BACKGROUND & AIMS: :We have constructed a physical map of a > 2-Mb region on mouse chromosome 6 that contains the natural killer gene complex (NKC). The map comprises a contig of 14 overlapping yeast artificial chromosomes onto which we positioned 25 NKC markers. NKC genetically linked genes encode > 17 proteins that directly control innate NK cell-mediated tumor lysis and disease resistance. Herein we show that Nkrp1 genes are clustered in a region flanked by A2m and Cd69 genes and that most Ly49 genes are clustered in a distal region -1 Mb distant. Importantly, syntenic intervals of mouse chromosome 6 and human chromosome 12p that include the NKC are conserved. NKC species conservation suggests that the human NKC may contain orthologues for the mouse viral disease resistance genes, Cmv1 and Rmp1. The high-resolution NKC map will facilitate investigation of NKC gene regulation and identification of phenotypically defined gene products that confer NK cell defense against viral pathogens.

背景与目标： ：我们在小鼠染色体6上构建了一个大于2-Mb区域的物理图，其中包含自然杀伤基因复合体（NKC）。该图包括14个重叠的酵母人工染色体的重叠群，我们在其上定位了25个NKC标记。 NKC遗传连锁基因编码> 17种蛋白质，这些蛋白质直接控制先天NK细胞介导的肿瘤溶解和疾病抵抗力。在本文中，我们显示Nkrp1基因聚集在A2m和Cd69基因侧翼的区域中，而大多数Ly49基因聚集在距离-1 Mb远的区域中。重要的是，保留了包含NKC的小鼠6号染色体和人类12p号染色体的同音间隔。 NKC物种保守性表明，人NKC可能含有小鼠病毒疾病抗性基因Cmv1和Rmp1的直向同源物。高分辨率的NKC图谱将有助于NKC基因调控的研究和表型定义的基因产物的鉴定，这些产物赋予NK细胞防御病毒病原体的能力。

BACKGROUND & AIMS: :6-Pyruvoyltetrahydropterin synthase (PTPS) catalyzes the second step of tetrahydrobiopterin (BH4) synthesis. We previously identified PTPS orthologs (bPTPS-Is) in bacteria which do not produce BH4. In this study we disrupted the gene encoding bPTPS-I in Synechococcus sp. PCC 7942, which produces BH4-glucoside. The mutant was normal in BH4-glucoside production, demonstrating that bPTPS-I does not participate in BH4 synthesis in vivo and bringing us a new PTPS ortholog (bPTPS-II) of a bimodular polypeptide. The recombinant Synechococcus bPTPS-II was assayed in vitro to show PTPS activity higher than human enzyme. Further computational analysis revealed the presence of mono and bimodular bPTPS-II orthologs mostly in green sulfur bacteria and cyanobacteria, respectively, which are well known for BH4-glycoside production. In summary we found new bacterial PTPS orthologs, having either a single or dual domain structure and being responsible for BH4 synthesis in vivo, thereby disclosing all the bacterial PTPS homologs.

背景与目标： ：6-丙酮酰基四氢蝶呤合成酶（PTPS）催化四氢生物蝶呤（BH4）合成的第二步。我们先前在不产生BH4的细菌中鉴定了PTPS直系同源物（bPTPS-Is）。在这项研究中，我们破坏了Synechococcus sp。中编码bPTPS-1的基因。 PCC 7942，生产BH4-葡萄糖苷。该突变体在BH4-葡萄糖苷生产中是正常的，表明bPTPS-I不参与体内BH4合成，并为我们带来了双模块多肽的新PTPS直向同源物（bPTPS-II）。在体外对重组Synechocooccus bPTPS-II进行了分析，结果表明PTPS活性高于人的酶。进一步的计算分析表明，分别在绿色硫细菌和蓝细菌中分别存在单和双模块bPTPS-II直系同源物，这对于BH4-糖苷的生产是众所周知的。总之，我们发现了新的细菌PTPS直系同源物，具有单域或双域结构并负责体内的BH4合成，从而揭示了所有细菌PTPS同源物。

BACKGROUND & AIMS: :Epithelial cells play an important role in orchestrating mucosal immune responses. In allergic-type inflammation, epithelial cells control the recruitment of eosinophils into the mucosa. Th2-type cytokine-driven release of eosinophil-active chemokines from epithelial cells directs eosinophil migration into the mucosal epithelium. CCR3, the main eosinophil chemokine receptor, regulates this process; however, the respective contribution of individual CCR3 ligands in eosinophil transepithelial migration is less well understood. Using an in vitro transepithelial chemotaxis system, we found that eotaxin-3 produced by IL-4-stimulated airway epithelial cells and CCR3 on eosinophils exclusively mediate eosinophil transepithelial migration. Eotaxin-3 protein levels were also increased in the nasal mucosal epithelium recovered from allergic patients as compared to non-allergic patients. Surprisingly, eotaxin-3 in IL-4-stimulated airway epithelial cells was predominantly cell surface bound, and the cell surface form was critical for eosinophil transepithelial migration. Eotaxin-3 cell surface association was partially glycosaminoglycan (GAG) dependent, but was completely protein dependent, suggesting that eotaxin-3 associates with both GAG and cell surface proteins. We thus provide evidence that cell surface-associated eotaxin-3 is the critical IL-4-dependent chemotactic signal mediating eosinophil transepithelial migration in the setting of allergic inflammation.

背景与目标： ：上皮细胞在协调粘膜免疫反应中起重要作用。在过敏型炎症中，上皮细胞控制嗜酸性粒细胞向粘膜的募集。 Th2型细胞因子驱动的嗜酸性粒细胞活性趋化因子从上皮细胞的释放引导嗜酸性粒细胞迁移到粘膜上皮中。 CCR3是主要的嗜酸性粒细胞趋化因子受体，调节这一过程。然而，单个CCR3配体在嗜酸性粒细胞上皮细胞迁移中的各自作用尚不清楚。使用体外经上皮趋化系统，我们发现由IL-4刺激的气道上皮细胞和嗜酸性粒细胞上的CCR3产生的eotaxin-3专门介导嗜酸性粒细胞跨上皮迁移。与非过敏患者相比，从过敏患者中回收的鼻粘膜上皮中的Eotaxin-3蛋白水平也有所增加。出人意料的是，IL-4刺激的气道上皮细胞中的eotaxin-3主要与细胞表面结合，并且细胞表面形式对于嗜酸性粒细胞跨上皮迁移至关重要。 Eotaxin-3细胞表面缔合部分依赖糖胺聚糖（GAG），但完全依赖蛋白质，这表明eotaxin-3与GAG和细胞表面蛋白缔合。因此，我们提供证据，在过敏性炎症的情况下，细胞表面相关的eotaxin-3是介导嗜酸性粒细胞跨上皮迁移的关键IL-4依赖性趋化信号。

BACKGROUND & AIMS: A primer extension (toeprinting) assay was used to monitor selection by ribosomes of the first versus the second AUG codon as a function of introducing mutations on the 3' side (positions +4, +5 and +6) of the first AUG codon. Six different flanking codons starting with G (GCG, GCU, GCC, GCA, GAU and GGA) strongly augmented selection of AUG#1 when compared with matched mRNAs that had A or C instead of G in position +4. Augmentation by G in position +4 failed only when it was combined with U in position +5, as in the sequence augGUA. In contrast with the usual enhancing effect of introducing G in position +4, most mutations in position +5 had no discernible effect, as shown with the series augANA (where N = C, A, G or U) and the series augCNA. AUG codon recognition was also unaffected by mutations in position +6, as shown by testing four mRNAs that had augCCN as the start site. Thus the primary sequence context that augments the recognition of AUG start codons does not appear generally to extend beyond G in position +4. When the toeprinting assay was used with mRNAs that initiate translation at CUG instead of AUG, cugGAU was not recognized better than cugGGU, contradicting the hypothesis that initiation at non-AUG codons might be favored by A instead of G in position +5.

背景与目标： 使用引物延伸（印迹）测定法来监测第一和第二AUG密码子的核糖体的选择，其作为在第一AUG密码子的3'侧（位置4、5和6）上引入突变的函数。与匹配的在位置4有A或C而不是G的mRNA相比，以G开头的六个不同的侧翼密码子（GCG，GCU，GCC，GCA，GAU和GGA）大大增强了对AUG＃1的选择。仅当它与位置5的U组合时失败，如序列augGUA所示。与在位置4引入G的通常增强作用相反，在位置5的大多数突变没有明显的作用，如augANA系列（其中N = C，A，G或U）和augCNA系列所示。 AUG密码子识别也不受位置6突变的影响，如测试以augCCN为起始位点的四个mRNA所显示的。因此，增强对AUG起始密码子识别的一级序列通常不会延伸到位置4的G范围之外。当将脚印法与在CUG而非AUG处起始翻译的mRNA一起使用时，cugGAU的识别度不如cugGGU，与以下假设相矛盾：在非AUG密码子处的起始可能会被位置5的A而不是G所偏爱。

BACKGROUND & AIMS: :IL-7 signals are crucial for the survival of naive and memory T cells, and the IL-7R is expressed on the surface of these cells. Following viral infection, the IL-7R is expressed on only a subset of effector CD8 T cells, and has been demonstrated to be important for the survival of these memory precursors. IL-7 message levels remain relatively constant during the T cell response to lymphocytic choriomeningitis virus, but a short-lived burst of GM-CSF is observed soon after infection. Retroviral expression of a chimeric GM-CSF/IL-7R, in which binding of GM-CSF by T cells leads to IL-7 signaling, allows for the delivery of an IL-7 signal in all effector T cells expressing the receptor. In mice infected with lymphocytic choriomeningitis virus, CD8 and CD4 T cells transduced with this chimeric receptor underwent an enhanced proliferative response compared with untransduced populations in the same host. Similarly, TCR transgenic CD8 cells expressing the chimeric receptor produced higher effector numbers during the peak of the T cell response to infection. Surprisingly, the enhanced proliferation did not lead to higher memory numbers, as the subsequent contraction phase was more pronounced in the transduced cell populations. These findings demonstrate that artificial IL-7 signaling during an infection leads to significantly increased Ag-specific effector T cell numbers, but does not result in increased numbers of memory progeny. The extent of contraction may be dictated by intrinsic factors related to the number of prior cell divisions.

背景与目标： ：IL-7信号对于幼稚和记忆T细胞的存活至关重要，而IL-7R在这些细胞的表面表达。病毒感染后，IL-7R仅在效应CD8 T细胞的一部分上表达，并已证明对这些记忆前体的存活很重要。在对淋巴细胞性脉络膜脑膜炎病毒的T细胞应答过程中，IL-7信息水平保持相对恒定，但是感染后不久就观察到了短暂的GM-CSF爆发。嵌合GM-CSF / IL-7R的逆转录病毒表达（其中T细胞与GM-CSF的结合导致IL-7信号传导）允许在表达该受体的所有效应T细胞中传递IL-7信号。在感染了淋巴细胞性脉络膜脑膜炎病毒的小鼠中，用该嵌合受体转导的CD8和CD4 T细胞与同一宿主中未转导的种群相比，具有增强的增殖反应。同样，表达嵌合受体的TCR转基因CD8细胞在感染的T细胞反应高峰期间产生更高的效应子数量。出人意料的是，增强的增殖并未导致更高的记忆数，因为随后的收缩期在转导的细胞群中更为明显。这些发现表明，感染期间的人工IL-7信号转导会导致Ag特异性效应T细胞数量显着增加，但不会导致记忆后代数量增加。收缩的程度可以由与先前细胞分裂数有关的内在因素决定。

BACKGROUND & AIMS: :Immunocompromise after a major injury is presumed to be a predisposing factor for sepsis. Mice subjected to sublethal cecal ligation and puncture (CLP) and challenged 5 days later with Pseudomonas aeruginosa had more bacterial growth in lung tissue, lower serum interferon gamma (IFN-gamma) and interleukin (IL) 12,and higher serum IL-10 when compared with sham CLP mice challenged with Pseudomonas. To test the functional significance of these alterations in cytokine production in the immune response to bacteria, we administered IFN-gamma and anti-IL-10 to post-CLP mice before the Pseudomonas challenge. Administration of IFN-gamma and anti-IL-10 did not improve bacterial clearance or mortality in post-CLP mice. In further studies, we administered IFN-gamma to IL-10 knockout mice before a challenge with P. aeruginosa. Our results showed no significant differences in bacterial clearance or mortality in IL-10 knockout mice with or without IFN-gamma treatment compared with wild-type controls. Finally, because most mortality occurred within 2 to 3 days of the Pseudomonas challenge in the aforementioned studies and was likely associated with a marked proinflammatory response, we investigated the effect of IFN-gamma and anti-IL-10 on clearance of Pseudomonas in C3H/HeJ mice, which do not mount an exaggerated proinflammatory response to endotoxin or Gram-negative bacteria. Neither clearance of the Pseudomonas bacteria nor mortality was improved in C3H/HeJ mice receiving anti-IL-10 and IFN-gamma. These results suggest that the suppressed IFN-gamma and IL-12 responses, in combination with an exaggerated IL-10 response to P. aeruginosa challenge after injury, do not correlate with bacterial clearance or survival.

背景与目标： ：大伤后的免疫功能低下被认为是败血症的诱因。进行半致死盲肠结扎和穿刺（CLP）并在5天后用铜绿假单胞菌攻击的小鼠的肺组织中细菌生长更多，血清干扰素-γ（IFN-γ）和白介素（IL）12更低，而血清IL-10更高。与假单胞菌攻击的假CLP小鼠相比。为了测试这些变化对细菌免疫反应中细胞因子产生的功能意义，我们在假单胞菌攻击之前向CLP后小鼠施用了IFN-γ和抗IL-10。在CLP后小鼠中，IFN-γ和抗IL-10的使用不能改善细菌清除率或死亡率。在进一步的研究中，我们在铜绿假单胞菌攻击之前向IL-10敲除小鼠施用了IFN-γ。我们的结果表明，与野生型对照相比，接受或未接受IFN-γ治疗的IL-10基因敲除小鼠的细菌清除率或死亡率无显着差异。最后，由于在上述研究中大多数死亡发生在假单胞菌攻击后的2到3天内，并且可能与明显的促炎反应有关，因此我们研究了IFN-γ和抗IL-10对C3H / 3中假单胞菌清除的影响HeJ小鼠，对内毒素或革兰氏阴性细菌没有过度的促炎反应。接受抗IL-10和IFN-γ的C3H / HeJ小鼠的假单胞菌细菌清除率和死亡率均未提高。这些结果表明，损伤后抑制的IFN-γ和IL-12反应，加上对损伤后铜绿假单胞菌攻击的过度IL-10反应，与细菌清除率或存活率无关。

BACKGROUND & AIMS: :Given the paucity of data on the distribution of serotonin (5-HT) receptors of type 6 (5-HT(6)) in the human brain, the aim of this study was to investigate their distribution in postmortem human prefrontal cortex, striatum and hippocampus by either immunohistochemical or immunofluorescence techniques. The brain samples were obtained from 6 subjects who had died for causes not involving primarily or secondarily the CNS. The 5-HT(6) receptor distribution was explored by the [(125)I]SB-258585 binding to brain membranes followed by immunohistochemical and immunofluorescence evaluations. A specific [(125)I]SB-258585 binding was detected in all the regions under investigation, whilst the content in the hippocampus and cortex being about 10-30 times lower than in the striatum. Immunohistochemistry and double-label immunofluorescence microscopy experiments, carried out in the prefrontal cortex and hippocampus only, since data in the striatum were already published, showed the presence of 5-HT(6) receptors in both pyramidal and glial cells of prefrontal cortex, while positive cells were mainly pyramidal neurons in the hippocampus. The heterogeneous distribution of 5-HT(6) receptors provides a preliminary explanation of how they might regulate different functions in different brain areas, such as, perhaps, brain trophism in the cortex and neuronal firing in the hippocampus. This study, taking into account all the limitations due to the postmortem model used, represents the starting point to explore the 5-HT(6) receptor functionality and its sub-cellular distribution.

背景与目标： ：鉴于缺乏6型血清素（5-HT）受体（5-HT（6））在人脑中的分布的数据，本研究的目的是研究它们在死后人类前额叶皮层，纹状体中的分布免疫组织化学或免疫荧光技术检测海马和海马。脑样本是从6名因主要或次要不涉及CNS的原因死亡的受试者中获得的。通过[（125）I] SB-258585与脑膜的结合，然后进行免疫组织化学和免疫荧光评估，探索了5-HT（6）受体的分布。在所有研究区域中均检测到特定的[（125）I] SB-258585结合，而海马和皮质中的含量比纹状体低约10-30倍。仅在前额叶皮层和海马体中进行的免疫组织化学和双标记免疫荧光显微镜实验，因为纹状体中的数据已经发表，显示在前额叶皮层的锥体细胞和神经胶质细胞中都存在5-HT（6）受体，而阳性细胞主要是海马中的锥体神经元。 5-HT（6）受体的异质分布为它们如何在不同的大脑区域调节不同的功能提供了初步的解释，例如，大脑皮层的营养和海马神经元的放电。这项研究，考虑到由于使用死后模型的所有限制，代表了探索5-HT（6）受体功能及其亚细胞分布的起点。

BACKGROUND & AIMS: :Accumulating evidence indicates that Toll-like receptor (TLR) signaling adapter protein interactions with Toll/Interleukin-1 Receptor (TIR) domains present in sensory neurons may modulate neuropathic pain states. Following ligand interaction with TLRs, TIR serves to both initiate intracellular signaling and facilitate recruitment of signaling adapter proteins to the intracytoplasmic domain. Although TLR TIR is central to a number of TLR signaling cascades, its role in sensory neurons is poorly understood. In this study we investigated the degree to which TLR TIR decoy peptide modified to include a TAT sequence (Trans-Activator of Transcription gene in HIV; TAT-4BB) affected LPS-induced intracellular calcium flux and excitation in sensory neurons, and behavioral changes due to TLR4 active metabolite, morphine-3-glucuronide (M3G) exposure in vivo. TAT-4BB inhibited LPS-induced calcium changes in a majority of sensory neurons and decreased LPS-dependent neuronal excitability in small diameter neurons. Acute systemic administration of the TAT-4BB reversed M3G-induced tactile allodynia in a dose-dependent manner but did not affect motor activity, anxiety or responses to noxious thermal stimulus. These data suggest that targeting TLR TIR domains may provide novel pharmacological targets to reduce or reverse TLR4-dependent pain behavior in the rodent.

背景与目标： ：越来越多的证据表明，Toll样受体（TLR）信号转接头蛋白与感觉神经元中存在的Toll / Interleukin-1受体（TIR）域相互作用可能会调节神经性疼痛状态。配体与TLR相互作用后，TIR既可以启动细胞内信号传导，也可以促进信号传导衔接子蛋白募集到胞质内域。尽管TLR TIR是许多TLR信号级联的核心，但对其在感觉神经元中的作用知之甚少。在这项研究中，我们调查了TLR TIR诱饵肽修饰为包含TAT序列（HIV转录基因的反式激活因子； TAT-4BB）在多大程度上影响LPS诱导的细胞内钙通量和感觉神经元的兴奋，以及行为改变对TLR4活性代谢产物吗啡-3-葡糖醛酸（M3G）的体内暴露。 TAT-4BB抑制了LPS诱导的大多数感觉神经元中钙的变化，并降低了小直径神经元中LPS依赖性神经元的兴奋性。 TAT-4BB的急性全身给药以剂量依赖的方式逆转了M3G诱导的触觉异常性疼痛，但并未影响运动活动，焦虑或对有害热刺激的反应。这些数据表明，靶向TLR TIR结构域可提供新的药理学靶标，以减少或逆转啮齿动物中TLR4依赖性的疼痛行为。

BACKGROUND & AIMS: BACKGROUND:Interleukin-17 (IL-17), which is secreted by Th17 cells, is a proinflammatory cytokine that is implicated in the pathogenesis of multiple sclerosis (MS) and plays a role in nonresponse of MS patients to interferon-β (IFN-β) therapy. OBJECTIVES:The purpose of this study was to establish a correlation between nonresponders (NR) and IL-17A serum titers and binding antibodies (BAbs) to IFN-β, as well as to find a correlation between IL-17A serum levels and other features of MS patients. METHODS:Our prospective study included 72 inactive relapsing-remitting multiple sclerosis (RRMS) patients that had been treated for at least 18 months with IFN-β and 15 healthy subjects. We determined the serum levels of IL-17A and of BAbs. IL-17A levels were considered elevated (IL-17A+) if the recorded value was greater than 1.6 pg/ml. RESULTS:Twenty-seven patients (37.5%) were NR and had a significantly higher serum IL-17A level compared to the responders group. Nineteen patients (26.4%) were IL-17A+ and had had a significantly higher number of relapses in the previous year and a higher Expanded Disability Status Score. The majority of IL-17A+ patients were NR and had a shorter MS duration. CONCLUSIONS:RRMS patients with high serum IL-17A levels do not respond well to IFN-β therapy and have shorter MS duration compared to patients with low IL-17A levels. This response is not influenced by the presence of BAbs.

背景与目标： 背景：Th17细胞分泌的白细胞介素17（IL-17）是一种促炎细胞因子，与多发性硬化症（MS）的发病机制有关，并且在MS患者对干扰素-β（IFN- β）疗法。
目的：本研究的目的是建立无应答者（NR）与IL-17A血清滴度和针对IFN-β的结合抗体（BAbs）之间的相关性，以及找出IL-17A血清水平与其他特征之间的相关性MS患者。
方法：我们的前瞻性研究包括72例接受IFN-β治疗至少18个月的非活动性复发缓解型多发性硬化症（RRMS）患者和15名健康受试者。我们确定了IL-17A和BAbs的血清水平。如果记录值大于1.6 pg / ml，则认为IL-17A水平升高（IL-17A）。
结果：27名患者（37.5％）为NR，与应答者组相比，血清IL-17A水平显着更高。 19名患者（26.4％）为IL-17A，在前一年中复发率明显更高，而“扩展残疾状态评分”更高。大多数IL-17A患者为NR，MS病程较短。
结论：与低IL-17A水平的患者相比，血清IL-17A水平高的RRMS患者对IFN-β治疗的反应不佳，MS病程较短。该反应不受BAbs的存在的影响。

BACKGROUND & AIMS: :IL-23 and IL-17 are pro-inflammatory cytokines. IL-23 is secreted by activated macrophages and dendritic cells, while IL-17 by Th17 cells. Serum IL-23 and IL-17 are known to be elevated in numerous inflammatory diseases including neurodegenerative diseases. The role of serum IL-23 and IL-17 in aneurysmal subarachnoid hemorrhage (aSAH) has still not been investigated. The present work investigates the serum IL-23 and IL-17 levels and their association with post hemorrhagic complications and clinical outcome in patients with aSAH. METHODS:In this study, 80 patients with aSAH (Hunt and Hess grade I-V) were prospectively recruited. We enrolled 24 control patients with lumbar spinal stenosis. Peripheral venous blood was withdrawn from controls and from aSAH patients at day 1 and day 7, allowed to clot and centrifuged to obtain serum. Enzyme linked immunoassay kits were employed to quantify the serum levels of IL-23 and IL-17 by applying 50µL of serum samples. Post hemorrhagic complications and clinical outcome were documented prospectively from patient's hospital record. RESULTS:Serum IL-23 and IL-17 levels were significantly elevated in aSAH patients at day 1 and day 7 (n=80) as compared to control patients (n=24). Further analysis after dichotomy of patients who suffered from post hemorrhagic complications including cerebral vasospasm, chronic hydrocephalus, seizures, cerebral ischemia, delayed neurological deficits showed differential correlations with different post hemorrhagic complications (Table 1). Serum IL-23 and IL-17 levels did not correlate with clinical outcome. CONCLUSION:Serum IL-23 and IL-17 levels were elevated in patients with aSAH showing upregulation of IL-23/IL-17 inflammatory axis after aSAH. Serum IL-23 and IL-17 showed differential correlations with post hemorrhagic complications and no correlation with clinical outcome.

背景与目标： ：IL-23和IL-17是促炎性细胞因子。 IL-23由活化的巨噬细胞和树突状细胞分泌，而IL-17由Th17细胞分泌。已知血清IL-23和IL-17在包括神经退行性疾病在内的许多炎性疾病中均升高。血清IL-23和IL-17在动脉瘤性蛛网膜下腔出血（aSAH）中的作用尚未进行研究。本工作调查了aSAH患者的血清IL-23和IL-17水平及其与出血后并发症和临床结局的关系。
方法：在这项研究中，前瞻性招募了80例aSAH患者（Hunt和Hess I-V级）。我们招募了24名腰椎管狭窄的对照患者。在第1天和第7天从对照和aSAH患者中抽取外周静脉血，使其凝结并离心以获得血清。采用酶联免疫分析试剂盒，通过施加50µL血清样品定量IL-23和IL-17的血清水平。从患者的住院记录中前瞻性地记录了出血后并发症和临床结局。
结果：与对照组患者（n = 24）相比，aSAH患者在第1天和第7天的血清IL-23和IL-17水平显着升高（n = 80）。二分法对患有出血后并发症（包括脑血管痉挛，慢性脑积水，癫痫发作，脑缺血，迟发性神经功能缺损）的患者进行二分法手术后的进一步分析显示，不同出血后并发症的相关性不同（表1）。血清IL-23和IL-17水平与临床结果无关。
结论：aSAH患者血清IL-23和IL-17水平升高，显示aSAH后IL-23 / IL-17炎症轴上调。血清IL-23和IL-17与出血后并发症呈差异相关，与临床结局无相关性。

BACKGROUND & AIMS: :Pneumocystis pneumonia (PCP) incidence was decreased in renal transplant thanks to prophylaxis, recommended during the first months after transplantation. However, many late PCP cases are observed after the first 6 months and recommendations to maintain or reintroduce prophylaxis are lacking. The objective of the study was to identify risk factors to guide the individual prescription of prophylaxis, 6 months after transplantation. Thirty-three late PCP cases were identified between 1995 and 2012 in Lille Hospital, France, and were compared to 72 randomized controls transplant recipients. In univariate analysis, age of donor (>48 years), retransplantation, a decrease glomerular filtration rate (≤45 mL/min), induction therapy mediated by anti-thymocyte globulin (ATG), steroid maintenance, high calcineurin inhibitors (CNI) doses (tacrolimus ≥0.5 mg/kg/day and cyclosporine ≥2.1 mg/kg/day), and cytomegalovirus (CMV) infection were significantly associated with PCP. In multivariate analysis, ATG (hazard ratio [HR]: 2.4 [1.1-5.4]), steroid therapy (HR: 3.1 [1.20-7.84], CNI (HR: 2.9 [1.28-6.38], and CMV (HR: 6.1 [2.74-16.33] remained associated with late PCP. In conclusion, we confirm that intensive immunosuppressive regimen and CMV infection are critical risk factors for late PCP and should be taken into account to decide on maintenance or reintroduction of a prophylactic treatment.

背景与目标： ：由于预防，建议在移植后的头几个月内，减少肾脏移植中肺囊虫性肺炎（PCP）的发生率。但是，在头6个月后观察到许多晚期PCP病例，缺乏维持或重新引入预防措施的建议。该研究的目的是确定危险因素，以指导移植后6个月进行预防的个体处方。在1995年至2012年之间，法国里尔医院确定了33例晚期PCP病例，并将其与72名随机对照移植受者进行了比较。在单因素分析中，供体年龄（> 48岁），再移植，肾小球滤过率降低（≤45mL / min），抗胸腺细胞球蛋白（ATG）介导的诱导治疗，类固醇维持，钙调神经磷酸酶抑制剂（CNI）剂量高他克莫司≥0.5mg / kg /天，环孢菌素≥2.1mg / kg /天）和巨细胞病毒（CMV）感染与PCP显着相关。在多变量分析中，ATG（危险比[HR]：2.4 [1.1-5.4]），类固醇治疗（HR：3.1 [1.20-7.84]，CNI（HR：2.9 [1.28-6.38]）和CMV（HR：6.1 [ [2.74-16.33]仍与晚期PCP相关，总的来说，我们确认强化免疫抑制方案和CMV感染是晚期PCP的关键危险因素，应考虑维持或重新引入预防性治疗。

BACKGROUND & AIMS: :An effective immune response to antigen requires professional antigen-presenting cell (APC), which not only present antigen, but also provide costimulation and cytokines (eg, IL-12) that drive T cell differentiation down the appropriate effector pathway (Tc1/TH1). For T cell-based immunotherapy protocols, the availability of large numbers of autologous professional APC is a major limitation because professional APC do not proliferate in vitro. T cells themselves can proliferate exponentially in vitro and have the ability to present antigen. They can also express costimulatory molecules after activation. Therefore, we hypothesized that if activated T cells were genetically modified to express proinflammatory cytokines required to polarize T cells toward a Tc1 response, they could fulfill the requirements for an abundant, autologous APC. To test this potential, T cells were activated by CD3/CD28 antibodies and pulsed with model HLA-A2+ peptides derived from CMVpp65, MAGE-3, and MART-1. Activated T-APC readily reactivated CD8 pp65 memory T cells from healthy CMV seropositive donors; however, the activation of MAGE-3 and MART-1-specific CD8 T cells required both IL-7 and IL-12, which could be provided either exogenously or by genetic modification of the T-APC. Responder T cells could be expanded to large numbers with subsequent stimulations using activated, peptide-pulsed T-APC and IL-2. Tumor antigen-specific T cell lines killed both peptide-pulsed target cells and tumor cell lines. Thus, T cells provide a platform for the generation of autologous APC that can be customized to express both antigens and therapeutic molecules for the induction of antigen-specific T cell immunity.

背景与目标： ：对抗原的有效免疫反应需要专业的抗原呈递细胞（APC），它不仅呈递抗原，而且还提供共刺激和细胞因子（例如IL-12），以驱动T细胞向适当的效应子途径（Tc1 / TH1）分化）。对于基于T细胞的免疫疗法方案，大量的自体专业APC的可用性是一个主要限制，因为专业APC不会在体外增殖。 T细胞本身可以在体外成倍增殖，并具有呈递抗原的能力。它们还可以在激活后表达共刺激分子。因此，我们假设，如果对活化的T细胞进行基因修饰以表达使T细胞朝Tc1反应极化所需的促炎细胞因子，那么它们就可以满足丰富的自体APC的要求。为了测试这种潜力，T细胞被CD3 / CD28抗体激活，并用衍生自CMVpp65，MAGE-3和MART-1的模型HLA-A2肽脉冲。激活的T-APC可以很容易地激活来自健康CMV血清反应阳性供体的CD8 pp65记忆T细胞。然而，MAGE-3和MART-1特异性CD8 T细胞的激活需要IL-7和IL-12，这可以通过外源或通过T-APC的遗传修饰来提供。可以使用激活的，肽脉冲的T-APC和IL-2在随后的刺激下将应答剂T细胞扩增为大量。肿瘤抗原特异性T细胞系杀死了肽脉冲的靶细胞和肿瘤细胞系。因此，T细胞提供了用于产生自体APC的平台，该平台可以被定制以表达抗原和治疗性分子以诱导抗原特异性T细胞免疫。

BACKGROUND & AIMS: :Peptidylarginine deiminase 6 (PAD6) is an enzyme that is uniquely expressed in male and female germ cells. To study the function of this enzyme in vivo we generated mice deficient for PAD6. Here we show that inactivation of the PAD6 gene in mice leads to female infertility whereas male fertility is not affected. The absence of the PAD6 protein and consequently absence of citrullination activity in oocytes results in dispersal of the cytoskeletal sheets in oocytes, indicating an essential role of these germ cell-specific structures in zygote/embryo development. PAD6 deficient mice do not show any other overt phenotype. Thus, we identify citrullination as a new regulator of fertility.

背景与目标： ：肽基精氨酸脱亚氨酶6（PAD6）是一种在雄性和雌性生殖细胞中独特表达的酶。为了研究该酶在体内的功能，我们产生了PAD6缺陷的小鼠。在这里，我们显示，小鼠中PAD6基因的失活导致女性不育，而男性不育不受影响。卵母细胞中缺乏PAD6蛋白，因此缺乏瓜氨酸化活性，导致卵母细胞中细胞骨架片的分散，表明这些生殖细胞特异性结构在合子/胚胎发育中起着至关重要的作用。 PAD6缺陷的小鼠没有显示任何其他明显的表型。因此，我们确定瓜氨酸化是生育的新调节剂。

BACKGROUND & AIMS: :The identification of germ-line mutations in 2 genes (BRCA1 and BRCA2) responsible for the majority of hereditary ovarian cancers has led an increasing number of women carriers of these mutations to undergo prophylactic oophorectomy (PO) to reduce their risk of subsequent ovarian carcinoma. A large number of unexpected, clinically occult neoplasms are thus being discovered. Up to December 2004, the Medical Genetics Service of the National Cancer Institute in Milan, Italy, has tested 756 probands from breast and/or ovarian cancer families for BRCA1 and BRCA2 germ-line mutations. Molecular screening of family members led to the identification of 344 female carriers of BRCA1 (239) or BRCA2 (105) germ-line mutations. Of the 186 potentially eligible women (37 of whom had tested positive for BRCA1 and 13 for BRCA2 mutation), 50 (26.8%) chose to undergo PO. Six clinically occult primary gynecologic malignancies (2 stage IIIC serous carcinomas of the ovary, 3 in situ serous carcinomas of the fallopian tube, and 1 stage IIB invasive serous carcinoma of the fallopian tube) and 1 occult ovarian metastasis from breast carcinoma were identified in the PO specimens of 7 women (all BRCA1 mutated). Four of the patients with occult primary gynecologic cancers are alive without disease 129, 87, 38, and 7 months after PO, respectively. One of the 2 patients with primary ovarian cancer and the single patient with tubal invasive carcinoma are alive with recurrent disease 83 and 20 months after PO, respectively. In addition, one of the patients whose PO specimen did not show any malignancy presented with stage IIIC tubal carcinoma 77 months after PO. The relatively high number of tubal neoplasms found at PO in this group of patients underlines the linkage between mutation and the risk of developing tubal cancer, and stresses the need to include removal of the entire tubes at the time of PO and of thoroughly evaluating the specimens at the microscopic level. The upstaging of all 3 invasive carcinomas after staging surgery, and the late recurrence and persistence of 2 of them despite treatment indicate that small size of the tumors should not preclude therapy.

背景与目标： ：对负责大多数遗传性卵巢癌的2个基因（BRCA1和BRCA2）的种系突变的鉴定，导致越来越多的女性携带这些突变的人进行了预防性卵巢切除术（PO），以降低其患上卵巢癌的风险。因此，发现了大量意想不到的临床隐匿性肿瘤。截至2004年12月，意大利米兰国家癌症研究所的医学遗传学服务已对756个来自乳腺癌和/或卵巢癌家族的先证者进行了BRCA1和BRCA2种系突变测试。家庭成员的分子筛查导致识别出BRCA1（239）或BRCA2（105）种系突变的344个雌性携带者。在186名可能符合条件的妇女中（其中37人的BRCA1测试呈阳性，13人的BRCA2突变检测为阳性），其中50人（26.8％）选择接受PO。在临床中确定了6例临床隐匿的原发性妇科恶性肿瘤（2例卵巢IIIC浆液性癌，3例输卵管原位浆液性癌和1例IIB输卵管浸润性浆液性癌）和1例隐匿性卵巢癌卵巢转移。 7名妇女的PO标本（所有BRCA1突变）。分别在PO后129、87、38和7个月，有四名患有隐匿性原发性妇科癌症的患者还活着而没有疾病。 2例原发性卵巢癌患者中的1例和输卵管浸润性癌的1例患者在PO后分别存活83个月和20个月。此外，其中一名PO标本未显示任何恶性肿瘤的患者在PO后77个月出现IIIC期输卵管癌。该组患者在PO中发现的相对较多的输卵管肿瘤强调了突变与发生输卵管癌的风险之间的联系，并强调需要在PO时切除整个管并彻底评估标本在微观层面上。分期手术后所有3种浸润性癌的分期升级，并且尽管有治疗，但其中2种仍较晚复发和持续存在，这表明较小的肿瘤不应排除治疗的可能性。

BACKGROUND & AIMS: :A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.

背景与目标： ：整联蛋白家族的跨膜细胞外基质受体，α6 beta 4，是半桥粒的成分，半桥粒是在上皮细胞结缔组织附着中重要的粘附复合物（Stepp，MA，S。Spurr-Michaud，A。Tisdale， J. Elwell和IK Gipson。1990.美国国家科学院院刊87：8970-8974； Jones，JCR，MA Kurpakus，HM Cooper和V.Quaranta。1991.《细胞调节》 2：427- 438）。血小体的胞质成分包括大疱性类天疱疮（BP）抗原，而细胞外成分包括锚定丝（CAF）和含VII型胶原的锚定纤丝的125 kD成分。我们已经监测了在体外伤口愈合的外植体模型中将α6β4整合素整合到形成hemidemosomes中。在最近从裸结缔组织上方未受伤部位边缘迁移的上皮细胞中，α6 beta 4首先沿整个细胞表面出现。在这一阶段，这些细胞几乎不含有或不含胞质半桥粒成分，至少可以通过使用BP自身抗体的免疫荧光检测到，而对于层粘连蛋白和CAF则已经呈阳性。在稍后的阶段，随着细胞对胞质半桥体成分（例如BP抗原）和VII型胶原呈阳性，α6β4沿着上皮细胞的基极集中，并与外植体的结缔组织邻接。针对β4的多克隆抗体不会干扰外植体中上皮细胞的迁移。然而，它们阻止了半桥粒复合物的组装并抑制了在伤口区域上迁移的细胞中VII型胶原的表达。另外，它们在外植体未受伤区域的非迁移细胞中诱导已建立的半染色体的破坏。针对α6的单克隆抗体具有更引人注目的效果，因为它们可以完全脱离外植体未受伤区域中的上皮细胞。 CAF抗体还可以在未受伤的区域分离上皮细胞，这显然是通过诱导基底膜区域的透明层上皮与结缔组织之间的分离来实现的。这些结果表明了一个模型，其中α6β4极化到细胞的基底表面（可能是由假定的锚定细丝相关配体诱导的）触发了半血球斑块的组装。