多发性硬化症脑脊液中活化T细胞抗原受体Vβ原因的表达

应用定量逆转录聚合酶链反应对４例多发性硬化症脑脊脑液细胞及未梢血单个核细胞中Ｔ细胞抗原受体Ｖβ基因表达进行了探讨。结果表明：未经刺激的ＣＳＦ细胞和ＰＢＭＣ表达广 Ｖβ基因，经ＴＣＧＦ／ＩＬ－２＝ＩＬ－４短期培养的ＣＳＦ细胞则表达限制性Ｖβ基因，而经同样培养的ＰＢＭＣ各Ｖβ基因表达的百分率同培养前ＰＢＭＣ相同。     

腺病毒、合胞病毒对外周血单个核细胞的IL-2受体表达及TNFα产生的影响

近年来国内外均注意到细胞因子及受体与病毒感染的关系。作者用１００ＴＣＩＤ５０的７型腺病毒（ＡＤＶ）及呼吸道合胞病毒（ＲＳＶ）Ｌｏｎｇ株刺激正常人体外培养的外周血淋巴细胞（ＰＢＭＣ），ＡＰＡＡＰ法检测淋巴细胞ＩＬ－２受体阳性率，酶联免疫法检测淋巴细胞上清液的ＴＮＦａ。初步观察了ＡＤＶ、ＲＳＶ对人ＰＢＭＣ的ＩＬ－２受体表达、ＴＮＦα产生的影响。经２００Ｕｇ／ｍｌ的ＰＨＡ活化的ＰＢＭＣ加入ＡＤＶ、ＲＳＶ后对照组ＩＬ－２＋细胞百分率为３４．３％，ＡＤＶ组为１７．３％，ＲＳＶ组为１７％，较对照组均显著降低…     

造血生长因子促进RV介导LacZ—Neo^R基因在人骨髓造血细胞中 …

目的：本文探索了不同组合的造血生长因子ＳＣＦ、ＩＬ３及ＩＬ６对逆转录病毒（ＲＶ）介导的ＬａｃＺ－Ｎｅｏ＾Ｒ双标志基因转染人骨髓造血细胞转染效率、表达水平的影响及其相关机理。方法：采用ＲＶ转染人骨髓非粘附造血细胞（ＮＡＢＭＣ）及经ＳＣＦ、ＩＬ３及ＩＬ６不同组合预激４８ｈ后的ＮＡＢＭＣ。经荧光素二－β－Ｄ－半乳糖呋喃苷脂（ＦＤＧ）标记的半乳糖苷酶、Ｇ４１８＾ＲＣＦＵ－ＧＭ及ＰＣＲ／Ｓｏｕｒｔｈｅｒｎ－     

亚急性重型肝炎患者TNF-α和IL-1表达及IL-4对其的调控

目的：分析ＩＬ－４对亚急性重型肝炎（亚重肝）患者外周血单个核细胞（ＰＢＭＣＳ）ＴＮＦ－α和ＩＬ－１的调控作用,评价ＩＬ－４在亚重肝治疗中的潜在价值。方法：应用细胞生物学、免疫组化和ＲＴ－ＰＣＲ等方面测定ＰＢＭＣＳ ＴＮＦ－α、ＩＬ－１的表达。结果：亚重肝患者ＰＢＭＣＳ ＴＮＦ－α、ＩＬ－１表达水平均较正常人显著增高血糖素含量和虽然ＩＬ－４ ＭＲＮＡ及蛋白质的表达与正常人比较一差异无但ＩＬ－４ ＭＲ     

T细胞受体Vβ基因在识别HSV—2过程中的表达水平和特点

本课题主要研究Ｔ细胞受体（ＴＣＲ）识别抗原后Ｖβ基因发生重排，ｍＲＮＡ表达水平改变。用紫外线处理的单纯疱疹病毒－２型（ＨＳＶ－２）、ＨＳＶ－２及ＰＨＡ分别感染或刺激正常人的ＰＢＬｓ，培养４～６天后，提取ｍＲＮＡ，并采用ＲＴ－ＰＣＲ、Ｓｏｕｔｈｅｒｎ杂交发现识别ＨＳＶ－２的ＴＣＲＶβ２、６、７、８基因在体外选择性扩增。在采用ＨＳＶ－２攻击皮肤病患者发作期、缓解期以及再发作期的ＰＢＬｓ时，发现ＴＣＲＶβ基因表达随着病程变化而改变，尤其Ｖβ７亚家族表达水平显著高于其它亚家族基因，这显示了ＴＣＲＶβ基因的变化是恒定特异性的。     

急性丙型肝炎患者免疫状况的研究

用间接免疫荧光法、酶联免疫吸附试验（ＥＬＩＳＡ）及ＬＤＨ释法，对１６例急性输血后丙型肝炎（抗－ＨＣＶ、ＨＣＶ－ＲＮＡ均阳性）患者，分别进行外周血单个核细胞（ＰＢＭＣ）的Ｔ细胞亚群计数、Ｔ４／Ｔ８比值、Ｔａｃ受体的检测及血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）ＮＫ细胞活性的测定。并与正常人组比较，经ｔ检验发现，急性丙型肝炎患者的Ｔ４亚群所占百分比、Ｔ４／Ｔ８比值及ＰＢＭＣＴａｃ受体表达均明显低于正常人组（Ｐ＜０．０５），而ＮＫ细胞活性、血清ｓＩＬ－２Ｒ明显高于正常人组（Ｐ＜０．０５）。患者的这些免疫状态改变，可能对其发病机理的研究有一定意义。     

造血生长因子促进RV介导LacZ-NeoR基因在人骨髓造血细胞中转染的机制

目的：本文探索了不同组合的造血生长因子ＳＣＦ、ＩＬ３及ＩＬ６对逆转录病毒（ＲＶ）介导的ＬａｃＺ－ＮｅｏＲ双标志基因转染人骨髓造血细胞转染效率、表达水平的影响及其相关机理。方法：采用ＲＶ转染人骨髓非粘附造血细胞（ＮＡＢＭＣ）及经ＳＣＦ、ＩＬ３及ＩＬ６不同组合预激４８ｈ后的ＮＡＢＭＣ。经荧光素二－β－Ｄ－半乳糖呋喃苷脂（ＦＤＧ）标记的半乳糖苷酶、Ｇ４１８ＲＣＦＵ－ＧＭ及ＰＣＲ／Ｓｏｕｒｔｈｅｒｎ－ｂｌｏｔ检测ＮｅｏＲ和ＬａｃＺ基因的表达。结果：早期造血生长因子（ＨＧＦｓ）的预激明显改善了ＲＶ介导的ＬａｃＺ－ＮｅｏＲ基因在人骨髓造血细胞中的转染效率与表达水平,ＳＣＦ＋ＩＬ３＋ＩＬ６＞ＳＣＦ＋ＩＬ３＞ＩＬ３＋ＩＬ６＞ＳＣＦ＋ＩＬ６。氚标记脱氧胸苷（３Ｈ－ＴｄＲ）自杀及５－溴脱氧尿苷－碘化丙啶（ＢｒｄＵ－ＰＩ）双标流式细胞仪（ＦＣＭ）检测显示,ＨＧＦｓ预激后人骨髓造血细胞及ＣＦＵ－ＧＭ的Ｓ期比例显著提高。结论：ＳＣＦ＋ＩＬ３＋ＩＬ６的联合预激可显著改善ＲＶ介导的外源基因在人骨髓造血细胞中的转染效率与表达水平,这可能与ＨＧＦｓ预激后明显提高人骨髓造血细胞及ＣＦＵ－ＧＭ的Ｓ期比例密切相关     

恶性疟原虫疫苗研究VⅡ.PfCMR基因与人白细胞介素—2基因…

用限制性内切酶ＥｃｏＲＩ和Ｓａｌｌ将恶性疟原虫复合抗原基因ＰｆＣＭＲ从质粒ＰＷＲ４５０－１／ＰｆＣＭＲ中切下，插入质粒ＰＢＶ２２０／ＩＬ－２中人白细胞介素－２（ＩＬ－２）基因的ＥｃｏＲＩ位点，重组质粒转化大肠杆菌ＤＨ５ａ，通过ＰＣＲ扩增和酶切鉴定，筛选出自向插入的重组载体ＰＢＶ２２０／ＰｆＣＭＲ－ＩＬ－２。为表达ＰｆＣＭＲ－ＩＬ－２融合蛋白打下基础。     

雷公藤单体T_4体外给药对IL－2R表达的影响

Ｔ_Ⅱ是中草药雷公藤的一类粗提物，临床治疗类风湿关节炎和系统性红斑狼疮等免疫性疾病效果显著。Ｔ_４是进一步从Ｔ_Ⅱ中提取出的单体成分。以正常人外周血单个核细胞（ＰＢＭＣ）为实验材料，测定了不同浓度的Ｔ_Ⅱ（０．０９８～３．１３Ｕ／ｍｌ）和Ｔ_４（０．０３９～５ｎｇ／ｍｌ）对于Ｔ细胞功能的影响。结果表明，Ｔ_Ⅱ和Ｔ_４对Ｔ细胞表面ＩＬ－２Ｒ的表达剂量相关的抑制作用。结果还显示，在药物作用下，ＩＬ－２对ＩＬ－２Ｒ表达的调节作用是很有限的。     

用肾综合征出血热病毒结构蛋白体外免疫正常人外周血淋巴细胞制备单克隆抗体

用亲和层析提取的肾综合征出血热病毒（ＨＦＲＳＶ）结构蛋白（５０ｋＤ和６７ｋＤ）为抗原，在体外免疫正常人外周血淋巴细胞（ＰＢＬ）。ＰＢＬ经亮氨酸甲基酯处理后，在含有ｓＰＷＭ－Ｔ、ＩＬ－２及不同浓度抗原的培养基中培养６ｄ，计数存活ＰＢＬ数量。结果表明，在上述体外免疫条件下存活ＰＢＬ的数量与抗原剂量呈正相关。将此体外免疫的ＰＢＬ与人－鼠杂交瘤细胞（Ｋ６Ｈ６／Ｂ５）融合，获得了３株分泌抗ＨＦＲＳＶ人单抗的杂交瘤细胞，表明在体外免疫条件下，正常人ＰＢＬ能够对ＨＦＲＳＶ结构蛋白发生特异性免疫应答。     

Regulatory T cells and T helper 17 cells in viral infection

CD4⁺ T cells are the central element of the adaptive immune responses and protect the body from a variety of pathogens. Starting from naive cells, CD4⁺ T cells can differentiate into various effector cell subsets with specialized functions including T helper (Th) 1, Th2, Th17, regulatory T (Treg) and T follicular helper (Tfh) cells. Among them, Tregs and Th17 cells show a strong plasticity allowing the functional adaptation to various physiological and pathological environments during immune responses. Although they are derived from the same precursor cells and their differentiation pathways are interrelated, the terminally differentiated cells have totally opposite functions. Studies have shown that Tregs and Th17 cells have rather complex interplays in viral infection: Th17 cells may contribute to immune activation and disease progression while Tregs may inhibit this process and play a key role in the maintenance of immune homoeostasis, possibly at the cost of compromised viral control. In this review, we take respiratory syncytial virus (RSV), hepatitis B virus (HBV)/hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections as examples to discuss these interplays and their impacts on disease progression in viral infection.     

Altered follicular regulatory T (Tfr)- and helper T (Tfh)-cell subsets are associated with autoantibody levels in microscopic polyangiitis patients

Antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) is an autoimmune disease characterized by B cells-derived ANCAs, and ANCA was proved to be a key factor in its pathogenesis. Follicular regulatory T (Tfr) and follicular helper T (Tfh) cells were T-cell subsets that play important roles in B-cell maturation and antibody production. However, their significances in microscopic polyangiitis (MPA) patients, one type of AAV, has not been thoroughly studied. In this study, comprehensive pattern analyses of circulating Tfr and Tfh were performed in MPA patients and healthy controls (HCs), and we found Tfr levels and Tfr/Tfh ratios were significantly decreased in MPA patients. Compared with HCs, Helios+, CD45RA-FoxP3hi, and Ki-67+ Tfr were lower in MPA patients, while CD226+ Tfr cells were higher. These phenotypes suggest that function and proliferation ability of Tfr cells were relatively impaired. Tfh subsets, including ICOS+PD-1+ and Ki-67+ Tfh, were significantly increased, suggesting that the function of Tfh was enhanced in MPA although the total Tfh levels did not change significantly. Circulating memory B cells and plasmablasts were significantly elevated and negatively correlated with Tfr levels and Tfr/Tfh ratios in MPA patients. In addition, Tfr levels and Tfr/Tfh ratios were negatively while Tfh was positively correlated with serum myeloperoxidase (MPO)-ANCA levels. Furthermore, Tfr and Tfr/Tfh ratio were also reversely associated with SCr, BUN, IL-4, and IL-21 levels. Our results suggest that the imbalance of Tfr and Tfh functional subsets is related to increased level of autoantibodies in MPA patients, and we propose a new mechanism for the pathogenesis of MPA.     

人体T细胞CD_8~+VV~+亚群在活动性SLE患者中显示反抑制活性

反抑制T 细胞(TCS)借助T 辅助细胞对抗T 抑制细胞的抑制效应,参与免疫调节。小鼠和人体的TCS 都表达长绒毛野豌豆凝集素受体(VV—R)。人体TCS 则主要是CD_8~+VV~+T 亚群,和临床多种疾病免疫功能异常密切相关。本文用抗VV 抗体单色和双色免疫荧光法和流式法细胞术(FCM)测知正常人VV~+T 亚群的比例是16.6±2.9%。用FCM 双色免疫荧光法检测到活动期SLE 患者CD_8VV~+亚群百分比高达23.63%和对照组(8.50%)及稳定期(14.49%)相比差异显著。然而CD_4~+VV~+亚群在三组SLE 中基本不变。进一步深入分析是做有关CD_8VV~+T 细胞亚群的功能测定,证实活动性SLE 患者VV~+T 细胞具有比正常个体高得多的反抑制活性,相对反应值从100%提高到485%和625%。提示CD_(?)VV~+细胞和SLE 免疫调节功能紊乱密切相关。     

Tocilizumab for the treatment of chimeric antigen receptor T cell-induced cytokine release syndrome

ABSTRACT

Introduction: Cancer-directed immunotherapies are transforming the landscape in oncology as new and exciting therapies move from the laboratory to the bedside. Chimeric antigen receptor T (CAR-T) cells are one of these novel therapies, demonstrating impressive efficacy against B-cell malignancies. With the development of new therapies, it is not uncommon to identify new and unanticipated toxicities. CAR-T cells cause unique toxicities not typically found with traditional cytotoxic chemotherapy or small molecule inhibitors.

Areas covered: CAR-T cell associated toxicities include cytokine release syndrome (CRS) and CAR-T cell-related encephalopathy syndrome (CRES), alternatively known as immune effector cell-associated neurotoxicity syndrome (ICANS). Prompt identification and management of CRS and CRES are imperative for the prevention of life-threatening complications of these innovative therapies. This literature review describes the seminal trials of CD19-directed immunotherapy and the pathophysiology and management of the toxicities found with CAR-T cells. In addition, the use of the interleukin-6 receptor antibody tocilizumab for CRS is reviewed.

Expert opinion: This review describes the recommended management of CRS and CRES and examines the current limitations in management. Alternative therapies for the treatment of CAR-T cell related toxicities are also explored. Furthermore, the review proposes future directions for research.     

Measuring renal tissue relaxation times at 7 T

As developments in RF coils and RF management strategies make performing ultra‐high‐field renal imaging feasible, understanding the relaxation times of the tissue becomes increasingly important for tissue characterization, sequence optimization and quantitative functional renal imaging, such as renal perfusion imaging using arterial spin labeling. By using a magnetization‐prepared single‐breath‐hold fast spin echo imaging method, human renal T₁ and T₂ imaging studies were successfully performed at 7 T with 11 healthy volunteers (eight males, 45 ± 17 years, and three females, 29 ± 7 years, mean ± standard deviation, S.D.) while addressing challenges of B₁⁺ inhomogeneity and short‐term specific absorption rate limits. At 7 T, measured renal T₁ values for the renal cortex and medulla (mean ± S.D.) from five healthy volunteers who participated in both 3 T and two‐session 7 T studies were 1661 ± 68 ms and 2094 ± 67 ms, and T₂ values were 108 ± 7 ms and 126 ± 6 ms. For comparison, similar measurements were made at 3 T, where renal cortex and medulla T₁ values of 1261 ± 86 ms and 1676 ± 94 ms and T₂ values of 121 ± 5 ms and 138 ± 7 ms were obtained. Measurements at 3 T and 7 T were significantly different for both T₁ and T₂ values in both renal tissues. Reproducibility studies at 7 T demonstrated that T₁ and T₂ estimations were robust, with group mean percentage differences of less than 4%. Copyright © 2014 John Wiley & Sons, Ltd.     

Increased brain perfusion contrast with T2‐prepared intravoxel incoherent motion (T2prep IVIM) MRI

The feasibility to measure brain perfusion using intravoxel incoherent motion (IVIM) MRI has been reported recently with currently clinically available technology. The method is intrinsically local and quantitative, but is contaminated by partial volume effects with cerebrospinal fluid (CSF). Signal from CSF can be suppressed by a 180° inversion recovery (180°‐IR) magnetization preparation, but this also leads to strong suppression of blood and brain tissue signal. Here, we take advantage of the different T₂ relaxations of blood and brain relative to CSF, and implement a T₂‐prepared IVIM (T2prep IVIM) inversion recovery acquisition, which permits a recovery of between 43% and 57% of arterial and venous blood magnetization at excitation time compared with the theoretical recovery of between 27% and 30% with a standard 180°‐IR. We acquired standard IVIM (IVIM), T2prep IVIM and dynamic susceptibility contrast (DSC) images at 3 T using a 32‐multichannel receiver head coil in eight patients with known large high‐grade brain tumors. We compared the contrast and contrast‐to‐noise ratio obtained in the corresponding cerebral blood volume images quantitatively, as well as subjectively by two neuroradiologists. Our findings suggest that quantitative cerebral blood volume contrast and contrast‐to‐noise ratio, as well as subjective lesion detection, contrast quality and diagnostic confidence, are increased with T2prep IVIM relative to IVIM and DSC. Copyright © 2014 John Wiley & Sons, Ltd.     

Adoptive transfer of all-trans-retinal-induced regulatory T cells ameliorates experimental autoimmune arthritis in an interferon-gamma knockout model

Maintaining an appropriate balance between subsets of CD4⁺ helper T cells and T regulatory cells (Tregs) is a critical process in immune homeostasis and a protective mechanism against autoimmunity and inflammation. To identify the role of vitamin A-related compounds, we investigated the regulation of interleukin (IL)-17-producing helper T cells (Th17 cells) and Tregs treated with all-trans-retinal (retinal). CD4⁺T cells or total cells from the spleens of C57BL/6 mice were stimulated under Treg-polarizing (anti-CD3/CD28 and TGF-β) or Th17-polarizing (anti-CD3/CD28, TGF-β, and IL-6) conditions in the presence or absence of retinal. To analyze their suppressive abilities, retinal-induced Tregs or TGF-β-induced Tregs were co-cultured with responder T cells. Collagen-induced arthritis (CIA) was established in interferon (IFN)-γ knockout mice. On day 13, retinal-induced Tregs were adoptively transferred to mice with established CIA after second immunizations. Compared with TGF-β-induced Treg cells, retinal-induced Tregs showed increased Foxp3 expression and mediated stronger suppressive activity. Under Th17-polarizing conditions, retinal inhibited the production of IL-17 and increased the expression of Foxp3.Retinal-induced Tregs showed therapeutic effects in IFN-γ knockout CIA mice. Thus, we demonstrated that retinal reciprocally regulates Foxp3⁺ Tregs and Th17 cells. These findings suggest that retinal, a vitamin A metabolite, can regulate the balance between pro- and anti-inflammatory immunity. A better understanding of the manipulation of Foxp3 and Tregs may enable the application of this tremendous therapeutic potential in various autoimmune diseases.     

T波检测和QT间期提取策略

提出一种T波检测和QT间期提取新策略,应用QRS波群起始点和终末点检测算法,检测到QRS波群的起始点和终末点;从QRS波群的终末点出发,向后求出16点线段参数的LS估计;根据LS估计确定窗口,在窗口内检测出T波的峰谷值位置,从而检出T波;从峰谷值位置向后根据LS估计确定R点和R回归直线,根据心电数据和R回归直线在R点前的偏离程度确定T波终末点,从而提取QT问期.应用具有广泛认可度的MIT-BIH数据库中QT数据库的所有具有T波终末点专家标记的数据文件来验证算法,在专家标记终末点的3 542个T波上获得98.2%的检出率,提取QT间期获得1.0 ms的平均误差,提取QT间期的准确率为97.2%.     

男性不育症患者血清IL-6、T、LH、FSH、PRL检测的临床意义

目的：探讨了男性不育症患者血清IL-6、T、LH、FSH和PRL水平的变化及临床意义。方法：应用放射免疫分析对68例男性不育症患者进行了血清IL-6、T、LH、FSH和PRL检测,并与35名男性正常人作比较。结果：男性不育症患者血清T水平非常显著地低于男性正常人组（P〈0.01）,而IL-6、FSH、LH和PRL水平又非常显著地高于男性正常人组（P〈0.01）。血清IL-6水平与T水平呈负相关（r=-0.4186,P〈0.01,与FSH、LH和PRL水平呈正相关（r=0.6018、0.5912、0.6214,P〈0.01）。结论：检测男性不育症患者血清IL-6、T、LH、FSH和PRL水平的变化对诊断、预后判断及治疗均有较大的实用价值。     

心电向量图T环改变对冠心病诊断价值的探讨

目的:探讨心电向量图T环改变对冠心病的临床诊断价值。方法:对56例冠心病患者及60例健康人进行心电向量T环改变统计分析。结果:冠心病组在QRS/T值、QRS-T夹角、T环的L/W比值、T环角度方面与对照组有显著性差异(P<0.01),而在T环转位方面无显著性差异(P>0.05)。结论:T向量环改变对冠心病诊断有重要价值。