GITR (glucocorticoid-induced TNF receptor) is a recently identified member of the TNF receptor superfamily. The receptor is preferentially expressed on CD4+CD25+ regulatory T cells and GITR signals break the suppressive activity of the subset. In this study, we wanted to reveal the in vivo function of GITR in herpes simplex virus type 1 (HSV-1) infection. A single injection of anti-GITR mAb (DTA-1) immediately after viral infection significantly increased the number of CD4+ and CD8+ T cells expressing CD25, an activation surface marker, and secreting IFN-gamma. We confirmed these in vivo observations by showing ex vivo that re-stimulation of CD4+ or CD8+ T cells with a CD4+ or CD8+ T-cell-specific HSV-1 peptide, respectively, induced a significant elevation in cell proliferation and in IFN-gamma secretion. Our results indicate that GITR signals play a critical role in the T-cell immunity to HSV-1.
Animals ; Antibodies, Monoclonal/pharmacology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cell Proliferation ; Female ; Glucocorticoids/*pharmacology ; Herpes Simplex/*immunology ; Herpesvirus 1, Human/pathogenicity ; *Immunity, Cellular ; Interferon Type II/secretion ; *Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Peptide Fragments/metabolism ; Receptors, Interleukin-2/metabolism ; Receptors, Nerve Growth Factor/genetics/immunology/*metabolism ; Receptors, Tumor Necrosis Factor/genetics/immunology/*metabolism ; Research Support, Non-U.S. Gov't ; T-Lymphocytes/*immunology/metabolism/virology

By searching an EST database, we identified two TNF receptor superfamily members (named mTNFRH1 and mTNFRH2). Amino acid sequences are highly conserved between the two receptors (78% identity). The chromosomal loci of mTnfrh1 and mTnfrh2 genes are found in distal chromosome 7 in the mouse. mTNFRH1 and mTNFRH2 do not contain the cytoplasmic domain, indicating that they might function as decoy receptors. Furthermore, an alternatively spliced form of mTNFRH1 was found which contains neither the transmembrane domain nor the cytoplasmic domain, thus presumably existing as a soluble form. Northern blot analysis showed that mTnfrh1 mRNA was negligibly expressed in tissues, while mTnfrh2 mRNA was strongly expressed in spleen, lung, liver, kidney, and testis. When the extracellular domains of mTNFRH1 and mTNFRH2 were expressed in bacteria, their molecular weight of extracellular region was approximately 15 kDa. Both of the soluble forms were effective in inhibiting T-cell proliferation stimulated by anti-CD3 monoclonal antibody. Our data suggest that mTNFRH1 and mTNFRH2 may be implicated in exerting a modulatory role in the immune response.
Alternative Splicing/genetics ; Amino Acid Sequence ; Animals ; Base Sequence ; Cell Division/*physiology ; Databases, Nucleic Acid ; Gene Expression/genetics ; Mice ; Molecular Sequence Data ; Receptors, Tumor Necrosis Factor/*biosynthesis ; Recombinant Proteins/*biosynthesis ; T-Lymphocytes/*cytology

BACKGROUND: To accelerate the healing of diabetic wounds, various kinds of growth factors have been employed. It is the short half-life of administered growth factors in hostile wound beds that have limited wide-spread clinical usage. To overcome this limitation, growth factor gene therapy could be an attractive alternative rather than direct application of factors onto the wound beds. We administered two growth factor DNAs, epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) into a cutaneous wound on diabetic mice. We compared the different characteristics of the healing wounds. METHODS: Streptozotocin was injected intraperitoneally to induce diabetes into C57BL/6J mice. The ultrasound micro-bubble destruction method with SonoVue as a bubbling agent was used for non-viral gene delivery of EGF828 and VEGF165 DNAs. Each gene was modified for increasing efficacy as FRM-EGF828 or minicircle VEGF165. The degree of neoangiogenesis was assessed using qualitative laser Doppler flowmetry. We compared wound size and histological findings of the skin wounds in each group. RESULTS: In both groups, accelerated wound closure was observed in the mice receiving gene therapy compared with non treated diabetic control mice. Blood flow detected by laser doppler flowmetry was better in the VEGF group than in the EGF group. Wound healing rates and histological findings were more accelerated in the EGF gene therapy group than the VEGF group, but were not statistically significant. CONCLUSION: Both non-viral EGF and VEGF gene therapy administrations could improve the speed and quality of skin wound healing. However, the detailed histological characteristics of the healing wounds were different.
Animals ; DNA ; Epidermal Growth Factor ; Genetic Therapy ; Half-Life ; Intercellular Signaling Peptides and Proteins ; Laser-Doppler Flowmetry ; Mice ; Phospholipids ; Skin ; Streptozocin ; Sulfur Hexafluoride ; Vascular Endothelial Growth Factor A ; Wound Healing

4-1BB, a transmembrane molecule, member of the tumor necrosis factor receptor superfamily, is an important costimulatory molecule in the immune response, plays a key role in the clonal expansion and survival of CD8(+)T cells. In this study, we investigated 4-1BB regulation of CD4(+)T cell responses using 4-1BB transgenic (TG) mice that constitutively expressed 4-1BB on mature T cells. We first showed that CD4(+)T cells of 4-1BB TG mice had more sustained proliferative capacity in response to TCR/4-1BB stimulation in vitro compared to WT mice. Secondly, 4-1BB TG mice exhibited a more elevated contact hypersensitivity (CHS) response mediated by CD4+ Th1 cells due to more vigorous expansion of and apoptotic inhibition of CD4(+)T cells. Finally, CD4(+)T cells of 4-1BB TG mice had a heightened capacity for T cell priming. Overall, our results demonstrate the involvement of 4-1BB in CD4(+)Th1 cell responses by regulating the clonal expansion and survival of CD4(+)T cells as seen in CD8(+)T cells.
Animals ; Antibodies/immunology ; Antigens, CD ; Antigens, CD137 ; CD4-Positive T-Lymphocytes/cytology/*immunology/*metabolism ; Cell Division ; Cell Lineage ; Dermatitis, Contact/genetics/immunology ; Flow Cytometry ; Gene Expression ; Mice ; Mice, Transgenic ; Receptors, Nerve Growth Factor/*genetics/*metabolism ; Receptors, Tumor Necrosis Factor/*genetics/*metabolism

BACKGROUND: Continuous renal replacement therapy (CRRT) is an important treatment modality for severe acute kidney injury. As such, the epidemiology of CRRT in Korea needs further investigation. METHODS: We conducted a nationwide, population-based study analyzing the claims data from National Health Insurance Service of Korea. All index intensive care unit admission cases of CRRT in government-designated tertiary referral hospitals in Korea from 2005 to 2016 were included. Patients with a history of renal replacement therapy or who were under 20 years old were not considered. In addition to baseline and treatment characteristics, patient outcomes, including all-cause mortality and renal survival rates, were investigated. We stratified the study patients according to 3-year time periods and major regions of the nation. RESULTS: We included 37,337 patients who received CRRT in Korea. The overall use of CRRT increased over time, and more than 80% of cases of acute renal replacement therapy were CRRT after 2014. Seoul was the region in which the majority of CRRT (45.0%) was performed. The clinical characteristics of CRRT patients were significantly different among time-intervals and regions. Both all-cause mortality and renal survival rates after CRRT were prominently improved in the recent time periods (P < 0.001). CONCLUSION: CRRT is a widely used treatment strategy for severe acute kidney injury in Korea. The prognosis of CRRT patients has improved compared to the past. This epidemiological study of CRRT in Korea revealed notable trends with regard to time period and geographic region.
Acute Kidney Injury ; Critical Care ; Dialysis ; Epidemiologic Studies ; Epidemiology* ; Humans ; Intensive Care Units ; Korea* ; Mortality ; National Health Programs* ; Prognosis ; Renal Replacement Therapy* ; Seoul ; Survival Rate ; Tertiary Care Centers

BACKGROUND: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. METHODS: Mouse AE II cell line MLE-12 were exposed to 1–10 µg/mL BLM and 0.01–100 µM baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. RESULTS: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor α, and transforming growth factor β1. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. CONCLUSION BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.

Radon is a naturally occurring radioactive material formed by the slow decay of uranium and thorium found in the earth's crust or construction materials. Internal exposure to radon accounts for about half of the natural background radiation dose to which humans are exposed annually. Radon is a carcinogen and is the second leading cause of lung cancer following smoking. An association between radon and lung cancer has been consistently reported in epidemiological studies on mine workers and the general population with indoor radon exposure. However, associations have not been clearly established between radon and other diseases, such as leukemia and thyroid cancer. Radiation doses are assessed by applying specific dose conversion coefficients according to the source (e.g., radon or thoron) and form of exposure (e.g., internal or external). However, regardless of the source or form of exposure, the effects of a given estimated dose on human health are identical, assuming that individuals have the same sensitivity to radiation. Recently, radiation exceeding the annual dose limit of the general population (1 mSv/yr) was detected in bed mattresses produced by D company due to the use of a monazite-based anion powder containing uranium and thorium. This has sparked concerns about the health hazards for mattress users caused by radiation exposure. In light of this event, this study presents scientific information about the assessment of radon and thoron exposure and its human implications for human health, which have emerged as a recent topic of interest and debate in society.
Background Radiation ; Beds ; Carcinogens ; Construction Materials ; Epidemiologic Studies ; Humans ; Korea ; Leukemia ; Lung Neoplasms ; Miners ; Radiation Exposure ; Radon ; Smoke ; Smoking ; Thorium ; Thyroid Neoplasms ; Uranium

Radon is a naturally occurring radioactive material formed by the slow decay of uranium and thorium found in the earth's crust or construction materials. Internal exposure to radon accounts for about half of the natural background radiation dose to which humans are exposed annually. Radon is a carcinogen and is the second leading cause of lung cancer following smoking. An association between radon and lung cancer has been consistently reported in epidemiological studies on mine workers and the general population with indoor radon exposure. However, associations have not been clearly established between radon and other diseases, such as leukemia and thyroid cancer. Radiation doses are assessed by applying specific dose conversion coefficients according to the source (e.g., radon or thoron) and form of exposure (e.g., internal or external). However, regardless of the source or form of exposure, the effects of a given estimated dose on human health are identical, assuming that individuals have the same sensitivity to radiation. Recently, radiation exceeding the annual dose limit of the general population (1 mSv/yr) was detected in bed mattresses produced by D company due to the use of a monazite-based anion powder containing uranium and thorium. This has sparked concerns about the health hazards for mattress users caused by radiation exposure. In light of this event, this study presents scientific information about the assessment of radon and thoron exposure and its human implications for human health, which have emerged as a recent topic of interest and debate in society.
Background Radiation ; Beds ; Carcinogens ; Construction Materials ; Epidemiologic Studies ; Humans ; Korea ; Leukemia ; Lung Neoplasms ; Miners ; Radiation Exposure ; Radon ; Smoke ; Smoking ; Thorium ; Thyroid Neoplasms ; Uranium

BACKGROUND: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. METHODS: Mouse AE II cell line MLE-12 were exposed to 1–10 µg/mL BLM and 0.01–100 µM baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. RESULTS: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor α, and transforming growth factor β1. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. CONCLUSION: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.
Animals ; Apoptosis ; Bisbenzimidazole ; Bleomycin ; Cell Cycle Checkpoints ; Cell Cycle ; Cell Line ; Cell Survival ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; Gene Expression ; Genes, vif ; Humans ; Idiopathic Pulmonary Fibrosis ; Interleukin-6 ; Lung ; Mice ; Propidium ; RNA, Messenger ; Transforming Growth Factors ; Tumor Necrosis Factor-alpha

Radon is a naturally occurring radioactive material formed by the slow decay of uranium and thorium found in the earth's crust or construction materials. Internal exposure to radon accounts for about half of the natural background radiation dose to which humans are exposed annually. Radon is a carcinogen and is the second leading cause of lung cancer following smoking. An association between radon and lung cancer has been consistently reported in epidemiological studies on mine workers and the general population with indoor radon exposure. However, associations have not been clearly established between radon and other diseases, such as leukemia and thyroid cancer. Radiation doses are assessed by applying specific dose conversion coefficients according to the source (e.g., radon or thoron) and form of exposure (e.g., internal or external). However, regardless of the source or form of exposure, the effects of a given estimated dose on human health are identical, assuming that individuals have the same sensitivity to radiation. Recently, radiation exceeding the annual dose limit of the general population (1 mSv/yr) was detected in bed mattresses produced by D company due to the use of a monazite-based anion powder containing uranium and thorium. This has sparked concerns about the health hazards for mattress users caused by radiation exposure. In light of this event, this study presents scientific information about the assessment of radon and thoron exposure and its human implications for human health, which have emerged as a recent topic of interest and debate in society.