地塞米松对MS的IFN—γ及IL—10分泌性T细胞的影响

目的 检测多发性硬化（ＭＳ）口才外周血单个核细胞在地塞米松（Ｄｅｘ）影响下的ＩＦＮ－γ和ＩＬ－１０的分汔细胞水平。方法 采用酶联免疫斑点技术（ＥＬＩＳＰＯＴ）检测体外培养的外周血单个核细胞（ＭＮＣ）在ＣＮＳ髓鞘素抗原ＭＢＰ下的地塞米松对照试验,检测ＩＦＮ－γ和ＩＬ－１０分泌笥Ｔ细胞水平,并与其他神经疾病（ＯＮＤ）组及健康对照组的检测结果进行对比。结果 显示ＭＳ患者ＩＦＮ－γ分泌细胞水平高于对照组,     

重症肌无力患者胸腺及外周血白细胞介素和干扰素—γ分泌细胞的检测

目的 研究重症肌无力（ＭＧ）胸腺以及外周血辅助性Ｔ细胞（Ｔｈ）,包括Ｔｈ１和Ｔｈ２亚群的免疫功能。方法 利用酶联免疫斑点（ＥＬＩＳＰＯＴ）法检测２０例ＭＧ患者胸腺及外周血乙酰胆碱受体（ＡＣｈＲ）特异反应性白细胞介素－２（ＩＬ－２）、白细胞介素－４（ＩＬ－４）、白细胞介素－１０（ＩＬ－１０）、干扰素－γ（ＩＦＮ－γ）分泌细胞数。结果 ＭＧ患者胸腺ＡＣｈＲ特异反应性ＩＬ－２、ＩＬ－４、ＩＬ－１０、ＩＦＮ－γ分泌细胞数均增高,外周血ＡＣｈＲ特异反应性ＩＬ－１０、ＩＦＮ－分泌细胞数增高,外周血与胸腺间ＡＣｈＲ特异反应性ＩＬ－４、ＩＬ－１０分泌细胞数呈正相关,３例ＭＧ患者在治疗后,外周血ＡＣｈＲ特异反应性ＩＬ－２、ＩＬ－４、ＩＦＮ－γ分泌细胞数均下降,ＩＬ－１０分泌细胞数则１例降低,２例增高。结论 ＭＧ胸腺及外周血存在着     

重症肌无力患者外周血乙酰胆碱受体特异性T细胞sIL-2R、IFN-γ、IL-4的表达和转录

目的了解ＭＧ患者的乙酰胆碱受体（ＡＣｈＲ）特异性细胞免疫应答。方法采用酶联免疫吸附试验（ＥＬＩＳＡ）检测３０例ＭＧ患者和２０名健康对照者经ＡＣｈＲ刺激后外周血单核细胞（ＰＢＭＣ），辅助性Ｔ细胞１（Ｔｈ１）相关的干扰素（ＩＦＮ）γ、辅助性Ｔ细胞２（Ｔｈ２）相关的白细胞介素（ＩＬ）４及与细胞免疫活化密切相关的可溶性白细胞介素２受体（ｓＩＬ２Ｒ）的分泌，用逆转录聚合酶链反应（ＲＴＰＣＲ）结合狭缝印迹杂交检测ＩＦＮγ、ＩＬ４的信息核糖核酸（ｍＲＮＡ）转录。结果总ＭＧ患者组ＩＦＮγ显著高于健康对照组，尤以急性ＭＧ患者组（１４例）更明显，且其升高与其ｍＲＮＡ转录水平一致。虽然这两组ＭＧ患者的ｓＩＬ２Ｒ亦明显高于健康对照组，但患者组的ＩＬ４表达及其ｍＲＮＡ转录与健康对照组相比差异无显著意义。结论ＭＧ患者有Ｔｈ１和Ｔｈ２的失衡，ＭＧ患者有ＡＣｈＲ特异性细胞免疫活化，Ｔｈ１的细胞因子ＩＦＮγ可能作为效应因子参与了ＭＧ的发病。     

急性脑梗死和Alzheimer病脑脊液高水平髓鞘素抗原B细胞免疫应答

目的确认急性脑梗死（ＡＣＩ）和Ａｌｚｈｅｉｍｅｒ病（ＡＤ）对髓鞘素碱性蛋白（ＭＢＰ）、髓鞘素结合糖蛋白（ＭＡＧ）和含脂质蛋白（ＰＬＰ）的Ｂ细胞免疫应答。方法采用酶联免疫斑点技术检测了ＡＣＩ、临床可能ＡＤ和其它神经疾病（ＯＮＤ）对照组患者外周血和脑脊液（ＣＳＦ）中ＭＢＰ、ＭＡＧ和ＰＬＰ抗体分泌细胞。结果ＡＣＩ、ＡＤ和ＯＮＤ患者外周血中均可检出ＩｇＧ、ＩｇＡ、ＩｇＭ三种表型ＭＢＰ、ＭＡＧ、ＰＬＰ抗体分泌细胞，无显著差异。但ＡＣＩ和ＡＤ患者ＣＳＦ中ＩｇＧ型ＭＢＰ、ＭＡＧ和ＰＬＰ抗体分泌细胞均呈显著性增高。结论ＡＣＩ急性脑缺血损伤和ＡＤ神经变性可能导致体内Ｂ细胞激活及ＣＮＳ内髓鞘素反应性Ｂ细胞免疫应答，其病理意义有待探讨。     

Alzheimer型痴呆的髓鞘素蛋白自身应答性T细胞免疫反应

通过计数分泌细胞因子γ－干扰素（ＩＦＮ－γ）、辅助性Ｔ细胞（Ｔｈ１），检查ＡＤ患者外周血和脑脊液（ＣＳＦ）中Ｔ细胞免疫应答。方法：将单个核细胞（ＭＮＣ）暴露于中枢神经系统（ＣＮＳ）髓鞘素抗原髓鞘碱性蛋白（ＭＢＰ）和含脂质蛋白（ＰＬＰ）中进行体外短时间培养，用酶联免疫斑点试验（Ｅｌｉｓｐｏｔ）检测ＩＦＮ－γ分泌性Ｔｈ１细胞链，同时检测急性脑血管病（ＣＶＤ）和其他神经疾病（ＯＮＤ）患者作为对照。结果显示ＡＤ患者外周血中呈高水平ＭＢＰ应答性Ｔｈ１细胞链，而其ＣＳＦ中尤为明显，外周血中ＰＬＰ应答性Ｔｈ１细胞亦显著高于ＯＮＤ对照组。结论认为ＡＤ患者存在高水平的髓鞘素蛋白自身应答性Ｔ细胞反应，且在与ＣＮＳ紧邻的ＣＳＦ中表现得尤为显著，其在ＡＤ发病机制中的作用还有待进一步深入探讨。     

重症肌无力患者外周血干扰素—γ和白细胞介素—10分泌性T细？…

目的 探讨重症肌无力（ＭＧ）患者乙酰胆碱受体（ＡＣｈＲ）特异性Ｔ细胞免疫应答及其国酰胆碱受体抗体间的关系。及该类Ｔ细胞在胸腺瘤切除前后的变化。方法 采用酶联免疫斑点技术在１５例ＭＧ患者和１０例其他神经疾病（ＯＮＤ）患者中,检测在ＡＣｈＲ等不同抗原特异干扰素－γ分泌性Ｔｈ１细胞和白细胞介素－１０（ＩＬ－１０）分泌笥ＺＴｈ２细胞数。结果 ＡＣｈＲＡｂ阳性组ＭＧ患者外周血ＡＣｈＲ特异性ＩＦＮ－γ、ＩＬ＿     

多发性硬化症髓鞘素组分特异性抗体分泌细胞

本文用酶联免疫斑点法（Ｅｌｉｓｐｏｔ）检测了２３例临床确诊多发性硬化症（ＭＳ）和１２例无菌性脑膜炎（ＡＭ）患者外周血（ＰＢ）和脑脊液（ＣＳＦ）中髓鞘素碱性蛋白（ＭＢＰ）、髓鞘素结合糖蛋白（ＭＡＧ）和含脂质蛋白（ＰＬＰ）特异性ＩｇＧ抗体分泌细胞。两组患者ＣＳＦ中该３种抗体分泌细胞均呈明显增多趋势，ＭＳ组尤著，但两组ＰＢ中该类细胞数均很少。指示对髓鞘素组分的Ｂ细胞免疫应答主要局限于与中枢神经系统（ＣＮ     

多发性硬化患者血清中可溶性白细胞介素─2受体（SIL—2R）的动态观察

采用ＥＬＩＳＡ双抗体夹心法对２３例多发性硬化（ＭＳ）患者在急性复发期和缓解期血清的ＳＩＬ—２Ｒ水平进行了动态检测。结果显示急性复发期患者的ＳＩＬ—２Ｒ水平明显高于正常对照组（Ｐ＜０．００１）。缓解期的ＳＩＬ—２Ｒ水平患较急性复发期明显降低（Ｐ＜０．０５），但仍显著高于正常对照组（Ｐ＜０．００１）。提示ＭＳ患者不论急性复发期还是缓解期其Ｔ淋巴细胞处于活化状态。我们认为对ＭＳ患者血清中ＳＩＬ—２Ｒ水平的动态观察可能有助于复发的预测，及预后的判断。     

急性脑梗死和Alzheimer病脑脊液高水平髓鞘素抗原B细胞免 …

目的 确认急性脑梗死对髓鞘素碱性蛋白（ＭＢＰ）,髓鞘素结合糖蛋白（ＭＡＧ）和含脂质蛋白（ＰＬＰ）的Ｂ细胞免疫应答。方法 采用酶联免疫斑点技术检测了ＡＣＩ,临床可能ＡＤ和其它神经疾病（ＯＮＤ）对照组患者外周血和脑脊液（ＣＳＦ）中ＭＢＰ,ＭＡＧ和ＰＬＰ抗体分泌细胞。     

急性脑梗塞患者分泌γ—干扰素的特异性髓鞘素记忆性T细胞的研究

通过计数分泌细胞因子γ－干扰素记忆性Ｔ细胞，观察急性脑梗塞（ＡＣＩ）患者的特异性Ｔ细胞免疫应答。方法采用酶联免疫斑点法（ＥＬＩＳＡ）检测了３２例ＡＣＩ、２４例炎症性神经疾病（ＯＩＮＤ）和２５例其他神经疾病（ＯＮＤ）患者髓鞘素抗原及其多肽应答性Ｔ细胞。结果ＡＣＩ患者外周血髓鞘素碱性蛋白（ＭＢＰ）及其多肽和含脂质蛋白（ＰＬＰ）自身抗原应答性Ｔ细胞数呈显著增高，脑脊液（ＣＳＦ）中ＭＢＰ应答性Ｔ细胞数约为外周血中的１１０倍。结论这种免疫应答可能是继发于急性缺血后的脑组织损伤，且在紧邻靶器官的ＣＳＦ中表现尤为明显，其病理意义还有待进一步探讨。     

ABO血型不合外周血造血干细胞移植10例☆

背景：ABO血型不合供受者之间进行外周血造血干细胞移植,面临受者血型的转变、移植后输血的选择等问题。 目的：观察ABO血型不合外周血造血干细胞移植治疗血液病的临床疗效和近远期并发症。 方法：回顾性分析了2005/2008武警总医院收治的10例ABO血型不合异基因外周血同胞供者造血干细胞移植患者的资料。总结植入效果,血型转变,移植物抗宿主病以及不良反应。 结果与结论：9例患者恢复造血功能,血型转变为完全供者型,1例患者白细胞血小板恢复,但红细胞植入延迟。所有患者在输注移植物时未出现短暂的血红蛋白尿,无严重急性溶血和迟发型溶血的发生,1例出现严重的移植物抗宿主病。可见ABO血型不合外周血造血干细胞移植对移植疗效无明显影响,红细胞植入延迟的预防和处理十分重要。     

Interleukin-10 inhibits endotoxin-induced pro-inflammatory cytokines in microglial cell cultures

Inflammation contributes to perinatal brain injury and can be induced by hypoxia-ischemia (HI) or exposure to infection (fetal inflammatory response). The anti-inflammatory cytokine interleukin-10 (IL10) has been shown to have neuroprotective effects following HI. To determine whether IL10 can reduce the inflammatory response to lipopolysaccharide (LPS) in microglial cell cultures, primary microglial (MG) and/or HAPI cells (new MG-like cell line) were treated with LPS (50 ng/ml) in the presence or absence of IL10 (20 ng/ml) for 0.5, 1, 4, and 8 h. TNFalpha, MIP-1alpha, and RANTES were assayed by ELISA. Chemokine receptors, CCR5, CXCR3, and CX3CR1 (fractalkine receptor) were assayed by semiquantitative RT-PCR. We found that in MG cell cultures TNFalpha, MIP-1alpha, and RANTES release after 8-h exposure to LPS was significantly higher compared to non-exposed MG cells (P < 0.001). In HAPI cell cultures similar stimulation of mRNA levels was found for TNFalpha, MIP-1alpha, CXCR3, and CX3CR1. IL10 inhibited TNFalpha, MIP-1alpha, and RANTES release of LPS-stimulated MG cells as well as TNFalpha, MIP-1alpha, and CXCR3 mRNA expression by HAPI cells after exposure to LPS (P < 0.05). In contrast to those inhibitory effects, there was no change in fractalkine, and a modest increase in CX3CR1 mRNA levels was found in the presence of IL10. We conclude that the inflammatory response induced in microglial cells by LPS can be markedly reduced by IL10. The increase in fractalkine receptor (CX3CR1) is also potentially protective. Our results suggest that treatment of damaging neuroinflammatory insults such hypoxia-ischemia, with IL10 may be protective for the immature brain.     

Interferon effects on interleukin-10 secretion Mononuclear cell response to interleukin-10 is normal in multiple sclerosis patients

The mechanism of action of recombinant Interferon β_1b (rIFNβ_1b/IFN β-1b), the approved therapy for multiple sclerosis (MS), is still unclear. Here we present evidence that part of the therapeutic effects of rIFNβ_1b in MS might result from the induction of the secretion of interleukin (IL)-10, a cytokine previously designated cytokine synthesis inhibitory factor (CSIF). We observed that rIFNβ_1b stimulated significant IL-10 secretion by monocytes from MS patients after brief incubation (18 h), whereas rIFNγ, an inducer of MS exacerbations, was unable to stimulate IL-10 production in similar conditions. To determine the role of IL-10 as CSIF in the disease, we have also investigated its effects on TNFα and IL-6 secretion by peripheral blood mononuclear cells from MS patients. Recombinant human IL-10 significantly inhibited tumor necrosis factor α and IL-6 secretion induced by rIFNγ, lipopolysaccharide (LPS), and rIFNγ + LPS in MS patients and in control subjects. The induction of IL-10 secretion by rIFNβ_1b and the IL-10 inhibitory activity o pro-inflammatory cytokine secretion induced by rIFNγ in MS make this cytokine a potential candidate to treat the disease.     

CXCL10-induced cell death in neurons: role of calcium dysregulation

Chemokines play a key role in the regulation of central nervous system disease. CXCL10 over-expression has been observed in several neurodegenerative diseases, including multiple sclerosis, Alzheimer's disease and HIV-associated dementia. More recent studies by others and us have shown that CXCL10 elicits apoptosis in fetal neurons. The mechanism of CXCL10-mediated neurotoxicity, however, remains unclear. In this study, we provide evidence for the direct role of Ca(2+) dysregulation in CXCL10-mediated apoptosis. We demonstrate that treatment of fetal neuronal cultures with exogenous CXCL10 produced elevations in intracellular Ca(2+) and that this effect was modulated via the binding of CXCL10 to its cognate receptor, CXCR3. We further explored the association of intracellular Ca(2+) elevations with the caspases that are involved in CXC10-induced neuronal apoptosis. Our data showed that increased Ca(2+), which is available for uptake by the mitochondria, is associated with membrane permeabilization and cytochrome c release from this compartment. The released cytochrome c then activates the initiator active caspase-9. This initiator caspase sequentially activates the effector caspase-3, ultimately leading to apoptosis. This study identifies the temporal signaling cascade involved in CXCL10-mediated neuronal apoptosis and provides putative targets for pharmaceutical intervention of neurological disorders associated with CXCL10 up-regulation.     

A retinal ganglion cell that can signal irradiance continuously for 10 hours

A recently discovered type of mammalian retinal ganglion cell encodes environmental light intensity and mediates non-image-forming visual behaviors, such as the pupillary reflex and circadian photoentrainment. These intrinsically photosensitive retinal ganglion cells (ipRGCs) generate endogenous, melanopsin-based photoresponses as well as extrinsic, rod/cone-driven responses. Because the ipRGCs' light responses and the behaviors they control are both remarkably tonic, these cells have been hypothesized to be capable of irradiance detection lasting throughout the day. I tested this hypothesis by obtaining multielectrode-array recordings from ipRGCs in a novel rat eyecup preparation that enhances the regeneration of rod/cone photopigments. I found that 10 h constant light could continuously evoke action potentials in these ganglion cells under conditions that stimulated (1) only melanopsin, (2) mainly the rod input, and (3) both intrinsic and extrinsic responses. In response to a 10 h stimulus with gradual intensity changes to simulate sunrise and sunset, ipRGC firing rates slowly increased during the "sunrise" phase and slowly decreased during the "sunset" phase. Furthermore, I recorded from putative ipRGCs of melanopsin-knock-out mice and found that these cells retained the ability to respond in a sustained fashion to 20 min light steps, indicating that melanopsin is not required for such tonic responses. In conclusion, ipRGCs can signal light continuously for at least 10 h and can probably track gradual irradiance changes over the course of the day. These results further suggest that the photoreceptors and ON bipolar cells presynaptic to ipRGCs may be able to respond to light continuously for 10 h.     

The IL-10-producing regulatory B cells (B10 cells) and regulatory T cell subsets in neuromyelitis optica spectrum disorder

B cells contribute to the pathogenesis of neuromyelitis optica (NMO) by producing Aquaporin 4-specific autoantibodies (AQP4-ab); on the other hand, there are certain B cells that suppress immune responses by producing regulatory cytokines, such as IL-10. In this study, we investigated the presence of IL-10-producing Breg cells among lymphocyte subsets. Twenty-two seropositive NMO spectrum disorder (NMOSD) patients (29 samples) and 13 healthy controls (HCs) (14 samples) were enrolled. All NMOSD patients have received one or more immunosuppressive drugs. The phenotype and frequency of B cell and T cell subsets in the peripheral blood were measured by flow cytometry. We defined Breg cells as IL-10-producing B (B10) cells, which are CD19⁺CD39⁺CD1d⁺IL-10⁺. The potential relations were evaluated between specific lymphocyte subsets and AQP4-ab intensity measured by the cell-based indirect immunofluorescence assay. The frequency of B10 cells was higher in patients with NMOSD regardless of the disease status than that in HCs (attack samples; p?=?0.009 and remission samples; p?<?0.001, respectively). In addition, the frequency of IL-17⁺ Treg cells among Treg cells was higher during remission than during an attack (uncorrected p?=?0.032). Among the lymphocyte subsets, B10 cells alone showed a positive correlation with the intensity of AQP4-ab positivity (ρ [rho]?=?0.402 and p?=?0.031). It was suggested that the suppressive subsets including B10 and IL-17⁺ Treg cells might have important roles in controlling disease status in NMOSD. Further functional studies may help to elucidate the immunological role of B10 and IL-17⁺ Treg cells in NMOSD.     

微囊化牛肾上腺髓质细胞蛛网膜下腔移植治疗癌症晚期疼痛10例临床初步观察

目的:观察海藻酸钠-壳聚糖-海藻酸钠(Alginate- Chitosan-Alginate,ACA) 微囊化牛肾上腺嗜铬细胞(Bovine Adrenachromaffin ells,BCC) 异种移植对慢性癌痛病人的镇痛效应、作用持续时间及不良反应。方法:用胶原酶加机械法消化分离BCC;二步法包裹于ACA微囊内。局麻下腰椎穿刺将5mL微囊化BCC悬液(约5～7×106) 缓慢注入10例使用止痛药物的中、晚期恶性肿瘤患者蛛网膜下腔。移植后观察病人疼痛的缓解程度,止痛作用持续时间及不良反应。结果: 微囊化BCC的镇痛效应:将微囊化BCC注入病人脊髓蛛网膜下第1～2d 开始,病人的疼痛便得到不同程度的缓解,一周内疼痛完全缓解2例,部分缓解1例,轻度缓解4例,3例无效。结论:ACA微囊化BCC异种移植于慢性癌痛病人脊髓蛛网膜下安全、能够迅速发挥镇痛作用,提示微囊化BCC异种移植是治疗人类慢性疼痛的一种极具希望的新途径。     

Mutations in the satellite cell gene MEGF10 cause a recessive congenital myopathy with minicores

We ascertained a nuclear family in which three of four siblings were affected with an unclassified autosomal recessive myopathy characterized by severe weakness, respiratory impairment, scoliosis, joint contractures, and an unusual combination of dystrophic and myopathic features on muscle biopsy. Whole genome sequence from one affected subject was filtered using linkage data and variant databases. A single gene, MEGF10, contained nonsynonymous mutations that co-segregated with the phenotype. Affected subjects were compound heterozygous for missense mutations c.976T?>?C (p.C326R) and c.2320T?>?C (p.C774R). Screening the MEGF10 open reading frame in 190 patients with genetically unexplained myopathies revealed a heterozygous mutation, c.211C?>?T (p.R71W), in one additional subject with a similar clinical and histological presentation as the discovery family. All three mutations were absent from at least 645 genotyped unaffected control subjects. MEGF10 contains 17 atypical epidermal growth factor-like domains, each of which contains eight cysteine residues that likely form disulfide bonds. Both the p.C326R and p.C774R mutations alter one of these residues, which are completely conserved in vertebrates. Previous work showed that murine Megf10 is required for preserving the undifferentiated, proliferative potential of satellite cells, myogenic precursors that regenerate skeletal muscle in response to injury or disease. Here, knockdown of megf10 in zebrafish by four different morpholinos resulted in abnormal phenotypes including unhatched eggs, curved tails, impaired motility, and disorganized muscle tissue, corroborating the pathogenicity of the human mutations. Our data establish the importance of MEGF10 in human skeletal muscle and suggest satellite cell dysfunction as a novel myopathic mechanism.