Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3⁺CCR4^-CD4⁺ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-γ, TNF-α, and IL-2 in CXCR3⁺CCR4^-CD4⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4⁺ or CXCR3⁺CCR4^-CD4⁺ T cells with DC (CD4⁺/DC or CXCR3⁺CD4⁺/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-α and LPS (mDC). Although there was no significant difference between the effects of the CXCR3⁺CCR4^-CD4⁺ and CD4⁺ T cells on DC phenotype expression, CXCR3⁺CD4⁺/DC in CTL culture were able to expand number of CD8⁺ T cells and increased frequencies of IFN-γ secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3⁺CCR4^-CD4⁺ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.     

Nuclear factor of activated T cells negatively regulates expression of the tumor necrosis factor receptor-related 2 gene in T cells

Tumor necrosis factor receptor-related 2 (TR2, HVEM or TNFRSF-14) plays an important role in immune responses, however, the mechanisms regulating its expression are unclear. To understand the control of TR2 gene expression, we studied the upstream region of the gene. Gel supershift assays revealed inducible binding of nuclear factor of activated T cells (NFAT) to a putative NFAT site within the TR2 promoter. Furthermore, cotransfection of a dominant negative NFAT construct, or siRNA for NFAT, resulted in increased expression of a TR2 reporter gene. Our findings demonstrate that NFAT negatively regulates TR2 expression in activated T cells.     

冻存人淋巴细胞凝集的成因分析

本文对长期冻存的人外周血淋巴细胞的细胞膜形态和功能变化及其与细胞凝集的关系进行了研究,并对凝集的成因作了分析,初步提出了降低凝集作用的综合措施. 采用含to劣二甲基亚矾和40}人AB型血清的RPMI 164。培养液作为冻存液,将淋巴细胞在液氮中冻存1年,复温后,检测其存活率,细胞凝集现象和细胞电泳速率,并观察了细胞表面超微结构的变化. 冻存淋巴细胞复苏后,部分细胞呈现凝集状态,这是由于细胞膜结构和功能受到损伤所致,表现在细胞表面净负电荷密度的降低,电泳率减缓,细胞轻度皱缩,膜表面微绒毛增粗,出现粗大的指状突起.二甲基亚讽是造成膜结构与功能变化的一个重要因素,在常温下更加明显,它具有较强的促凝集作用.此外,淋巴细胞悬液内混杂血小板,也是促进细胞凝集的因素.     

富硒米对大鼠细胞免疫功能影响的实验研究

为探讨富硒米的剂量对大鼠免疫功能的影响和富硒米用于保健食疗提供有意义的实验依据,选用纯种ＳＤ大鼠４０只,随机分为高中低三个剂量组,分别给予富硒米和常规饲料喂养３０ｄ,称重,取血做Ｔ淋巴细胞转化和自然杀伤细胞活性试验。结果表明：①富硒米能显著提高大鼠Ｔ淋巴细胞转化率,中剂量组与对照组比较有显著性差异（ｔ检验,Ｐ〈０．０５）。②富硒米大鼠ＮＫ活性对照比较,没有显著性差异 。结论是富硒米能显著提高大鼠Ｔ     

MS-Untersuchung der Leberfunktion mit 15NH3

Dose response curves for lymphocyte chromosome aberration frequencies using X- and gamma radiations became an important and reliable indicators as biological dosimeters especially in radiation accidents and professional over exposures. Nowadays electron beams therapy are frequently used for their advantages in cases of tumours under or near to the body surface. Dose-response curves for these electron beams are rarely published. Human peripheral blood lymphocytes were in vitro irradiated with various low and high doses (0.1 Gy to 4.9 Gy) of 18 MeV electron beam to utilize such a dose-response curve using chromosome aberration frequencies as a biological marker. Then we compared this biological curve with physically obtained curves normally used in planning for radiotherapy treatment. It is interesting to find a significant difference between both of them. The biological curve is generally higher in value and that the aberrations induced by 93% of a dose is significantly higher and deeper in site than those aberrations induced by the 100% dose calculated physically. If the above observation is confirmed by detailed studies, it would be of importance to the radiotherapist to plan for isodose curves according to biological determinations.

Dosis-Wirkungs-Kurven der Mutationsraten von Lymphozyten Chromosomen bei dcr Anwendung von Röntgen- und Gammastrahlen sind wichtige und verläβliche Indikatoren in Form von biologischen Dosimetern, insbesondere bei Strahlenunfällen und bei berufsbedingten Überbestrahlungen. Gegenwärtig werden häufig auch Elektronenstrahlen wegen ihrer Vorteile bei der Therapie von Tumoren unter oder in der Nähe der Körperoberfläche eingesetzt. Dosis- Wirkungs-Kurven für diese Elektronenstrahlen werden nur selten veröffentlicht. Peripherisehe Bhutlymphozyten des Menschen wurden in vitro mit verschiedenen niedrigen und hohen Dosen (0,1 Gy … 4,9 Gy) von IS MeV-Elektronen bestrahlt, um solche Dosis-Wirkungs-Kurven als biologische Anzeiger für die Chromosomen Mutationsraten zu erhalten. Wir haben dann diese biologische Kurve mit physikalisch erhaltenen Kurven verglichen, die normalerweise bei der Planung radiotherapeutischer Behandlungsmaβnahmen verwendet werden. Es ist interessant festzustellen, daβ zwischen diesen beiden Kurven signifikante Unterschiede bestehen. Die biologische Kurve ist im allgemeinen in ihren Werten hölier und die induzierten Mutationsraten bei 93% einer Dosis sind signifikant höher und tiefer am Ort als jene induzierten Mutationsraten, die durch einer 100%ige Dosis physikalisch berechnet wurde. Sollte sich diese Beobachtung durch weitere dedaillierte Studien erhärten, so ist dies von groβer Bedeutung für Radiotherapeuten, die Isodosenkurven nach biologischen Bestimmungen zu planen haben.     

甜菜碱对小鼠脾淋巴细胞的增殖作用

研究甜菜碱对小鼠脾淋巴细胞的增殖作用.应用MTT法观察体外甜菜碱在不同浓度、不同时间对小鼠脾淋巴细胞增殖的影响,应用流式细胞仪（FCM）测小鼠脾淋巴细胞在不同时间周期的变化.结果表明,甜菜碱终浓度0.4、4、20 mmol/L均可促淋巴细胞增值,但终浓度4 mmol/L效果最为明显,在甜菜碱刺激24、48、72 h后,淋巴细胞均可显著增值,但随时间的加倍,增值幅度并不明显,综合考虑,24 h效果更好.终浓度4 mmol/L甜菜碱不同时间均可促进淋巴细胞由G0/G1期进入S期,且作用18 h效果最明显,其G0/G1期由97.03%下降到89.99%,S期由2.58%升高到9.08%.到24 h后,其诱导作用反而有些下降.因此,甜菜碱4 mmol/L作用24 h促淋巴细胞增殖最为理想.4 mmol/L的甜菜碱18 h促进淋巴细胞由G0/G1期进入S期的作用效果最好.     

绒白乳菇中提取的乳菇菌素-D免疫抑制作用的研究

研究从绒白乳菇菌中分离出的一种新型倍半萜类化合物-乳菇菌素D的免疫抑制作用及机制。运用体外培养法取BALB/C小鼠脾细胞,研究乳菇菌素D对静止脾细胞、Con A(刀豆蛋白A)和LPS(脂多糖)刺激后的脾细胞(含B淋巴细胞和T淋巴细胞)增殖的抑制作用;运用MTT法经酶标仪检测吸光度,并计算刺激指数(stimulating index,SI);体外培养小鼠脾细胞,运用ELISA检测法观察乳菇菌素D对脾脏中T淋巴细胞分泌IL-2的抑制作用。结果显示该化合物对静止的脾细胞无抑制作用;对Con A和LPS刺激后的脾细胞增殖有显著抑制作用(P0.05),最大浓度药物组刺激指数SI低至0.28;高浓度的乳菇菌素D对Con A刺激的小鼠脾细胞产生的IL-2,具有显著抑制作用(P0.05)。结果显示乳菇菌素D通过抑制抗原刺激后的T淋巴细胞和B淋巴细胞的增殖、抑制其分泌细胞因子,从而发挥免疫抑制作用。研究结果将为开发以乳菇菌素D为母体、经结构修饰、安全低毒的免疫抑制剂提供理论基础。     

Proteomics Analysis of T Lymphocytes Damage Induced by Ionizing Irradiation

Human T lymphocytes were found to be highly radiosensitive and complex cellular responses including apoptosis could be induced upon exposure to X‐ray irradiation. However, the mechanism of apoptosis associated with irradiation was not clear. In this study, a proteomic method was applied to investigation on alteration of proteome of human T‐lymphocyte cells after irradiation. The Jurkat cells were irradiated with 4 Gy X‐ray and the cell lysates were collected at different times after irradiation (6, 12, 18, 24 and 48 h). The whole proteins were separated and quantified by two‐dimensional fluorescence difference gel electrophoresis, and then the differentially expressed proteins were identified by mass spectrometry. 4 proteins exhibited significant irradiation‐induced difference in abundance, including L‐plastin, bifunctional purine biosynthesis protein, tubulin beta chain, beta‐actin. Differentially expressed proteins were reported to be directly or indirectly involved in the function of human T lymphocyte. Thus, this study might provide clues to identify proteins with biological significance related to irradiation.     

Time‐lapse Raman imaging of single live lymphocytes

We present time‐lapse Raman imaging (TLRI) of living cells as a new approach in label‐free chemical imaging through non‐electronic resonant, spontaneous Raman microspectroscopy. Raman hyperspectral datacubes of individual live peripheral blood lymphocytes were successively acquired. The Raman imaging time per voxel, with a volume of 0.3 fl, was 100 ms and the total image time of a 32 × 32 pixels image was less than 2 min. Multiple images of an individual cell have been obtained. A full series of TLRI images typically resulted in more than 1.6 million data points per image. We analyzed the datasets using hierarchical cluster analysis. A fingerprint of molecular changes was observed before the cell was blebbing. The molecular fingerprint was related to a gradual disappearance of the Raman signal from carotenoids. Concomitant changes occurred in the C H stretch high wavenumber region, presumably due to a change in the protein and lipid environment of carotenoids. These changes were smaller than 5% of the total signal at 2937 cm⁻¹. We hypothesize that the lipid environment of the carotenoids changes as a result of the photophysics in the carotenoid molecules. The detectability of carotenoids was shown to be 2.3 µMper voxel, which corresponds to 415 molecules. TLRI enables high‐speed chemical imaging not only in the intense high wavenumber region of the Raman spectrum, but particularly in the more informative fingerprint region between 500 and 1800 cm⁻¹. Copyright © 2010 John Wiley & Sons, Ltd.     

红车轴草提取物对小鼠T淋巴细胞体外活化与增殖的影响

目的:探讨红车轴草提取物(Trifoliumpratense Leguminosaeextract,TLE)对小鼠T淋巴细胞的体外活化和增殖的影响.方法:MTT比色法检测TLE对细胞的毒副作用及增殖抑制情况;双色荧光抗体染色结合流式细胞术分析TLE对小鼠T淋巴细胞在多克隆刺激剂(ConA)刺激下体外活化抗原CD69表达的影响;羧基荧光素乙酰乙酸(carboxyfluorescein diacetate,succinimidyl ester,CFDA-SE)标记技术结合流式细胞术,分析CD3 T淋巴细胞增殖相关指数.结果:TLE对小鼠淋巴结来源的淋巴细胞微毒副作用;终质量浓度为10、20、30、40 mg/L的TLE对ConA刺激下的T淋巴细胞体外活化抗原CD69表达具有明显的抑制作用(P＜001),40 mg/L抑制达到峰值;MTT法和CFDA-SE染色法都显示,10、20、30、40 mg/L TLE对ConA诱导的T淋巴细胞增殖具有显著的抑制作用(P＜001).