Interleukin-4-triggered, STAT6-dependent production of a factor that induces mouse mast cell apoptosis

IL-4 can suppress mast cell development from mouse spleen, bone marrow and peritoneal cells by an indirect process that is dependent on the presence of macrophages. Mast cells undergo apoptosis when exposed to supernatants collected from cultures of IL-4-stimulated peritoneal cells due to the IL-4-induced production of an apoptosis-inducing factor in the cultures. This effect of IL-4 is shown to be dependent on STAT6 signaling, because IL-4 and IL-13 do not suppress mast cell development from the spleen and peritoneal cells of STAT6-/- mice. Moreover, supernatants from cultures of IL-4- and IL-13-stimulated peritoneal cells of STAT6-/- mice do not exhibit apoptosis-inducing activity. We confirm, by using deficient mice, neutralizing antibodies and recombinant cytokines, that IL-4-induced apoptosis is not related to the well-known apoptosis-inducing factors Fas, Fas ligand, TNF-alpha, TRAIL, TGF-beta or perforin. These results demonstrate a novel mechanism whereby IL-4 and IL-13 can suppress mast cell development by inducing the production of an apoptosis-inducing factor from macrophages.     

Hapten-specific tolerance induced by acute, low-dose ultraviolet B radiation of skin requires mast cell degranulation

The deleterious effects of ultraviolet B radiation (UVR) on cutaneous immunity are mediated in part by cytokines released from cutaneous cells following radiation exposure. On the one hand, TNF-alpha has been advocated as the primary mediator of failed contact hypersensitivity induction, and, on the other hand, IL-10 has been held responsible for tolerance. While keratinocytes exposed to UVR have been found to produce both TNF-alpha and IL-10, there is reason to question whether these major cellular constituents of the epidermis are the relevant source of immunomodulatory cytokines after UVR. Dermal mast cells also produce TNF-alpha and IL-10, and we have recently reported that mast cell-derived TNF-alpha is required for UVR-induced impairment of CH induction. In this study, we have examined whether mast cells are also a relevant source of IL-10 in UVR-dependent tolerance. We found that (a) UVR fails to induce tolerance in mast cell-deficient mice, and (b) that tolerance occurs if mast cells are triggered to degranulate after ligation of the IgE receptor. Both types of tolerance were neutralized with anti-IL-10 antibodies, are hapten specific, and are associated with regulatory lymphoid cells. We conclude that mast cells are required in UVR-induced tolerance and may be one of the major sources of IL-10 that mediates the tolerance induced by acute, low-dose UVR.     

Inhibition of substance P-induced histamine release from rat peritoneal mast cells by ultraviolet light B irradiation: decreased intracellular calcium as a partial mechanism

Background Ultraviolet light irradiation has been shown to suppress immunological reactions of irradiated individuals, however, less attention has been paid to the effects on neurogenic inflammation. Objective We have investigated the effect of ultraviolet light B (UVB) irradiation on substance P (SP)-induced histamine release from rat mast cells. Methods Rat peritoneal mast cells were treated with UVB irradiation and challenged by SP. Histamine release from the cells and intracellular calcium concentrations ([Ca²⁺]i) in the cells were measured. Results UVB irradiation at doses used in the present study did not induce histamine release from mast cells. UVB irradiation at doses from 25 to 200mJ/cm² inhibited 10^–5mol/L SP-induced histamine release in a dose-dependent manner. On the other hand, SP-induced increase of [Ca²⁺]_i was inhibited by UVB irradiation only at doses of 100 and 200 mJ/cm². Conclusion These data suggest that UVB irradiation inhibits histamine release from SP-activated rat peritoneal mast cells partially through the suppression of an increase in [Ca²⁺]_i.     

Molecular mechanism of monoamine toxicity in Parkinson's disease: hypothetical cell death model

Although there have been experimental approaches to understanding the etiology of Parkinson's disease, the cause of cell degeneration in this neurological disorder remains a mystery. Herein, a hypothetical model is proposed to explain the mechanism leading neurons to die. The model is based on recent experimental evidence and it attempts to dissect the actions of dopamine and metal ions as potential triggers for the activation of an ordered cascade of events of the cell death machinery.     

Ultraviolet B irradiation induces skin accumulation of plasmacytoid dendritic cells: A possible role for chemerin

Abstract

Photosensitivity represents a common feature for most forms of lupus erythematosus (LE) including cutaneous LE. Skin inflammatory infiltrates in response to ultraviolet (UV) exposure are closely involved in the development of skin lesions of LE patients. Skin-infiltrating plasmacytoid dendritic cells (pDCs), considered as a hallmark of cutaneous LE, contribute to its pathogenesis via the production of type I interferons (IFNs). Chemerin, a recently identified chemoattractant for pDCs through its functional receptor chemR23, was found to be elevated in skin lesions of LE patients. The aim of this study was to investigate the effect of UVB irradiation on skin pDC recruitment and chemerin expression. We found that UVB irradiation induced a rapid but transient influx of pDCs as well as a persistent infiltration of neutrophils and macrophages in the mouse skin. The mRNA expression levels of IRF-7, IFN-α and chemR23 were increased in UVB-irradiated skin. Furthermore, UVB irradiation up-regulated skin chemerin production and pDC accumulation in parallel, both of which reached their peaks simultaneously (24?h post-irradiation). Dermal fibroblasts seemed to be the major source of chemerin as evidenced by significantly increased chemerin secretion by UVB-irradiated dermal fibroblasts. More importantly, LE-prone MRL/lpr mouse exhibited greatly increased skin pDC accumulation and chemerin production in response to UVB irradiation, indicating their contributions to increased susceptibility of photosensitivity in the MRL/lpr mouse. Thus, our findings demonstrated that elevated chemerin expression positively correlates with pDC accumulation in UVB-irradiated skin, suggesting a role of chemerin in mediating skin recruitment of pDCs in response to UVB exposure.     

IL-17 induces apoptosis of vascular endothelial cells: a potential mechanism for human acute coronary syndrome

Th17 cells producing IL-17 are involved in the pathogenesis of atherosclerosis, but the underlying mechanisms remain unclear. In this study, we investigated the effects of IL-17 on human vascular endothelial cells and showed that IL-17 induced cell death of the vascular endothelial cells, which played a pivotal role in plaque destabilization triggering acute coronary syndrome (ACS). We showed that circulating Th17 cells and IL-17 increased in patients with ACS compared to the patients with stable angina or health individuals; the plasma levels of IL-6 increased but TGF-β decreased in ACS patients, exhibiting a positive and negative correlation with that of IL-17, respectively. Importantly, we uncovered that IL-17 promoted the production of von Willebrand factor by endothelial cells and induced endothelial apoptosis by activating caspase-3, caspase-9 and up-regulating the ratio of Bax/Bcl-2, indicating the function of IL-17 in vascular endothelial damage as a potential mechanism for the pathogenesis of human ACS.     

Lifestyle factors and cancer of the pancreas: a hypothetical mechanism

The interaction of a genetically determined protease inhibitor, the enzymes whose functions are modified by that inhibitor and lifestyle factors, such as cigarette smoking, high lipid diet and alcohol consumption, are considered key factors in a proposed protease-antiprotease imbalance mechanism for pancreatic oncogenesis. Epidemiologic and experimental laboratory evidence in support of the mechanism is presented along with a discussion of suggested research initiatives to further test the hypothesis.     

Men considering a hypothetical treatment for prostate cancer: a comparison to patients

BACKGROUND: In facilitating informed decision making about PSA screening, men are often asked to consider the potential consequences of the test, including a diagnosis of prostate cancer and how they would want to be treated. However, there is no empirical evidence thus far demonstrating that men are able to consider this hypothetical situation in a realistic manner. PURPOSE: To compare the features (attributes) of treatments that are important to non-patient men considering a hypothetical diagnosis of prostate cancer with those deemed important to men actually diagnosed with early-stage disease. METHODS: Two groups of men went through a decision aid interview to help them choose between treatment options for early-stage prostate cancer: non-patient men who imagined themselves to be diagnosed with the disease, and newly diagnosed patients. During the interview participants identified features of the treatment and disease that were important to their decisions. RESULTS: The percentage of non-patients and patients that thought particular attributes were important was correlated: r (33) = 0.77, p < 0.01. The effects on bladder and bowel functioning were considered important to >or=50% of each group. In addition to the 22 attributes initially presented, 49% of non-patients and 67% of patients identified additional attributes as being important to their decision. Eight (42%) of the 19 additional attributes were identified by non-patients and patients alike. CONCLUSIONS: The group of non-patient men provided a close approximation to the group of newly diagnosed men with respect to the attributes identified as being important to their treatment decisions for early-stage prostate cancer, suggesting that the consideration of what is important to the decision by non-patient men is realistic.     

羟基喜树碱通过激活NF-κB诱导人乳腺癌Bcap-37细胞凋亡

 目的: 探讨NF-κB信号转导途径在羟基喜树碱诱导人乳腺癌Bcap-37细胞凋亡中所起的作用。方法: 突变IκB-α-pcDNA3.1(-)质粒转染Bcap-37细胞，并用G418进行阳性克隆选择；采用MTT法测定细胞的增殖活力，琼脂糖凝胶电泳观察细胞凋亡，流式细胞仪进行细胞周期分析，Western blotting法和DIG-EMSA研究细胞凋亡途径。结果: 羟基喜树碱对Bcap-37有明显的生长抑制作用，琼脂糖凝胶电泳检测到典型的DNA梯状条带；羟基喜树碱可通过降解IκBα 蛋白进而激活NF-κB，并使NF-κB发生核移位；羟基喜树碱没有激活突变IκB-α-pcDNA3.1(-)质粒转染细胞的NF-κB，并且转染细胞凋亡受到抑制。结论: 羟基喜树碱对Bcap-37有明显的生长抑制和诱导凋亡作用；NF-κB 信号转导途径在羟基喜树碱诱导人乳腺癌Bcap-37细胞凋亡中起着促进凋亡的作用。     

Bevacizumab induces A549 cell apoptosis through the mechanism of endoplasmic reticulum stress in vitro

Aims: To observe the effect of bevacizumab on human A549 cells and explore its mechanism. Methods: After different concentrations (0 μM, 1 μM, 5 μM, 25 μM) of bevacizumab treating in A549 cells, CCK8 assay detect the impact of bevacizumab on A549 cell proliferation and flow cytometry determine the effect of bevacizumab on human A549 cells apoptosis. Real-time PCR and Western blotting detect the changing expression of the target gene (CHOP, caspase-4, IRE1, XBP-1) on mRNA and Protein level. Results: Treatment with bevacizumab for 24-hr have induced cell death in a does-dependent manner dramatically (P<0.05). In terms of the mRNA level, expression of XBP-1 has increased obviously in each group (1 μM, 5 μM, 25 μM) (P<0.01); the expression of CHOP (25 μM) and caspase-4 (5 μM) have increased slightly (P<0.05). In terms of the protein level, the expression of CHOP has increased obviously in each group (1 μM, 5 μM, 25 μM) when compared with the control group (0 μM) (P<0.05). As for caspase-4 (5 μM, 25 μM), the expression have increased slightly when compared with the control group (0 μM) (P<0.05). Conclusion: Bevacizumab can induce A549 cell apoptosis through the mechanism of endoplasmic reticulum stress.     

A blood-borne antigen induces rapid T-B cell contact: a potential mechanism for tolerance induction

Understanding the difference between the development of a productive T‐cell response and tolerance is central to discerning how the immune system functions. Intravenous injection of soluble protein is thought to mimic the presentation of self‐serum and orally introduced antigens. It is generally toleragenic. The current view is that this outcome reflects the failure of ‘immunogenic’ dendritic cells to relocate to the T‐cell zone of the secondary lymphoid tissues. Here, using a peptide/I‐E^k tetramer and antibodies to stain splenic sections, we showed that antigen‐specific T cells were activated in the spleen within hours of injection or feeding of protein. The activated T cells were found to be located at the T–B junction, the bridging zone and the B‐cell area, interacting directly with B cells. In addition, B cells gain the ability to present antigen. Our results suggest a way for T cells to be stimulated by blood‐borne antigen presented by naïve B cells, a potential mechanism of tolerance induction.     

纳米银抗菌作用机制：剂量依赖性促进细菌凋亡

 背景：纳米银具有高效、广谱抗菌性及不易产生耐药性等优点,已成为目前抗菌材料的研究热点,但目前尚不明确其抗菌的确切机制。目的：研究纳米银抵抗细菌生长的机制。方法：采用抑菌环实验检测Ti、TiO₂及载纳米银TiO₂对单克隆大肠杆菌和金黄色葡萄球菌的抑制作用;将大肠杆菌接种于LB液体培养基中,分别加入0,5,10 mg/L的纳米银溶液,检测24 h内的菌液A值,进行琼脂糖凝胶电泳,分析细菌DNA的变化,应用Annexin V和PI双染色,采用流式细胞术检测细菌凋亡。结果与结论：Ti和TiO₂对金黄色葡萄球菌和大肠杆菌无明显抑制现象,在TiO₂管载纳米银材料片周围出现了明显抑菌环。加入纳米银溶液后,大肠杆菌菌液A值明显降低,并且随着加入纳米银质量浓度的增加,降低趋势更明显。加入纳米银溶液后,大肠杆菌的DNA含量明显减少,并且减少程度随加入纳米银溶液质量浓度的增加而增加。加入纳米银后,细菌Annexin V阳性率增加,并且这种增加趋势与加入纳米银的质量浓度呈正比。表明纳米银可通过促进细菌凋亡这一机制来影响细菌生长。 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

Knockdown of KDM1B inhibits cell proliferation and induces apoptosis of pancreatic cancer cells

Pancreatic cancer (PC) is one of the common malignant tumors in digestive tract with a high fatality rate. The oncogenic role of lysine-specific demethylase1 (LSD1/KDM1?A) has been well recognized in PC. While, the role of its homolog LSD2 (KDM1B) in regulating PC progression is poorly understood. In this study, we attempted to evaluate the functional role of KDM1B in PC cells. The expression of KDM1B was detected by immunohistochemistry and immunoblotting in PC tissues and cells. Lentivirus-mediated shRNA was applied to silence KDM1B in PANC-1 and SW1990 cells. Cell proliferation was measured by MTT and Celigo assay. Cell apoptosis was determined by both Caspase-Glo^®3/7 assay and Flow cytometry. Intracellular signaling molecules were detected using a PathScan intracellular signaling array kit. In this study, we found KDM1B was highly expressed in PC tissues compared to paracancerous tissues. Moreover, elevated expression of KDM1B was detected in PC cell lines (BxPC-3, CFPAC-1, PANC-1 and SW1990) as compared with a normal human pancreatic duct epithelial cell line (HPDE6-C7). Further investigations revealed that KDM1B knockdown significantly inhibited PC cell proliferation. Furthermore, the apoptosis of PANC-1 and SW1990 cells was significantly increased after KDM1B knockdown. Notably, the activations of p-ERK1/2, p-Smad2, p-p53, cleaved PARP, cleaved Caspase-3, cleaved Caspase-7, p-eIF2a and Survivin were promoted by KDM1B knockdown, while IkBa was suppressed. Taken together, our findings provided new insights into the critical and multifaceted roles of KDM1B in the regulation of cell proliferation and apoptosis, and offered a potentially novel target in preventing the progression of PC.     

Nitric oxide induces murine thymocyte apoptosis by oxidative injury and a p53-dependent mechanism.

Previously, we showed that NO induces thymocyte apoptosis via a caspase-1-dependent mechanism [(1) ]. In the present study, we investigated the role of heme oxygenase, catalase, bax, and p53 in this process. The NO donor, S-nitroso-N-acetyl penicillamine (SNAP), induced DNA fragmentation in thymocytes in a time- and concentration-dependent way. SNAP (100 microM) induced 50--60% apoptosis; higher doses did not increase the rate of apoptosis significantly. SNAP decreased catalase and heme iron (Fe) levels without affecting superoxide dismutase, glutathione, or total Fe stores in thymocytes. SNAP significantly increased the expression of heme oxygenase 1 (HSP-32), p53, and bax but not bcl-2. Treatment with the heme oxygenase inhibitor, tin protoporphyrin IX inhibited SNAP-induced thymocyte apoptosis. Furthermore, thymocytes from p53 null mice were resistant to NO-induced apoptosis. Our data suggest that NO may induce its cytotoxic effects on thymocytes by modulating heme oxygenase and catalase activity as well as up-regulating pro-apoptotic proteins p53 and bax.     

Granzyme B leakage-induced apoptosis is a crucial mechanism of cell death in nasal-type NK/T-cell lymphoma

This study aims to investigate the role of granzyme B in the apoptosis of nasal-type NK/T-cell lymphoma. Twenty-four nasal-type NK/T-cell lymphomas were examined by TdT-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling (TUNEL) assay and immunohistochemical staining for active caspase 3, poly(ADP-ribose) polymerase (PARP-1/p85)/p85, and Bcl-2. In addition, HANK-1 and NKL cell lines were analyzed using Western blot analysis. Immunoprecipitation was performed to identify the binding of granzyme B and intrinsic serpin proteinase inhibitor 9 (PI-9). To localize granzyme B, immunogold labeling and immunofluorescence staining were performed. The expression level of granzyme B in tumor tissue was correlated with the apoptosis rate (P=0.015), degree of necrosis (P=0.002), and the levels of active caspase 3 (P=0.036) and poly ADP-ribose polymerase (PARP)-1/p85 (P=0.040). The granzyme B-positive HANK-1 cell line showed increased spontaneous cell death compared to the granzyme B-negative NKL cell line. The untreated HANK-1 cells released cytochrome c into the cytosol with cleavage of caspase 3 and PARP-1. Treatment with granzyme B inhibitor and caspase inhibitor decreased the cleavage of PARP-1. By performing immunogold labeling, granzyme B was identified within the cytolytic granules as well as in the cytosol. Confocal microscopy and immunoprecipitation assays confirmed the colocalization of PI-9 and granzyme B, which formed an SDS-resistant complex. These results suggested that granzyme B leakage induces cell death in NK/T-cell lymphomas via both caspase-dependent and -independent mechanisms, and this leads to the extensive necrosis that is commonly seen in NK/T-cell lymphoma.     

Codeine induces human mast cell chemokine and cytokine production: involvement of G-protein activation

BACKGROUND: Activation of mast cells and the systemic release of histamine are common side effects of opiates such as codeine and morphine. In some individuals, codeine not only elicits a sizable early response due to mast cell degranulation, but can also lead to late cutaneous allergic inflammation possibly through the production of chemokines. However, individuals who exhibit a late phase reaction to codeine often do not react to its synthetic analog, meperidine. The goal of this study was to test whether codeine and meperidine induce secretion of inflammatory mediators in human mast cells. METHODS: To characterize opiate activation of human mast cells, we stimulated cultured human (LAD2 cell line and CD34+-derived) mast cells with codeine and meperidine and measured degranulation and chemokine production. RESULTS: Codeine, but not meperidine, activated human mast cell degranulation within 30 min in a dose-dependent manner. Degranulation was blocked by the phosphoinositol-3 kinase (PI3K) inhibitor, wortmannin, and pertussis toxin but not by Ro-31-8220, a PKC inhibitor or forskolin, a cyclic adenylyl cyclase activator. After 3 and 8 h of stimulation, codeine, but not meperidine, activated human mast cells to release monocyte chemoattractant protein-1 (CCL2), regulated on activation, normal T expressed and secreted (RANTES, CCL5) and interleukin-8 (CXCL 8) but not inducible protein-10 (CXCL10). CONCLUSIONS: Codeine activates human mast cell degranulation and chemokine production by activating protein kinase A and PI3 kinase, possibly leading to NF-kappaB activation. Therefore, opiates may regulate late phase allergic inflammation by activating chemokine production by human mast cells.     

Burkholderia pseudomallei induces cell fusion and actin-associated membrane protrusion: a possible mechanism for cell-to-cell spreading

Burkholderia pseudomallei, a facultative intracellular bacterium, is the causative agent of a broad spectrum of diseases collectively known as melioidosis. Its ability to survive inside phagocytic and nonphagocytic cells and to induce multinucleated giant cell (MNGC) formation has been demonstrated. This study was designed to assess a possible mechanism(s) leading to this cellular change, using virulent and nonvirulent strains of B. pseudomallei to infect both phagocytic and nonphagocytic cell lines. We demonstrated that when the cells were labeled with two different cell markers (CMFDA or CMTMR), mixed, and then infected with B. pseudomallei, direct cell-to-cell fusion could be observed, leading to MNGC formation. Staining of the infected cells with rhodamine-conjugated phalloidin indicated that immediately after the infection, actin rearrangement into a comet tail appearance occurred, similar to that described earlier for other bacteria. The latter rearrangement led to the formation of bacterium-containing, actin-associated membrane protrusions which could lead to a direct cell-to-cell spreading of B. pseudomallei in the infected hosts. Results from 4', 6'-diamidine-2-phenylindole dihydrochloride (DAPI) nuclear staining, poly-ADP ribose polymerase cleavage, staining of infected cells for phosphatidylserine exposure with annexin V, and electrophoresis of the DNA extracted from these infected cells showed that B. pseudomallei could kill the host cells by inducing apoptosis in both phagocytic and nonphagocytic cells.