gamma-induced intracellular adhesion molecule (ICAM)-1 in normal human fibroblasts irradiated in vitro with a broad-spectrum UVB lamp or with a Philips 'Performance' tanning lamp. RESULTS: With broad-spectrum UVB irradiation, upregulation of IL-6 and TNF-alpha mRNA was detected 6 h after irradiation, and a dose-dependent increase of cytokines in the supernatants of irradiated cells was found 24 h after irradiation. In contrast, there was no cytokine secretion and little evidence for mRNA upregulation following exposure to a tanning lamp. When cells were exposed first to broad-spectrum UVB, then the tanning lamp, UVB-induced cytokine secretion was inhibited, although mRNA levels were upregulated to a level close to that observed with UVB alone. By using a Schott WG 320 nm filter to attenuate the level of UVB relative to UVA emitted by the tanning lamp, the inhibition of cytokine secretion was shown to be associated with UVA exposure. Both UV sources inhibited IFN-gamma-induced ICAM-1 mRNA expression in a dose-dependent fashion. By using a Schott WG 335 nm filter, inhibition of ICAM-1 mRNA expression by the tanning lamp was shown to be associated with UVB exposure. CONCLUSIONS: These results suggest that UV sources emitting different levels of UVA and UVB have differential effects on the modulation of different immunoregulatory molecules, and indicate that there are potential interactions between these wavelengths.
Assuntos
Molécula 1 de Adesão Intercelular/efeitos da radiação , Interleucina-6/efeitos da radiação , Pele/efeitos da radiação , Fator de Necrose Tumoral alfa/efeitos da radiação , Raios Ultravioleta , Indústria da Beleza , Técnicas de Cultura de Células , Relação Dose-Resposta à Radiação , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Tolerância Imunológica/efeitos da radiação , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Queratinócitos/imunologia , Queratinócitos/efeitos da radiação , Dímeros de Pirimidina/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Pele/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos da radiaçãoRESUMO
Cell adhesion molecules (CAMs) are proteins which anchor cells to each other and to the extracellular matrix (ECM), but whose functions also include signal transduction, differentiation, and apoptosis. We are testing a hypothesis that particle radiations modulate CAM expression and this contributes to radiation-induced lens opacification. We observed dose-dependent changes in the expression of beta 1-integrin and ICAM-1 in exponentially-growing and confluent cells of a differentiating human lens epithelial cell model after exposure to particle beams. Human lens epithelial (HLE) cells, less than 10 passages after their initial culture from fetal tissue, were grown on bovine corneal endothelial cell-derived ECM in medium containing 15% fetal bovine serum and supplemented with 5 ng/ml basic fibroblast growth factor (FGF-2). Multiple cell populations at three different stages of differentiation were prepared for experiment: cells in exponential growth, and cells at 5 and 10 days post-confluence. The differentiation status of cells was characterized morphologically by digital image analysis, and biochemically by Western blotting using lens epithelial and fiber cell-specific markers. Cultures were irradiated with single doses (4, 8 or 12 Gy) of 55 MeV protons and, along with unirradiated control samples, were fixed using -20 degrees C methanol at 6 hours after exposure. Replicate experiments and similar experiments with helium ions are in progress. The intracellular localization of beta 1-integrin and ICAM-1 was detected by immunofluorescence using monoclonal antibodies specific for each CAM. Cells known to express each CAM were also processed as positive controls. Both exponentially-growing and confluent, differentiating cells demonstrated a dramatic proton-dose-dependent modulation (upregulation for exponential cells, downregulation for confluent cells) and a change in the intracellular distribution of the beta 1-integrin, compared to unirradiated controls. In contrast, there was a dose-dependent increase in ICAM-1 immunofluorescence in confluent, but not exponentially-growing cells. These results suggest that proton irradiation downregulates beta 1-integrin and upregulates ICAM-1, potentially contributing to cell death or to aberrant differentiation via modulation of anchorage and/or signal transduction functions. Quantification of the expression levels of the CAMs by Western analysis is in progress.
Assuntos
Integrina beta1/metabolismo , Integrina beta1/efeitos da radiação , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Cristalino/efeitos da radiação , Prótons , Adesão Celular/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Células Cultivadas , Relação Dose-Resposta à Radiação , Células Epiteliais/efeitos da radiação , Fator 2 de Crescimento de Fibroblastos/farmacologia , Humanos , Integrina beta1/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/efeitos dos fármacosRESUMO
PURPOSE: The purpose of our investigation was to describe the dose- and time-dependent histomorphologic alterations of the irradiated tissue, the composition of the infiltrate, and the expression patterns of various adhesion molecules. METHODS AND MATERIALS: We analyzed immunohistochemically alterations in oral mucosa in 13 head and neck cancer patients before radiotherapy and with 30 Gy and 60 Gy. All had oral mucosa irradiation, with a final dose of 60 Gy using conventional fractionation. Snap-frozen specimens were stained using the indirect immunperoxidase technique. Histomorphology was studied in paraffin-embedded sections. In addition, we determined the clinical degree of oral mucositis. RESULTS: Histomorphologic evaluation showed no vascular damage. Irradiation caused a steep increase of beta2-integrin-bearing cells (p < 0.01), whereas the percentage of beta1-integrin-positive cells remained at low levels. Additionally we found an increase in the expression of endothelial intercellular adhesion molecule-1 (ICAM-1) (p < 0.01) and E-selectin (p < 0.05), while endothelial vascular cell adhesion molecule-1 (VCAM-1) expression remained at very low levels. CONCLUSION: Our findings indicate that in radiation-induced oral mucositis there is no marked vascular damage until the end of radiotherapy. For recruitment of leukocytes, beta2 is more involved than beta1. Pharmaceuticals that block leukocyte adhesion to E-selectin or ICAM-1 may prevent radiation-mediated inflammation in oral mucosa.
Assuntos
Moléculas de Adesão Celular/efeitos da radiação , Neoplasias de Cabeça e Pescoço/radioterapia , Lesões por Radiação/metabolismo , Estomatite/metabolismo , Idoso , Moléculas de Adesão Celular/metabolismo , Relação Dose-Resposta à Radiação , Selectina E/metabolismo , Selectina E/efeitos da radiação , Humanos , Integrina alfa4beta1 , Integrinas/metabolismo , Integrinas/efeitos da radiação , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Antígeno-1 Associado à Função Linfocitária/metabolismo , Antígeno-1 Associado à Função Linfocitária/efeitos da radiação , Antígeno de Macrófago 1/metabolismo , Antígeno de Macrófago 1/efeitos da radiação , Pessoa de Meia-Idade , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia , Mucosa Bucal/efeitos da radiação , Estudos Prospectivos , Lesões por Radiação/patologia , Receptores de Retorno de Linfócitos/metabolismo , Receptores de Retorno de Linfócitos/efeitos da radiação , Estomatite/etiologia , Estomatite/patologia , Fatores de Tempo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Molécula 1 de Adesão de Célula Vascular/efeitos da radiaçãoRESUMO
OBJECTIVES: Radiation treatment is one of the most standardized and effective modalities for contemporary cervical cancer therapy. In addition, the radiation-potentiating effects of retinoic acid have been recently described. In order to investigate whether enhanced immunogenicity might be responsible for such potentiation, we have evaluated the effects of retinoic acid combined with high doses of gamma-irradiation on the expression of major histocompatibility complex (MHC) Class I and II and intercellular adhesion molecule-1 (ICAM-1) in human cervical carcinoma cell lines. METHODS: The expression of surface antigens (MHC Class I and II and ICAM-1) was evaluated by FACS analysis in untreated control cells and in cells following their exposure to retinoic acid, high doses of gamma-irradiation (i.e., 5000 and 10,000 cGy), or the combination of the two procedures. RESULTS: HT-3 and SiHa cervical cancer cells expressed variable levels of MHC Class I and ICAM-1 antigens while Class II surface antigens were not detectable. Exposure to either 5000 or 10,000 cGy completely inhibited cell replication in both cell lines and significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation. Irradiation was unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). In a similar fashion, retinoic acid was also able to significantly increase the expression of MHC Class I and ICAM-1 antigens when compared to untreated tumor cells but was not able to induce the expression of HLA Class II surface antigens. Exposure to the combination of radiation plus retinoic acid significantly upregulated HLA Class I and ICAM-1 molecules in an additive manner when compared to the levels obtainable with the exposure to radiation or retinoic acid alone. CONCLUSIONS: These data indicate that the combination of these two treatments could induce an additive effect on the expression of immunologically important surface antigens in human cervical cancer cells. These findings, together with the powerful antiproliferative effect of retinoids and irradiation on tumor cells, suggest that the combined regimen may be a promising and more effective combination for the treatment of cervical cancer.
Assuntos
Antineoplásicos/farmacologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Antígenos de Histocompatibilidade Classe I/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Tretinoína/farmacologia , Neoplasias do Colo do Útero/imunologia , Feminino , Antígenos de Histocompatibilidade Classe I/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe I/efeitos da radiação , Antígenos de Histocompatibilidade Classe II/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe II/efeitos da radiação , Humanos , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/efeitos da radiação , Doses de Radiação , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/efeitos da radiaçãoRESUMO
PURPOSE: To investigate the changes in antigenic expression of intercellular adhesion molecule-1 (ICAM-1) caused by ionizing radiation of cultured human adenocarcinoma cells. METHODS AND MATERIALS: Human colonic BM314 and gastric MKN45 adenocarcinoma cells were irradiated to investigate the expression of ICAM-1 on the cell membrane and in the supernatant. In addition, the ICAM-1 gene expression (mRNA) was analyzed using a ICAM-1 cDNA as a probe. RESULTS: The expression of ICAM-1 on the membrane was found to increase by irradiation. This effect was also observed in the supernatant. In addition, the irradiated cell population showed slight, but clear increases in ICAM-1 mRNA expression. CONCLUSIONS: These results show that the enhancement of expression of ICAM-1 by radiation takes place at the ICAM-1 gene expression (mRNA) level. The results suggest that the low dose radiation may be useful for accumulating LFA-1 positive cytotoxic T lymphocytes (CTL) at the local tumor tissue, by which tumor cells may be attacked.
Assuntos
Antígenos de Neoplasias/efeitos da radiação , Molécula 1 de Adesão Intercelular/efeitos da radiação , Adenocarcinoma/imunologia , Antígenos de Neoplasias/efeitos dos fármacos , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Antineoplásicos/farmacologia , Citometria de Fluxo , Humanos , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Interferons/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , RNA Mensageiro/efeitos da radiação , Doses de Radiação , Neoplasias Gástricas/imunologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/efeitos da radiaçãoRESUMO
PURPOSE: We initiated studies to analyze the effects of high doses of gamma irradiation on the surface antigen expression of MHC Class I, Class II, and ICAM-1 on human cervical carcinoma cell lines. METHODS AND MATERIALS: The expression of surface antigens (MHC Class I, Class II, and ICAM-1) was evaluated by FACS analysis on two cervical cell lines at different time points, following their exposure to high doses of gamma irradiation (i.e., 25.00, 50.00, and 100.00 Gy). RESULTS: The CaSki and SiHa cervical cancer cells we analyzed in this study expressed variable levels of MHC Class I and ICAM-1 antigens, while Class II surface antigens were not detectable. Whereas irradiation doses of 25.00 Gy were not sufficient to totally block cell replication in both cell lines, exposure to 50.00 or 100.00 Gy was able to completely inhibit cell replication. Range doses from 25.00 to 100.00 Gy significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation but were unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). Importantly, such upregulation was shown to be dose dependent, with higher radiation doses associated with increased antigen expression. Moreover, when the kinetic of this upregulation was studied after 2 and 6 days after irradiation, it was shown to be persistent and lasted until all the cells died. CONCLUSIONS: These findings may partially explain the increased immunogenicity of tumor cells following irradiation and may suggest enhanced immune recognition in tumor tissue in patients receiving radiation therapy.
Assuntos
Antígenos de Neoplasias/efeitos da radiação , Fracionamento da Dose de Radiação , Antígenos de Histocompatibilidade Classe II/efeitos da radiação , Antígenos de Histocompatibilidade Classe I/efeitos da radiação , Molécula 1 de Adesão Intercelular/efeitos da radiação , Neoplasias do Colo do Útero/imunologia , Antígenos de Neoplasias/metabolismo , Feminino , Antígenos de Histocompatibilidade Classe I/metabolismo , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Células Tumorais Cultivadas/imunologia , Células Tumorais Cultivadas/efeitos da radiação , Regulação para Cima , Neoplasias do Colo do Útero/radioterapiaRESUMO
Inflammatory cell infiltration of the lung is a predominant histopathological change that occurs during radiation pneumonitis. Emigration of inflammatory cells from the circulation requires the interaction between cell adhesion molecules on the vascular endothelium and molecules on the surface of leukocytes. We studied the immunohistochemical pattern of expression of cell adhesion molecules in lungs from mice treated with thoracic irradiation. After X-irradiation, the endothelial leukocyte adhesion molecule 1 (ELAM-1; E-selectin) was primarily expressed in the pulmonary endothelium of larger vessels and minimally in the microvascular endothelium. Conversely, the intercellular adhesion molecule 1 (ICAM-1; CD54) was expressed in the pulmonary capillary endothelium and minimally in the endothelium of larger vessels. Radiation-mediated E-selectin expression was first observed at 6 h, whereas ICAM-1 expression initially increased at 24 h after irradiation. ICAM-1 and E-selectin expression persisted for several days. P-selectin is constitutively expressed in Weibel-Palade bodies in the endothelium, which moved to the vascular lumen within 30 min after irradiation. P-selectin was not detected in the pulmonary endothelium at 6 h after irradiation. The radiation dose required for increased cell adhesion molecule expression within the pulmonary vascular endothelium was 2 Gy, and expression increased in a dose-dependent manner. These data demonstrate that ICAM-1 and E-selectin expression is increased in the pulmonary endothelium following thoracic irradiation. The pattern of expression of E-selectin, P-selectin, and ICAM-1 is distinct from one another.
Assuntos
Selectina E/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Pulmão/metabolismo , Pulmão/efeitos da radiação , Selectina-P/metabolismo , Animais , Relação Dose-Resposta à Radiação , Selectina E/imunologia , Selectina E/efeitos da radiação , Endotélio/metabolismo , Endotélio Vascular/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/efeitos da radiação , Camundongos , Camundongos Endogâmicos C3H , Selectina-P/imunologia , Selectina-P/efeitos da radiaçãoRESUMO
UVA radiation is the major component of the UV solar spectrum that reaches the earth, and the therapeutic application of UVA radiation is increasing in medicine. Analysis of the cellular effects of UVA radiation has revealed that exposure of human cells to UVA radiation at physiological doses leads to increased gene expression and that this UVA response is primarily mediated through the generation of singlet oxygen. In this study, the mechanisms by which UVA radiation induces transcriptional activation of the human intercellular adhesion molecule 1 (ICAM-1) were examined. UVA radiation was capable of inducing activation of the human ICAM-1 promoter and increasing ICAM-1 mRNA and protein expression. These UVA radiation effects were inhibited by singlet oxygen quenchers, augmented by enhancement of singlet oxygen life-time, and mimicked in unirradiated cells by a singlet oxygen-generating system. UVA radiation as well as singlet oxygen-induced ICAM-1 promoter activation required activation of the transcription factor AP-2. Accordingly, both stimuli activated AP-2, and deletion of the putative AP-2-binding site abrogated ICAM-1 promoter activation in this system. This study identified the AP-2 site as the UVA radiation- and singlet oxygen-responsive element of the human ICAM-1 gene. The capacity of UVA radiation and/or singlet oxygen to induce human gene expression through activation of AP-2 indicates a previously unrecognized role of this transcription factor in the mammalian stress response.
Assuntos
Proteínas de Ligação a DNA/genética , Molécula 1 de Adesão Intercelular/efeitos da radiação , Queratinócitos/efeitos da radiação , Oxigênio/metabolismo , Fatores de Transcrição/genética , Ativação Transcricional/efeitos da radiação , Linhagem Celular , Proteínas de Ligação a DNA/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Queratinócitos/metabolismo , Regiões Promotoras Genéticas/genética , Fator de Transcrição AP-2 , Fatores de Transcrição/metabolismo , Raios UltravioletaRESUMO
We have assessed the pattern of dermal endothelial adhesion molecule expression following broadband UVB irradiation in vivo and in vitro. Skin biopsies were taken from 4 human volunteers at baseline and at 4, 8 and 24 h post-irradiation with 2.5 minimal erythema doses of UVB. Sections were stained immunohistochemically for E-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1). CD31 and neutrophil elastase. The effect of direct UVB irradiation on E-selectin, ICAM-1 and VCAM-1 was examined in a human dermal microvascular endothelial cell line, HMEC-1. Cultured HMEC-1 were irradiated with 2.5-40 mJ/cm2 of UVB, and assessed for adhesion molecule expression by immunofluorescence microscopy and fluorescence-activated cell sorter analysis. In vivo, E-selectin was minimally expressed on EC at baseline and was induced by 4 h following irradiation, P < 0.01. ICAM-1 was moderately expressed at baseline and appeared mildly induced at 24 h, although this did not reach statistical significance. VCAM-1 was weakly expressed in unirradiated skin while CD31 was moderately expressed, but neither was induced by UVB irradiation. A significant neutrophilic infiltrate appeared by 8 h and was maximal at 24 h, P < 0.05. Neutrophil infiltration correlated with E-selectin expression, r = 0.96. In HMEC-1, ICAM-1 was upregulated at 24 h post-irradiation, with an increase in mean channel fluorescence from 100% at baseline to 145 (SD12)% at 24 h, P < 0.05. No change was seen in expression of E-selectin, VCAM-1 or CD31. These studies support the involvement of endothelial adhesion molecules E-selectin and ICAM-1 in UVB-induced inflammation. Whereas ICAM-1 is upregulated by direct irradiation of endothelial cells, E-selectin stimulation appears to be an indirect effect.
Assuntos
Moléculas de Adesão Celular/metabolismo , Pele/metabolismo , Pele/efeitos da radiação , Raios Ultravioleta , Adulto , Moléculas de Adesão Celular/efeitos da radiação , Linhagem Celular , Selectina E/metabolismo , Selectina E/efeitos da radiação , Endotélio/metabolismo , Endotélio Vascular/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/metabolismo , Molécula 1 de Adesão Intercelular/efeitos da radiação , Masculino , Microcirculação/metabolismo , Microscopia de Fluorescência , Pessoa de Meia-Idade , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Pele/irrigação sanguínea , Regulação para Cima , Molécula 1 de Adesão de Célula Vascular/metabolismo , Molécula 1 de Adesão de Célula Vascular/efeitos da radiaçãoRESUMO
PURPOSE: To investigate the in vivo acute phase molecular response of the brain to ionizing radiation. METHODS AND MATERIALS: C3Hf/Sed/Kam mice were given midbrain or whole-body irradiation. Cerebral expression of interleukins (IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6), interferon (IFN-gamma), tumor necrosis factors (TNF-alpha and TNF-beta), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthetase (iNOS), von Willebrand factor (vWF), alpha 1-antichymotrypsin (EB22/5.3), and glial fibrillary acidic protein (GFAP) was measured at various times after various radiation doses by ribonuclease (RNase) protection assay. The effects of dexamethasone or pentoxifylline treatment of mice on radiation-induced gene expression were also examined. RESULTS: Levels of TNF-alpha, IL-1 beta, ICAM-1, EB22/5.3 and to a lesser extent IL-1 alpha and GFAP, messenger RNA were increased in the brain after irradiation, whether the dose was delivered to the whole body or only to the midbrain. Responses were radiation dose dependent, but were not found below 7 Gy; the exception being ICAM-1, which was increased by doses as low as 2 Gy. Most responses were rapid, peaking within 4-8 h, but antichymotrypsin and GFAP responses were delayed and still elevated at 24 h, by which time the others had subsided. Pretreatment of mice with dexamethasone or pentoxifylline suppressed radiation-induced gene expression, either partially or completely. Dexamethasone was more inhibitory than pentoxifylline at the doses chosen. CONCLUSIONS: The initial response of the brain to irradiation involves expression of inflammatory gene products, which are probably responsible for clinically observed early symptoms of brain radiotherapy. This mechanism explains the beneficial effects of the clinical use of steroids in such circumstances.