Adrenergic and glucocorticoid modulation of the sterile inflammatory response

Exposure to an intense, acute stressor, in the absence of a pathogen, alters immune function. Exposure to a single bout of inescapable tail shock increases plasma and tissue concentrations of cytokines, chemokines, and the danger associated molecular pattern (DAMP) Hsp72. Although previous studies have demonstrated that adrenergic receptor (ADR) and glucocorticoid receptor (GCR)-mediated pathways alter pathogen or microbial associated molecular pattern (MAMP)-evoked levels of cytokines, chemokines, and Hsp72, far fewer studies have tested the role of these receptors across multiple inflammatory proteins or tissues to elucidate the differences in magnitude of stress-evoked sterile inflammatory responses. The goals of the current study were to (1) compare the sterile inflammatory response in the circulation, liver, spleen, and subcutaneous (SQ) adipose tissue by measuring cytokine, chemokine, and DAMP (Hsp72) responses; and (2) to test the role of alpha-1 (α₁), beta-1 (β₁), beta-2 (β₂), and beta-3 (β₃) ADRs, as well as GCRs in signaling the sterile inflammatory response. The data presented indicate plasma and SQ adipose are significantly more stress responsive than the liver and spleen. Further, administration of ADR and GCR-specific antagonists revealed both similarities and differences in the signaling mechanisms of the sterile inflammatory response in the tissues studied. Finally, given the selective increase in the chemokine monocyte chemotactic protein-1 (MCP-1) in SQ tissue, it may be that SQ adipose is an important site of leukocyte migration, possibly in preparation for infection as a consequence of wounding. The current study helps further our understanding of the tissue-specific differences of the stress-induced sterile inflammatory response.     

Functional characterization of histamine H4 receptor on human mast cells

Among the four different types of histamine receptors (H1-H4), H4R is predominantly expressed in immune cells and involved in immunomodulatory response. Here, in this study we determined the expression of H4R in human mast cells (HMC-1, LAD-2 and primary cord blood derived CD34+ human mast cells) and characterized its functional properties. Interestingly, we found that human mast cells responded to both histamine (natural ligand) and 4-methylhistamine (selective H4R agonist) for sustained intracellular calcium mobilization, degranulation and cytokine production. However, only histamine induced the release of cAMP, but 4-methylhistamine down regulates cAMP indicating that H4R mediates its effect through Gαi/o protein and H1R via Gαq protein. Furthermore, both histamine and 4-methylhistamine induced the production of cysteinyl leukotrienes and LTB4. Using human inflammation antibody array membrane, we found that H4R induced the expression of various inflammatory proteins, involving pro-inflammatory cytokines and chemokines and these are TGF-β1, TNF-α, TNF-β, PDGF-BB, TIMP-2, M-CSF, IP-10, IL-16, IL-6, IL-3, IL-10, MIP-1α, IL-1α, ICAM-1, Eotaxin-2, RANTES, IL-8, MCP-1, and IL-6sR. We also quantified the level of various inflammatory cytokines produced by human mast cells through H4R. It was observed that, the production level of Th2 cytokines IL-4(401.34 pg/ml), IL-5 (64.21 pg/ml) and IL-13 (1044 pg/ml) and classical proinflammatory cytokines IL-6 (221.27 pg/ml) and IL-1β (34.24 pg/ml) and chemokines MCP-1(106 pg/ml) and IL-8 (818.32 pg/ml). Furthermore, activation of H4R caused the phosphorylation of ERK and PI3 K in a time dependent manner. Taken together these data demonstrate that, the activation of H4R in human mast cells produced not only inflammatory mediators that are associated with allergic reactions but also other inflammatory conditions.     

牙龈卟啉单胞菌感染对血管内皮细胞与单核细胞黏附的影响

目的：研究Pg感染对人脐静脉内皮细胞（HUVEC）与单核细胞黏附的影响。方法：建立Pg感染HUVEC细胞株EA．hy926的体外模型,虎红染色法观察Pg381和Pg33277菌株感染HUVEC 6 h和24 h后,HUVEC与人单核细胞株THP-1细胞黏附量的变化。结果：培养6 h时Pg381感染组的THP-1细胞黏附量相对值为0．210±0．025,高于Pg33277感染组（0．078±0．024,P＜0．05）和空白对照组（0．062±0．022,P＜0．05）,Pg33277感染组和空白对照组无差异。24 h时3组间无差异（P＞0．05）。结论：Pg381感染HUVEC后能显著促进其与单核细胞的黏附。     

A three-dimensional in vitro model to quantify inflammatory response to biomaterials

In vivo models are the gold standard for predicting the clinical biomaterial–host response due to the scarcity of in vitro model systems that recapitulate physiological settings. However, the simplicity, control and relatively lower cost of in vitro models make them more appropriate to quantify the contribution by each cell, material and molecule within the healing environment. In this study, human fibroblasts and monocytes are co-cultured in a three-dimensional (3-D) tissue model to study foreign body response by observing morphological changes and monitoring inflammatory cytokine production with multiplex quantitative protein analysis. While control monocultures of either cell type alone produced low levels of cytokines, their interactions in co-culture led to morphological changes and increased release of inflammatory cytokines. When challenged with a well-characterized biopolymer, poly(lactic-co-glycolic acid), the co-cultured human cells secreted elevated levels of IL-1β, IL-6, GM-CSF and TNF-α. This 3-D in vitro co-culture model may serve as a building block towards a versatile platform to study mechanisms of material–host interactions by co-culturing cells with engineered phenotypes and reporter systems, or predict patient-specific biocompatibility by using the individual patients’ cells.     

The role of monocytes in models of infection by protozoan parasites

The confirmation of developmental differences between tissue macrophages and peripheral monocytes has changed our view of the functions and dynamics of these two important components of the innate immune system. It has been demonstrated conclusively that homeostasis of tissue resident macrophages is maintained by a low proliferative turn over. During an inflammatory response, bone marrow derived monocytes enter the tissue in large numbers and take part in the defense against the pathogens. After the destruction of invading pathogens, these cells disappear and tissue resident macrophages can be detected again. This new appreciation of the innate immune response has not only answered many outstanding questions regarding the role of the different myeloid cell types in inflammation, but also opened up new areas of research relating to the tissue- and pathogen-specific fate of the inflammatory macrophages or dendritic cells (DCs), and the transfer of this knowledge from mouse models to the human immune system. Nevertheless, there is still confusion in infection models, and especially in studies of human infections, as to what extent these recent observations and findings influence previous interpretations of data. This review will focus on insights from mouse models, summarize the literature on the ontogeny of macrophages and monocytes, explain the role of frequently used monocyte markers and effector molecules, and finally, discuss the role of inflammatory monocytes/macrophages/DCs in two experimental parasitic diseases.     

硬膜外腔注射芬太尼对胃癌根治术病人单核细胞、TNF-α表达的影响

目的 观察硬膜外芬太尼复合全麻较单纯全麻对胃癌根治术病人单核细胞TNF—α表达的影响。方法 择期手术病人60例，随机分为两组。Ⅰ组（n=30）术前芬太尼0．1mg注入硬膜外腔，术中每1．5h重复注入与术前相同剂量，同时复合全麻。Ⅱ组（n=30）单纯使用全身麻醉。于麻醉前、术终、术后24h采血，观察单核细胞TNF—α阳性细胞百分比。结果 两组病人单核细胞TNF—α阳性率术终和术后24h均较术前明显增高（P〈0．01）。术终单核细胞TNF—α阳性反应组Ⅰ低于组Ⅱ（P〈0．05）。结论 硬膜外芬太尼复合全身麻醉较单纯全身麻醉能明显抑制开腹手术病人单核细胞TNF—α阳性反应。     

双歧杆菌对单核细胞来源的树突状细胞成熟及功能影响的体外研究

目的 研究双歧杆菌对正常成年人外周血单核细胞来源的树突状细胞(dendritic cell, DC)刺激淋巴细胞增殖功能及分泌细胞因子的影响.方法 以GM-CSF、IL-4联合诱导单核细胞生成未成熟DC,加入不同剂量热灭活的双歧杆菌,观察DC形态,检测混合淋巴细胞反应,ELISA法测IL-12、IFN-γ分泌.结果 经双歧杆菌死菌刺激后,可获得具有典型树突状突起形态的DC;诱导后的DC刺激同种异体T淋巴细胞增殖的能力增强(P＜0.01),分泌IL-12、IFN-γ的水平提高(P＜0.05),呈剂量依赖型.结论 双歧杆菌能影响单核细胞来源的DC的分化、成熟及功能的发挥,且不同剂量的双歧杆菌对DC的成熟程度影响有差异.     

Effect of filtration leukocytapheresis therapy: modulation of white blood cell enzyme activities in patients with rheumatoid arthritis

We treated 12 patients with rheumatoid arthritis by filtration leukocytapheresis (FLCP) and evaluated its effect on leukocyte enzyme activities. We calculated the number of leukocytes removed and assessed the clinical response. We also evaluated the cellular enzyme activities of elastase and dipeptidylpeptidase IV (DPP IV). Out of 12 patients, 7 patients achieved 20% improvement for 4 weeks following FLCP. The FLCP treatment resulted in removal of 96% of granulocytes, 98% of monocytes, and 61% of lymphocytes. Granulocytes and monocytes with high elastase activity were effectively removed by FLCP. The elastase activity of granulocytes was increased 4 weeks after the last FLCP only in responders. On the other hand, the DPP IV activity of lymphocytes was low at 4 weeks after the last FLCP in responders. Modulation of leukocyte enzyme activities is one of the main effects of FLCP therapy and alteration of granulocytes, monocytes, and lymphocytes.