RESUMO
Exposure to 2.45 GHz electromagnetic radiation (EMR) emitted from commonly used devices has been reported to induce oxidative stress in several experimental models. Our study aims to evaluate the efficacy of sulforaphane, a well-known natural product, in preventing radiation-induced toxic effects caused by a 24 h exposure of SH-SY5Y neuronal-like cells and peripheral blood mononuclear cells (PBMCs) to 2.45 GHz EMR. Cells were exposed to radiation for 24 h in the presence or absence of sulforaphane at different concentrations (5-10-25 µg/mL). Cell viability, mitochondrial activity alterations, the transcription and protein levels of redox markers, and apoptosis-related genes were investigated. Our data showed a reduction in cell viability of both neuronal-like cells and PBMCs caused by EMR exposure and a protective effect of 5 µg/mL sulforaphane. The lowest sulforaphane concentration decreased ROS production and increased the Mitochondrial Transmembrane Potential (Δψm) and the NAD+/NADH ratio, which were altered by radiation exposure. Sulforaphane at higher concentrations displayed harmful effects. The hormetic behavior of sulforaphane was also evident after evaluating the expression of genes coding for Nrf2, SOD2, and changes in apoptosis markers. Our study underlined the vulnerability of neuronal-like cells to mitochondrial dysfunction and oxidative stress and the possibility of mitigating these effects by supplementation with sulforaphane. To our knowledge, there are no previous studies about the effects of SFN on these cells when exposed to 2.45 GHz electromagnetic radiation.
Assuntos
Radiação Eletromagnética , Isotiocianatos , Leucócitos Mononucleares , Potencial da Membrana Mitocondrial , Neurônios , Estresse Oxidativo , Sulfóxidos , Isotiocianatos/farmacologia , Humanos , Sulfóxidos/farmacologia , Leucócitos Mononucleares/efeitos da radiação , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Neurônios/efeitos da radiação , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/genética , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/efeitos da radiação , Mitocôndrias/metabolismo , Linhagem Celular TumoralRESUMO
Small fractions of patients suffer from radiotherapy late severe adverse events (AEs Grade ≥ 3), which are usually irreversible and badly affect their quality of life. A novel functional DNA repair assay characterizing several steps of double-strand break (DSB) repair mechanisms was used. DNA repair activities of peripheral blood mononuclear cells were monitored for 1 week using NEXT-SPOT assay in 177 breast and prostate cancer patients. Only seven patients had Grade ≥ 3 AEs, 6 months after radiotherapy initiation. The machine learning method established the importance of variables among demographic, clinical and DNA repair data. The most relevant ones, all related to DNA repair, were employed to build a predictor. Predictors constructed with random forest and minimum bounding sphere predicted late Grade ≥ 3 AEs with a sensitivity of 100% and specificity of 77.17 and 86.22%, respectively. This multiplex functional approach strongly supports a dominant role for DSB repair in the development of chronic AEs. It also showed that affected patients share specific features related to functional aspects of DSB repair. This strategy may be suitable for routine clinical analysis and paves the way for modelling DSB repair associated with severe AEs induced by radiotherapy.
Assuntos
Algoritmos , Quebras de DNA de Cadeia Dupla , Reparo do DNA , Humanos , Masculino , Feminino , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Idoso , Pessoa de Meia-Idade , Radioterapia/efeitos adversos , Neoplasias da Próstata/radioterapia , Neoplasias da Mama/radioterapia , Leucócitos Mononucleares/efeitos da radiação , Aprendizado de Máquina , Lesões por Radiação/etiologiaRESUMO
This work reports on a model that describes patient-specific absorbed dose-dependent DNA damage response in peripheral blood mononuclear cells of thyroid cancer patients during radioiodine therapy and compares the results with the ex vivo DNA damage response in these patients. Blood samples of 18 patients (nine time points up to 168 h post-administration) were analyzed for radiation-induced γ-H2AX + 53BP1 DNA double-strand break foci (RIF). A linear one-compartment model described the absorbed dose-dependent time course of RIF (Parameters: c characterizes DSB damage induction; k1 and k2 are rate constants describing fast and slow repair). The rate constants were compared to ex vivo repair rates. A total of 14 patient datasets could be analyzed; c ranged from 0.012 to 0.109 mGy-1, k2 from 0 to 0.04 h-1. On average, 96% of the damage is repaired quickly with k1 (range: 0.19-3.03 h-1). Two patient subgroups were distinguished by k1-values (n = 6, k1 > 1.1 h-1; n = 8, k1 < 0.6 h-1). A weak correlation with patient age was observed. While induction of RIF was similar among ex vivo and in vivo, the respective repair rates failed to correlate. The lack of correlation between in vivo and ex vivo repair rates and the applicability of the model to other therapies will be addressed in further studies.
Assuntos
Quebras de DNA de Cadeia Dupla , Reparo do DNA , Neoplasias da Glândula Tireoide , Humanos , Neoplasias da Glândula Tireoide/radioterapia , Neoplasias da Glândula Tireoide/sangue , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/genética , Pessoa de Meia-Idade , Masculino , Feminino , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Adulto , Idoso , Dano ao DNA , Radioisótopos do Iodo/uso terapêutico , Proteína 1 de Ligação à Proteína Supressora de Tumor p53/metabolismo , Histonas/metabolismo , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/efeitos da radiação , Modelos BiológicosRESUMO
PURPOSE: In case of a nuclear accident, individuals with high-dose radiation exposure (>1-2 Gy) should be rapidly identified. While ferredoxin reductase (FDXR) was recently suggested as a radiation-responsive gene, the use of a single gene biomarker limits radiation dose assessment. To overcome this limitation, we sought to identify reliable radiation-responsive gene biomarkers. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from mice after total body irradiation, and gene expression was analyzed using a microarray approach to identify radiation-responsive genes. RESULTS: In light of the essential role of the immune response following radiation exposure, we selected several immune-related candidate genes upregulated by radiation exposure in both mouse and human PBMCs. In particular, the expression of ACOD1 and CXCL10 increased in a radiation dose-dependent manner, while remaining unchanged following lipopolysaccharide (LPS) stimulation in human PBMCs. The expression of both genes was further evaluated in the blood of cancer patients before and after radiotherapy. CXCL10 expression exhibited a distinct increase after radiotherapy and was positively correlated with FDXR expression. CONCLUSIONS: CXCL10 expression in irradiated PBMCs represents a potential biomarker for radiation exposure.
Assuntos
Leucócitos Mononucleares , Exposição à Radiação , Humanos , Camundongos , Animais , Leucócitos Mononucleares/efeitos da radiação , Relação Dose-Resposta à Radiação , Regulação para Cima , Triagem , Exposição à Radiação/efeitos adversos , Biomarcadores/metabolismo , Quimiocina CXCL10/genética , Quimiocina CXCL10/metabolismoRESUMO
The high level natural radiation areas (HLNRA) of Kerala coast provide unique opportunity to study the biological effect of chronic low dose ionizing radiation (LDIR) on human population below 100 mSv. The radiation level in this area varies from < 1.0-45 mGy /year due to patchy distribution of monazite in the sand, which contains 232Th (8-10%), 238U (0.3%), and their decay products. Telomere length attrition has been correlated to DNA damage due to genotoxic agents. The objective of the present study is to evaluate the effect of natural chronic LDIR exposure on telomere length and transcriptional response of telomere specific and DNA damage repair genes in peripheral blood mononuclear cells (PBMCs) of individuals from normal level natural radiation areas (NLNRA) and HLNRA of Kerala coast, southwest India. Blood samples were collected from 71 random male donors (24-80 years) from NLNRA (≤1.50 mGy/year; N = 19) and two HLNRA dose groups [1.51-10 mGy/year (N = 17); > 10 mGy/year, (N = 35)]. Genomic DNA was isolated from PBMCs and relative telomere length (RTL) was determined using real time q-PCR. Radio-adaptive response (RAR) study was carried out in PBMCs of 40 random males from NLNRA (N = 20) and HLNRA (>10 mGy/year; N = 20), where PBMCs were given a challenged dose of 2.0 Gy gamma radiation at 4 h. Transcriptional profile of telomere specific (TRF1, TRF2, POT1, TIN2, TPP1, RAP1), DNA damage response (RAD17, ATM, CHEK1) and base excision repair pathway (BER) (OGG1, XRCC1, NTH1, NEIL1, MUTYH, MBD4) genes were analysed at basal level and after a challenge dose of 2.0 Gy at 4 h. Our results did not show any significant effect of chronic LDR on RTL among the individuals from NLNRA and two HLNRA groups (p = 0.195). However, influence of age on RTL was clearly evident among NLNRA and HLNRA individuals. At basal level, TRF1, TRF2, TIN2, MBD4, NEIL1 and RAD17 showed significant up-regulation, whereas XRCC1 was significantly down regulated in HLNRA individuals. After a challenge dose of 2.0 Gy, significant transcriptional up-regulation was observed at telomere specific (TRF2, POT1) and BER (MBD4, NEIL1) genes in HLNRA individuals as compared to NLNRA suggesting their role in RAR. In conclusion, elevated level of natural chronic LDR exposure did not have any adverse effect on telomere length in Kerala coast. Significant transcriptional response at TRF2, MBD4 and NEIL1 at basal level and with a challenge dose of 2.0 Gy suggested their active involvement in efficient repair and telomere maintenance in individuals from HLNRA of Kerala coast.
Assuntos
DNA Glicosilases , Exposição à Radiação , Humanos , Masculino , Complexo Shelterina , Leucócitos Mononucleares/efeitos da radiação , Radiação de Fundo , Telômero/genética , Telômero/metabolismo , Exposição à Radiação/efeitos adversos , Proteína 1 Complementadora Cruzada de Reparo de Raio-X/metabolismo , Proteínas de Ciclo Celular/metabolismo , DNA Glicosilases/metabolismoRESUMO
The treatment of chronic inflammatory and degenerative diseases by low dose radiation therapy (LDRT) is promising especially for patients who were refractory for classical therapies. LDRT aims to reduce pain of patients and to increase their mobility. Although LDRT has been applied since the late 19th century, the immunological mechanisms remain elusive. Within the prospective IMMO-LDRT01 trial (NCT02653079) the effects of LDRT on the peripheral blood immune status, as well as on pain and life quality of patients have been analyzed. Blood is taken before and after every serial irradiation with a single dose per fraction of 0.5Gy, as well as during follow-up appointments in order to determine a detailed longitudinal immune status by multicolor flow cytometry. Here, we report the results of an interim analysis of 125 patients, representing half the number of patients to be recruited. LDRT significantly improved patients' pain levels and induced distinct systemic immune modulations. While the total number of leukocytes remained unchanged in the peripheral blood, LDRT induced a slight reduction of eosinophils, basophils and plasmacytoid dendritic cells and an increase of B cells. Furthermore, activated immune cells were decreased following LDRT. Especially cells of the monocytic lineage correlated to LDRT-induced improvements of clinical symptoms, qualifying these immune cells as predictive biomarkers for the therapeutic success. We conclude that LDRT improves pain of the patients by inducing systemic immune modulations and that immune biomarkers could be defined for prediction by improved patient stratification in the future.
Assuntos
Subpopulações de Linfócitos B/imunologia , Eosinófilos/imunologia , Leucócitos Mononucleares/patologia , Monócitos/imunologia , Osteoartrite/radioterapia , Dor/radioterapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Contagem de Células , Feminino , Seguimentos , Humanos , Imunomodulação , Leucócitos Mononucleares/efeitos da radiação , Masculino , Pessoa de Meia-Idade , Osteoartrite/imunologia , RadioterapiaRESUMO
Our laboratory has demonstrated that captopril, an angiotensin converting enzyme inhibitor, mitigates hematopoietic injury following total body irradiation in mice. Improved survival in mice is correlated with improved recovery of mature blood cells and bone marrow, reduction of radiation-induced inflammation, and suppression of radiation coagulopathy. Here we investigated the effects of captopril treatment against radiation injuries in the Göttingen mini pig model of Hematopoietic-Acute Radiation Syndrome (H-ARS). Minipigs were given captopril orally (0.96 mg/kg) twice daily for 12 days following total body irradiation (60Co 1.79 Gy, 0.42-0.48 Gy/min). Blood was drawn over a time course following irradiation, and tissue samples were collected at euthanasia (32-35 days post-irradiation). We observed improved survival with captopril treatment, with survival rates of 62.5% in vehicle treated and 87.5% in captopril treated group. Additionally, captopril significantly improved recovery of peripheral blood mononuclear cells, and a trend toward improvement in recovery of red blood cells and platelets. Captopril significantly reduced radiation-induced expression of cytokines erythropoietin and granulocyte-macrophage colony-stimulating factor and suppressed radiation-induced acute-phase inflammatory response cytokine serum amyloid protein A. Using quantitative-RT-PCR to monitor bone marrow recovery, we observed significant suppression of radiation-induced expression of redox stress genes and improved hematopoietic cytokine expression. Our findings suggest that captopril activities in the Göttingen minipig model of hematopoietic-acute radiation syndrome reflect findings in the murine model.
Assuntos
Síndrome Aguda da Radiação/tratamento farmacológico , Captopril/farmacologia , Sistema Hematopoético/efeitos dos fármacos , Lesões Experimentais por Radiação/tratamento farmacológico , Síndrome Aguda da Radiação/patologia , Animais , Modelos Animais de Doenças , Eritropoetina/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Sistema Hematopoético/lesões , Sistema Hematopoético/patologia , Sistema Hematopoético/efeitos da radiação , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/efeitos da radiação , Camundongos , Oxirredução/efeitos dos fármacos , Lesões Experimentais por Radiação/patologia , Suínos , Porco Miniatura , Irradiação Corporal Total/efeitos adversosRESUMO
Radiation therapy-mediated salivary gland destruction is characterized by increased inflammatory cell infiltration and fibrosis, both of which ultimately lead to salivary gland hypofunction. However, current treatments (e.g., artificial saliva and sialagogues) only promote temporary relief of symptoms. As such, developing alternative measures against radiation damage is critical for restoring salivary gland structure and function. One promising option for managing radiation therapy-mediated damage in salivary glands is by activation of specialized proresolving lipid mediator receptors due to their demonstrated role in resolution of inflammation and fibrosis in many tissues. Nonetheless, little is known about the presence and function of these receptors in healthy and/or irradiated salivary glands. Therefore, the goal of this study was to detect whether these specialized proresolving lipid mediator receptors are expressed in healthy salivary glands and, if so, if they are maintained after radiation therapy-mediated damage. Our results indicate that specialized proresolving lipid mediator receptors are heterogeneously expressed in inflammatory as well as in acinar and ductal cells within human submandibular glands and that their expression persists after radiation therapy. These findings suggest that epithelial cells as well as resident immune cells represent potential targets for modulation of resolution of inflammation and fibrosis in irradiated salivary glands.
Assuntos
Tolerância a Radiação , Receptores de Quimiocinas/genética , Receptores de Formil Peptídeo/genética , Receptores Acoplados a Proteínas G/genética , Receptores do Leucotrieno B4/genética , Receptores de Lipoxinas/genética , Glândula Submandibular/efeitos da radiação , Células Acinares/citologia , Células Acinares/metabolismo , Células Acinares/efeitos da radiação , Adulto , Idoso , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Células Endoteliais/efeitos da radiação , Feminino , Raios gama , Expressão Gênica , Humanos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/efeitos da radiação , Masculino , Pessoa de Meia-Idade , Receptores de Quimiocinas/metabolismo , Receptores de Formil Peptídeo/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores do Leucotrieno B4/metabolismo , Receptores de Lipoxinas/metabolismo , Glândula Submandibular/citologia , Glândula Submandibular/metabolismoRESUMO
PURPOSE: For triage purposes following a nuclear accident or a terrorist event, gene expression biomarkers in blood have been demonstrated to be good bioindicators of ionizing radiation (IR) exposure and can be used to assess the dose received by exposed individuals. Many IR-sensitive genes are regulated by the DNA damage response pathway, and modulators of this pathway could potentially affect their expression level and therefore alter accurate dose estimations. In the present study, we addressed the potential influence of temperature, sample transport conditions and the blood cell fraction analyzed on the transcriptional response of the following radiation-responsive genes: FDXR, CCNG1, MDM2, PHPT1, APOBEC3H, DDB2, SESN1, P21, PUMA, and GADD45. MATERIALS AND METHODS: Whole blood from healthy donors was exposed to a 2 Gy X-ray dose with a dose rate of 0.5 Gy/min (output 13 mA, 250 kV peak, 0.2 mA) and incubated for 24 h at either 37, 22, or 4 °C. For mimicking the effect of transport conditions at different temperatures, samples incubated at 37 °C for 24 h were kept at 37, 22 or 4 °C for another 24 h. Comparisons of biomarker responses to IR between white blood cells (WBCs), peripheral blood mononuclear cells (PBMCs) and whole blood were carried out after a 2 Gy X-ray exposure and incubation at 37 °C for 24 hours. RESULTS: Hypothermic conditions (22 or 4 °C) following irradiation drastically inhibited transcriptional responses to IR exposure. However, sample shipment at different temperatures did not affect gene expression level except for SESN1. The transcriptional response to IR of specific genes depended on the cell fraction used, apart from FDXR, CCNG1, and SESN1. CONCLUSION: In conclusion, temperature during the incubation period and cell fraction but not the storing conditions during transport can influence the transcriptional response of specific genes. However, FDXR and CCNG1 showed a consistent response under all the different conditions tested demonstrating their reliability as individual biological dosimetry biomarkers.
Assuntos
Regulação da Expressão Gênica/efeitos da radiação , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/efeitos da radiação , Radiometria/métodos , Temperatura , Adulto , Relação Dose-Resposta à Radiação , Humanos , MasculinoRESUMO
The coronavirus disease-2019 (COVID-19) pandemic caused by SARS-CoV-2 has exacted an enormous toll on healthcare systems worldwide. The cytokine storm that follows pulmonary infection is causally linked to respiratory compromise and mortality in the majority of patients. The sparsity of viable treatment options for this viral infection and the sequelae of pulmonary complications have fueled the quest for new therapeutic considerations. One such option, the long-forgotten idea of using low-dose radiation therapy, has recently found renewed interest in many academic centers. We outline the scientific and logistical rationale for consideration of this option and the mechanistic underpinnings of any potential therapeutic value, particularly as viewed from an immunological perspective. We also discuss the preliminary and/or published results of prospective trials examining low-dose radiation therapy for COVID-19.
Assuntos
COVID-19/radioterapia , Doses de Radiação , SARS-CoV-2/efeitos da radiação , Internalização do Vírus/efeitos da radiação , Replicação Viral/efeitos da radiação , COVID-19/epidemiologia , COVID-19/virologia , Citocinas/metabolismo , Relação Dose-Resposta à Radiação , Interações Hospedeiro-Patógeno/efeitos da radiação , Humanos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/efeitos da radiação , Leucócitos Mononucleares/virologia , Pandemias , Dosagem Radioterapêutica , SARS-CoV-2/fisiologiaRESUMO
PURPOSE: In suspected radiation exposures, cytokinesis-block micronucleus (CBMN) assay is used for biodosimetry by detecting micronuclei (MN) in binucleated (BN) cells in whole blood and isolated peripheral blood mononuclear cell (PBMC) cultures. Standardized harvest protocols for whole blood were published by the International Atomic Energy Agency (IAEA) in 2001 (Technical report no. 405) and 2011 (EPR-Biodosimetry). For isolated PBMC harvest, cytocentrifugation of fresh cells is recommended to preserve cytoplasmic boundaries for MN scoring. However, cytocentrifugation utilizes specialized equipment and long-term cell suspension storage is difficult. In this study, an alternative CBMN harvest protocol is proposed for laboratories interested in culturing PBMCs and storing fixed cells with routine biodosimetry methods. MATERIALS AND METHODS: Peripheral blood from 4 males (24, 34, 41, 51 y.o.) and females (26, 37, 44, 56 y.o.) was irradiated with 0 and 2 Gy X-rays. For cells harvested with IAEA 2001 and 2011 protocols, whole blood was used. For cells harvested with our protocol (CRG), isolated PBMCs were used. CRG protocol was validated in DAPI, acridine orange and Giemsa stain, and in three other laboratories. Cytoplasm status, nuclear division index (NDI) and induced MN frequency (MN frequency at 2 Gy - background MN frequency at 0 Gy) (MN/1000 BN) of Giemsa-stained BN cells were compared in IAEA 2001, IAEA 2011, IAEA 2011 + formaldehyde (FA) and CRG protocols. Effects of low and high humidity spreading were evaluated. RESULTS: >94% of 1000 BN cells were scorable with clear cytoplasmic boundaries in all donors harvested with CRG protocol. FA addition in IAEA 2011 protocol reduced cell rupture in whole blood cultures, but cell rupture was affected by age, sex and humidity. Almost all cells harvested with IAEA 2001 protocol had cytoplasm loss. PBMCs harvested with CRG protocol stained well in DAPI, acridine orange and Giemsa, and showed high scorable BN frequency in all laboratories. A higher NDI and a lower induced MN frequency were seen in 2 Gy isolated PBMC than whole blood cultures. CONCLUSION: This quick CBMN harvest protocol for isolated PBMCs is a viable alternative to cytocentrifugation, as many scorable BN cells were obtained with routine biodosimetry reagents and equipment. IAEA 2011 + FA protocol should be used to improve CBMN harvest in whole blood cultures. Humidity during spreading should be optimized depending on the harvest protocol. NDI and MN frequency should be separately evaluated for whole blood and isolated PBMC cultures.
Assuntos
Separação Celular/métodos , Leucócitos Mononucleares/efeitos da radiação , Testes para Micronúcleos/métodos , Adulto , Citocinese , Feminino , Humanos , Umidade , Leucócitos Mononucleares/ultraestrutura , Masculino , Pessoa de Meia-Idade , Doses de RadiaçãoRESUMO
BACKGROUND: Extracorporeal photopheresis (ECP) is one of the most widely used and effective cell-based therapies for the treatment of T-cell-mediated diseases. The patients' white blood cells (WBCs) are collected by apheresis and exposed to the photosensitizer 8-methoxypsoralen (8-MOP) and ultraviolet A (UVA) light before retransfusion. The UVA/8-MOP combination has been in use in ECP for more than 4 decades; however, whether ECP can be simplified by UVA light irradiation only has never been analyzed. STUDY DESIGN AND METHODS: Peripheral blood mononuclear cells were treated with classical ECP or different UVA light doses only (UVAonly ). Treatment efficacy was investigated by apoptosis induction in WBC subsets, inhibition of T-cell proliferation, and the ability of monocytes to induce allogeneic T-cell expansion and to respond to lipopolysaccharide and interferon-γ stimulation in vitro. RESULTS: High-dose UVAonly treatment (5 J/cm2 ) was as efficient as ECP to induce apoptosis within 48 hours. UVAonly treatment modulated the composition of the surviving cells by improving monocyte survival and promoting CD8+ T-cell apoptosis. Both ECP and UVAonly treatment inhibited anti-CD3/anti-CD28 triggered T-cell proliferation. Interestingly, whereas ECP-treated monocytes exhibited a markedly reduced capacity to respond to stimulation and to induce allogeneic T-cell proliferation, UVAonly treatment preserved monocyte functionality to some degree. CONCLUSIONS: High-dose UVAonly and standard ECP showed comparable efficacy in inducing apoptosis and inhibiting direct T-cell proliferation. Hence, UVAonly treatment can be a simplified alternative to ECP therapy. Furthermore, increased monocyte survival with partially preserved functionality after UVAonly treatment may provide a novel method for immunoregulation.
Assuntos
Apoptose/efeitos da radiação , Proliferação de Células/efeitos da radiação , Leucócitos Mononucleares/efeitos da radiação , Fotoferese/métodos , Linfócitos T/efeitos da radiação , Apoptose/efeitos dos fármacos , Antígenos CD28/metabolismo , Complexo CD3/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Humanos , Técnicas In Vitro , Interferon gama/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Metoxaleno/farmacologia , Monócitos/efeitos dos fármacos , Monócitos/efeitos da radiação , Linfócitos T/efeitos dos fármacos , Raios UltravioletaRESUMO
OBJECTIVE: to determine the association between the expression of lipoprotein lipase (LPL) and c-MYC genes inperipheral blood cells of chronic lymphocytic leukemia (CLL) patients affected by the Chornobyl catastrophedepending on the mutational status of IGHV genes. METHODS: Analysis was performed in the group of 69 CLL patients irradiated due to the Chornobyl NPP accident (58clean-up workers of 1986 year, 6 inhabitants of radionuclide contaminated areas, and 5 evacuees). The IGHV genemutational status was studied by polymerase chain reaction (PCR) followed by direct sequencing. LPL and c-MYCexpression was evaluated by Quantitative Real-time PCR. Data were analyzed with the SPSS software package, version 20.0. RESULTS: Relative LPL expression levels in CLL samples ranged from 0 to 1663.5 (mean 138.47 ± 30.69, median 26.1).A strong correlation between individual LPL expression levels and IGHV mutational status was found (r = 0.684;p < 0.0001). The average relative c-MYC expression level was 5.7 ± 0.87 (median 2.86; range 0-48.5). No association between c-MYC expression and IGHV mutational status was found. Among unmutated IGHV cases, a correlationbetween LPL and c-MYC gene expression levels was identified: r = 0.351; p = 0.013. CONCLUSIONS: Our data confirm the dominant concept that unmutated IGHV CLL cases are more sensitive to the actionof proliferative stimuli compared to mutated IGHV CLL cases. This is manifested by an increase in the expression ofa functionally significant LPL gene, is one for the strongest negative prognostic markers in CLL.
Assuntos
Acidente Nuclear de Chernobyl , Genes de Cadeia Pesada de Imunoglobulina , Leucemia Linfocítica Crônica de Células B/genética , Lipase Lipoproteica/genética , Proteínas Proto-Oncogênicas c-myc/genética , Exposição à Radiação/efeitos adversos , Lesões por Radiação/genética , Idoso , Poluentes Radioativos do Ar/efeitos adversos , Socorristas , Feminino , Contaminação Radioativa de Alimentos , Regulação da Expressão Gênica , Humanos , Leucemia Linfocítica Crônica de Células B/etiologia , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Linfocítica Crônica de Células B/patologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Leucócitos Mononucleares/efeitos da radiação , Lipase Lipoproteica/imunologia , Masculino , Pessoa de Meia-Idade , Mutação , Prognóstico , Proteínas Proto-Oncogênicas c-myc/imunologia , Lesões por Radiação/etiologia , Lesões por Radiação/imunologia , Lesões por Radiação/patologia , Radioisótopos , Poluentes Radioativos do Solo/efeitos adversos , UcrâniaRESUMO
OBJECTIVE: To study proliferative potential of peripheral blood lymphocytes of Chornobyl clean-up workers by levelof expression of cyclin D1 and quantitative parameters of cell cycle at a remote period after radiation exposure. MATERIALS AND METHODS: The research subject was the peripheral blood lymphocytes (PB) of Chornobyl clean-upworkers 30-33 years after radiation exposure. A total of 207 men were surveyed, 164 of them were clean-up workers exposed in the dose range 10.43-3623.31 mSv and 43 persons of the control group. Analysis of proliferationpotential (cell cycle initiation) and cyclin D1 expression in PB lymphocytes were performed in vitro by a micro methodof whole blood leukocytes culture with phytohemagglutinine-P (PHA). Sample preparation was performed by a standard immunofluorescent assay for intracellular proteins using the FITC labelled Mouse Anti-Human Cyclin D1Antibody Set. Cell distribution by cell cycle phases studied by propidium iodide DNA staining and analysis onFACSCalibur laser flow cytometer in histogram mode with separation of G0/G1-, S- and G2/M-regions and Sub-G0/G1-region (apoptotic cells). RESULTS AND CONCLUSIONS: An increase in the level of spontaneous Ñyclin D1 expression and disturbance of ÑyclinD1-dependent regulation of cell cycle of PB lymphocytes after mitogen activation were determined in a remote period after radiation exposure. An increase in the level of cyclin D1 expression was accompanied by increase in pool ofcells in the S- and G2/M-phases of cell cycle which characterizes the high proliferative potential of PB lymphocytes.Mitogen-induced delay of cell cycle of lymphocytes in G1/S check point and reduction of S-phase was revealed.These changes are a manifestation of genomic instability caused by the effect of radiation and depend on the radiation dose. The results confirm the hypothesis about the significance of levels of cyclin D1 expression, as a criterion for manifestations of genome instability and risks of oncogenesis in a remote period after irradiation.
Assuntos
Ciclo Celular/efeitos da radiação , Acidente Nuclear de Chernobyl , Ciclina D1/genética , Leucócitos Mononucleares/efeitos da radiação , Exposição à Radiação/efeitos adversos , Lesões por Radiação/genética , Idoso , Estudos de Casos e Controles , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Ciclina D1/metabolismo , Relação Dose-Resposta à Radiação , Socorristas , Expressão Gênica , Instabilidade Genômica , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Fito-Hemaglutininas/farmacologia , Cultura Primária de Células , Lesões por Radiação/etiologia , Lesões por Radiação/metabolismo , Lesões por Radiação/patologia , Radioisótopos , Radiometria , UcrâniaRESUMO
OBJECTIVE: to establish the connection of radiation-induced changes in gene expression with the realized pathology of the broncho-pulmonary and cardiovascular systems in Chornobyl clean-up workers. MATERIALS AND METHODS: We examined 314 male Chornobyl clean-up workers (main group; age (58.94 ± 6.82) years(M ± SD); min 33, max 79 years; radiation dose (411.82 ± 625.41) mSv (M ± SD); min 1.74, max 3600 mSv) with various nosological forms of cardiovascular and broncho-pulmonary pathology (BPP) and 50 subjects of the controlgroup: age (50.50 ± 5.73) years (M ± SD); min 41, max 67 years. The relative level of BCL2, CDKN2A, CLSTN2, GSTM1,IFNG, IL1B, MCF2L, SERPINB9, STAT3, TERF1, TERF2, TERT, TNF, TP53, CCND1, CSF2, VEGFA genes expression was determined inperipheral blood leukocytes by real-time PCR (7900 HT Fast Real-Time PCR System (Applied Biosystems, USA)). The«gene-disease¼ association was determined on statistical models stratified separately for each disease and gene.Logistic regression was used to calculate the odds ratio. RESULTS: Increased GSTM1 gene expression and no changes in angiogenesis-related VEGFA gene expression werefound in the main group of patients with coronary heart disease (CHD). It was established overexpression of TP53,VEGF and IFNG genes in the group of patients with arterial hypertension (AH). At combination of these diseases anincrease of expression of СSF2, TERF1, TERF2 genes was established. The detected changes demonstrate an activationof the antioxidative defense system in patients with CHD, while AH is associated with the expression of genes ofangiogenesis and immune inflammation. It was shown an increase in the expression of genes associated with apoptosis and kinase activity (BCL2, CLSTN2, CDKN2), immune inflammation (CSF2, IL1B, TNF) in Chornobyl clean-upworkers with BPP. Expression of TP53 and GSTM1 (gene, associated with the glutathione system) was significantlyupregulated in the group of individuals with chronic bronchitis, whereas in patients with chronic obstructive pulmonary disease, no increase was detected; the expression of SERPINB9 and MCF2L genes was downregulated. CONCLUSIONS: Changes in the expression of genes, associated with the development of somatic pathology in theremote period after irradiation, in particular the genes of the immune response and inflammatory reactions CSF2,IFNG, IL1B, TNF; expression of genes that regulate cell proliferation, aging and apoptosis TP53, BCL2, MCF2L, CDKN2A,SERPINB9, TERF1, TERF2, TERT; genes that regulate cell adhesion and angiogenesis CLSTN2, VEGF.
Assuntos
Doenças Cardiovasculares/genética , Acidente Nuclear de Chernobyl , Expressão Gênica/efeitos da radiação , Pneumopatias/genética , Exposição à Radiação/efeitos adversos , Lesões por Radiação/genética , Idoso , Poluentes Radioativos do Ar/efeitos adversos , Proteínas Angiogênicas/genética , Proteínas Angiogênicas/metabolismo , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/patologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Fatores Estimuladores de Colônias/genética , Fatores Estimuladores de Colônias/metabolismo , Socorristas , Contaminação Radioativa de Alimentos , Humanos , Interferons/genética , Interferons/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Leucócitos Mononucleares/efeitos da radiação , Pneumopatias/etiologia , Pneumopatias/metabolismo , Pneumopatias/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Lesões por Radiação/etiologia , Lesões por Radiação/metabolismo , Lesões por Radiação/patologia , Radioisótopos , Poluentes Radioativos do Solo/efeitos adversos , Proteínas de Ligação a Telômeros/genética , Proteínas de Ligação a Telômeros/metabolismo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , UcrâniaRESUMO
BACKGROUND: Application of the most advanced radiation technologies of brachytherapy featuring the high dose ratesources i.e. 60Co and 192Ir within contemporary management protocols for gynecological cancer provides maximum dosedistribution in the clinical target along with minimal radiation exposure on surrounding organs and tissues. It involvesirradiation of large spaces with delivery of high therapeutic doses at the tolerance bound of «critical¼ organs (bladder,rectum) and tissues. Thus minimization of the early and late radiation complications, life span extent and quality oflife increase remain just the issues in contemporary radiation oncology requiring therefore the elaboration of radiobiological criteria along with substantiation of physiÑо-engineering properties of the radiation sources. Taking intoaccount the basic radiobiological patterns will ensure a definitive further progress in the field of radiation oncology. OBJECTIVE: to study and compare the biological effects of 192Ir with the effects of the reference gamma radiation 60Coand increase the effectiveness of brachytherapy using a 192Ir source. MATERIALS AND METHODS: Radiobiological dosimetry on the basis of a test system of peripheral blood lymphocytesfrom the gynecological cancer patients with subsequent cytogenetic analysis of radiation-induced chromosomeaberrations was performed to study and compare the biological effects of 192Ir and reference 60Со γ-radiation, and toenhance the efficiency of 192Ir brachytherapy. RESULTS: Radiation markers, i.e. dicentric chromosomes with an accompanying paired fragment prevailed in thespectrum of radiation-induced damage. Variability of individual cytogenetic parameters of peripheral lymphocytesupon the first fraction of irradiation at the same dose of 5 Gy indicated an individual sensitivity of patients to the192Ir γ-irradiation. Comprehensive conservative treatment with adjuvant radiotherapy was applied to the patients(n = 98) having got secondary vaginal cancer stage II-III, T2-3N0-1M0. The high dose-rate (HDR) brachytherapy using 192Ir radiation sources was applied in the main study group (n = 37), HDR brachytherapy using 60Co radiation sourceswas applied in the control group (n = 35). CONCLUSION: The HDR brachytherapy with 192Ir and 60Co sources on the up-to-date technology intensive devices provides a high accuracy of dose distributions when irradiating the malignant neoplasms with minimized radiationexposure to the «critical¼ tissues. Treatment results are improved therefore. The use of 192Ir radiation sources compared with 60Co ones resulted in an increased throughput of treatment, enhanced tumor regression, and reduced incidence of radiation effects on the critical organs. Currently we perform the radiobiological studies on somatic cellsfrom cancer patients at the genetic, biochemical, biophysical, and cytological levels in order to receive a biologicalindication of radiation damage under the impact of 192Ir isotope. Continuation of clinical trials with radiobiologicalsupport will provide an opportunity to predict the early and late radiation complications and thus to provide a personalized approach in brachytherapy of cancer patients using the 192Ir sources of γ-rays.
Assuntos
Braquiterapia/métodos , Aberrações Cromossômicas/efeitos da radiação , Neoplasias dos Genitais Femininos/radioterapia , Radioisótopos de Irídio/uso terapêutico , Leucócitos Mononucleares/efeitos da radiação , Braquiterapia/instrumentação , Radioisótopos de Cobalto/uso terapêutico , Relação Dose-Resposta à Radiação , Feminino , Neoplasias dos Genitais Femininos/genética , Neoplasias dos Genitais Femininos/patologia , Humanos , Leucócitos Mononucleares/patologia , Estadiamento de Neoplasias , Cultura Primária de Células , Radioterapia (Especialidade)/instrumentação , Radioterapia (Especialidade)/métodos , Radiometria , Microambiente Tumoral/efeitos da radiaçãoRESUMO
Our immune system can be exposed to neutral ionizing radiation. This work proposed to investigate the impacts of low-dose fast neutrons on the molecular structure of the lipids of peripheral blood mononuclear cells (PBMCs) as the primary immune cells. The effects have been followed at the time of 0, 2, 4, and 8 days after neutron exposure. Sixty-four female Wistar rats were used in this work. Thirty-two of them were irradiated using a low-dose fast neutron (241Am-Be, 0.2 mGy/h). The other thirty-two were utilized as a control group. The changes in their lipids molecular structure were investigated using Fourier transform infrared (FTIR) spectroscopy. Besides, lipids were extracted, and the total phospholipids contents were estimated. At the time of 2 and 4 days after irradiation, the results showed significant changes in the molecular structure of lipids in those irradiated samples compare with their control samples, and multivariate analysis succeeded in differentiating between control and irradiated rats. In contrast, no alterations between control and irradiated were detected at the time of 0 and 8 days after. The low-dose fast neutron could induce free radicals, which indirectly affect the molecular structure of the lipids and could damage the phospholipids molecules, and the damage could be repaired.
Assuntos
Nêutrons Rápidos/efeitos adversos , Leucócitos Mononucleares/efeitos da radiação , Fosfolipídeos/análise , Fosfolipídeos/efeitos da radiação , Animais , Feminino , Estrutura Molecular , Fosfolipídeos/química , Análise de Componente Principal , Radiação Ionizante , Ratos , Ratos Wistar , Fatores de Risco , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Dose assessment is an important issue for radiation emergency medicine to determine appropriate clinical treatment. Hematopoietic tissues are extremely vulnerable to radiation exposure. A decrease in blood cell count following radiation exposure is the first quantitative bio-indicator using hematological techniques. We further examined induction of oxidative stress biomarkers in residual lymphocytes to identify new biomarkers for dosimetry. In vivo whole-body radiation to mice exposed to 5 Gy significantly induces DNA double-strand breaks, which were visualized by γ-H2AX in mouse blood cells. Mouse blood smears and peripheral blood mononuclear cells (PBMC) isolated from irradiated mice were used for immunostaining for oxidative biomarkers, parkin or Nrf2. Parkin is the E3 ubiquitin ligase, which is normally localized in the cytoplasm, is relocated to abnormal mitochondria with low membrane potential (ΔΨm), where it promotes clearance via mitophagy. Nrf2 transcription factor controls the major cellular antioxidant responses. Both markers of oxidative stress were more sensitive and persistent over time than nuclear DNA damage. In conclusion, parkin and Nrf2 are potential biomarkers for use in radiation dosimetry. Identification of several biological markers which show different kinetics for radiation response is essential for radiation dosimetry that allows the assessment of radiation injury and efficacy of clinical treatment in emergency radiation incidents. Radiation-induced oxidative damage is useful not only for radiation dose assessment but also for evaluation of radiation risks on humans.
Assuntos
Leucócitos Mononucleares/efeitos da radiação , Fator 2 Relacionado a NF-E2/análise , Estresse Oxidativo/efeitos da radiação , Ubiquitina-Proteína Ligases/análise , Irradiação Corporal Total/efeitos adversos , Animais , Biomarcadores/análise , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Leucócitos Mononucleares/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: This study aimed at evaluating the association between the speed of nuclear factor-kappa B (NF-κB) activation in peripheral blood mononuclear cells (PBMCs) and late skin and subcutaneous fibrosis in patients with head and neck squamous cell carcinoma (HNSCC) after radiotherapy. METHODS: The speed of NF-κB activation was represented by the nuclear p65 expression ratio before and after irradiation. The optimal time point to measure the ratio was determined by Western blot in the PBMCs from healthy outpatients ranging from 0 to 12 hours after ex vivo irradiation. We recruited patients with HNSCC who had received ratiotherapy and who were under regular follow-up care. We assessed the association between the risk of developing ≥grade 2 late fibrosis and the nuclear p65 expression ratio in the PBMCs after ex vivo irradiation in these patients. RESULTS: The maximum nuclear p65 ratio was observed at 1 hour after ex vivo irradiation in the PBMCs from the healthy outpatients. The speed of NF-κB activation was then represented by the nuclear p65 ratio in the PBMCs before and 1 hour after ex vivo irradiation. A total of 200 patients with HNSCC were recruited, 32.50% (n = 65) of which presented with ≥grade 2 late fibrosis. There was a significant association between the speed of NF-κB activation in the PBMCs and an increased risk of developing ≥grade 2 late fibrosis in these patients (P = 0.004). Subgroup analysis suggested that this finding was independent of the known clinical characteristics. CONCLUSIONS: The speed of NF-κB activation might be a potential predictor of late toxicity in cancer patients after radiotherapy. Prospective studies are needed for validation.
Assuntos
Fibrose/metabolismo , Leucócitos Mononucleares/metabolismo , NF-kappa B/metabolismo , Radioterapia/efeitos adversos , Dermatopatias/metabolismo , Pele/metabolismo , Adulto , Idoso , Núcleo Celular/metabolismo , Feminino , Humanos , Leucócitos Mononucleares/efeitos da radiação , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/fisiologia , Transdução de Sinais/efeitos da radiação , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/radioterapia , Fator de Transcrição RelA/metabolismoRESUMO
BACKGROUND: Although there have been many studies investigating the effects of electromagnetic fields on humans cells and tissues, the effects of radiofrequency electromagnetic fields exposure on the cells of the immune system are still controversial. OBJECTIVE: To investigate the effects of 1800 MHz RF-EMF exposure on peripheral blood mononuclear cells by measuring T helper cells count and the cytokine profile under different conditions of durations and distances. METHODS: The peripheral blood mononuclear cells (PBMCs) from healthy human subjects were exposed to 1800 MHz RF-EMF, with durations of 15, 30, 45, and 60 minutes and distances of 5 and 25 cm. The effects of RF-EMF exposure on the number of CD4+ T cells, and the expression of IL-2, IL-10, and IL-17a after 48 hours of culture were evaluated using flow cytometry. RESULTS: Our findings indicated that closer distance and longer exposure inducedlower number of CD4+ T cells. Similarly the percentagesof IL-2, IL-10 and IL-17a expressing CD4+ T cells weredecreased significantly. The number of IL-2 expressing CD4+T cells wasincreased significantly as the duration of exposure was increased, but the number was decreased after 60 minutes exposure when compared with control group with no exposure. CONCLUSION: Exposure to RF-EMF for 60 minutes at 5 cm distance causes a significant reduction in the number of CD4+ T cells, IL-2, IL-10 and IL-17a expressing T cells.