微小RNA-7敲减对小鼠脾脏中T淋巴细胞体外功能的影响

观察微小RNA-7(microRNA-7,miR-7)敲减(Knock down,KD)对小鼠脾脏T淋巴细胞体外功能的影响并探讨其意义。常规分离野生型(wild type,WT)小鼠脾脏T淋巴细胞,经CD3和CD28抗体刺激后,Real-time PCR检测不同时间点(0h;24h;48h)细胞中miR-7的表达变化;进一步用Con A、CD3和CD28抗体刺激miR-7KD小鼠脾脏T淋巴细胞,CCK8检测细胞增殖率;Real-time PCR检测miR-7KD小鼠脾脏T淋巴细胞IL-12、IL-4、IL-6、TNF-α、IFN-γ和IL-10表达的变化;FACS检测CD4+T和CD8+T细胞的数量变化及CD4+T细胞膜分子CD44、CD62L和IL-4、IFN-γ的表达变化。结果显示,WT小鼠脾脏T淋巴细胞活化后,miR-7的表达水平显著上调(P0.05);与WT小鼠相比,在Con A、CD3和CD28抗体作用下miR-7KD小鼠脾脏T淋巴细胞增殖明显增加(P0.05);miR-7KD小鼠脾脏T淋巴细胞IL-12、IL-4、IL-6、TNF-α和IFN-γ水平均明显上调(P0.05),而IL-10表达显著下调(P0.05);FACS检测结果显示CD4+T细胞比例明显上调(P0.05),而CD8+T细胞的比例变化不显著(P0.05);CD4+T细胞膜分子CD62L水平显著下降,CD69及IL-4、IFN-γ的表达水平均显著上调(P0.05)。结果表明,miR-7敲减以后可显著影响小鼠脾脏T淋巴细胞的功能,本实验为后续深入探讨其在T淋巴细胞功能调控中的作用提供实验依据。     

白藜芦醇对小鼠CD4+T淋巴细胞活化的影响

目的 研究白藜芦醇对小鼠CD4+T细胞活化的影响.方法 将BALB/c鼠(SPF,雌性,9～10周龄)脾脏单个核细胞与不同浓度(0、10、20、40 mmol/L)的白藜芦醇共同孵育30 min后,分别采用刀豆蛋白A(ConA)、抗CD3抗体、抗CD3/抗CD28抗体刺激继续培养;分别共培养6h、36 h后收集细胞采用单克隆抗体抗CD4-PerCP和抗CD69-FITC或抗CD71-FITC染色后,流式细胞术检测CD4+T细胞CD69、CD71的表达.结果 白藜芦醇(≥10 mmol/L)可显著抑制抗CD3抗体引起的CD4+T淋巴细胞CD69表达(P＜0.01),其抑制作用呈剂量依赖关系.白藜芦醇(≥20 mmol/L)可显著抑制由ConA、抗CD3/CD28抗体引起的CD4+T淋巴细胞CD69表达(P＜0.05),其抑制作用呈剂量依赖关系.白藜芦醇(≥10 mmol/L)可显著抑制ConA引起的CD4+T淋巴细胞CD71表达(P＜0.05),其抑制作用呈剂量依赖关系.白藜芦醇(≥20 mmol/L)可显著抑制由抗CD3抗体、抗CD3/抗CD28抗体引起的CD4+T淋巴细胞CD71表达(P＜0.05),其抑制作用呈剂量依赖关系.结论 白藜芦醇具有抑制小鼠CD4+T细胞活化的作用,其抑制作用呈剂量依赖关系.     

NOD小鼠人源化后CD4~+T和CD8~+T细胞及其分泌IFN-γ与IL-17的频率变化及意义

目的 观察分析NOD小鼠和NOD.β2mnullHHD小鼠Ⅰ型糖尿病(Type1 diabetes,T1D)发病情况以及脾T细胞亚群的频率及功能差异,揭示CD4+T和CD8+T细胞亚群在HLA-A*0201转基因NOD小鼠和NOD小鼠的T1D发病中作用的异同。方法采用测量血糖的方法观察2种小鼠的发病情况,采用流式细胞术分析小鼠脾淋巴细胞CD3+CD4+T、CD3+CD8+T细胞亚群频率以及这2群细胞分泌IL-17和IFN-γ频率的差异。结果 NOD.β2mnullHHD小鼠较NOD小鼠发病早且严重;NOD.β2mnullHHD小鼠脾淋巴细胞的CD3+CD4+T细胞亚群显著高于NOD小鼠,NOD.β2mnullHHD小鼠脾淋巴细胞的CD3+CD8+T细胞亚群显著低于NOD小鼠;2种小鼠的脾淋巴细胞中CD3+CD4+IL-17+T细胞亚群与CD3+CD8+IL17+T细胞频率无差异;NOD.β2mnullHHD小鼠脾淋巴细胞的CD3+CD4+IFN-γ+T和CD3+CD8+IFN-γ+T细胞亚群频率显著高于NOD小鼠。结论 HLA-2.1分子转入NOD小鼠后HLA-2.1分子加速了HLA-A*0201转基因NOD小鼠T1D的发病进程;NOD.β2mnullHHD小鼠相对NOD小鼠的T1D发病更早、病情更重,这与CD3+CD4+T和CD3+CD8+T细胞分泌的IFN-γ显著相关,而与IL-17无关,为T1D防治的临床转化基础研究提供实验数据。     

GPIF对免疫损伤小鼠T淋巴细胞膜表面共刺激分子表达的影响

目的：分析羊胎盘免疫调节因子(GPIF)对BALB/c小鼠T淋巴细胞共刺激表面抗原分子表达及其细胞因子分泌的影响，探讨羊胎盘免疫调节因子免疫促进作用机理。方法: 60Coγ-ray辐射所致免疫抑制小鼠连续7 d腹腔注射GPIF，流式细胞分析术分析BALB/c小鼠脾细胞表达CD28+、CD152+单阳性细胞百分率，表达CD4+CD28+、CD8+CD28+、CD4+CD152+、CD8+CD152+双阳性细胞百分率；ELISA法检测小鼠血清IL-2、IFN-γ分泌水平。结果: 羊胎盘免疫调节因子显著提高免疫损伤小鼠脾淋巴细胞CD28+、CD4+CD28+、CD8+CD28+阳性细胞百分率(P<0.05，P<0.01)，降低CD152+、CD4+CD152+阳性细胞百分率(P<0.05，P<0.01)，提高小鼠血清IL-2、IFN-γ分泌水平(P<0.01)。结论: 羊胎盘免疫调节因子的免疫促进作用与其调节T淋巴细胞CD28、CD152共刺激分子通路的活化信号传递，降低T淋巴细胞的功能抑制，促进T淋巴细胞的活化有关。活化的T淋巴细胞分泌细胞因子IL-2、IFN-γ，参与细胞因子介导的免疫网络调节。     

香加皮杠柳苷对荷瘤小鼠的免疫调节作用

目的:研究香加皮杠柳苷(CPP)对荷瘤小鼠的免疫调节作用及其作用机制.方法:建立BALB/c小鼠H22皮下移植瘤,观察低、中、高剂量CPP(0.25、0.50、1.00 mg/kg)对荷瘤小鼠免疫器官的影响,应用流式细胞术检测各组小鼠脾T淋巴细胞亚群的分布,应用MTT法检测ConA诱导的小鼠脾淋巴细胞增殖活性,用ELISA试剂盒测定各组荷瘤小鼠血清中细胞因子TNF-α、IL-2和IL-12的含量.结果:对照组荷瘤小鼠的胸腺指数及脾脏指数均明显低于未荷瘤正常小鼠(P<0.05),而CPP组荷瘤小鼠的胸腺指数及脾脏指数较对照组明显增高,甚至高于正常对照组(P<0.05).CPP对荷瘤小鼠体内CD8+T细胞数量无影响,但可明显上调CD3+、CD4+T细胞数及CD4+/CD8+比值,其中CD3+、CD4+细胞百分率与正常小鼠无差别(P>0.05),CD4+/CD8+比值高于正常小鼠(P<0.05).CPP可明显增强ConA诱导的荷瘤小鼠脾淋巴细胞的增殖能力,SI甚至超过正常对照组(P<0.05).不同剂量CPP组小鼠血清中TNF-α、IL-2和IL-12水平均较对照组明显增高(P<0.05),并随剂量增加呈升高趋势,至接近或超过正常小鼠水平.结论:CPP可保护荷瘤小鼠的免疫器官不受损害,并可明显提高CD4+T细胞百分率和CD4+/CD8+比值,增强荷瘤小鼠T细胞增殖能力,促进细胞因子TNF-α、IL-2和IL-12的产生,表明CPP具有显著的免疫增强作用.     

原发免疫性血小板减少症患者CD4+T细胞CD28的表达及其与IFN-γ/IL-10比率的关系

目的 研究原发免疫性血小板减少症(ITP)患者治疗前、后,外周血CD4+的T淋巴细胞中共刺激分子CD28的表达及与IFN-γ/IL-10比率的关系.方法 采用流式细胞术分析30例ITP患者糖皮质激素治疗前、后和26例正常对照外周血CD4+T细胞上CD28表达率,用ELISA双夹心法检测外周血血清中IFN-γ和IL-10的水平,评价CD4+ CD28+与IFN-γ/IL-10平衡状态、血小板计数之间的关系.结果 ITP患者治疗前CD4+ CD28+的表达显著高于正常对照组(P＜0.05),治疗后与正常对照组比较,差异无统计学意义(P>0.05);ITP患者治疗前IFN-γ水平较正常对照组显著增高,IL-10水平显著降低,IFN-γ/IL-10比值显著增高(P＜0.01),治疗后与正常对照组比较,差异无统计学意义(P>0.05);ITP患者治疗前CD4+ CD28+与IFN-γ/IL-10比值呈正相关(P＜0.05),与血小板计数呈负相关(P＜0.05).结论 共刺激分子CD28与ITP免疫紊乱密切相关,可能通过参与Th1优势状态形成而在ITP的发病中发挥一定作用.     

微小RNA-126基因敲减对CD4~+T细胞体外功能的影响

目的研究微小RNA-126(microRNA-126,miR-126)基因敲减(knock down,KD)后对CD4~+T细胞体外功能的影响并探讨其意义。方法 MACS分选miR-126 KD小鼠脾脏CD4~+CD62L~+T细胞,ConA刺激48h后,Real-time PCR技术检测细胞内miR-126成熟体表达水平;FACS分别检测CD4~+T细胞表面活化分子CD69、CD62L、CD44和增殖相关分子Ki-67,以及细胞因子IFN-γ、IL-4、IL-17表达变化;Annexin V/PI双染色法检测CD4~+T细胞的凋亡情况。结果与野生型(wide-type,WT)小鼠相比,miR-126KD小鼠的CD4~+T细胞中miR-126成熟体表达显著下调(P0.001);miR-126 KD小鼠CD4~+T细胞活化后CD62L的表达比例显著减少(P0.01),而CD69、CD44的比例显著增加(P0.05);同时Ki-67~+的比例也明显增加(P0.05);此外,miR-126 KD小鼠CD4~+T细胞的INF-γ表达比例显著上调(P0.001),IL-4的比例显著下调(P0.01),而IL-17的比例没有明显变化(P0.05);最后,CD4~+T细胞的凋亡比例显著减少(P0.05)。结论 miR-126基因敲减后CD4~+T细胞的活化、增殖和相关细胞因子分泌均明显增强,为后续深入探讨miR-126在免疫应答中的作用及机制提供前期实验基础。     

小鼠CD4~+ CD25~+ Foxp3~+调节性T细胞体外扩增

目的:本研究旨在探讨CD4+CD25+Foxp3+调节性T细胞体外扩增的方法。方法:采用磁珠分选小鼠CD4+T细胞,αCD3单克隆抗体包被24孔板,加入αCD28单克隆抗体、雷帕霉素、rhIL-2,培养3周后,流式细胞仪测定培养细胞中CD4+CD25+T细胞的含量,实时定量PCR检测CD4+CD25+T细胞Foxp3 mRNA的表达;单向混合淋巴细胞反应和增殖抑制试验测定扩增的CD4+CD25+T细胞的增殖及其抑制功能;ELISA检测培养上清中IL-10和TGF-β1的含量。结果:小鼠CD4+T细胞培养3周后,CD4+CD25+T细胞达(76.05±2.73)%,高于未加雷帕霉素组(52.17±1.36)%(P<0.001),磁珠分选的CD4+CD25+T细胞Foxp3 mRNA的表达是未加雷帕霉素组的5倍(P<0.001),增殖能力是未加雷帕霉素组的0.29倍(P<0.001),对CD4+T细胞增殖抑制能力是未加雷帕霉素组的3.6倍(P<0.001),培养上清中IL-10和TGF-β1分别是对照组的1.8倍和1.6倍(P<0.001)。结论:小鼠CD4+T细胞在含有1μg/ml的αCD28、rhIL-2 100 U/ml和终浓度为10 nmol/L雷帕霉素的培养体系中培养3周后能有效扩增CD4+CD25+Foxp3+调节性T细胞。     

妊娠浓度的雌激素对诱导CD4~+CD25~-naive T细胞转化为CD4~+CD25~+Treg细胞的影响

目的 体外观察妊娠浓度的雌激素能否诱导CD4+CD25-naiveT细胞转化为CD+CD25+Treg细胞,并探讨其相关性.方法 以CD4+CD25-T细胞作为反应细胞,实验组加入妊娠水平的雌激素(E2)及CD3/CD28单抗作为刺激原培养72 h,设阴性对照组(仅加入CD3/CD28单抗)和空白对照组.72 h后检测各组中CD4+CD25+T细胞和CD4+Foxp+T细胞比例变化及Foxp3 mRNA表达.结果 1)阴性对照组CD+CD25+T细胞比例较空白对照组显著增高(P＜0.001),而实验组CD4+CD25+T细胞比例进一步升高(P＜0.001).2)阴性对照组不能诱导CD4+Foxp+T细胞比例增高,但实验组CD4+Foxp3+T细胞的比例则较其它2组均显著升高(P＜0.001).3)RT-PCR提示阴性对照组Foxp3 mRNA的表达量较空白对照组无显著差异(P＞0.05);而实验组F0xp3 mRNA的表达昔较其它2组均显著升高(P＜0.001).结论 体外淋巴细胞刺激实验提示妊娠状态下雌激素的高水平与CD4+CD25+Foxp3+Treg细胞比例的升高密切相关.     

IL-37通过作用树突状细胞调节CD8~+T细胞功能

将小鼠骨髓细胞诱导分化为DC,用IL-37处理后,流式细胞术检测细胞表面共刺激分子(CD86、CD80),RT-PCR法检测TNF-α、IFN-γ、IL-6、IL-12和TGF-βmRNA的表达,多重液相蛋白定量CBA试剂盒及ELISA试剂盒检测细胞上清中TNF-α、IL-6、IL-12、IFN-γ、MCP-1和TGF-β蛋白的表达。Western blotting方法检测DC中磷酸化蛋白的表达。将DC与T细胞共培养,流式细胞术检测CD8+T细胞增殖及活化程度(CFSE、CD69)。结果显示,IL-37能够降低表达CD80+/CD86+的DC数量。促炎因子TNF-α、IL-12和IL-6的表达被明显抑制,而T淋巴细胞抑制因子TGF-β明显增高。IL-37预处理的DC明显降低T淋巴细胞的增殖和活化能力。IL-37能够降低DC中磷酸化ERK和NF-κB的表达。IL-37处理的DC对CD8+T细胞产生明显抑制作用。结果表明,IL-37能够通过影响ERK和NF-κB依赖的信号通路抑制DC的成熟和免疫反应,从而抑制CD8+T细胞的活化及增殖。     

Chronic lymphocytic leukemia with peripheral T lymphocytes expressing CD 2+, CD 3+, CD 4-, CD 8-, CD 16+, and CD 56+ and lymph-node lymphocytes expressing CD 2+, CD 3-, CD 4-, CD 8-, CD 16+, CD 38+, and CD 56+]

A 33-year-old man was hospitalized because of thrombocytopenia and severe splenomegaly. On admission 78% of peripheral lymphoid cells were abnormally large, with pale cytoplasm. Flow cytometry of the abnormal lymphocytes showed that they expressed CD 2, CD 3, CD 11, CD 16, and CD 56, but not CD 4 nor CD 8, so they were T-cell large granular lymphocytes (T-LGL). Abnormal lymphocytes obtained from a lymph node expressed CD 2, CD 16, CD 38, and CD 56, but not CD 3, CD 4, and CD 8, so they were natural killer(NK) cells. Splenectomy was performed and the operative specimen showed diffuse infiltration of pleomorphic lymphocytes, probably chronic lymphocytic leukemia cells. After splenectomy, the platelet count returned to normal but the lymphocytosis continued. Two years after discharge, chemotherapy was done because of thrombocytopenia and hepatomegaly. The patient died of disseminated intravascular coagulation arising from sepsis. The differences and similarities between peripheral and lymph-node lymphocytes suggest that LGL and NK cells may be differentiated from the same kind of cell, somewhat differentiated from stem cells.     

A CD44 monoclonal antibody differentially regulates CD11a/CD18 binding to intercellular adhesion molecules CD54, CD102 and CD50

We have made a monoclonal anti-CD44 antibody which is able to activate the leukocyte integrin CD11a/CD18. Activated T cells strongly aggregated, and the aggregation was shown to be intercellular adhesion molecule (ICAM)-1 (CD54) and ICAM-2 (CD102) dependent. Using purified ICAM coated on plastic, only binding to ICAM-1 was increased by the CD44 antibody, whereas activation by phorbol ester increased binding to both ICAM-1 and ICAM-3. The binding to ICAM-2 was not affected by either treatment. These findings show that the CD11a/CD18 integrin can be activated in a ligand-specific manner by engagement of CD44.     

Reduction in CD16/CD56 and CD16/CD3/CD56 Natural Killer Cells in Coronary Artery Disease

Background: Natural killer (NK) cells are the potential modulators of inflammatory reactions that exert several unique biological effects and could lead to future adverse events of coronary artery disease (CAD).

Hypothesis: The purpose of this study was to find out the possible association of modulation in NK cell, TNK cells, T cells, B cells, and tumor necrosis factor alpha (TNF-α) in CAD patients and various forms of myocardial infarction.

Methods: The present study included total 190 subjects (98 confirmed CAD patients both men and women and 92 healthy control individuals). Serum concentration of TNF-α was measured by ELISA method. For the measurement of various immune cells, viz., NK cell, TNK cells, T cells, and B cells, flow-cytometric analysis was performed.

Results: A significant reduction by 15% (P < 0.001) in CD16/CD56 NK cells was observed in CAD patients. Moreover, non-ST segment elevation myocardial infarction (NSTEMI), ST segment elevation myocardial infarction (STEMI), unstable angina (UA), and combined UA + NSTEMI group also showed a significant decline in NK cells compared with control individuals. CD16/CD56/CD3 TNK cells showed a significant reduction in CAD, NSTEMI, STEMI, and UA categories. However, UA + NSTEMI group did not show any significant change in TNK cells. On the other hand, the level of TNF-α was found to be significantly elevated in CAD, STEMI, and UA groups. NSTEMI and combined UA + NSTEMI group did not show any significant change in TNF-α level.

Conclusion: Current study provides an insight toward the association of immune cells and inflammation with CAD.     

CD28/CTLA-4 and CD80/CD86 families

T cell stimulation in the absence of a second, costimulatory signal can lead to anergy or the induction of cell death. CD28 is a major T cell costimulatory receptor, the coengagement of which can prevent anergy and cell death. The CD28 receptor is a member of a complex family of polypeptides that includes at least two receptors and two ligands. Cytotoxic lymphocyte-associated molecule-4 (CTLA-4, CD152) is the second member of the CD28 receptor family. The ligands or counterreceptors for these two proteins are the B7 family members, CD80 (B7-1) and CD86 (B7-2). This article reviews the CD28/CTLA4 and CD80/CD86 families, and outlines the functional outcomes and biochemical signaling pathways recruited after CD28 ligation.     

Interferons alpha/beta inhibit IL-7-induced proliferation of CD4- CD8- CD3- CD44+ CD25+ thymocytes, but do not inhibit that of CD4- CD8- CD3- CD44- CD25- thymocytes.

Type 1 interferons (IFN-alpha/beta) have recently been shown to inhibit interleukin-7 (IL-7)-induced growth and survival of early B-lineage cells. The CD3- CD4- CD8- (triple negative; TN) thymocytes from normal mice strongly proliferated upon stimulation with IL-7 in suspension, culture. Such an IL-7-induced proliferation was suppressed by the addition of IFN-alpha/beta, but a fraction of the TN thymocytes still showed proliferation. The IL-7-induced growth of TN thymocytes from acid mice, which lack the CD44- CD25- subpopulation, was completely inhibited by the addition of IFN-alpha/beta. The IL-7 induced proliferation of CD4- CD8- thymocytes from T-cell receptor (TCR) transgenic mice, the majority of which are CD3+ CD44- CD25-, was resistant to IFN-alpha/beta-mediated suppression. In fetal thymus organ cultures (FTOC), the addition of IL-7 greatly increased the population of CD4- CD8- CD44+ CD25+ thymocytes and IFN-alpha/beta inhibited this IL-7-driven expansion. In contrast, the addition of IL-7 markedly decreased the percentages of CD4- CD8- CD3- CD44- CD25- cells, and IFN-alpha/beta reversed the effect and increased the subpopulations of CD44- CD25+ and CD44- CD25-. Finally, IFN-beta mRNA was found to be expressed in the thymus. The data suggest that type I interferons inhibit IL-7-driven proliferation of TN thymocytes, but do not block the normal differentiation process.     

Regulatory function of cytomegalovirus-specific CD4CD27CD28 T cells

CMV infection is characterized by high of frequencies of CD27⁻CD28⁻ T cells. Here we demonstrate that CMV-specific CD4⁺CD27⁻CD28⁻ cells are regulatory T cells (T_R). CD4⁺CD27⁻CD28⁻ cells sorted from CMV-stimulated PBMC of CMV-seropositive donors inhibited de novo CMV-specific proliferation of autologous PBMC in a dose-dependent fashion. Compared with the entire CMV-stimulated CD4⁺ T-cell population, higher proportions of CD4⁺CD27⁻CD28⁻ T_R expressed FoxP3, TGFβ, granzyme B, perforin, GITR and PD-1, lower proportions expressed CD127 and PD1-L and similar proportions expressed CD25, CTLA4, Fas-L and GITR-L. CMV-CD4⁺CD27⁻CD28⁻ T_R expanded in response to IL-2, but not to CMV antigenic restimulation. The anti-proliferative effect of CMV-CD4⁺CD27⁻CD28⁻ T_R significantly decreased after granzyme B or TGFβ inhibition. The CMV-CD4⁺CD27⁻CD28⁻ T_R of HIV-infected and uninfected donors had similar phenotypes and anti-proliferative potency, but HIV-infected individuals had higher proportions of CMV-CD4⁺CD27⁻CD28⁻ T_R. The CMV-CD4⁺CD27⁻CD28⁻ T_R may contribute to the downregulation of CMV-specific and nonspecific immune responses of CMV-infected individuals.     

A Combination of CD15/CD10, CD64/CD33, CD16/CD13 or CD11b Flow Cytometric Granulocyte Panels is Sensitive and Specific for Diagnosis of Myelodysplastic Syndrome

Flow cytometry (FCM) is a reproducible and objective technique that may be useful in the diagnosisof myelodysplastic syndrome (MDS) by detecting abnormal immunophenotypes specific to MDS. We investigated 5 granulocyte/monocyte panels by FCM to find a sensitive and specific combination of panels in order to discriminate MDS from non-clonal hematologic disorders. Bone marrow aspirates from 35 patients with MDS and 25 patients with non-clonal hematologic disorders were studied. We performed FCM using 5 granulocyte/monocyte panels (CD15/CD10/CD45, CD64/CD33/CD45, CD16/CD13/CD45, CD16/CD11b/CD45, and CD56/CD19/CD7/CD45) to examine the positive rate in MDS and controls, and to find an optimal combination that maximizes the detection rate of MDS. In MDS, the number of abnormal immunophenotypes per 5 granulocytic and 5 monocytic panels were 2.1±1.2 and 2.2±1.4. The rates were higher than the controls (P< 0.001, respectively). As the number of employed panels increased, the percent values of abnormal immunophenotypes increased (P=0.002). The maximum rate of abnormal immunophenotype was 89.7% in MDS patients, especially 100.0% in normal karyotype, when a combination of three panels, CD15/CD10/CD45, CD64/CD33/CD45, and either CD16/CD13/CD45 or CD16/CD11b/CD45 was used. This study demonstrates that a combination of CD15/CD10, CD64/CD33, CD16/CD13 or CD11b granulocyte panels in FCM is sensitive and specific for diagnosis of MDS.     

Kikuchi病中检测出全T细胞相关抗原丢失

目的 观察全T细胞相关抗原CD2、CD3、CD5、CD7在Kikuchi病中是否存在丢失,探讨将全T细胞相关抗原丢失作为鉴别T细胞良、恶性病变辅助诊断依据的局限性.方法 收集33例明确诊断为Kikuchi病和15例淋巴组织反应性增生病例,通过复习HE切片并应用免疫组织化学EliVision法检测病变中CD2、CD3、CD5、CD7的表达情况.结果 72.7％ (24/33)的Kikuchi病患者中存在一种或几种全T细胞相关抗原的丢失,其中13例仅CD5丢失,1例仅CD7丢失,1例仅CD2丢失,2例CD2和CD7丢失,4例CD5和CD7丢失,2例CD2和CD5丢失,1例CD2、CD5和CD7丢失.以CD5丢失最多见(60.6％,20/33),其次为CD7(24.2％,8/33)和CD2( 18.2％,6/33).抗原的丢失多见于增生型及坏死型.经随访,抗原的丢失与Kikuchi病的预后无明显相关性.15例淋巴组织反应性增生病例中无明显抗原丢失现象.结论 Kikuchi病中存在一种或几种全T细胞相关抗原的丢失.因此,将全T细胞相关抗原丢失作为T细胞淋巴瘤辅助诊断依据不适用于Kikuchi病.     

Lower proportion of CD19+IL-10+ and CD19+CD24+CD27+ but not CD1d+CD5+CD19+CD24+CD27+ IL-10+ B cells in children with autoimmune thyroid diseases

Abstract

Introduction: As it is generally known, regulatory B cells (Bregs) control inflammation and autoimmunity. The significance of Bregs in the population of children with autoimmune thyroid diseases (AITD) still offers plenty of potential to explore. The aim of this study was to estimate the expression of Bregs (phenotype CD19⁺CD24⁺CD27⁺IL-10⁺, CD19⁺IL-10⁺, CD1d⁺CD5⁺CD19⁺IL-10⁺ and CD1d⁺CD5⁺CD19⁺CD24⁺CD27⁺) in a paediatric cohort with AITD and in health controls.

Materials and methods: A total of 100 blood samples were obtained from 53 paediatric patients with Graves’ disease (GD) (N?=?12 newly diagnosed, mean age 12.5?±?3.5 and N?=?17 during methimazole therapy, mean age 12.7?±?4.4), Hashimoto’s thyroiditis (HT) (N?=?10 newly diagnosed, mean age 13.3?±?2.9 and N?=?10 during L-thyroxine therapy, mean age 13.7?±?3.4) and compared with healthy controls (C) (N?=?15, mean age 13.1?±?3.1). The expressions of the immune cell populations were analysed by four-color flow cytometry using a FASC Canto II cytometer (BD Biosciences).

Results: There was a decreasing tendency in the number of lymphocytes B producing IL-10 (B10) cells among all B lymphocytes and more widely, also among all lymphocytes, in each study group, as compared to C. We reported a reduction in IL-10 production in Bregs with the expression of CD19⁺CD24⁺CD27⁺IL-10 and CD1d⁺CD5⁺CD19⁺IL-10⁺ in both untreated and treated AITD.

Conclusions: Our data demonstrate that the reduction in the number of Bregs with CD19⁺CD24⁺CD27⁺IL-10⁺ and CD19⁺IL-10⁺ expression could be responsible for breaking immune tolerance and for AITD development in children.