视黄酸对淋巴细胞膜IL—2Ra,TfR表达及淋巴细胞功能活性影…

用流式细胞仪观测一定剂量视黄酸对淋巴细胞蛋白介素２受体ａ链及铁传递蛋白受体表达及淋巴细胞细胞周期相细胞分布特征的影响，同时测定淋巴细胞增殖功能和＾４５Ｃａ内流，结果显示：１＞ＲＡ在２．５×１０＾－５－２．５×１０＾７ｍｏｌ／Ｌ剂量范围内能促进人外周血淋巴细胞膜ＩＬ－２Ｒａ，ＴｆＲ表达，２．５×１０＾６ｍｏｌ／Ｌ作用最明显，２．５×１０＾－４ｍｏｌ／ｌ     

人外周血淋巴细胞视黄酸受体基因的表达与调控

目的：研究维生素Ａ增强人体免疫功能的机理。方法：采用ＲＴ－ＰＣＲ技术，检测分析人视黄酸受体（ｈＲＡＲｓ）基因在脐血和成人外周血淋巴细胞（ＰＢＬ）的表达，及丝裂原和视黄酸（ＲＡ）对其的调节。结果：人外周血淋巴细胞主要表达ｈＲＡＲα亚型基因，ｈＲＡＲγ亚型基因也有低水平转录；ＰＨＡ刺激２４小时脐血淋巴细胞（ＣＢＬ）ｈＲＡＲα和ｈＲＡＲγｍＲＮＡ都较刺激前显著升高（Ｐ＜０．０５）；在ＰＨＡ或ＳＡＣ刺激同时加入生理浓度ＲＡ，ＣＢＬｈＲＡＲαｍＲＮＡ较单用ＰＨＡ（Ｐ＜０．０１）或ＳＡＣ刺激明显增高（Ｐ＜０．０５）；ＰＢＬ的ｈＲＡＲｓ基因表达受丝裂原和ＲＡ的调节变化不大。结论：维生素Ａ促进免疫功能的机制中，调节免疫细胞中ＲＡＲｓ的表达可能是一条重要途径     

视黄酸对淋巴细胞跨膜电位影响的研究

用流式细胞仪检测大剂量视黄酸（Ｒｅｔｉｎｏｉｃａｃｉｄ，ＲＡ）及生理剂量ＲＡ对淋巴细胞跨膜电位的影响。结果显示ＲＡ能诱导静止淋巴细胞起极化，尤以大剂量ＲＡ作用明显、ＲＡ与淋巴细胞３７℃温育５～３０ｍｉｎ后用ＣｏｎＡ活化．２．５×１０－４ｍｏｌ／ＬＲＡ诱导的细胞超极化反应较ＲＡ单独作用时减弱，而２．５×１０－６ｍｏｌ／ＬＲＡ先诱导一弱的超极化（温育５ｍｉｎ），然后细胞进入轻微去极化状态；另外ＲＡ与细胞温育３０ｍｉｎ后，用ＴＰＡ活化，２．５×１０－６ｍｏｌ／ＬＲＡ诱导的膜电位变化与ＴＰＡ对照无明显差异，但２．５×１０－４ｍｏｌ／ＬＲＡ的作用则使细胞进一步超极化。     

高压氧环境对小鼠脾淋巴细胞Thy—1及IL2R（α）的影响

C_(57)BL/6小鼠在0.25MPa下吸99.2％氧作用1.5h,其脾淋巴细胞Thy-1及IL-2R(α)的阳性细胞百分率明显低于正常组小鼠;而去肾上腺组和去肾上腺髓质组明显高于高压氧组与假手术组。结果提示:高压氧环境可使细胞免疫功能降低。在这一机制中,肾上腺起着重要作用。     

视黄酸受体特异性激动剂对脐血T淋巴细胞细胞因子的影响

[目的]研究体外培养中视黄酸受体(RAR)特异性激动剂四氢-四甲基萘丙烯苯甲酸(TTNPB)对脐血T淋巴细胞IFN-γ和IL-4产生的影响,初步探究其作用机制,为脐血运用于今后临床研究奠定基础.[方法]采用体外细胞培养体系,从足月正常分娩新生儿脐血中分离T淋巴细胞,加入不同浓度TTNPB(10-4 mol/L,100 mol/L,10-6 mol/L,10-7 mol/L,10-8 mol/L和空白对照组)体外培养,提取上清液,用EUSA法测定IFN-γ、IL-4的OD值,用单因素方差检验分析各剂量对IFN-γ、IL-4产生的影响.[结果]TTNPB在10-4 mol/L,IFN-γ的浓度为(12.11±5.57)pg/ml(P<0.05),IL-4的浓度为(2.937 1±0.076 6)pg/ml(P<0.05).[结论]TTNPB在10-4 mol/L下对IFN-γ有明显抑制作用,对IL-4有明显促进作用,这种调节可能是通过RAR选径起作用.     

视黄酸对幼儿淋巴结B细胞发育的影响

目的:研究视黄酸在B细胞分化发育中的作用,探讨维生素A增加抗体产生的机制。方法:取幼儿正常淋巴结细胞体外培养,加入视黄酸或视黄酸受体拮抗剂干预前后,采用流式细胞术检测分析细胞表面标志的变化,观察B细胞的成熟分化。结果:体外培养的幼儿淋巴结B细胞在无刺激剂存在时,CD19+IgM+的成熟B细胞百分比随培养时间逐渐增加,而相对不成熟的CD19+IgM-B细胞百分比逐渐减少,以48h明显(P<0.05);体外培养加入视黄酸可使CD19+IgM+B细胞百分比进一步升高,在培养24h和48h均显著高于对照组(P均<0.05);同时加入视黄酸拮抗剂则能完全拮抗该促进作用。结论:视黄酸通过视黄酸受体介导促进体外培养的淋巴结B细胞的分化发育。     

视黄酸对肠道树突状细胞成熟分化及其功能的影响

目的研究视黄酸对粘膜树突状细胞(DC)的成熟分化及其功能的影响,从免疫应答的初始环节探讨维生素A影响肠粘膜免疫的作用及其机制。方法大鼠肠粘膜体外培养,加入全反式视黄酸(RA)和/或视黄酸受体α(RARα)拮抗剂(Ro41-5253),于培养24h、48h收获后,1.采用流式细胞仪检测大鼠DC表面分化标志OX62、OX6和CD86的荧光强度,观察RA对DC成熟分化的影响;2.抽提RNA,采用荧光定量PCR法测定细胞因子和RARαmRNA表达水平,分析其对粘膜免疫的作用及途径。结果体外培养中加入RA,DC数目未受影响,但可明显促进DC成熟,并且RARαmRNA水平上调,该作用于培养24h显著。RA还可下调Th1细胞因子IL-12和IFN-γmRNA表达,对Th2细胞因子IL-4的产生无明显影响,可促进调节性细胞因子IL-10的mRNA表达。Ro41-5253可逆转RA的上述作用。结论对DC的调节作用可能是维生素A影响肠道粘膜免疫的重要机制之一,RARα参与介导了RA的调节作用。     

视黄酸受体抑动剂对正常人体T淋巴细胞增殖的影响

[目的]评价视黄酸受体RARs特异性抑动剂BIBn和视黄酸受体RXRs特异性抑动剂HX603对正常成年人体T淋巴细胞的增殖的影响,确定剂量水平,建立一套通过受体途径研究维生素A功能的实验方法。[方法]用密度梯度分离法和磁性细胞分选法从健康人的外周血分离T淋巴细胞,进行体外培养,采用MTT法,绘制人T淋巴细胞的体外生长曲线,并测定在10-4mol/L、10-5mol/L、10-6mol/L、10-7mol/L4个不同浓度时的OD值,用方差检验分析各剂量对细胞的影响。[结果]在培养24h时1×10-4mol/L、1×10-5mol/L、1×10-6mol/L、1×10-7mol/L浓度的BIBn剂量组结果OD值分别为0.538±0.128、0.692±0.068、0.627±0.136、0.730±0.157(P﹤0.05)。[结论]在培养24h时BIBn对T淋巴细胞增殖有抑制作用。HX603对T淋巴细胞增殖可能有抑制作用。     

TTNPB对体外培养的脐血T淋巴细胞分化的影响

[目的]研究视黄酸受体特异性激动剂TTNPB对体外培养的脐血T淋巴细胞表型分化的影响.[方法]采用体外人脐血淋巴细胞培养体系,以视黄酸受体(RARs)特异性激动剂TYNPB刺激细胞,TTNPB设立阴性对照组及10-4 mol/L、10-5 mol/L、10-6 mol/L、10-7 mol/L、10-8 mol/L 5个剂量组,培养72 h后收集淋巴细胞,采用流式细胞术分析T淋巴细胞表型分化情况.[结果]体外人脐血淋巴细胞培养体系能支持脐血T淋巴细胞的分化成熟.TTNPB剂量在10-4mol/L时,CDS+细胞比例为21.10%±0.10%(P=0.024),CD4+/CD8+比值为2.01±0.01(P=0.004).[结论]在体外TTNPB能抑制脐血CD4+CD8+胸腺细胞向CD8+细胞转化,TTNPB能升高CD4+/CD8+比值.视黄酸受体途径可能是维生素A调节免疫功能的机制之一.     

铁负荷过重对淋巴细胞转铁蛋白受体表达的影响

本研究采用体外细胞培养、ＡＢＣ－ＲＬＩＳＡ法、ＲＮＡ斑点杂交和原子吸收光谱分析等技术方法，观察了高浓度柠檬酸铁对健康人外周血淋巴细胞转铁蛋白受体（ＴｆＲ）及其基因表达的影响。结果显示：高铁负荷组比正常对照组ＴｆＲ减少了２２％、３８％和６１％（Ｐ＜０．０１），且ＴｆＲｍＲＮＡ水平也随着基质中铁浓度的增高而降低。细胞内铁浓度与ＴｆＲ数量变化呈显著负相关（ｒ＝－０．９７６８，ｎ＝６，Ｐ＜０．０５），说明铁负荷过重可下向调节淋巴细胞ＴｆＲ及其基因表达。     

IL-2（rs6822844）和IL-2Ra（rs2104286）单核苷酸多态性与原因不明复发性流产的相关性研究

目的：探讨白细胞介素2基因（IL-2）及其受体α（IL-2Rα）单核苷酸多态性（SNPs）与原因不明复发性流产（URSA）的相关性。方法：采用病例对照研究,应用实时荧光定量聚合酶链反应（qRT-PCR）技术,对中国浙江地区145例URSA患者及220例健康对照IL-2（rs6822844）G/T及IL-2Rα（rs2104286）A/G进行分析。结果：URSA组IL-2Rα（rs2104286）AA基因型及A等位基因频率均高于对照组,差异均有统计学意义（P＜0.05）。A等位基因携带者URSA患病风险是G等位基因携带者的2倍（OR=2.06,95%CI：1.34～3.16,P=0.001）,AA基因型较AG+GG基因型URSA患病风险增加约1倍（OR=2.27,95%CI：1.40～3.68,P=0.001）。URSA组及对照组IL-2（rs6822844）GG基因型频率均为100%,差异无统计学意义（P＞0.05）。结论：IL-2Rα（rs2104286）A等位基因可能是浙江地区URSA的遗传易感基因,IL-2（rs6822844）基因多态性与浙江地区URSA无相关性。     

维甲酸加化疗和小剂量肝素治疗急性早幼粒细胞白血病15例临床观察

目的总结维甲酸、化疗、小剂量肝素合用治疗急性早幼粒细胞白血病（APL）并预防弥漫性血管内凝血（DIC）的疗效。方法15例APL患者,采用全反式维甲酸（ATRA）、适量化疗诱导分化至完全缓解（CR）,同时予以小剂量肝素。结果APL的完全缓解率达86．7％,平均缓解时间达37．4d,无一例DIC发生。结论维甲酸、化疗、小剂量肝素合用治疗急性早幼粒细胞白血病并预防DIC效果满意。     

硒对人外周血单个核细胞IL-2系统的影响

为探讨硒、脂质过氧化反应与人外周血单个核细胞 (PBMC)功能的关系 ,离体观察了人PBMC经硒和脂质过氧化诱导剂叔丁基过氧化氢 (tBHP)作用后分泌IL - 2活性、表达IL - 2R阳性百分率和培养液中过氧化脂质 (MDA)含量变化。结果 ,PBMC产生IL - 2活性及其膜上IL - 2R表达百分率随tBHP剂量的增加而降低 ,培养液中MDA含量呈相反变化。直线相关分析表明 ,培养液中MDA含量与PBMC产生IL - 2活性水平呈负相关 (r =- 0 .978,P <0 .0 1)。而经硒预保护作用 6h的PBMC仍保持较高的IL - 2分泌和IL - 2R表达能力。同时发现硒对正常人PBMC分泌IL - 2和表达IL - 2R无显著影响。提示脂质过氧化反应可影响人PBMC的功能 ,硒对此具有保护性调节作用     

镉暴露大鼠血中白细胞介素-2和皮质醇水平及白细胞免疫功能的变化

目的探讨急性镉暴露对大鼠白细胞免疫功能及血浆白细胞介素-2(IL-2)和皮质醇水平的影响.方法取36只雄性成年SD大鼠,随机分为3组,即对照组、实验Ⅰ组、实验Ⅱ组,每组12只,分别按0、0.5、1 0 mg/kg腹腔注射CdC12,连续7 d.分别在注射后第4天和第7天宰杀大鼠,采集血样.结果在整个实验期内,Ⅰ、Ⅱ组大鼠体重均非常显著地低于对照组(P＜0.001);白细胞总数均高于对照组;Ⅰ、Ⅱ组的嗜中性粒细胞和淋巴细胞分别在染毒第7天明显高于和低于对照组(P＜0.05),而单核细胞、嗜酸性粒细胞、嗜碱性粒细胞则变化无显著性(P＞0.05);Ⅰ、Ⅱ组血中T淋巴细胞和IL-2水平低于对照组,B淋巴细胞高于对照组(P＜0.05);Ⅰ、Ⅱ组血浆皮质醇水平在染毒第7天高于对照组(P＜0 05).结论急性镉暴露可影响白细胞免疫功能,引起细胞免疫功能低下,肾上腺皮质内分泌活动显著变化.     

Mutual enhancement of IL-2 and IL-7 on DNA vaccine immunogenicity mainly involves regulations on their receptor expression and receptor-expressing lymphocyte generation

Our previous study showed that IL-2 and IL-7 could mutually enhance the immunogenicity of canine parvovirus VP2 DNA vaccine, although the underlying mechanism remained unknown. Here, we used the OVA gene as a DNA vaccine in a mouse model to test their enhancement on DNA vaccine immunogenicity and to explore the molecular mechanism. Results showed that both IL-2 and IL-7 genes significantly increased the immunogenicity of OVA DNA vaccine in mice. Co-administration of IL-2 and IL-7 genes with OVA DNA significantly increased OVA-specific antibody titers, T cell proliferation and IFN-γ production compared with IL-2 or IL-7 alone, confirming that IL-2 and IL-7 mutually enhanced DNA vaccine immunogenicity. Mechanistically, we have shown that IL-2 significantly stimulated generation of IL-7 receptor-expressing lymphocytes, and that IL-7 significantly induced IL-2 receptor expression. These results contribute to an explanation of the mechanism of the mutual effects of IL-2 and IL-7 on enhancing DNA vaccine immunogenicity and provided a basis for further investigation on their mutual effects on adjuvant activity and immune regulation.     

Effect of Retinoic Acid on Ferritin H Expression During Brain Development and Neuronal Differentiation

Abstract

We have previously shown that brain ferritin H expression, which has been associated with iron utilization, is developmentally regulated. Because retinoic acid (RA) regulates gene expression and is involved in cellular differentiation, we tested the hypothesis that RA regulates ferritin H during brain development and neuronal differentiation. RA, administered to rats on postnatal day 1, produced a 4-fold increase in brain ferritin H mRNA (p<0.01) after 24 h. To examine whether RA-stimulated neuronal differentiation contributed to this up-regulation, ferritin and ferritin H mRNA were measured in human neuronal precursor cells (NTera-2, NT2) before and after 4-weeks of RA-stimulated differentiation into post-mitotic neurons. Differentiation resulted in a 2-fold increase in both ferritin and ferritin H mRNA (p<0.05). Immunocytochemistry and Northern analysis showed significant elevations in ferritin expression that began as early as 24 h after RA treatment. While there was also a significant increase in the labile iron pool after RA treatment, this did not occur until 72 h. These data show that RA regulates ferritin H expression during rat brain development and neuronal differentiation and suggests a new role for RA in brain iron metabolism.     

Expression of IL-2α and IL-2β Receptors on the Membrane Surface of Human Sperm

Cytokines are secreted proteins that act as local immunological mediators. Increased seminal cytokine concentrations are associated with fertility problems. The purpose of the present study was to investigate the presence of IL-2 &#102, and IL-2 &#103 receptors on fresh and isolated sperm by flow cytometry and transmission electron microscopy. Twenty sperm samples from oligospermic men were incubated with CD25, a mouse monoclonal antibody specific for IL-2 &#102 -chain receptor, and CD122, a mouse monoclonal antibody specific for IL-2 &#103 -chain receptor. The strong initial fluorescence intensity and, subsequently, a labeling index yielded by CD25 and CD122 decreased in sperm centrifuged on a Percoll gradient ( p <. 05). The expression of CD25 and CD122 correlated negatively with fresh sperm concentration, but in sperm centrifuged on a Percoll gradient there was no correlation. Labeling with CD25 and CD122 antibody was evident on the head and the middle piece in fresh sperm, while in sperm centrifuged on a Percoll gradient a weak labeling was observed only on the principal piece. The authors have identified and localized cytokine receptors on human sperm for the first time. Cytokine receptors may be involved in the regulation of pathophysiological events in sperm cell functions and male infertility. The exact pathway involved in modulation of these receptors requires further investigation. These results contribute to the understanding of cytokine-sperm relationships.     

乳腺癌患者血清转化生长因子β_1与白细胞介素6和白细胞介素2水平及临床意义

目的:探讨乳腺癌患者血清中转化生长因子β1(TGFβ1)、白细胞介素6(IL-6)和白细胞介素2(IL-2)的变化及其临床意义。方法:采用放射免疫分析(RIA)法测定28例乳腺癌患者、11例乳腺良性疾病患者和30例健康妇女血清TGFβ1、IL-6和IL-2的水平。结果:乳腺癌患者血清TGFβ1和IL-6活性水平均高于乳腺良性病变组及对照组(P<0.05),而IL-2活性水平则明显降低(P<0.05)。结论:乳腺癌患者血清中TGFβ1和IL-6活性水平升高及IL-2活性水平降低可能与乳腺癌发生、发展有关,测定其水平可作为临床病情观察和评估疗效的辅助手段。