粒细胞集落刺激因子动员对供者外周血细胞树突状细胞亚群的影响

目的：研究粒细胞集落刺激因子(G-CSF)动员对外周血单个核细胞（PBMNC）中树突状细胞（DC）亚群及其功能的影响。方法：以CD34^-Lin^- HLA-DR⁺细胞群设门4色荧光分析方法检测25例无关供者G-CSF动员前、后外周血细胞DC亚群；ELISA检测其血清相关细胞因子IL-12p40、IL-10、IFN-γ、IL-4活性，并分析DC₁/DC₂（CD11c⁺CD123^- /CD11c^-CD123⁺）比值与CD34⁺细胞含量的相关性。结果：G-CSF动员后, 供者PBMNC 的DC₁/DC₂比值显著低于动员前（P<0.05）；DC₁/DC₂与CD34⁺细胞含量呈负相关（r=-0.438, P<0.05），CD34⁺细胞/MNC≥0.4%时， DC₁/DC₂倒置；而血清相关细胞因子IL-12p40、IL-10、IFN-γ、IL-4水平无显著改变（P>0.05）； DC₂ HLA-DR表达上调而CD83不表达。结论： G-CSF动员后，供者外周血CD34⁺细胞数量增加的同时，DC₂数量也增加，这些DC₂尽管HLA-DR表达上调，但仍处前体细胞状态，不直接分泌或调节Th2细胞分泌免疫抑制因子。     

粒细胞巨噬细胞集落刺激因子调控树突状细胞发育和功能的研究进展

树突状细胞(DC)是重要的抗原提呈细胞,专门负责抗原捕获、加工并提呈给下游T淋巴细胞从而诱导免疫应答或者免疫耐受。维持DC亚群在体内的动态平衡是保证机体免疫系统正常运转的基础。无论机体处于炎症或静息状态,粒细胞巨噬细胞集落刺激因子(GM-CSF)对DC亚群发育、分布、功能及维持DC亚群动态平衡均具有重要的调控作用。本文主要综述了GM-CSF对DC亚群发育与功能的调控作用。阐明DC发育与功能的调控机制,有助于开发新的基于DC的免疫治疗方案。     

粒细胞集落刺激因子对外周血单个核细胞功能的体外调节作用

本研究收集了15例慢性重型乙型肝炎患者和10例健康对照者的外周血单个核细胞（PBMC），在体外环境下观察G-CSF对其释放几种细胞因子TNF-α、白介素-6（IL-6）、干扰素．了（IFN-γ）、白介素．10（IL-10）的调节作用。     

新生儿感染性疾病粒细胞集落刺激因子水平的测定

采用酶联免疫吸附试验（ELISA）测定180例新儿感染性疾病血清粒细胞集落刺激因子（G－GSF）水平，同时作血培养、C反应蛋白（CRP）、白细胞计数。结果表明：阳性率分别为：G－CSF73．9％、CRP50．5％、血培养21．1％。G－CSF阳性率明显高于后二者，差异显著（P均＜0．05）。败血症组、其他感染组、肺炎组的G－CSF阳性率分别为93．9％、85．1％、51．7％。粒细胞值＞10×109／L，G－CSF阳性组达81．2％，阴性组仅18．8％，尤其在抗感染治疗以前意义更大。提示应用ELISA技术测定G－CSF阳性率高，G－CSF水平对鉴定新生儿细菌感染具有较高的敏感性和实用价值。     

国产重组人粒细胞集落刺激因子临床应用研究

利用基因技术生产的重组人粒细胞集落刺激因子（ｒｈＧ－ＣＳＦ）能作用于中性粒细胞系，具有显著的刺激中性粒细胞生成，释放并增强成熟中性粒细胞的功能，且没有明显毒副作用．但其产品美国的生白能（ＧＭ－ＣＳＦ），价格昂贵，应用受到限制．在国内首家开发出中国 ｒｈＧ－ ＣＳＦ：“吉粒芬”得到卫生部批准试用．由于价格适中，临床已较普遍应用，现把我们近两年使用情况报告如下．１资料与方法１．１对象本组使用吉粒芬患者４２例次，均是本院的住院病人：男２６例次，女１６例次，年龄５～６５岁．其中急性白血病１２例次，慢粒１０…     

粒细胞集落刺激因子在干细胞移植中的应用进展

粒细胞集落刺激因子(G-CSF)是一种促粒细胞增殖的细胞因子,主要应用于各种原因引起的粒细胞减少症。近年来大量国内外的基础及临床研究表明,G-CSF能够动员骨髓间充质及外周血中的造血干细胞,在干细胞移植领域中为干细胞的释放、富集、动员、促进迁移、诱导分化等一系列问题的解决提供了一种新思路。本文旨在对G-CSF在干细胞移植领域中的新进展进行综述。     

鼠抗粒细胞集落刺激因子单克隆抗体的研制

本文运用杂交瘤技术，成功地建立了６株能稳定分泌小鼠抗重组人粒细胞集落刺激因子（Ｇ－ＣＳＦ）单克隆抗体的杂交瘤细胞１Ｂ９Ｄ１０、１Ｇ５Ｆ１０、２Ｃ８Ｃ１、２Ｅ４Ｆ６、３Ｃ６Ｅ１１、７Ｄ２Ｃ７）。试验结果表明，６株单抗均为ｌｇＧ类，且特异性强，能分别识别不同的抗原结合位点，相对亲和力２Ｅ４Ｆ６单抗最高，１Ｇ５Ｆ１０最低，为今后Ｇ－ＣＳＦ单抗的应用打下了基础。     

两种不同HLA-B分子对NK细胞表面受体表达的研究

目的：探讨两种HLA-B分子(HLA-B51和HLA—B39)对NK细胞表面活化性受体CDl6和抑制性受体KIR3DL1表达的调节。方法：采用流式细胞仪检测NK细胞分别与转染HLA-BSl和HLA-B39分子的K562细胞相互作用后，CD16和KIR3DL1的表达情况。结果：外周血淋巴细胞与K562细胞作用24小时后，CD56 CD16 细胞数、KIR3DL1 细胞数均明显增加。与表达HLA-B39的K562细胞相比，表达HLA-B51的K562细胞与外周血淋巴细胞作用后，CD56 CD16 细胞数、KIR3DL1 细胞数均明显下降。结论：NK细胞杀伤靶细胞时，活化性受体CD16表达上调后会伴有抑制性受体KIR3DL1的上调；HLA-B51分子表达在K562细胞后，表达外源性HLA-B51分子的K562细胞与NK细胞作用，NK细胞表面受体KIR3DL1的表达可下调，同时伴有CD16的表达下调。     

Selective disbalances of peripheral blood T lymphocyte subsets in human CD3γ deficiency

The selection of T lymphocytes in the thymus and their activation upon the encounter with foreign antigens in the periphery require the aggregation and signals of the Tcell receptor (TcR)/CD3 complex and several surface molecules termed coreceptors (notably CD4 or CD8 and CD45). The spatial arrangement and interactions of the different molecules in the resulting multimolecular recognition structure are mostly unknown. Here we report, from studies on a healthy human CD3γ deficiency, that the lack of the CD3γ component of the TcR/CD3 complex is associated with a long-term severe defect of peripheral blood CD4⁺CD45RA⁺ and CD8+ lymphocytes, whereas CD4⁺CD45RO⁺, B and natural killer lymphocytes are unaffected. These results suggest that the CD3y site of the TcR/CD3 complex is required for the peripheral representation of certain Tcell types.     

强直性脊柱炎患者外周血T细胞亚群极化状态的研究

目的:分析强直性脊柱炎患者外周血中T细胞亚群的极化状态,进一步阐明强直性脊柱炎的免疫学发病机制.方法:运用流式细胞(FCM)技术,通过CD3/CD4或CD3/CD8设门,检测早、晚期AS患者和健康对照者外周血淋巴细胞的CD3 CD4 CD30 、CD3 CD4 CD30ˉ、CD3 CD8 CD30 、CD3 CD8 CD30ˉT细胞的百分率,分别表达Th1、Th2、Tc1、Tc2细胞,并进行比较分析.结果:在早期AS患者CD4 T细胞和CD4 T/CD8 T细胞的比值均明显低于健康对照组(P＜0.05),Th1细胞百分率及Th1/Th2比值明显降低(P＜0.05),CD8 T细胞的百分率明显高于健康对照组(P＜0.05),Tc2细胞明显升高(P＜0.05),且Tc1/Tc2比值明显降低(P＜0.05);在晚期AS患者Tc2细胞明显升高(P＜0.05),Tc1/Tc2比值明显降低(P＜0.05).结论:早期AS患者外周血CD4 T细胞、CD8 T细胞、CD4 T/CD8 T的变化与健康对照比较具有显著性意义,阐明了AS早期患者体内的免疫功能紊乱,呈现一种免疫功能低下的状态.重要的是Th1细胞、Th1/Th2、Tc1/Tc2的明显减少,提示在早期AS患者体内Th1/Th2、Tc1/Tc2的失调,且以Th2、Tc2占有优势,而晚期AS患者因Tc1/Tc2明显降低导致比例失衡,这些显示Th1/Th2、Tc1/Tc2的比例失衡可能是导致AS发病的重要因素,是AS发病机制之一.     

Molecular signatures induced by interleukin-2 on peripheral blood mononuclear cells and T cell subsets

Experimentally, interleukin-2 (IL-2) exerts complex immunological functions promoting the proliferation, survival and activation of T cells on one hand and inducing immune regulatory mechanisms on the other. This complexity results from a cross talk among immune cells which sways the effects of IL-2 according to the experimental or clinical condition tested. Recombinant IL-2 (rIL-2) stimulation of peripheral blood mononuclear cells (PBMC) from 47 donors of different genetic background induced generalized T cell activation and anti-apoptotic effects. Most effects were dependent upon interactions among immune cells. Specialized functions of CD4 and CD8 T cells were less dependent upon and often dampened by the presence of other PBMC populations. In particular, cytotoxic T cell effector function was variably affected with a component strictly dependent upon the direct stimulation of CD8 T cells in the absence of other PBMC. This observation may provide a roadmap for the interpretation of the discrepant biological activities of rIL-2 observed in distinct pathological conditions or treatment modalities.     

Quantitative T cell subsets profile in peripheral blood from patients with idiopathic inflammatory myopathies: tilting the balance towards proinflammatory and pro-apoptotic subsets

The role of T cells in idiopathic inflammatory myopathies (IIM) is not yet clear. Some alterations in certain subsets have been reported in inflamed muscle cells. However, a broad quantitative assessment of peripheral T cell subsets has not been evaluated. The aim of this study was to address the quantitative profile of potential pathogenic T cell subsets, namely follicular helper T cells (Tfh), T helper type 17 (Th17), CD28^null and regulatory T cells (T_regs) in peripheral blood from IIM patients. Thirty IIM patients and 30 age- and gender-matched healthy donors were included. Peripheral blood mononuclear cells were isolated. T cell subsets were evaluated by flow cytometry, as follows: Tfh (CD4⁺CXCR5⁺) and its subsets Tfh1 (CXCR3⁺CCR6⁻), Tfh2 (CXCR3⁻CCR6⁻), Tfh17 (CXCR3⁻CCR6⁺), Th17 (CD4⁺IL17A⁺), CD28^null (CD4⁺CD28⁻CD244⁺) and T_regs (CD4⁺CD25^highforkhead box protein 3 (FoxP3⁺); CD8⁺CD25^highFoxP3⁺). Percentage, absolute numbers and mean fluorescence intensity were analysed. We found increased numbers of total Tfh cells (28 ± 8·16 versus 6·64 ± 1·29, P = 0·031) in IIM patients when compared to healthy controls. Moreover, this increment was dependent upon Tfh2 and Tfh17 (Tfh2:9·49 ± 2·19 versus 1·66 ± 0·46, P = 0·005; Tfh17 9·48 ± 2·83 versus 1·18 ± 0·21, P = 0·014). Also, IIM patients showed higher numbers of Th17 cells (30·25 ± 6·49 versus 13·46 ± 2·95, P = 0·031) as well as decreased number of T_regs (5·98 ± 1·61 versus 30·82 ± 8·38, P = 0·009). We also found an expansion of CD28^null cells (162·88 ± 32·29 versus 64 ± 17·35, P = 0·015). Our data suggest that IIM patients are characterized by an expansion of peripheral proinflammatory T cells, such as Tfh and Th17, as well as pro-apoptotic CD28 null cells and a deficiency of suppressor populations of T_regs (CD4⁺ and CD8⁺).     

Identification of two distinct CD5- B cell subsets from human tonsils with different responses to CD40 monoclonal antibody

This study investigated the response of different CD5^? B cell subsets to CD40 monoclonal antibody (mAb) in various combinations with interleukin (IL)-4 or rabbit anti-human μ chain antibody (a-μ-Ab). The different CD5 B cell subsets were isolated from tonsillar B cell suspensions depleted of CD5⁺ B cells and subsequently fractionated on Percoll density gradients. While resting CD5⁺ B cells proliferated and produced IgM molecules in response to a-μ-Ab, IL-4 and CD40 mAb as well as to Staphylococcus aureus Cowan strain I (SAC) and IL-2, resting CD5^? B cells, which were co-purified in the same 60% Percoll fractions, consistently failed to respond. These cells were, however, activated by the stimuli employed, as demonstrated by their capacity to express the surface activation markers CD69, CD25 and CD71. Resting CD5⁺ B cells had the typical phenotype of mantle zone B cells (IgM⁺ IgD⁺ CD39+ CD38^? CD10^? CDw75^dim), whereas resting CD5^? B cells were CD38^? CD39^? CD 10^? CDw75 intermediate and expressed surface IgM but relatively little surface IgD and could not be classified as mantle zone or germinal center cells. The finding that purified germinal center cells (CD38⁺ CD10⁺ CD39^? CDw75^bright, IgG⁺) responded to CD40 mAb and IL-4 and also to SAC plus IL-2 further underlined the differences to resting CD5^? B cells. However, some of the data collected suggest possible relationships between CD5^? B cells and germinal center cells. The CD5^? B cells isolated from the 50 % Percoll fraction proliferated in response to a-μ-Ab, CD40 mAb and IL-4 as well as to SAC and IL-2. These cells had the same mantle zone B cell phenotype as the CD5⁺ B cells, but their capacity to respond to the stimuli in vitro was unrelated to a possible contamination with CD5⁺ B cells, as documented by the appropriate controls. Furthermore, upon exposure to SAC or phorbol esters, the large majority of CD5^? B cells from the 50 % Percoll fraction did not express surface CD5 and there was very little if any accumulation of CD5 mRNA. Finally, most of the cycling cells in the stimulated CD5^? B cells did not express CD5. The CD5^? B cells from the 50 % Percoll fraction were comprised of a consistent proportion of cells that expressed surface activation markers. The removal of these cells abrogated the capacity of the suspensions to respond to the stimuli in vitro, possibly suggesting that these cells additional activation signals in vivo which were essential to acquire the capacity to respond and that could not be reproduced in vitro. The present study underlines the phenotypic and functional heterogeneity of CD5^? B cells and contributes to the identification of two subsets of these cells which differ in phenotype, tissue distribution and in vitro responses to different stimuli.     

协同刺激分子4-1BB和CD28对人外周血不同T细胞亚群的激活作用

目的 :探讨 4 1BB/ 4 1BBL协同刺激信号在CD4 和CD8 T细胞活化、增殖中的作用 ,并与CD2 8/B7信号作比较。方法 :用抗CD3单抗 (mAb)刺激人外周血单个核细胞 (PBMC)。用阻断型抗 4 1BBLmAb和抗CD80mAb ,分别阻断 4 1BB/ 4 1BBL和CD2 8/B7 1协同刺激信号。利用流式细胞术 (FCM)检测CD4 T细胞、CD8 T细胞的增殖率、CD8/CD4T细胞的比值变化和细胞分泌IFN γ的情况。结果 :用抗 4 1BBLmAb和抗CD80mAb阻断相应的协同刺激途径后 ,CD4 和CD8 T细胞的增殖和细胞分泌IFN γ的水平均明显下降。培养 8d,抗CD3mAb单独刺激组CD8/CD4T细胞的比值为 1.98± 0 .0 6 ;抗 4 1BBLmAb阻断组CD8/CD4T细胞的比值下降为 0 .96±0 .0 3;而在抗CD80mAb阻断组 ,其比值上升为 2 .6 9± 0 .16。结论 :4 1BB分子可在CD4 T细胞和CD8 T细胞的活化、增殖中提供协同刺激信号。 4 1BB分子所介导的协同刺激信号 ,在CD8 T细胞活化及增殖中发挥了更为重要的作用 ;而CD2 8分子所介导的协同刺激信号则更有利于CD4 T细胞的活化     

慢性乙型肝炎患者外周血CD45RA+、CD45RO+T淋巴细胞亚群的特点及临床意义

目的：探讨慢性乙型肝炎患者外周血CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群的特点及其与肝病病情的关系。方法：采集46例轻中度慢性乙型肝炎患者、58例重度慢性乙型肝炎患者和30例健康人的外周抗凝血，应用流式细胞技术三色荧光分析法对其外周血中CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO＋T淋巴细胞亚群进行检测。结果：轻中、重度慢性乙型肝炎患者与正常人相比，其外周血中CD4^＋、CD8^＋T细胞均无明显改变；CD8^＋CD45RA^＋T细胞均明显降低，CD8^＋CD45RO^＋T细胞均明显增高，而CD4^＋CD45RA^＋、CD4^＋CD45RO^＋T细胞均无明显改变；重度慢性乙型肝炎患者与轻中度慢性乙型肝炎患者相比，CD8^＋CD45RA^＋T细胞明显降低（P〈0.05），CD8^＋CD45RO^＋T细胞明显升高（P〈0.05）。结论：乙型肝炎慢性化过程中，CD8^＋CD45RO^＋T细胞起重要作用且与慢性乙型肝炎患者病情的进展呈正相关；检测CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群比检测CD4^＋和CD8^＋T细胞亚群更能正确、充分、全面地了解慢性乙型肝炎的发病机制和预后，从而有效地指导临床治疗。     

CD30在肾综合征出血热患者T细胞表达的检测及其临床意义

目的 :研究CD3 0 在肾综合征出血热 (HFRS)患者急性期外周血T淋巴细胞亚群的表达及意义。方法 :应用免疫荧光抗体双重标记和流式细胞仪技术分析CD3 0 在HFRS患者急性期外周血CD 4,CD 8T细胞表达的水平。结果 :CD 4CD-3 0 细胞在重型组 ,中轻型组和正常组三组间均无显著差异 (P >0 0 5 ) ;患者CD 4CD 3 0 ,CD 8CD-3 0 细胞明显增加 ,且重型组 ,中轻型组和正常组三组间均有极显著差异 (P <0 0 1)。结论 :HFRS患者急性期存在体液免疫和细胞免疫功能亢进 ,各种T淋巴细胞亚群的比例失衡与该病免疫发病机制及病情的严重程度密切相关。