商陆皂甙辛对小鼠脾脏细胞产生IL-3和IL-6的影响

利用MTT法测定了商陆皂甙辛 (EH )对ConA诱导的小鼠脾淋巴细胞IL 3和IL 6活性的影响 ,采用地高辛标记斑点杂交法测定了IL 3mRNA和IL 6mRNA的表达水平。结果显示EH在 1 0～ 1 0 0 μg/ml浓度范围内能增强ConA诱导的小鼠脾淋巴细胞IL 3和IL 6活性 ,伴随mRNA水平的上升。提示EH通过提高IL 3和IL 6基因转录水平 ,从而使其产物活性增强 ,是其免疫调节和提高造血功能的分子机制之一     

Andrographolide Suppresses IL-6/Stat3 Signaling in Peripheral Blood Mononuclear Cells from Patients with Chronic Rhinosinusitis with Nasal Polyps

The mechanisms underlying the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remain largely unknown. CRSwNP has garnered considerable public health concern owing to its high incidence and unsatisfactory treatment outcomes. Herbal remedies are promising candidates for the treatment of CRSwNP. We examined the utility of andrographolide, a diterpenoid lactone extracted from the Chinese herb Andrographis paniculata, an anti-inflammatory agent for CRSwNP treatment by evaluating interleukin (IL)-6 and IL-17 production and monitoring T helper 17 (Th17) differentiation of peripheral blood mononuclear cells (PBMCs) isolated from 20 Chinese CRSwNP patients and 11 control subjects. All CRSwNP patients exhibited clinical features of CRSwNP. Andrographolide significantly inhibited IL-6 and IL-17 production, suppressed p-Stat3 expression, and inhibited Th17 differentiation of PBMCs in vitro. These findings suggested that andrographolide has useful anti-inflammatory properties and could be used for the treatment of CRSwNP.     

Enrichment of IL-2-Producer T Cells from Mouse Spleen by Use of Bauhiniapurpurea Lectin

Bauhinia purpurea agglutinin (BpA) was used to enrich the interleukin 2 (IL 2)-producing T-cell subset from mouse spleen. IL 2 was found to be produced by the BpA-non-agglutinated (BpA-) T cells of mouse spleen on stimulation with concanavalin A (Con A). The degree of IL 2 production by BpA-agglutinated (BpA+) T cells was significantly low. The proliferative response to Con A of BpA- T cells was not influenced by the addition of exogenous IL 2, whereas BpA+ T cells were partially dependent on the exogenous addition of IL 2 in the proliferative response.     

Interleukin-4 Receptor–Stat6 Signaling in Murine Infections with a Tissue-Dwelling Nematode Parasite

Interleukin-4 (IL-4) has been shown to be crucial in parasite expulsion in several gastrointestinal nematode infection models. Data from both epidemiological studies with humans and experimental infections in animals imply a critical role for the type II helper response, dominated by IL-4, in host protection. Here we utilized inbred mice on two distinct backgrounds to document the involvement of IL-4 in the clearance of a primary infection of Brugia from the murine host. Our data from infections of IL-4 receptor(-/-) and Stat6(-/-) mice further indicate that IL-4 exerts its effects by activating the Stat6 molecule in host target cells, a finding which links clearance requirements of a gastrointestinal tract-dwelling nematode with those of a tissue-dwelling nematode. Additionally, we show that the requirements for IL-4 receptor binding and Stat6 activation extend to accelerated clearance of a secondary infection as well. The data shown here, including analysis of cell populations at the site of infection and infection of immunoglobulin E (IgE)(-/-) mice, lead us to suggest that deficiencies in eosinophil recruitment and isotype switching to IgE production may be at least partially responsible for slower parasite clearance in the absence of IL-4.     

IL-4 and IL-13 Stimulate Human Bronchial Epithelial Cells to Release IL-8

Cytokine networks are important in regulating the traffic of inflammatory cells in the airways. Interleukin-8 (IL-8) released by human bronchial epithelial cells (HBECs) is thought to be of particular importance in attracting neutrophils and monocytes to sites of inflammation. Increased release of IL-8 by HBECs in response to Th-1 cytokines such as TNF alpha and IL-1 beta may be an important pathophysiologic pathway. The present study was designed to explore the role of the Th2 cytokine IL-4 and the functionally related interleukins IL-10, and IL-13 on the regulation of IL-8 release by HBECs. HBECs (passage 4–6) were cultured in LHC9/RPMI and when confluent cells were stimulated in unsupplemented medium LHCD/RPMI by IL-4, IL-10, and IL-13 at 10 ng/ml concentration for all cytokines. TNF alpha stimulation was used as a positive control. After 24 hours supernatants were collected and tested for IL-8 by a sandwich ELISA. Unstimulated HBECs spontaneously released limited amounts of IL-8 (11 ± 1 pM) and significantly increased cytokine production in response to IL-4 (42 ± 1 pM), IL-13 (30 ± 1 pM) and TNF (128 ± 11 pM). Stimulation with IL-10 (11 ± 1 pM) did not change basal production of IL-8. When HBECs were co-stimulated with IL-4 plus TNF, the production of IL-8 was further increased (204 ± 5 pM). In contrast, IL-10 attenuated the effect of TNF during co-stimulation (82 ± 5 pM). IL-13 did not affect the release of IL-8 induced by TNF (111 ± 9 pM). Northern blot analysis of IL-8 mRNA levels showed the highest induction of IL-8 mRNA in HBECs co-stimulated with TNF and IL-4. We conclude from our study that IL-4 directly induces IL-8 release from HBECs and amplifies the release of IL-8 in response to TNF alpha. IL-13 is less active and IL-10 has an inhibitory effect. Airway epithelial cells are able to interact, therefore, with products of both Th1 and Th2 cells with respect to modulating release of IL-8.     

白细胞介素6促人脾NK细胞活性及其机制初探

本文采用~(51)Cr释放试验测定人重组白细胞介素6(HrIL-6)处理正常成人睥单个核细胞(MNC)24小时后,NK细胞抗K562靶细胞的杀伤活性。结果证实,IL-6能明显增强脾NK细胞的抗瘤活性(P<0.05,与对照组比较),且可诱导MNC产生IL-2,NK活性的增强效应和MNCIL-2的合成水平均与IL-6的处理剂量呈依赖关系,提示IL-6促人脾NK活性的机制可能是通过诱导脾细胞产生IL-2而实现的。     

Increased Susceptibility of Periodate-Treated Tumour Cells to Human but Not to Mouse or Rat Natural Killer Cells

Treatment of intact cells in the cold with low concentrations (1 mM) of sodium meta periodate (PI) selectively oxidizes the surface-exposed sialic acid residues to the corresponding aldehydes. Such treated tumour cells show greatly enhanced sensitivity to lysis by fresh human NK cells but not to mouse or rat NK cells. Reduction of the PI-treated cells with sodium borohydride (NaBH4) reduced their NK sensitivity to that of untreated cells. In target conjugate formation assays PI-treated tumour cells displayed a higher binding capacity than control cells or PI+NaBH4-treated cells to both mouse and human effector cells. Neuraminidase treatment of K562 and Molt-4 increased target susceptibility to human NK cells but not to mouse, whereas the susceptibility of Yac-1 cells was left unchanged using both human and mouse effector cells. The same pattern of reactivity is shown in the target binding assay. These findings indicate that subtle molecular changes in the surface-exposed carbohydrates of target cells might have a fundamental impact on their sensitivity to lysis by NK cells from certain species, and that in cross species effector-target combinations a higher binding capacity is not sufficient for increased lysis to occur.     

Stat6-independent GATA-3 autoactivation directs IL-4-independent Th2 development and commitment

The initial source of IL-4-inducing Th2 development and the mechanism of stable Th2 commitment remain obscure. We found the reduced level of IL-4 production in Stat6-deficient T cells to be significantly higher than in Th1 controls. Using a novel cell surface affinity matrix technique, we found that IL-4-secreting Stat6-deficient T cells stably expressed GATA-3 and Th2 phenotype. Introducing GATA-3 into Stat6-deficient T cells completely restored Th2 development, inducing c-Maf, Th2-specific DNase I hypersensitive sites in the IL-4 locus, and Th2 cytokine expression. The fact that GATA-3 fully reconstitutes Th2 development in Stat6-deficient T cells indicates it is a master switch in Th2 development. Finally, GATA-3 exerts Stat6-independent autoactivation, creating a feedback pathway stabilizing Th2 commitment.     

Hypoxia Modulates Melanoma Cells Proliferation and Apoptosis via miRNA-210/ISCU/ROS Signaling

Bulletin of Experimental Biology and Medicine - In this study, luciferase reporter assay was used to establish the relationship between miR-210 and ISCU. This research was performed on both cell...     

Tissue-Specific Expression of Splice Variants of Human IL-4 and IL-6 Gene mRNA

The spectrum of alternatively spliced IL-4 and IL-6 gene mRNA was studied in peripheral blood mononuclears from healthy donors and in human fetal tissues. It was found that the expression of alternatively spliced IL-4 and IL-6 gene mRNA in fetal tissues is tissue specific and that hemopoiesis- and immunopoiesis-related tissues differ by the amount of IL-4 and IL-4δ2 mRNA. An mRNA variant IL-4alt3 carrying partial exon 3 deletion was for the first time identified in human mononuclear cells.     

In Vitro Bactericidal and Associated Metabolic Activities of Mouse Spleen Cells

Spleen cell suspensions from AKR and CD-1 mice are able to kill Escherichia coli in vitro. The optimal ratio of splenocytes to bacteria for this activity is 1: 1. Incubation of these cells with inert polystyrene latex spherules (0.81 mum diameter) results in a fourfold increase in glucose-1-(14)C oxidation. Under these conditions, there is also a 2.5-fold increase in both reduced nicotinamide adenine dinucleotide phosphate oxidase activity and formate oxidation. Spleen cell fractions have been shown to have significant peroxidase activity. This has been quantitated by the guaiacol oxidation method. The 20,000 x g pellet fraction of spleen cell homogenate can kill E. coli when H(2)O(2) and chloride ions are added and the reaction is carried out at pH 5.5 and 37 C.     

Baicalin Induced Apoptosis of Human Cholangiocarcinoma Cell through Activating AMPK/mTORC1/p70S6K Signaling Pathway

Bulletin of Experimental Biology and Medicine - Baicalin (naturally bioactive flavone compound isolated from Scutellaria baicalensis) has been demonstrated to exert strong anticancer activity...     

BMP4/Smad Signaling Pathway Induces the Differentiation of Mouse Spermatogonial Stem Cells via Upregulation of Sohlh2

Spermatogonial stem cells (SSCs) capable of self‐renewal and differentiation are the foundation for spermatogenesis. Although several factors that govern these processes have been investigated, the underlying molecular mechanisms have not been fully elucidated. Here, we investigated the role of BMP4 in mouse SSC differentiation, and found that SSCs cultured in the presence of BMP4 underwent differentiation, characterized by downregulation of SSC self‐renewal markers, Plzf, and upregulation of SSC differentiation marker, c‐kit. Smad1/5/8 proteins were phosphorylated during BMP4‐induced differentiation. The effects of BMP4 on SSCs were blocked by BMP4 inhibitor (Dorsomorphin). The activation of BMP4/Smad signaling pathway in SSCs increased the expression of Sohlh2, which is involved in the early differentiation of spermatogonia. Knockdown sohlh2 expression by RNA interference abolished the effect of BMP4 on SSC differentiation and the upregulation of c‐kit expression. Overall, our results suggest that BMP4 plays an important role during the early differentiation of SSCs via upregulation of sohlh2. Anat Rec, 297:749–757, 2014. © 2014 Wiley Periodicals, Inc.     

应用牛乳头瘤病毒载体产生人IL-4哺乳类细胞株的建立

将PHA活化的人外周血白细胞cDNA文库与人工合成的人IL-4探针杂交,解离一段cDNA序列,将其插入πH3M质粒并导入COS猿猴细胞;经FACS测定证实细胞培养上清中具有人IL-4的生物学活性;再把经改建的pdBhIL-4载体转染小鼠C127细胞,建立人IL-4的高表达细胞株。     

Membrane Antigenic Changes Associated with Pha Transformation of Mouse Spleen Cells in Vitro

The changes in lymphocyte surface antigens associated with blast transformation have been studied by membrane immunofluorescence. When mouse spleen cells were stimulated in vitro with phytohaemagglutinin for 4 days virtually all of the resulting blast cells were shown to have histocompatibility (H-2^d), theta(θ)and mouse-specific T lymphocyte (MTLA) antigens; approximately 50% had detectable surface immunoglobulin and 70% mouse-specific B lymphocyte antigen (MBLA). The intensity of the staining reactions on blast cells was increased for the anti-MTLA serum and reduced for the anti-immunoglobulin and anti-MBLA sera when compared with the staining of fresh spleen cells. No difference was detected for the anti-θ and anti-H-2^d sera. Cap formation was detected on fresh spleen cells labelled with any of the antisera, but after stimulation only anti-immunoglobulin-treated blasts showed the cap pattern.

The simultaneous expression of B-and T-cell markers on PHA transformed cells suggests a convergence of the lymphocyte differentiation pathways.