BACKGROUND & AIMS: :CD28, described as a T cell costimulatory molecule so far, is expressed on human peripheral blood neutrophils, as shown by cell surface staining and immunoprecipitation with anti-CD28 monoclonal antibody, and by reverse transcription PCR. The phorbol 12-myristate 13-acetate-augmented expression of CD28 on these cells can be blocked by actinomycin D, an RNA transcription inhibitor, and staurosporin, a protein kinase inhibitor. Cross-linking of CD28 results in an early increase in IL-8 receptor A (IL-8RA or CXCR-1) expression and a concurrent increase in IL-8-induced chemotaxis. The expression of CXCR-1 is down-regulated by receptor internalization 3 h after CD28 cross-linking with concurrent decrease in IL-8-induced chemotactic migration. Thus, our results demonstrate for the first time that CD28 is expressed on human peripheral blood neutrophils and that CD28 may play an important role in the regulation of IL-8RA expression and migration of neutrophils in response to IL-8.

背景与目标： ：迄今为止，被描述为T细胞共刺激分子的CD28在人外周血中性粒细胞中表达，如细胞表面染色和用抗CD28单克隆抗体进行的免疫沉淀以及逆转录PCR所示。这些细胞上CD28的佛波12-肉豆蔻酸酯13-醋酸酯增强表达可以被放线菌素D（一种RNA转录抑制剂）和星形孢菌素（一种蛋白激酶抑制剂）阻断。 CD28的交联导致IL-8受体A（IL-8RA或CXCR-1）表达的早期增加，并同时引起IL-8诱导的趋化性增加。 CD28交联后3小时，受体内化作用下调了CXCR-1的表达，并同时降低了IL-8诱导的趋化性迁移。因此，我们的结果首次证明了CD28在人外周血中性粒细胞上表达，并且CD28可能在调节IL-8RA表达和中性粒细胞对IL-8的迁移中起重要作用。

BACKGROUND & AIMS: :In this study we investigated the effect of adrenomedullin (AM) on fMLP-mediated activation of human neutrophils. AM partially, but significantly, suppressed fMLP-induced upregulation of CD11b expression. The inhibitory effects of AM upon fMLP-induced upregulation of CD11b expression were completely blocked by CGRP [8-37], a CGRP receptor antagonist. AM significantly increased cAMP content in neutrophils and SQ-22,536, an adenylate cyclase inhibitor, and KT-5720, a PKA inhibitor, significantly blocked the inhibitory effects of AM upon fMLP-induced upregulation of CD11b expression. This study indicates that binding of AM to the CGRP receptor suppresses fMLP-induced upregulation of CD11b expression of human neutrophils by increasing intracellular cAMP levels. AM may play an important role in the regulation of inflammatory processes, especially in the binding of neutrophils to vascular endothelial cells and subsequent neutrophil emigration evident in acute pulmonary inflammation.

背景与目标： ：在这项研究中，我们研究了肾上腺髓质素（AM）对fMLP介导的人类嗜中性粒细胞活化的影响。 AM部分但显着抑制fMLP诱导的CD11b表达上调。 CGRP [8-37]是一种CGRP受体拮抗剂，完全抑制了AM对fMLP诱导的CD11b表达上调的抑制作用。 AM显着增加了中性粒细胞和腺苷酸环化酶抑制剂SQ-22,536和PKA抑制剂KT-5720中的cAMP含量，从而显着阻断了AM对fMLP诱导的CD11b表达上调的抑制作用。这项研究表明，AM与CGRP受体的结合可通过增加细胞内cAMP水平来抑制fMLP诱导的人类嗜中性粒细胞CD11b表达的上调。 AM在炎症过程的调节中可能起重要作用，特别是在嗜中性粒细胞与血管内皮细胞的结合以及随后在急性肺部炎症中明显的嗜中性粒细胞迁移中。

BACKGROUND & AIMS: :Receptors involved in the phagocytosis of microorganisms under nonopsonic conditions have been little studied in neutrophils. Complement receptor type 3 (CR3) is a pattern recognition receptor able to internalize zymosan and C3bi-coated particles. We report that Abs directed against CR3 strongly inhibited nonopsonic phagocytosis of Mycobacterium kansasii in human neutrophils. In these cells CR3 has been found associated with several GPI-anchored proteins localized in cholesterol-rich microdomains (rafts) of the plasma membrane. Cholesterol sequestration by nystatin, filipin, or beta-cyclodextrin as well as treatment of neutrophils with phosphatidylinositol phospholipase C to remove GPI-anchored proteins from the cell surface markedly inhibited phagocytosis of M. kansasii, without affecting phagocytosis of zymosan or serum-opsonized M. kansasii. Abs directed against several GPI-anchored proteins inhibited phagocytosis of M. kansasii, but not of zymosan. N:-acetyl-D-glucosamine, which is known to disrupt interactions between CR3 and GPI proteins, also strongly diminished phagocytosis of these mycobacteria. In conclusion, phagocytosis of M. kansasii involved CR3, GPI-anchored receptors, and cholesterol. In contrast, phagocytosis of zymosan or opsonized particles involved CR3, but not cholesterol or GPI proteins. We propose that CR3, when associated with a GPI protein, relocates in cholesterol-rich domains where M. kansasii are internalized. When CR3 is not associated with a GPI protein, it remains outside of these domains and mediates phagocytosis of zymosan and opsonized particles, but not of M. kansasii.

背景与目标： ：在中性粒细胞中很少研究非调理条件下参与微生物吞噬作用的受体。 3型补体受体（CR3）是一种模式识别受体，能够内化酵母聚糖和C3bi包被的颗粒。我们报道，针对CR3的Abs强烈抑制人嗜中性粒细胞中堪萨斯分枝杆菌的非调理吞噬作用。在这些细胞中，已发现CR3与定位在质膜富含胆固醇的微区（筏）中的几种GPI锚定蛋白有关。制霉菌素，菲利普林或β-环糊精的胆固醇隔离以及用磷脂酰肌醇磷脂酶C处理嗜中性白细胞从细胞表面去除GPI锚定的蛋白显着抑制了堪萨斯支原体的吞噬作用，而又不影响酵母聚糖的吞噬作用或血清调理的M.堪萨斯州。针对几种GPI锚定蛋白的Abs抑制了堪萨斯分枝杆菌的吞噬作用，但抑制了酵母聚糖的吞噬作用。众所周知，N：-乙酰基-D-葡萄糖胺会破坏CR3和GPI蛋白之间的相互作用，也大大降低了这些分枝杆菌的吞噬作用。总之，堪萨斯分枝杆菌的吞噬作用涉及CR3，GPI锚定受体和胆固醇。相反，酵母聚糖或调理过的颗粒的吞噬作用涉及CR3，但不涉及胆固醇或GPI蛋白。我们建议CR3，当与GPI蛋白相关时，重新定位在坎萨西分枝杆菌被内化的富含胆固醇的域中。当CR3与GPI蛋白不相关时，它将保留在这些结构域之外，并介导酵母聚糖和调理过的颗粒的吞噬作用，而不是堪萨斯分枝杆菌的吞噬作用。

BACKGROUND & AIMS: :Asbestos exposure causes diffuse interstitial pulmonary fibrosis. Since alveolar epithelial cell injury is hypothesized to precede the fibrotic response in asbestosis, we investigated whether asbestos, either alone or in conjunction with neutrophils (PMNs), injured cultured human pulmonary epithelial cells (HPECs). HPEC cytotoxicity was assessed with a standard 51chromium release assay after a 16-hour incubation with asbestos and PMNs. Negligible HPEC cytotoxicity was observed after incubation with either amosite asbestos (500 micrograms/ml) or PMNs alone in serum-free media. However, incubation with both asbestos and PMNs caused significant HPEC injury, which was asbestos dose-dependent; causing 25% +/- 4% detachment and 52% +/- 8% 51chromium release with 500 micrograms/ml asbestos. The cytotoxic effects of asbestos plus PMNs were nearly completely attenuated with serum (20%) or catalase (100 micrograms/ml) but were not prevented with scavengers of superoxide anion, hydroxyl radical, or hypochlorous acid. A role for hydrogen peroxide (H2O2) in mediating HPEC injury was also suggested by the demonstration of asbestos-induced generation of H2O2 by PMNs. Furthermore, H2O2 alone (10(-4)mol/L) caused significant HPEC damage. Intimate contact between asbestos-activated PMNs and HPECs was a necessary requirement for PMN-mediated HPEC cytotoxicity. These data suggest that pulmonary epithelial cell injury is mediated in part by H2O2 release from asbestos-activated PMNs as well as intimate contact between the epithelial cell, PMNs, and asbestos.

背景与目标： ：石棉暴露引起弥漫性间质性肺纤维化。由于假设肺泡上皮细胞损伤先于石棉沉滞中的纤维化反应，所以我们调查了石棉（单独还是与中性粒细胞（PMN）联合使用）是否损伤了培养的人肺上皮细胞（HPEC）。与石棉和PMN孵育16小时后，使用标准的51铬释放分析法评估HPEC的细胞毒性。在无血清培养基中与铁石棉（500微克/毫升）或单独的PMN孵育后，观察到的HPEC细胞毒性可忽略不计。但是，与石棉和PMNs一起孵育会引起严重的HPEC损伤，这是石棉剂量依赖性的；导致25％/-4％的脱离和52％/-8％的51铬释放，含500微克/毫升的石棉。血清（20％）或过氧化氢酶（100微克/毫升）几乎完全减弱了石棉和PMN的细胞毒性作用，但用超氧阴离子，羟基自由基或次氯酸清除剂却无法阻止。 PMN在石棉诱导的H2O2生成中的作用也暗示了过氧化氢（H2O2）在介导HPEC损伤中的作用。此外，仅H2O2（10（-4）mol / L）会对HPEC造成严重损害。石棉激活的PMN和HPEC之间的紧密接触是PMN介导的HPEC细胞毒性的必要条件。这些数据表明，肺部上皮细胞损伤部分是由石棉激活的PMN释放的H2O2以及上皮细胞，PMN和石棉之间的紧密接触所介导的。

BACKGROUND & AIMS: :Peripheral blood polymorphonuclear neutrophils (PMN) suppressed the induction of PBL lymphokine-activated killer (LAK) function by rIL-2 in vitro. The suppression depended on the concentration of PMN in the IL-2 culture, and required intact PMN. However, PMN did not require treatment with immunoregulators such as IL-2, LPS, or TNF to express the suppressive activity, and no direct contact with PBL was needed for the suppression. Addition of anti-TNF antibodies had no effect on the suppression, suggesting that no endogenous TNF in the culture was involved in the suppression. PMN did not inhibit LAK function by preventing utilization of IL-2 by PBL or by selective depletion of NKH-1+ cells which constitute the majority of LAK precursors in PBL. The suppression was reversed by superoxide dismutase but not by catalase, suggesting that superoxide anion, not hydrogen peroxide, was involved in the suppression. No other suppressive factor was detectable in PMN culture supernates. Our results of PMN regulating LAK induction in vitro suggest that PMN may have a role in determining the outcome of immunotherapy with IL-2 in vivo.

背景与目标： ：外周血多形核中性粒细胞（PMN）在体外抑制了rIL-2对PBL淋巴因子激活的杀手（LAK）功能的诱导。抑制取决于IL-2培养物中PMN的浓度，并需要完整的PMN。但是，PMN不需要用免疫调节剂（例如IL-2，LPS或TNF）进行治疗即可表达抑制活性，并且不需要直接与PBL接触即可进行抑制。添加抗TNF抗体对抑制没有影响，表明培养物中没有内源性TNF参与抑制。 PMN不能通过阻止PBL或通过选择性消耗NKH-1细胞（构成PBL中大部分LAK前体）来利用IL-2来抑制LAK功能。抑制作用被超氧化物歧化酶逆转，但不被过氧化氢酶逆转，表明抑制作用涉及超氧阴离子而不是过氧化氢。在PMN培养上清液中未检测到其他抑制因子。我们的PMN在体外调节LAK诱导的结果表明，PMN可能在体内确定IL-2免疫疗法的结果中具有作用。

BACKGROUND & AIMS: :Nonopsonic interaction of host immune cells with pathogens is an important first line of defense. We hypothesized that nonopsonic recognition between type III group B streptococcus and human neutrophils would occur and that the interaction would be sufficient to trigger neutrophil activation. By using a serum-free system, it was found that heat-killed type III group B streptococci bound to neutrophils in a rapid, stable, and inoculum-dependent manner that did not result in ingestion. Transposon-derived type III strain COH1-13, which lacks capsular polysaccharide, and strain COH1-11 with capsular polysaccharide lacking terminal sialic acid demonstrated increased neutrophil binding, suggesting that capsular polysaccharide masks an underlying binding site. Experiments using monoclonal antibodies to complement receptor 1 and to the I domain or lectin site of complement receptor 3 did not inhibit binding, indicating that the complement receptors used for ingestion of opsonized group B streptococci were not required for nonopsonic binding. Nonopsonic binding resulted in rapid activation of cellular p38 and p44/42 mitogen-activated protein kinases. This interaction was not an effective trigger for superoxide production but did promote release of the proinflammatory cytokine interleukin-8. The release of interleukin-8 was markedly suppressed by the p38 mitogen-activated protein kinase inhibitor SB203580 but was only minimally suppressed by the mitogen-activated protein/extracellular signal-regulated kinase inhibitor PD98059. Thus, nonopsonic binding of type III group B streptococci to neutrophils is sufficient to initiate intracellular signaling pathways and could serve as an arm of innate immunity of particular importance to the immature host.

背景与目标： ：宿主免疫细胞与病原体的正典相互作用是重要的第一道防线。我们假设III型B组链球菌与人类嗜中性粒细胞之间会发生非调理性识别，并且这种相互作用足以触发嗜中性粒细胞的活化。通过使用无血清系统，发现热灭活的III型B组链球菌以快速，稳定和接种物依赖性的方式与嗜中性粒细胞结合，而不会导致摄入。缺乏荚膜多糖的转座子衍生的III型菌株COH1-13，以及缺少末端唾液酸的荚膜多糖的COH1-11菌株显示嗜中性粒细胞结合增加，表明荚膜多糖掩盖了潜在的结合位点。使用针对补体受体1以及补体受体3的I结构域或凝集素位点的单克隆抗体进行的实验不会抑制结合，这表明用于非调理结合不需要摄入调理B组链球菌所使用的补体受体。非光子结合导致细胞p38和p44 / 42丝裂原活化蛋白激酶的快速活化。这种相互作用不是产生超氧化物的有效触发器，但确实促进了促炎细胞因子白细胞介素8的释放。 p38丝裂原活化的蛋白激酶抑制剂SB203580明显抑制了白细胞介素8的释放，但丝裂原活化的蛋白/细胞外信号调节的激酶抑制剂PD98059仅最小程度地抑制了白介素8的释放。因此，III型B组链球菌与嗜中性粒细胞的非调理结合足以启动细胞内信号传导途径，并可作为对未成熟宿主特别重要的先天免疫的一部​​分。

BACKGROUND & AIMS: 1. Inositol hexakisphosphate (InsP6) is a ubiquitous and abundant cytosolic inositol phosphate that has been reported to prime human neutrophils for enhanced agonist-stimulated superoxide anion generation. This led to the proposal that the release of InsP6 from necrotic cells may augment the functional responsiveness of neutrophils at an inflammatory focus. The aim of this study was to examine whether the functional effects of InsP6 in neutrophils are receptor-mediated and establish the magnitude of this priming effect relative to other better characterized priming agents. 2. Analysis of [3H]-InsP6 binding to human neutrophil membranes in 20 mM Tris, 20 mM NaCl, 100 mM KCl, 5 mM EDTA (pH 7.7) buffer using 0.1 mg ml-1 membrane protein and 2.5 nM [3H]-InsP6 (90 min, 4 degrees C), demonstrated specific low affinity [3H]-InsP6 binding that was non-saturable up to a radioligand concentration of 10 nM. 3. [3H]-InsP6 displacement by InsP6 gave a Hill coefficient of 0.55 and best fitted a two-site logistic model (53% KD 150 nM, 47% KD 5 microM). [3H]-InsP6 binding also displayed low (3 fold) selectivity for InsP6 over Ins(1,3,4,5,6)P5. 4. The specific [3H]-InsP6 binding displayed a pH optimum of 8, was abolished by pre-boiling the membranes, and was enhanced by Ca2+, Mg2+ and Na+. 5. In incubations with intact neutrophils, where high levels of specific [3H]-LTB4 binding was observed, no [3H]-InsP6 binding could be identified. 6. Preincubation of neutrophils with 100 microM InsP6 had no effect on resting cell morphology, but caused a minor and transient (maximal at 30 s) enhancement of (0.1 nM) fMLP-induced shape change (% cells shape changedfMLP 53 +/- 3%, fMLP+InsP6 66 +/- 4%).

Similarly, InsP6 (100 microM, 30 s) had no effect on basal superoxide anion generation and, compared to lipopolysaccharide (LPS, 100 ng ml-1, 60 min), tumour necrosis factor-alpha (TNF alpha, 200 u ml-1, 30 min) or platelet-activating factor (PAF, 100 nM, 5 min) caused only a small enhancement of 100 nM fMLP-stimulated superoxide anion generation (fold-increase in superoxide anion generation over fMLP aloneInsP6 1.8 +/- 0.3, LPS 6.8 +/- 0.6, TNF alpha 5.2 +/- 0.7, PAF 5.8 +/- 0.6). 7. While these data support the presence of a specific, albeit low affinity, [3H]-InsP6 binding site in human neutrophil membrane preparations, the lack of binding to intact cells implies that the functional effects of InsP6 (ie. enhanced fMLP-stimulated superoxide anion generation and shape change) are not receptor-mediated.

背景与目标： 1.肌醇六磷酸肌醇（InsP6）是一种普遍存在且富含胞质的肌醇磷酸肌醇，据报道可引发人类嗜中性粒细胞，以增强激动剂刺激的超氧阴离子生成。这导致了一个提议，即从坏死细胞释放InsP6可能会增强炎症中心的嗜中性粒细胞的功能反应性。这项研究的目的是检查InsP6在嗜中性粒细胞中的功能作用是否是受体介导的，并确定这种引发作用相对于其他更好表征的引发剂的强度。 2.在使用0.1 mg ml-1膜蛋白和2.5 nM [3H]-的20 mM Tris，20 mM NaCl，100 mM KCl，5 mM EDTA（pH 7.7）缓冲液中分析[3H] -InsP6与人嗜中性白细胞膜的结合。 InsP6（90分钟，4摄氏度）显示出特定的低亲和力[3H] -InsP6结合，直到10nM的放射性配体浓度，该结合都是不饱和的。 3.用InsP6置换[3H] -InsP6的希尔系数为0.55，最适合两点逻辑模型（53％KD 150 nM，47％KD 5 microM）。与Ins（1,3,4,5,6）P5相比，[3H] -InsP6结合对InsP6的选择性也低（3倍）。 4.特定的[3H] -InsP6结合表现出最适的pH值为8，已通过预煮膜来消除，并被Ca2，Mg2和Na增强。 5.在与完整的中性粒细胞温育中，观察到高水平的特异性[3H] -LTB4结合，无法鉴定到[3H] -InsP6结合。 6.中性粒细胞与100 microM InsP6的预温育对静息细胞形态没有影响，但引起（0.1 nM）fMLP诱导的形状变化的轻微和短暂的增强（最大30 s）（％细胞形状变化fMLP 53 /-3％ ，fMLP InsP6 66 /-4％）。

同样，InsP6（100 microM，30 s）对基础超氧阴离子的生成没有影响，并且与脂多糖（LPS，100 ng ml-1，60分钟） ），肿瘤坏死因子-α（TNFα，200 u ml-1，30分钟）或血小板活化因子（PAF，100 nM，5分钟）仅引起100 nM fMLP刺激的超氧阴离子生成的少量增强（折叠-仅通过fMLP可以增加超氧阴离子的产生-InsP6 1.8 /-0.3，LPS 6.8 /-0.6，TNFα5.2 /-0.7，PAF 5.8 /-0.6）。 7.虽然这些数据支持人嗜中性粒细胞膜制剂中存在特定的，尽管亲和力很低的[3H] -InsP6结合位点，但缺乏与完整细胞结合的能力暗示了InsP6的功能作用（即增强的fMLP刺激超氧阴离子的产生和形状变化）不是受体介导的。

BACKGROUND & AIMS: :The MAPK family member JNK/stress-activated MAPK (SAPK) is involved in extracellular stress and proinflammatory cytokine responses, including production of cytokines such as IL-12. The JNK1 and 2 isoforms are widely expressed, but JNK3 is largely restricted to tissues of the brain, testis, and heart. In this study, we focus on mouse neutrophils, a cell type in which JNK/SAPK expression and activity has been given little study. We used Western blot analysis to examine expression patterns of JNK/SAPK in wild-type and JNK2-/- polymorphonuclear leukocytes (PMN). Surprisingly, neutrophils displayed a major deficiency in JNK1 expression, in contrast to macrophages that expressed high levels of both JNK1 and JNK2 MAPK. JNK1 expression was steadily reduced during the neutrophil maturation in bone marrow. We used PMN infection with the protozoan parasite Toxoplasma gondii to determine whether neutrophil JNK2 was functional. The parasite induced rapid JNK2 phosphorylation and intracellular FACS staining demonstrated preferential activation in infected neutrophils. Use of JNK2-/- neutrophils revealed that this MAPK family member was required for PMN IL-12p40 and CCL2/MCP-1 production. The chemotactic response displayed a minor JNK2 dependence but phagocytosis and oxidative burst activity did not require this MAPK. These findings are important because they demonstrate 1) a previously unrecognized unusual JNK expression pattern in mouse neutrophils, 2) JNK2 in PMN is activated by Toxoplasma invasion, and 3) a requirement for JNK2 in PMN IL-12p40 and CCL2/MCP-1 production in response to a microbial pathogen.

背景与目标： MAPK家族成员JNK /应激激活MAPK（SAPK）与细胞外应激和促炎性细胞因子反应有关，包括细胞因子如IL-12的产生。 JNK1和2亚型广泛表达，但JNK3在很大程度上限于大脑，睾丸和心脏的组织。在这项研究中，我们集中于小鼠嗜中性粒细胞，这是一种细胞类型，其中对JNK / SAPK表达和活性的研究很少。我们使用蛋白质印迹分析来检查JNK / SAPK在野生型和JNK2-/-多形核白细胞（PMN）中的表达模式。出乎意料的是，嗜中性粒细胞在JNK1表达中表现出主要缺陷，这与表达高水平JNK1和JNK2 MAPK的巨噬细胞相反。在骨髓中性粒细胞成熟过程中，JNK1表达稳定降低。我们使用原生动物寄生虫弓形虫PMN感染来确定中性粒细胞JNK2是否起作用。寄生虫诱导的快速JNK2磷酸化和细胞内FACS染色显示在感染的中性粒细胞中优先激活。使用JNK2-/-中性粒细胞表明该MAPK家族成员是PMN IL-12p40和CCL2 / MCP-1生产所必需的。趋化反应显示出较小的JNK2依赖性，但吞噬作用和氧化爆发活性不需要此MAPK。这些发现很重要，因为它们证明了1）小鼠嗜中性粒细胞中以前无法识别的异常JNK表达模式，2）PMN中的JNK2被弓形虫入侵激活，3）PMN IL-12p40和CCL2 / MCP-1生产中对JNK2的要求对微生物病原体的反应。

BACKGROUND & AIMS: :The expression and production of cytokines by cells of the innate immune system, including monocytes/macrophages, dendritic and NK cells, play a critical role not only in defensive and inflammatory but also in immunoregulatory and anti-/pro-tumoral processes. Studies performed in the last years have well ascertained that polymorphonuclear neutrophils can also be induced to express and produce chemokines, proinflammatory, anti-inflammatory, immunoregulatory, angiogenic and fibrogenic cytokines, as well as ligands belonging to the TNF superfamily. Among the latter group of molecules, B-cell-activating factor (BAFF)/B lymphocyte stimulator (BLyS), known to be essential for B lymphocyte homeostasis and related pathologies, has recently been identified as one of the factors potentially expressed by human neutrophils. The addition of this novel TNF superfamily member, and more recently also of the closely related "A Proliferation-Inducing Ligand" (APRIL), to the list of cytokines produced by neutrophils not only testifies to the continuous growth of this area of investigation, but also implies the involvement of neutrophils in B-cell-dependent autoimmune diseases and tumors.

背景与目标： 天然免疫系统细胞（包括单核细胞/巨噬细胞，树突状细胞和NK细胞）表达和产生细胞因子，不仅在防御和炎症中起着关键作用，而且在免疫调节和抗肿瘤/促肿瘤过程中也起着至关重要的作用。近年来进行的研究已经确定，多形核中性粒细胞也可以被诱导表达和产生趋化因子，促炎，抗炎，免疫调节，血管生成和纤维生成细胞因子，以及属于TNF超家族的配体。在后一组分子中，B细胞活化因子（BAFF）/ B淋巴细胞刺激物（BLyS）是已知的B淋巴细胞稳态和相关病理必不可少的，最近已被确定为人类嗜中性粒细胞潜在表达的因子之一。在嗜中性粒细胞产生的细胞因子列表中增加了这种新的TNF超家族成员，以及最近与之密切相关的“增殖诱导配体”（APRIL），不仅证明了这一研究领域的持续发展，而且证明了这一领域的持续发展。还暗示嗜中性粒细胞参与B细胞依赖性自身免疫性疾病和肿瘤。

BACKGROUND & AIMS: :The majority of peripheral serotonin is stored in platelets, which secrete it on activation. Serotonin releases Weibel-Palade bodies (WPBs) and we asked whether absence of platelet serotonin affects neutrophil recruitment in inflammatory responses. Tryptophan hydroxylase (Tph)1–deficient mice, lacking non-neuronal serotonin, showed mild leukocytosis compared with wild-type (WT), primarily driven by an elevated neutrophil count. Despite this, 50% fewer leukocytes rolled on unstimulated mesenteric venous endothelium of Tph1(-/-) mice. The velocity of rolling leukocytes was higher in Tph1(-/-) mice, indicating fewer selectin-mediated interactions with endothelium. Stimulation of endothelium with histamine, a secretagogue of WPBs, or injection of serotonin normalized the rolling in Tph1(-/-) mice. Diminished rolling in Tph1(-/-) mice resulted in reduced firm adhesion of leukocytes after lipopolysaccharide treatment. Blocking platelet serotonin uptake with fluoxetine in WT mice reduced serum serotonin by > 80% and similarly reduced leukocyte rolling and adhesion. Four hours after inflammatory stimulation, neutrophil extravasation into lung, peritoneum, and skin wounds was reduced in Tph1(-/-) mice, whereas in vitro neutrophil chemotaxis was independent of serotonin. Survival of lipopolysaccharide-induced endotoxic shock was improved in Tph1(-/-) mice. In conclusion, platelet serotonin promotes the recruitment of neutrophils in acute inflammation, supporting an important role for platelet serotonin in innate immunity.

背景与目标： ：大多数外周血清素都储存在血小板中，血小板在激活时会分泌出来。血清素释放Weibel-Palade体（WPB），我们询问是否缺乏血小板血清素会影响中性粒细胞在炎症反应中的募集。缺乏非神经元5-羟色胺的色氨酸羟化酶（Tph）1缺陷小鼠与野生型（WT）相比，表现出轻度白细胞增多，主要是由于嗜中性白细胞计数升高所致。尽管如此，Tph1（-/-）小鼠未经刺激的肠系膜静脉内皮上滚动的白细胞减少了50％。 Tph1（-/-）小鼠中滚动白细胞的速度较高，表明较少的选择素介导的与内皮的相互作用。组胺刺激，外周血白蛋白的促分泌素或血清素注射刺激正常化Tph1（-/-）小鼠的滚动。 Tph1（-/-）小鼠的滚动减少导致脂多糖治疗后白细胞牢固粘附减少。氟西汀在野生型小鼠中阻断血小板对血清素的摄取，使血清素减少了80％以上，同样降低了白细胞的滚动和粘连。炎症刺激后四个小时，Tph1（-/-）小鼠中性粒细胞向肺，腹膜和皮肤伤口的渗出减少，而体外中性粒细胞趋化性独立于5-羟色胺。 Tph1（-/-）小鼠中脂多糖诱导的内毒素休克的存活率得到改善。总之，血小板5-羟色胺可促进中性粒细胞在急性炎症中的募集，支持血小板5-羟色胺在先天免疫中的重要作用。

BACKGROUND & AIMS: :Superoxide production by neutrophils triggered with a chemotactic peptide or a phorbol ester is inhibited by the protein kinase antagonists staurosporine or 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7). We evaluated the effects of these antagonists on the protein tyrosine kinases and protein kinase C activities of neutrophils. Staurosporine completely inhibited all of these enzymes, whereas 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine was only substantially effective against protein kinase C. Thus, if a protein tyrosine kinase is involved in superoxide production, it is likely to function with a second kinase sensitive to 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine.

背景与目标： ：由趋化肽或佛波醇酯触发的嗜中性粒细胞产生的超氧化物被蛋白激酶拮抗剂星形孢菌素或1-（5-异喹啉基磺酰基）-2-甲基哌嗪（H-7）抑制。我们评估了这些拮抗剂对嗜中性粒细胞蛋白酪氨酸激酶和蛋白激酶C活性的影响。星形孢菌素完全抑制所有这些酶，而1-（5-异喹啉基磺酰基）-2-甲基哌嗪仅对蛋白质激酶C有效。对1-（5-异喹啉基磺酰基）-2-甲基哌嗪敏感的激酶。

BACKGROUND & AIMS: :Hypersensitivity pneumonitis (HP) is mediated by Th1 immune response. NKT cells regulate immune responses by modulating the Th1/Th2 balance. Therefore, we postulated that NKT cells play a critical role in the development of the HP by modulating the Th1/Th2 response. To address this issue, we explored the functional roles of NKT cells in Saccharopolyspora rectivirgula (SR)-induced HP. In CD1d(-/-) mice, the HP was worse in terms of histological changes, hydroxyproline levels, the CD4:CD8 ratio in bronchoalveolar lavage fluid, and SR-specific immune responses than in control mice. CD1d(-/-) mice showed elevated IFN-gamma production in the lung during the HP, and this was produced mainly by Gr-1+ neutrophils. The blockade of IFN-gamma in CD1d(-/-) mice attenuated the HP, whereas the injection of rIFN-gamma aggravated it. Moreover, the depletion of Gr-1+ neutrophils reduced CD8+ T cell numbers in bronchoalveolar lavage fluid during the HP. The adoptive transfer of IL-4(-/-) mouse NKT cells did not attenuate the HP, whereas wild-type or IFN-gamma(-/-) mouse NKT cells suppressed the HP. In conclusion, NKT cells producing IL-4 play a protective role in SR-induced HP by suppressing IFN-gamma-producing neutrophils, which induce the activation and proliferation of CD8+ T cells in the lung.

背景与目标： 超敏性肺炎（HP）是由Th1免疫反应介导的。 NKT细胞通过调节Th1 / Th2平衡来调节免疫反应。因此，我们推测NKT细胞通过调节Th1 / Th2反应在HP的发展中起关键作用。为了解决此问题，我们探讨了NKT细胞在糖多孢菌直肠病毒（SR）诱导的HP中的功能作用。在CD1d（-/-）小鼠中，在组织学变化，羟脯氨酸水平，支气管肺泡灌洗液中CD4：CD8比值和SR特异性免疫反应方面，HP较对照组小鼠差。 CD1d（-/-）小鼠在HP期间在肺中显示升高的IFN-γ产生，这主要是由Gr-1中性粒细胞产生的。 CD1d（-/-）小鼠中IFN-γ的阻滞减弱了HP，而rIFN-γ的注射加重了HP。此外，在HP期间，Gr-1中性粒细胞的消耗减少了支气管肺泡灌洗液中CD8 T细胞的数量。 IL-4（-/-）小鼠NKT细胞的过继转移不会减弱HP，而野生型或IFN-γ（-/-）小鼠NKT细胞则抑制HP。总之，产生IL-4的NKT细胞通过抑制产生IFN-γ的嗜中性粒细胞在SR诱导的HP中起保护作用，该嗜中性粒细胞诱导肺中CD8 T细胞的活化和增殖。

BACKGROUND & AIMS: :During the resolution phase of inflammation, the 'corpses' of apoptotic leukocytes are gradually cleared by macrophages. Here we report that during the resolution of peritonitis, the CCR5 chemokine receptor ligands CCL3 and CCL5 persisted in CCR5-deficient mice. CCR5 expression on apoptotic neutrophils and activated apoptotic T cells sequestered and effectively cleared CCL3 and CCL5 from sites of inflammation. CCR5 expression on late apoptotic human polymorphonuclear cells was downregulated by proinflammatory stimuli, including tumor necrosis factor, and was upregulated by 'proresolution' lipid mediators, including lipoxin A4, resolvin E1 and protectin D1. Our results suggest that CCR5+ apoptotic leukocytes act as 'terminators' of chemokine signaling during the resolution of inflammation.

背景与目标： 在炎症消退阶段，凋亡白细胞的“尸体”被巨噬细胞逐渐清除。在这里，我们报告在解决腹膜炎期间，CCR5趋化因子受体配体CCL3和CCL5在CCR5缺陷型小鼠中持续存在。 CCR5在凋亡中性粒细胞和活化的凋亡T细胞上的表达被隔离并有效地从炎症部位清除了CCL3和CCL5。晚期凋亡的人多形核细胞上的CCR5表达被促炎刺激（包括肿瘤坏死因子）下调，并被“促分辨率”脂质介质（包括脂蛋白A4，resolvin E1和保护素D1）上调。我们的结果表明，CCR5凋亡性白细胞在炎症消退过程中充当趋化因子信号传导的“终止子”。

BACKGROUND & AIMS: :Neutrophils have been reported to acquire surface expression of MHC class II and co-stimulatory molecules as well as T-cell stimulatory activities when cultured with selected cytokines. However, cellular identity of those unusual neutrophils showing antigen presenting cell (APC)-like features still remains elusive. Here we show that both immature and mature neutrophils purified from mouse bone marrow differentiate into a previously unrecognized "hybrid" population showing dual properties of both neutrophils and dendritic cells (DCs) when cultured with granulocyte macrophage-colony-stimulating factor but not with other tested growth factors. The resulting hybrid cells express markers of both neutrophils (Ly6G, CXCR2, and 7/4) and DCs (CD11c, MHC II, CD80, and CD86). They also exhibit several properties typically reserved for DCs, including dendritic morphology, probing motion, podosome formation, production of interleukin-12 and other cytokines, and presentation of various forms of foreign protein antigens to naïve CD4 T cells. Importantly, they retain intrinsic abilities of neutrophils to capture exogenous material, extrude neutrophil extracellular traps, and kill bacteria via cathelicidin production. Not only do our results reinforce the notion that neutrophils can acquire APC-like properties, they also unveil a unique differentiation pathway of neutrophils into neutrophil-DC hybrids that can participate in both innate and adaptive immune responses.

背景与目标： ：嗜中性粒细胞据报道在与选定的细胞因子一起培养时会获得II类MHC和共刺激分子的表面表达以及T细胞的刺激活性。但是，那些表现出抗原呈递细胞（APC）样特征的异常嗜中性粒细胞的细胞特性仍然难以捉摸。在这里，我们显示从小鼠骨髓中纯化的未成熟和成熟的嗜中性粒细胞分化为先前无法识别的“杂种”种群，与粒细胞巨噬细胞集落刺激因子一起培养时，嗜中性粒细胞和树突状细胞（DC）具有双重性质，但未与其他测试的嗜中性粒细胞一起培养生长因子。所得的杂交细胞表达中性粒细胞（Ly6G，CXCR2和7/4）和DC（CD11c，MHC II，CD80和CD86）的标志物。它们还表现出通常保留给DC的几种特性，包括树突形态，探测运动，足小体形成，白介素12和其他细胞因子的产生，以及将各种形式的外源蛋白抗原呈递给幼稚的CD4 T细胞。重要的是，它们保留了嗜中性粒细胞捕获外源物质，挤出嗜中性粒细胞胞外捕获物并通过cathelicidin产生杀死细菌的内在能力。我们的结果不仅增强了中性粒细胞可以获得类似APC的特性的观念，而且还揭示了中性粒细胞向中性粒细胞-DC杂交体的独特分化途径，该途径可以参与先天性和适应性免疫应答。

BACKGROUND & AIMS: :We performed a comparative analysis of functional activity of neutrophils in patients with type 2 diabetes mellitus with and without symptoms of CHD. Enhanced H2O2 production by neutrophils in response to N-formyl-Met-Leu-Phe (fMLP) was found in patients with type 2 diabetes mellitus. In patients with type 2 diabetes mellitus associated with CHD, fMLP-induced release of myeloperoxidase from azurophilic granules of neutrophils was reduced and plasma myeloperoxidase level was elevated. Increased peroxidase activity of myeloperoxidase, reduced plasma catalase activity, and increased levels of TBA-reactive lipid peroxidation products and oxidized glutathione were detected in patients of both groups. Since myeloperoxidase is an important neutrophilic mediator of oxidative stress, its increased activity in the blood can be an additional marker of oxidative stress and cardiovascular risk in patients with diabetes mellitus.

背景与目标： ：我们对患有和不患有冠心病症状的2型糖尿病患者中性粒细胞的功能活性进行了比较分析。在2型糖尿病患者中，嗜中性粒细胞对N-甲酰-Met-Leu-Phe（fMLP）的反应增强了H2O2的产生。在患有CHD的2型糖尿病患者中，fMLP诱导的嗜中性粒细胞嗜酸性颗粒中髓过氧化物酶的释放减少，血浆髓过氧化物酶水平升高。两组患者均检测到髓过氧化物酶的过氧化物酶活性增加，血浆过氧化氢酶活性降低，TBA反应性脂质过氧化产物和氧化型谷胱甘肽水平升高。由于髓过氧化物酶是氧化应激的重要嗜中性介质，因此其血液中活性的增加可能是糖尿病患者氧化应激和心血管风险的另一个标志。