类风湿关节炎患者CD4~+CD25~+调节性T细胞和趋化因子受体表达及相关性

目的探讨类风湿关节炎(RA)患者外周血中CD4+CD25+表达率及调节性T细胞上趋化因子受体表达水平及相关性。方法检测活动期RA组和健康对照组调节性T细胞的表达频率;判断T细胞表达率、T细胞上趋化因子受体CCR4的表达水平与RA疾病活动性指标的相关性;判断血清肿瘤坏死因子(TNF)-α、IL-10、IL-17及干扰素(IFN)-γ等与CD4+上趋化因子受体CCR4+表达水平的相关性。结果活动期RA患者T细胞表达率与趋化因子受体表达水平较健康对照组低;T细胞表达率和趋化因子受体表达水平与RA疾病活动度存在显著相关;与血清IL-10、IL-17水平呈显著正相关,与IFN-γ呈显著负相关,而与血清TNF-α则无明显相关。结论 T细胞表达率和趋化因子受体表达水平与RA疾病活动度存在显著相关,CD4+CD25+T细胞上趋化因子受体表达参与RA发病过程。     

晚期血吸虫病患者外周血CD4~+T细胞Tim-3分子表达及与肝功能损伤指标的相关性

目的检测T细胞免疫球蛋白域黏蛋白结构域分子3(T cell immunoglobulin and mucin domain protein 3,Tim-3)在晚期血吸虫病患者外周血CD4+T细胞表面的表达,并探讨其与肝功能损伤指标的关系。方法以28例晚期血吸虫病患者作为研究对象,同时选择20例慢性血吸虫病患者和30例健康人群作为对照,采用流式细胞术检测CD4+T细胞Tim-3表达水平,用ELISA法检测血清干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)水平,以日立7600型生化分析仪检测肝功能指标丙氨酸氨基转移酶(ALT)、γ-谷氨酰转移酶(γ-GT)和总胆红素(TBIL)浓度。结果晚期血吸虫病、慢性血吸虫病患者以及健康人群外周血CD4+T细胞表面Tim-3表达水平差异具有统计学意义(F=4.578,P0.05),晚期血吸虫病患者外周血CD4+T细胞Tim-3表达水平为(8.33±2.28)%,显著高于健康对照人群的(6.57±1.99)%(t=3.015,P0.01)。Spearman相关分析显示,晚期血吸虫病患者外周血CD4+T细胞Tim-3表达水平与患者血清ALT(rs=0.746,P0.01)、γ-GT(rs=0.656,P0.01)、IL-4水平(rs=0.672,P0.01)呈正相关,与IFN-γ水平(rs=-0.404,P0.05)呈负相关。结论晚期血吸虫病患者外周血CD4+T细胞Tim-3表达上调,其可能通过调节CD4+T细胞功能参与晚期血吸虫病肝损伤过程。     

红薯叶黄酮对老龄糖尿病模型大鼠的免疫调节作用

目的 观察红薯叶黄酮(FIBL)对老龄糖尿病(DM)大鼠脾脏淋巴细胞亚群及血清细胞因子的影响.方法 采用链脲佐菌素(STZ,65 mg/kg)腹腔注射建立老龄糖尿病大鼠模型,实验分为正常对照组、DM组和FIBL低、中和高(L、M、H)剂量组.经FIBL处理8 w后,光镜观察胰腺组织形态学变化,采用流式细胞术检测脾细胞中CD4+、CD8+T细胞百分数及CD4+/CD8+T细胞比值;应用酶联免疫技术检测各组大鼠血清细胞因子的含量.结果 FIBL处理8 w后,FIBL组使胰腺炎性细胞浸润程度较DM对照组明显减轻.与DM组相比,FIBL组大鼠脾CD4+T细胞百分数明显降低(P＜0.01),CD8+T细胞百分数明显增加(P＜0.01),CD4+/CD8+T细胞比值显著降低(P＜0.01);FIBL组IFN-γ、IL-2和TNF-α的水平明显降低,IL-4水平明显升高,且呈剂量依赖性.结论 FIBL能够调节老龄DM大鼠的免疫功能,纠正其免疫失衡状态,从而达到保护机体的作用.     

衰老成血管T细胞与高血压患者血管内皮功能障碍及炎症状态的相关性

目的 探讨高血压患者成血管T(Tang)细胞的水平及其免疫衰老状态和促炎能力与内皮功能障碍及全身炎症状态的相关性.方法 连续纳入2019年9月-2020年3月于中山大学附属第一医院高血压血管病科因高血压住院、未经降压治疗的原发性高血压患者120例,通过血流介导的肱动脉舒张功能(FMD)评估血管内皮功能,根据FMD≤7％和＞7％将入组的高血压患者分为合并和不合并血管内皮功能障碍两组.流式细胞技术检测患者外周血Tang细胞[CD3+ CD31+ CXC趋化因子受体-4(CXCR4)+]CD4+/CD8+亚群中CD28null和CD28+细胞比例、细胞内γ-干扰素(IFN-γ),肿瘤坏死因子α(TNF-α),白细胞介素6(IL-6)和白细胞介素10(IL-10)的阳性率及衰老标志物CD57、CD27、趋化因子受体7(CCR7)和人端粒酶逆转录酶(hTERT)的表达.采用酶联免疫吸附试验(ELISA)法检测患者血清中炎症因子IL-6、IL-17、IFN-γ和TNF-α的水平,比较两组患者一般资料、血生化指标、Tang细胞亚群、血清炎症因子的差异,Pearson相关分析Tang细胞亚群与FMD和血清炎症因子的相关性,采用多元logistic回归和受试者工作特征曲线分析Tang细胞亚群对血管内皮功能障碍的影响和诊断效能.结果 高血压伴内皮功能障碍患者CD4+ Tang细胞中CD28null亚群比例显著升高,与FMD呈负相关(r=-0.643,P＜0.01).多元logistic回归分析显示,CD28nullCD4+Tang细胞比例升高是内皮功能障碍的影响因素(OR=3.428,95％CI 1.423～5.012,P＜0.001),对内皮功能障碍具有良好的诊断效能(曲线下面积=0.885,P＜0.001).免疫分型显示,CD28null CD4+Ta.的CD57表达升高,而CCR7、CD27及hTERT表达下降,呈现衰老特征,同时细胞内IL-6、IFN-γ和TNF-α阳性比例升高.CD28nullCD4+ Tang细胞数与高血压患者血清中IL-6、IL-17、IFN-γ和TNF-α水平显著相关.结论 具有衰老和促炎表型的CD28nullCD4+ Tang细胞亚群与FMD和全身性炎症反应相关,有望成为评估高血压患者内皮功能障碍的生物学标志物.     

重症肌无力患者外周血CD4+T细胞功能的变化及相关机制

目的探讨重症肌无力(MG)患者外周血CD4~+T细胞功能的变化对其机制。方法抽取初治及治疗病情稳定的MG患者及同期入院进行健康体检者各20例的静脉血10 ml。流式细胞术(FCM)测定其外周血白细胞介素(IL)-17~+CD4~+T细胞、干扰素(IFN)-γ~+CD4~+T细胞、肿瘤坏死因子(TNF)-α~+CD4~+T细胞、IL-2~+CD4~+T细胞和CD4~+CD25high CD127~-Treg细胞比例。CD4~+T细胞与CD4~+CD25high CD127~- Treg细胞共孵育后,FCM术测定CD4 T细胞分泌IL-17、IFN-γ、TNF-α及IL-2变化情况。结果与对照组及治疗病情稳定的MG患者相比,初治MG患者外周血IL-17~+CD4~+T细胞、IFN-γ+CD4~+T细胞、TNF-α~+CD4~+T细胞、IL-2~+CD4~+T细胞比例均显著升高(P<0.05)。MG患者与HC外周血CD4~+Foxp3~+Treg细胞比例比较差异无统计学意义(P>0.05)。对照组外周血CD4~+CD25high CD127~-Treg细胞可显著抑制CD4 T细胞分泌IL-17、IFN-γ、TNF-α和IL-2,而MG患者外周血CD4~+CD25high CD127~-Treg细胞抑制CD4+TT细胞分分泌细胞因子的能力显著降低。结论 MG患者外周血CD4~+Foxp3~+Treg细胞功能的丧失,致使外周血CD4~+T细胞细胞因子分泌能力增强,从而导致MG的发生和发展。     

激素抵抗性哮喘患者外周血CD4+CD25+Foxp3+调节性T细胞及IL-10、TGF-β1的变化及意义

目的 观察激素抵抗性哮喘(SRA)患者外周血CD4+CD25+Foxp3+调节性T细胞(Treg)及白介素10(IL-10)、转化生长因子β1(TGF-β1)的变化,分析其在SRA发病机制中的作用.方法 采用流式细胞术检测40例SRA患者(激素抵抗组)外周血单个核细胞CD4+CD25+Foxp3+ Treg数目,并计算CD4+CD25+Foxp3+ Treg占CD4+T淋巴细胞的百分比;酶联免疫吸附试验(ELISA)法检测其血清IL-10、TGF-β1水平,并与激素敏感性患者(激素敏感组,46例)及正常体检者(正常组,30例)进行对比.结果 激素抵抗组患者外周血CD4+CD25+Foxp3+Treg占CD4+T淋巴细胞的百分比、CD4+CD25+Foxp3+Treg绝对值及血清IL-10、TGF-β1水平均明显低于激素敏感组与正常组(P＜0.01,P＜0.05);激素敏感组患者外周血CD4+CD25+Foxp3+Treg占CD4+T淋巴细胞的百分比、CD4+CD25+Foxp3+Treg绝对值及血清TGF-β1水平明显低于正常组(P＜0.01,P＜0.05),血清IL-10无明显差异(P＞0.05);CD4+CD25+Foxp3+Treg/CD4+T及CD4+CD25+Foxp3+Treg绝对数均与血清IL-10、TGF-β1水平呈明显正相关(P＜0.01).结论 SRA患者外周血CD4+CD25+Foxp3+ Treg数目减少及IL-10、TGF-β1含量减低可能与SRA的发生、发展有关.     

活动性肺结核患者外周血CD4~+T细胞中TLR4与血清IFN-γ、IL-4的关系

目的探究活动性肺结核(PTB)患者外周血CD4~+T细胞中Toll样受体4(TLR4)与γ干扰素(IFN-γ)、白细胞介素4(IL-4)的关系。方法选取我院收治的95例活动性PTB患者为研究对象,将本组患者根据疾病严重程度分为轻度组、中度组和重度组,同期选取30例体检健康者为对照组。分别检测各组患者外周血CD4~+T细胞中TLR4表达情况和血清IFN-γ、IL-4水平。结果 PTB轻度、中度、重度组CD4~+T细胞TLR4 mRNA表达量明显高于对照组,重度组也明显高于轻度和重度组(P0.05);轻中度和重度组血清IFN-γ水平均显著低于对照组,其中重度组明显低于其他两组(P0.05);轻中度和重度组血清IL-4水平均显著高于对照组,其中重度组明显高于其他两组(P0.05);Pearson相关性分析,肺结核患者CD4~+T细胞TLR4mRNA表达情况与血清IFN-γ水平呈负相关(P0.05);与IL-4水平呈正相关(P0.05)。结论 PTB患者外周血CD4~+T细胞TLR4表达显著升高,且与血清IFN-γ、IL-4水平有显著相关性,推测外周血CD4~+T细胞TLR4表达可能参与PTB的发生发展。     

激素抵抗性哮喘患者外周血CD4+CD25+Foxp3+调节性T细胞及IL-10、TGF-β1的变化及意义

目的 观察激素抵抗性哮喘(SRA)患者外周血CD4+ CD25+ Foxp3+调节性T细胞(Treg)及白介素10(IL-10)、转化生长因子β1(TGF-β1)的变化,分析其在SRA发病机制中的作用.方法 采用流式细胞术检测40例SRA患者(激素抵抗组)外周血单个核细胞CD4+ CD25+ Foxp3+ Treg数目,并计算CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比;酶联免疫吸附试验(ELISA)法检测其血清IL-10、TGF-β1水平,并与激素敏感性患者(激素敏感组,46例)及正常体检者(正常组,30例)进行对比.结果 激素抵抗组患者外周血CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比、CD4+ CD25+Foxp3+ Treg绝对值及血清IL-10、TGF-β1水平均明显低于激素敏感组与正常组(P＜0.01,P＜0.05);激素敏感组患者外周血CD4+ CD25+ Foxp3+ Treg占CD4+T淋巴细胞的百分比、CD4+ CD25+ Foxp3+Treg绝对值及血清TGF-β1水平明显低于正常组(P＜0.01,P＜0.05),血清IL-10无明显差异(P＞0.05);CD4+ CD25+ Foxp3+ Treg/CD4+T及CD4+ CD25+ Foxp3+ Treg绝对数均与血清IL-10、TGF-β1水平呈明显正相关(P＜0.01).结论 SRA患者外周血CD4+ CD25+ Foxp3+ Treg数目减少及IL-10、TGF-β1含量减低可能与SRA的发生、发展有关.     

CD4+CD25+ T细胞在溃疡性结肠炎中的表达及其临床意义

目的 研究溃疡性结肠炎(UC)患者外周血CD 4CD 25调节性T细胞比例的变化,探讨其在UC病理机制中的意义.方法 选择UC患者33例,对照组20例.用流式细胞仪检测外周血CD 4CD 25T细胞阳性率.用RT-PCR检测外周单个核细胞(PBMC)中Foxp3 mRNA的表达.用酶联免疫吸附试验检测血清中IL-10和TGF-β的浓度.结果 UC患者CD 4CD 25T细胞占CD 4T细胞的比例明显低于对照组(P＜0.01),并且与疾病的活动指数及血沉水平均呈显著负相关(R值分别为-0.660和-0.572,P值均＜0.01).UC患者Foxp3 mRNA的表达也明显低于对照组(P＜0.01).两组患者血清IL-10和TGF-β的浓度比较无明显差异(P＞0.05).结论 UC患者外周血CD 4CD 25 调节性T细胞明显降低,与疾病活动性相关,提示这类细胞可能在UC的病理机制中发挥作用.Foxp3表达降低可能是导致CD 4CD 25 T细胞发育障碍的重要因素.     

慢性丙型肝炎患者CD4+CD25+调节性T细胞表达增加

目的:探讨CD4+CD25+调节性T(Treg)细胞在慢性丙型肝炎患者免疫下调中的意义.方法:流式细胞仪检测慢性丙型肝炎患者外周血中CD4+CD25+Treg细胞的数量;与CD4+CD25-T细胞共同培养,检测其抑制功能;流式细胞仪检测其对CD4+CD25-T细胞合成IFN-γ和IL-4的影响;RT-PCR检测CD4+CD25+Treg细胞中Foxp3的mRNA表达.结果:CD4+CD25+Treg细胞约占慢性丙型肝炎患者外周血中CD4+T细胞的14.1±1.6%,显著高于正常对照5.3±0.8%(P＜0.01),显著抑制CD4+T细胞的增殖(P=0.002),以及合成IFN-γ.CD4+CD25+Treg 细胞高表达Foxp3.结论:持续性HCV感染患者CD4+CD25+Treg细胞表达增加,特异性抑制Th1细胞反应.     

Synthesis and antitumor activity of 4-demethoxyadriamycin and 4-demethoxy-4' -epiadriamycin

Two new analogs of adriamycin have been obtained by chemical synthesis, 4-demethoxyadriamycin and 4-demethoxy-4' -epiadriamycin. Both compounds were highly effective against experimental mouse tumors at doses about ten times lower than those effective for adriamycin. At the optimal dose, 4-demethoxyadriamycin displayed antitumor activity similar to that of adriamycin in solid Sarcoma 180 (S180), L1210, P388, and Gross leukemias, and mammary carcinoma, while it did not markedly inhibit the growth of Moloney sarcoma virus-induced sarcoma in mice treated before the virus infection. At the optimal dose, 4-demethoxy-4' -epiadriamycin was as active as adriamycin against L1210, P388,and Gross leukemias, and less active against solid S180. The results show that anthracycline derivatives characterized by the absence of the methoxyl group at the C-4 position are markedly more potent than the parent compound, and may exhibit a differential antitumor effect on a number of mouse tumors.     

Expressions of 2H4 and 4B4 antigens on ATL cells

We studied the expression of 2H4 and 4B4 on the surfaces of leukemia cells from 17 patients with adult T-cell leukemia (ATL) as well as of cells belonging to 2 T-cell lines derived from ATL patients. The effects of the supernatants obtained from culture fluids of the ATL cells and the T-cell lines on IgG production of a human B-cell line, CESS cells, were also examined. On the surfaces of the ATL cells from 15 out of 17 cases and of the cells of 2 T-cell lines 4B4 obviously existed at higher percentage than 2H4 and more than 80% of ATL cells from 16 out of these 17 cases showed the expression of T4 (CD4). These findings revealed that the most of ATL cells had a helper-inducer phenotype. Supernatants (Sups) of culture fluids of ATL cells from 4 patients and those of 2 T-cell lines were added at various concentrations to the CESS cells. In only 1 Sup from ATL patient enhanced the IgG production of the CESS cells at lower concentration. However, other 5 Sups suppressed the IgG production of the CESS cells in proportion to the increase of Sup added. These results showed that phenotypical type of ATL cells does not always correspond to their functions, and the ATL cells may produce humoral factors that regulate B cell functions.     

Interleukin-4

Summary Since its discovery in 1982, numerous biological activities of interleukin-4 (IL-4) have been described. Like other cytokines, IL-4 is highly pleiotropic, both with respect to the number of different target cells that are responsive to it and with respect to the number of different biological responses it elicits. Interleukin-4 was initially described as a costimulant for the proliferation of B lymphocytes stimulated with anti-IgM antibody. Synonyms for this cytokine are B cell growth factor-1 (BCGF-1) and B cell stimulatory factor-1 (BSF-1). After cloning of both the murine and human IL-4, the use of recombinant IL-4 enabled detailed studies of its biological functions. Many cell types, mainly of hematological origin, express receptors for IL-4. Accordingly, effects of IL-4 have been described on B lymphocytes, T lymphocytes, NK cells, mononuclear phagocytes, mast cells, fibroblasts and hematopoietic progenitor cells (Fig. 1). Currently, there are three major areas in which IL-4 appears to play an important role: 1) regulation of B cell growth and of antibody isotype expression. In this context, a possible role for IL-4 in allergic reactions is of special interest. 2) Stimulation of T cell growth and the generation of cytotoxic T lymphocytes. In addition to the suppressive effects on the induction of non HLA-restricted cellular cytotoxicity by natural killer- (NK) and lymphokine-activated killer (LAK) cells, this suggests a role for IL-4 in the regulation of cellular immune responses. 3) Regulation of the growth and differentiation of hematopoietic bone marrow stem cells. IL-4 itself does not induce proliferation of hematological progenitor cells but it can modulate the growth-factor dependent proliferation of these cells. In this review the biological functions of IL-4, reported until present, are discussed.Abbreviations CSF colony stimulating factor - EPO erythropoietin - IFN interferon - IL interleukin - G-CSF granulocyte-CSF - GM-CSF granulocyte-macrophage-CSF - M-CSF macrophage-CSF - LAK lymphokine activated killer - NK natural killer - SA Staphylococcus aureus strain Cowan I - TIL tumor infiltrating lymphocyte - TNF- tumor necrosis factor alpha - tPA tissue plasminogen activator     

DNA A-tract bending in three dimensions: solving the dA4T4 vs. dT4A4 conundrum

DNA A-tracts have been defined as four or more consecutive A.T base pairs without a TpA step. When inserted in phase with the DNA helical repeat, bending is manifested macroscopically as anomalous migration on polyacrylamide gels, first observed >20 years ago. An unsolved conundrum is why DNA containing in-phase A-tract repeats of A(4)T(4) are bent, whereas T(4)A(4) is straight. We have determined the solution structures of the DNA duplexes formed by d(GCAAAATTTTGC) [A4T4] and d(CGTTTTAAAACG) [T4A4] with NH(4)(+) counterions by using NMR spectroscopy, including refinement with residual dipolar couplings. Analysis of the structures shows that the ApT step has a large negative roll, resulting in a local bend toward the minor groove, whereas the TpA step has a positive roll and locally bends toward the major groove. For A4T4, this bend is nearly in phase with bends at the two A-tract junctions, resulting in an overall bend toward the minor groove of the A-tract, whereas for T4A4, the bends oppose each other, resulting in a relatively straight helix. NMR-based structural modeling of d(CAAAATTTTG)(15) and d(GTTTTAAAAC)(15) reveals that the former forms a left-handed superhelix with a diameter of approximately 110 A and pitch of 80 A, similar to DNA in the nucleosome, whereas the latter has a gentle writhe with a pitch of >250 A and diameter of approximately 50 A. Results of gel electrophoretic mobility studies are consistent with the higher-order structure of the DNA and furthermore depend on the nature of the monovalent cation present in the running buffer.