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1.
目的 探讨口腔鳞癌患者外周血CD57+细胞亚群变化及临床意义。方法 入组2018年11月至2019年12月就诊的可手术的口腔鳞癌患者,根据临床分期分为早期组和局部中晚期组。采用流式细胞仪检测外周血CD3+淋巴细胞、CD57+细胞、CD57+CD8+T细胞、CD57+CD4+T细胞、CD57+CD56+CD16+和CD57+CD56-CD16+NK细胞计数;以CD57+CD8+T细胞比例水平中位数为界值,绘制Kaplan-Meier生存曲线,分析CD57+CD8+T细胞低水平组和高水平组生存风险。结果与早期患者相比,局部进展期的口腔鳞癌患者CD57+淋巴细胞(P=0.001)和CD57+<...  相似文献   

2.
背景:课题组已建立胎儿骨髓基质细胞联合细胞因子的造血细胞体外培养体系,该培养体系能否有效扩增各个发育阶段的造血细胞有待验证。 目的:观察骨髓基质细胞联合细胞因子培养体系对脐血单个核细胞表面抗原CD133、CD34表达的影响。 方法:将从脐血标本中分离出来的单个核细胞接种于无血清培养体系,实验分为3组:①F组:干细胞因子+Flt3配体+促血小板生成素+单个核细胞。②S组:基质细胞+单个核细胞。③SF组:基质细胞+干细胞因子+Flt3配体+促血小板生成素+单个核细胞。在第0,6,10,14天检测有核细胞总数、CD133+、CD34+、CD133+CD34+细胞数以及集落形成单位数。 结果与结论:SF组有核细胞总数在各个检测时间点均比其他两组高;除了第14天外,第6、10天两个时间点SF组中CD133+、CD34+、CD133+CD34+细胞及集落形成单位数均高于其他组;含骨髓基质细胞的S组和SF组中CD133+细胞/有核细胞、CD34+细胞/有核细胞、CD133+CD34+细胞/有核细胞的比例保持在较高的水平。结果说明骨髓基质细胞联合细胞因子能有效的扩增脐血单个核细胞及其中的CD133+、CD34+、CD133+CD34+细胞,基质细胞对维持造血干细胞的原始性具有重要的作用。  相似文献   

3.
背景:上调CD4+CD25+CD127low/-调节性T细胞是目前治疗哮喘的新靶点。骨髓间充质干细胞在体外可上调正常人外周血的调节性T细胞,但机制尚未明确。 目的:观察血红素加氧酶1对间充质干细胞上调哮喘患者外周血CD4+CD25+CD127low/-调节性T细胞的影响。 方法:分别加入0,15,30,45,60 μmol/L氯化高铁血红素和0,5,10,15,20 μmol/L锌-原卟啉对骨髓间充质干细胞进行预处理,荧光定量PCR检测各组间充质干细胞中血红素加氧酶1 mRNA的表达量。密度梯度离心法分离哮喘急性发作期患者和健康对照者的外周血单个核细胞,分别与经氯化高铁血红素预处理、经锌-原卟啉预处理、不经预处理的间充质干细胞共孵育,加入植物血凝素反应72 h,流式细胞仪检测各组CD4+CD25+CD127low/-调节性T细胞占CD4+T细胞的比例。 结果与结论:人骨髓间充质干细胞中有血红素加氧酶1表达,其表达在体外可被诱导和抑制。间充质干细胞中血红素加氧酶1 mRNA的表达量随氯化高铁血红素浓度增加而增加(P < 0.05),随锌-原卟啉浓度增加而减少(P < 0.05),具有剂量依赖性。间充质干细胞在体外可提高哮喘患者和健康对照者CD4+CD25+CD127low/-调节性T细胞的水平(P < 0.01),诱导间充质干细胞中血红素加氧酶1的表达可使共孵育后CD4+CD25+CD127low/-调节性T细胞水平提高(P < 0.01),抑制间充质干细胞中血红素加氧酶1的表达可使共孵育后CD4+CD25+CD127low/-调节性T细胞的水平降低(P < 0.01),但仍高于对照组(P < 0.01)。提示骨髓间充质干细胞部分通过血红素加氧酶1上调哮喘患者外周血CD4+CD25+CD127low/-调节性T细胞。  相似文献   

4.
背景:通过促进调节性T细胞的产生及增强其功能的发挥已成为维持移植物免疫耐受的有效手段。 目的:探讨他克莫司和西罗莫司对行肝移植的肝癌患者Foxp3+调节性T细胞产生及肝癌复发的影响。 方法:纳入符合米兰标准的肝癌肝移植患者40例,随机分为西罗莫司组和他克莫司组,每组20例,移植后第2~12个月间每月抽取受试者外周血检测Foxp3+调节性T细胞,并行彩超和外周血检测甲胎蛋白,必要时肝穿刺活检观察排斥反应及肿瘤的复发情况。 结果与结论:流式细胞仪检测结果显示,西罗莫司组外周血Foxp3+调节性T细胞阳性率明显高于他克莫司组(P < 0.05),移植肝穿刺活检证实西罗莫司组排斥反应与他克莫司组差异无显著性意义(P > 0.05),而移植肝彩超、外周血甲胎蛋白检测及移植肝穿刺活检或手术亦证实在肝癌复发率方面西罗莫司组明显低于他克莫司组(P < 0.05)。说明西罗莫司在肝癌肝移植中对肿瘤复发的抑制作用方面优于他克莫司,且排斥反应较他克莫司并未增加,甚至有更好的免疫耐受效果。  相似文献   

5.
背景:有实验表明CXCR3拮抗剂Genistein可减轻胰腺移植大鼠的急性排斥反应。 目的:观察Genistein联合小剂量他克莫司在胰腺移植抗排斥反应中的作用。 方法:以Wistar大鼠为供体,SD大鼠为受体,建立胰腺移植模型,随机分成5组:对照组(未进行药物治疗)、大剂量他克莫司组、小剂量他克莫司组、Genistein组、Genistein+小剂量他克莫司组。术后第7天行病理学,肝肾功能,血清中CD3+、CD4+、CD8+T淋巴细胞、干扰素γ、白细胞介素2浓度检测。 结果与结论:与对照组、小剂量他克莫司组、Genistein组比较,Genistein+小剂量他克莫司组移植胰腺组织损伤减轻,淋巴细胞浸润减少,急性排斥反应减轻,血清CD3+、CD4+、CD8+ T淋巴细胞明显减少,干扰素γ、白细胞介素2水平降低(P < 0.05);并且避免了大剂量他克莫司对肝肾功能的损害。说明Genistein联合小剂量他克莫司能有效减轻胰腺移植急性排斥反应而不增加肝肾毒性。  相似文献   

6.
 目的:建立分析CD3+、CD4+和CD8+ T细胞Vβ亚家族中免疫抑制性受体程序性细胞死亡蛋白1(PD-1)分子免疫表型的方法,从而了解人体外周血T细胞谱系中PD-1表型细胞的分布频率。方法:收集健康个体(HI)外周血10例,利用抗CD45、CD3、CD4、CD8、PD-1等多色荧光流式单抗,以及T细胞受体(TCR)Vβ谱系试剂盒[IOTest® Beta Mark TCR Vβ Repertoire Kit,共8管,每一管均为FITC和PE偶联的复合抗体(A~H),每一管复合抗体包含3个TCR Vβ亚家族的抗体],检测T细胞亚群TCR Vβ亚家族中PD-1的分布情况。结果:在10例HI的检测中,CD3+、CD3+CD4+和CD3+CD8+ T细胞亚群中检测的24个TCR Vβ亚家族总和符合试剂盒所提供的Quick Reference Card数据,初步结果显示,HI中CD3+ T细胞主要优势TCR谱系是Vβ2、Vβ3、Vβ8、Vβ9、Vβ5.1、Vβ13.1和Vβ13.2;而在CD3+CD4+ T细胞中的优势利用主要集中在Vβ2、Vβ3、Vβ8、Vβ9、Vβ5.1和Vβ13.1;在CD3+CD8+ T细胞中则主要为Vβ1、Vβ2、Vβ3、Vβ9、Vβ13.1和Vβ13.2等亚家族。进一步分析显示PD-1+细胞在HI的CD3+、CD3+CD4+和CD3+CD8+ T细胞24个TCR Vβ亚家族中均可以检测到,PD-1+细胞在T细胞各亚群中分布频率有个体差异,在CD4+ T细胞中,以Vβ5.2+ T细胞的分布频率较高,而在CD8+ T细胞中,以Vβ11+和Vβ13.6+ T细胞的分布频率较高。结论:本项工作利用健康人样本成功建立了多色荧光流式细胞术检测T细胞亚群TCR Vβ谱系中免疫抑制性受体PD-1分子免疫表型的方法,为进一步应用于分析白血病等病人TCR Vβ谱系的免疫抑制特点提供了新方法。  相似文献   

7.
背景:他克莫司应用于同种异体肝移植的免疫耐受已多见报道。 目的:探索他克莫司在异体肢体移植时的最佳应用剂量。 方法:建立同种异体肢体移植大鼠模型,移植造模后设立给予不同他克莫司剂量的0.5,1,2 mg/(kg•d)组及对照组,对大鼠进行大体观察、组织学、淋巴细胞亚群测定。 结果与结论:移植后出现排斥反应的时间分别是对照组(3.43±0.79) d、0.5 mg/(kg•d)组(5.68±0.97) d、1 mg/(kg•d)组(9.13±1.17) d、2 mg/(kg•d) 组(9.61±2.38) d,排斥反应的病理分级4组分别为(2.61±0.38),(1.57±0.43),(0.85±0.24),(0.71±0.19)级。移植后各给药组CD4+、CD8+检测值均明显下降,CD4/CD8比值轻度增加或在正常值范围内;对照组CD4+、CD8+无明显变化,CD4/CD8比值明显增加。提示,他克莫司的应用剂量在1 mg/(kg•d)时就能达到理想的抑制免疫排斥反应,提高用量后意义不大。  相似文献   

8.
目的:建立一种检测HIV-1特异性CD4+T细胞亚群功能的活化诱导标记法(activation-induced markers,AIM),从而更有效地评价HIV-1抗原特异性CD4+T细胞免疫应答水平。方法:选取12例未经抗病毒治疗的HIV-1慢性感染者及6例未感染HIV-1的健康人,分别以基于多色流式细胞术的AIM法和细胞内因子染色法(intracellular cytokine staining,ICS)检测抗原特异性T淋巴细胞功能,并探讨两种方法用于评价HIV-1感染者抗原特异性T细胞免疫应答的能力。结果:AIM法检测HIV-1慢性感染者中HIV-1抗原特异性PD-1+CD25+CD4+T、CD69+CD200+CD4+T、CD69+ICOS+CD4+T细胞阳性的比例为11/12、8/12和7/12,检测CD69+ICOS+CD8+T、CD137+CD69+CD8+T、PD-1+CD25+CD8+、OX40+PD-1+CD8+T细胞阳性的比例为8/12、8/12、7/12、7/12。ICS法检测HIV-1抗原特异性IL-2+CD4+T、IFN-γ+CD4+T、TNF-α+CD4+T细胞阳性的占比为2/12、2/12、0;IFN-γ+CD8+T、TNF-α+CD8+T、IL-2+CD8+T细胞阳性的占比为12/12、10/12、5/12。结论:AIM法在评价CD4+T细胞功能方面更为敏感,可作为ICS法的补充,两种方法联合使用可更全面地评估抗原特异性T淋巴细胞反应。  相似文献   

9.
背景:肝移植后受者体内绝大部分病毒负荷被清除,植入新肝后其复发性肝炎病原体从肝外进入肝内的途径及其复制规律目前尚无定论。 目的:检测肝移植前后外周血单个核细胞和肝组织中乙型肝炎病毒cccDNA及血清中乙型肝炎病毒DNA的表达。 方法:采用淋巴细胞分离液从乙肝病毒相关终末期肝病37例患者外周血中分离出单个核细胞,采用荧光定量PCR检测肝移植前后及移植后乙肝复发3个时期外周血单个核细胞和肝组织中cccDNA及血清乙型肝炎病毒DNA表达。 结果与结论:肝移植前,单个核细胞cccDNA阳性12例,肝组织cccDNA阳性6例,检出率分别为32%和16%,单个核细胞、肝组织中cccDNA拷贝范围分别为(3.028~6.508)×104,(4.158~6.234)×104 拷贝/mL。肝移植后,单个核细胞cccDNA阳性1例,无血清乙型肝炎病毒DNA检测阳性病例。6例肝移植后乙肝复发病例中外周血单个核细胞cccDNA阳性4例,肝组织活检cccDNA阳性1例,6例血清乙型肝炎病毒 DNA均为阳性。提示乙肝病毒相关终末期肝病患者肝移植后乙肝复发途径可能是残留乙肝病毒在外周单个核细胞中以cccDNA为模板复制,然后再迁移到肝脏。  相似文献   

10.
背景:急性心肌梗死后具有动员骨髓和外周血中的CD34+干细胞的潜能,参与坏死心肌的自身修复,但外周血CD34+细胞与其他相关指标的关系尚不明确, 目的:观察急性心肌梗死患者外周血单个核CD34+细胞的动态变化规律,并进一步分析其与肌酸激酶同工酶、N末端B型利钠肽原、左室射血分数的相关性。 方法:采集急性心肌梗死患者(实验组)发病第2,3,4,5,7,30,90,120天的外周静脉血及入院后相应时间点非冠状动脉粥样硬化性心脏病患者(对照组)外周静脉血,采用流式细胞仪测定外周血单个核细胞CD34+的百分率。同时对心肌梗死患者进行肌酸激酶同工酶、N末端B型利钠肽原及心脏超声检测。 结果与结论:①实验组单个核细胞CD34+细胞百分率于心肌梗死后第3天开始升高,第4天达到高峰,第90天回降至基线水平。对照组单个核细胞CD34+细胞百分率无明显变化。②实验组外周血单个核CD34+细胞与左室射血分数呈正相关关系,与N末端B型利钠肽原呈负相关关系;与肌酸激酶同工酶无相关性。提示急性心肌梗死后4~8 d内给予干细胞治疗可适度改善心脏收缩功能,并且较高的CD34+干细胞表达有益于急性心肌梗死后心肌功能的恢复。  相似文献   

11.
背景:凋亡细胞能够主动调节机体的免疫功能,并能通过调节机体细胞免疫和体液免疫的途径诱导免疫耐受,但这些结果只在大鼠肝脏移植模型中证实。 目的:探讨通过60Co γ射线体外处理后的供体淋巴细胞预输注诱导猪肝移植特异性免疫耐受的作用中,对淋巴细胞亚群的影响。 方法:建立非转流小型猪原位肝移植模型。将受体猪随机摸球法均分为2组:空白对照组,受体猪无特殊处理,行肝移植;淋巴细胞组:受体猪在肝移植前7 d经耳静脉注射60Co γ射线处理过的5×108个供体淋巴细胞。观察两组受体猪移植后的存活时间,移植后T淋巴细胞亚型CD4+T、CD8+T、CD4+CD25+Tr变化及病理。 结果与结论:移植后3 d,两组病理活检均呈急性中、重度排斥反应;移植后6 d,两组均呈急性重度排斥反应。移植后1,3,6 d CD4+T、CD8+T、CD4+CD25+Tr升降趋势,两组间差异无显著意义(P > 0.05)。提示,60Co γ射线体外处理过的淋巴细胞预输注未能够诱导猪同种异体肝移植特异性免疫耐受,未能引起T淋巴细胞亚群变化有关。  相似文献   

12.
BACKGROUND:Tumor stem cells are the root of cancer recurrence and metastasis, so clinical researches should focus on the effects of different treatments on tumor stem cells. OBJECTIVE:To explore the effects of endocrine therapy and chemotherapy on stem cells in patients with breast cancer. METHODS:After recovery and cultivation of estrogen receptor-positive human breast cancer cell lines MCF-7, passage 3 cells in logarithmic phase were selected and divided into three groups containing control, estradiol and estradiol with tamoxifen groups. The estradiol group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol was added into the medium, respectively; the estradiol with tamoxifen group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol with 10-6 mol/L tamoxifen were added into the medium, respectively. The same amount of absolute ethyl ethanol was added into the medium of control group. Fifteen female patients with late recurrence and metastasis of breast cancer received chemotherapy as recurrence and metastasis group. Another 15 healthy volunteers were selected as healthy control group. RESULTS AND CONCLUSION:The proportion of CD44+CD24-/low cell subsets in the estradiol and estradiol with tamoxifen groups was significantly higher than that of the control group (P < 0.05), and the proportion of CD44+CD24-/low cell subsets in the estradiol group was significantly higher than that of the estradiol with tamoxifen group at the same concentration (P < 0.05). The proportion of CD44+CD24-/low cell subsets had no significant differences among groups at 10 and 20 days of culture (P < 0.05). The proportion of CD44+CD24-/low cell subsets significantly increased in MCF-7 cells after 24-hour intervention with different chemotherapy drugs. But only the proportion of CD44+CD24-/low cell subsets in the paclitaxel and doxorubicin groups was significantly higher than that of the control group after 20-day intervention (P < 0.05). Besides, the proportion of CD44+CD24-/low cell subsets in the peripheral blood of healthy volunteers was significantly lower than that of the recurrence and metastasis group (P < 0.05). Among 15 patients with late recurrence and metastatic of breast cancer, 9 had stable disease, 5 had partial remission, 1 had failed chemotherapy and cancer progression. Moreover, the proportion of CD45-CD44+CD24-/low cell subsets in the peripheral blood of patients sensitive for chemotherapy was significantly lower than that before treatment (P < 0.05). In conclusion, both endocrine therapy and chemotherapy exert a certain effect on the CD44+CD24-/low cell subsets of breast cancer positive for estrogen receptor. Given that CD44+CD24-/low cell subsets in MCF-7 cells resist chemotherapy drugs, the proportion of CD45-CD44+CD24-/low cells in the peripheral blood of patients sensitive for chemotherapy is decreased.  相似文献   

13.
Infections are a major cause of illness and death amongst elderly people. Peripheral blood CD8+ T lymphocytes -which play a crucial role in host defence against viral infections-, are divided in subsets based upon the expression of several cell and activation markers. Since in senescence changes in peripheral blood CD8+ T lymphocyte compartment have been described, studies were performed to determine whether in aging there are variations in the peripheral blood CD8+CD38+, CD8+CD57+, CD8+HLA-DR+, CD8+CD45RA+ and CD8+CD45RO+ cell subset. A decrease in the CD8+CD45RA+ lymphocytes was observed, indicating that variations in the CD8+ compartment can take place with ageing.  相似文献   

14.
CD28CD4+ T-cell subpopulation is expanded in kidney allograft patients with long graft survival. To seek for the roles of CD28CD4+ T cells in the long-term acceptance of kidney allografts, we characterized this population by analyzing cell surface molecules, TCR Vβ repertoire, mixed lymphocyte reaction (MLR), and cytokine production. The number of CD28CD4+ T cells increased correlatively with time after transplantation in this group of patients. The CD28CD4+ T cells did not express detectable levels of CD25, CD69, V24, or CTLA-4 but expressed heterogeneous amounts of CD45 RA on the surface. Freshly sorted CD28CD4+ T cells revealed a restricted Vβ repertoire, whereas the Vβ usage of CD28+CD4+ T cells from the same patients was much diversified. Expression levels of TGF-β and IFNγ gene were significantly higher in the CD28 CD4+ T cells than in the CD28+CD4+ T cells from the kidney allograft patients. These findings suggest that an oligoclonal CD28 CD4+ T-cell population is continuously activated in patients with long allograft survival, which may be linked with the long-term acceptance.  相似文献   

15.
BACKGROUND:Heart transplantation is an effective method for treatment of end-stage heart failure, but immune rejection that seriously impact therapeutic effacicy is easy to occur after transplantation. OBJECTIVE:To investigate the regulatory effect of bone marrow mesenchymal stem cells on the immune function of rats undergoiong heart transplantation. METHODS:Twenty Lewis rats were enrolled as donors, and 20 Wistar rats as recipients. Heart transplantation models were established in the Wistar rats. These 20 model rats were randomized into cell transplantation and control group with 10 rats in each group. Forty-eight hours after heart transplantation, rats in the cell transplantation group were given bone marrow mesenchymal stem cell suspension (1 mL, 2×108 cells/L) via the tail vein, while rats in the control group were given normal saline in the same dose. Then, the expression levels of serum interleukin-2, interleukin-10 and percentage of CD4+, CD8+, CD4+/CD8+, CD4+CD25high, CD4+CD25high Foxp3+ T cells in the venous blood were detected in the two groups at 7 days after cell transplantation. Additionally, rat myocardial tissues were taken and observed pathologically. RESULTS AND CONCLUSION:The survival time of the cell transplantation group was significantly longer than that of the control group (P < 0.05). The expression level of interleukin-2 showed no significant difference between the two groups (P > 0.05), but the level of interleukin-10 in the cell transplantation group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the percentage of CD4+/CD8+, CD4+CD25high, CD4+CD25high Foxp3+ and CD4+ T cells was significantly higher, and the percentage of CD8+ T cells was significantly lower in the cell transplantation group (P < 0.05). Histopathological findings showed that there were a small amount of infiltrated lymphocytes in the cell transplantation group with the presence of slight bleeding and edema, and these inflammatory reactions were milder than those in the control group. These findings indicate that bone marrow mesenchymal stem cell transplantation can effectively reduce the rejection in rats undergoing heart transplantation.  相似文献   

16.
Presence of functional immune system is critical for any attempt aimed at improving survival of breast cancer patients by strategies based on immune system manipulation. We evaluated by flow cytometry the phenotype of peripheral blood leukocyte of 43 breast cancer patients. In 11 patients, the phenotype was evaluated before and during the chemotherapy by combination of doxorubicin and paclitaxel (AT). Compared with controls breast cancer patients had significantly higher relative and absolute numbers of CD3-HLADR+, CD3-CD69+ and CD14+CD16-, and significantly lower percentages of CD3- and CD8-CD28+ cells. After one cycle of AT, the absolute numbers of CD3+, CD3-CD4-, CD3+CD8+ and CD8-CD28+ cells increased significantly. Present data show a presence of T-cell activation in breast cancer patients. Administration of AT may lead to an increase in functional T-cells in peripheral blood, indicating a potential for combining chemotherapy with immunotherapy in the treatment of breast cancer patients.  相似文献   

17.
目的 探讨当归多糖(ASP)对人白血病干细胞(LSCs)增殖及体内移植的影响.方法 1.ASP 对CD34+CD38-人LSCs体外增殖的影响.免疫磁性法分选正常人和髓系白血病患者骨髓中CD34+CD38-细胞,分为正常CD34+CD38-对照组、CD34+CD38-LSCs对照组、正常CD34+CD38-ASP组和C...  相似文献   

18.
Graft-versus-host disease is a major complication of allogeneic hematopoietic stem cells transplantation, leading to serious morbidity and mortality. Mesenchymal stem cells(MSC)from bone marrow cause immunoregulation in vitro and in vivo. They also have the potential to protect from lethal GVHD after both autologous and allogeneic Hematopoietic Stem Cells Transplantation (HSCT). In this study, we investigated the mechanisms responsible for GVHD in the allo-HSCT co-transplantation with MSC condition. The model of acute GVHD in Rats was established using allogeneic HSC with donor-derived T cells transplantation, with or without additional donor-derived MSC co-transplantation. The degrees of GVHD were compared, the differentiation of CD4+, CD8+, Th1/Th2 and CD4+CD25+ T cells in vivo were assessed by flow cytometry and RT-PCR analyses. We found that MSC inhibited lethal GVHD after allo-HSCT. The value of CD8+ and CD4+ T cells and the ratio of Th1/Th2 T cell subsets decreased, at the same time the proportion of CD4+CD25+ T cells increased both in spleen lymphocytes and thymocytes in vivo after allo-HSCT with MSC co-transplantation compared with conventional allo-HSCT. Our results strongly suggested that BM-derived MSC has the function of preventing lethal GVHD after allo-HSCT by means of homeostasis of T subsets in vivo.  相似文献   

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