巨细胞病毒感染患儿细胞免疫应答反应及临床意义

目的:探讨巨细胞病毒(CMV)感染患儿细胞免疫应答反应及临床意义.方法:选择2015年6月至2019年5月在本院确诊的97例CMV感染患儿,根据其有无临床症状分为症状组和无症状组,选择同期健康体检婴儿48例为对照组.采用RT-PCR反应检测所有患儿尿液标本中CMV DNA载量;采用ELISA法检测所有纳入研究对象IL-...     

小儿慢性丙型肝炎外周血T细胞亚群和TH1/TH2型细胞因子的研究

目的通过研究小儿慢性丙型肝炎外周血T细胞亚群及TH1/TH2型细胞因子的表达,进一步探讨小儿慢性丙型肝炎的免疫发病机制。方法(1)流式细胞仪(FACS)检测16例慢性丙型肝炎患儿及10例正常对照外周血T细胞亚群。(2)将慢性丙型肝炎患儿和正常对照外周血单个核细胞(PBMC)体外培养72h后,用ELISA法检测培养上清中TH1型细胞因子(IFN-γ、IL-2、IL-12和TNF-γ)和TH2型细胞因子(IL-4、IL-10)的浓度。结果(1)CD4 细胞无明显变化。CD8 细胞与正常对照比较明显升高(P<0.05)。CD3 细胞升高,CD4 /CD8 比值下降,但与正常对照比较无统计学意义(P>0.05)。(2)PBMC培养上清中IFN-γ、IL-10和TNF-α的水平明显升高(P<0.01),而没有检测到IL-2、IL-4、IL-12的基础分泌。结论慢性丙型肝炎患儿体内T淋巴细胞存在数量和功能的异常,CD8 细胞数升高,CD4 细胞功能异常,表现在以TH2型细胞因子的分泌为主。这可能与丙肝病毒(HCV)感染的慢性化有关。     

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.     

黄芪复方对重症肌无力患者T淋巴细胞亚群及IFN－γ、TNF—α、IL-4和AChRAb的影响

目的探讨黄芪复方对重症肌无力（MG）患者的细胞免疫调节机制及疗效。方法30例患者服用黄芪复方,疗程12周,观察症状改善情况及不良反应。应用流式细胞仪检测MG患者治疗前后外周血T淋巴细胞亚群分布的变化情况;应用酶联免疫吸附试验（ELISA）法测定患者治疗前后外周血血清细胞因子IFN-γ、TNF—α、IL4及乙酰胆碱受体抗体（AChRAb）的变化情况：结果经治疗12周后,临床显效率60％（18／30）,总有效率为80％（24／30）。CD4^＋T细胞、CD4^＋／CD8^＋比值均有明显下降,差异有统计学意义（P〈0．01）。治疗后CD8^＋T细胞明显增加,与治疗前比较,差异有统计学意义（P〈0．05）;治疗前血清IFN-γ、IL-4、TNF-α和AChRAb水平显著高于健康组,治疗后IFN-γ、IL-4、TNF-α和AChRAb水平显著降低,与治疗前比较,差异均有统计学意义（P〈0．01）。结论通过调节淋巴细胞亚群比例分布以及细胞因子水平的变化,可能是黄芪复方发挥免疫调节作用机制之一。     

支气管肺炎患儿机体免疫功能变化的分析

目的通过对支气管肺炎患儿T细胞亚群、血清免疫球蛋白、血清补体及细胞因子变化的分析,探讨小儿支气管肺炎机体免疫功能状态及免疫学发病机制,为临床了解病情、选择适当的治疗方案提供理论依据。方法支气管肺炎组65例,男39例,女26例,年龄2个月～13岁,平均年龄4.1岁。对照组20例,男12例,女8例,年龄1～12岁,平均年龄4.3岁。应用多色流式细胞术检测T细胞亚群(CD3+、CD3+CD4+Th、CD3+CD8+Ts、CD3+CD4+Th/CD3+CD8+Ts)、应用免疫散射比浊法检测血清免疫球蛋白(IgG、IgA、IgM)、血清补体(C3、C4)、应用ELISA法检测血清细胞因子(IFN-γ、IL-4)。结果支气管肺炎组T细胞亚群改变明显,CD3+、CD3+CD4+Th显著低于对照组,有统计学意义(P<0.01);CD3+CD8+Ts显著高于对照组,有统计学意义(P<0.01);CD3+CD4+Th/CD3+CD8+Ts比值降低,与对照组相比,有统计学意义(P<0.05)。支气管肺炎组血清IgA、IgG水平显著低于对照组,有统计学意义(P<0.01);支气管肺炎组血清补体C3低于对照组,有统计学意义(P<0.05)。支气管肺炎组血清细胞因子IFN-γ浓度及Th1/Th2比值高于对照组,有统计学意义(P<0.05)。结论支气管肺炎患儿存在细胞免疫和体液免疫功能紊乱,对支气管肺炎患儿检测免疫功能有助于判断病情、指导治疗。     

肝移植患者血清白细胞介素-6、细胞间粘附分子-1和P-选择素高水平表达分析

目的：研究白细胞介素-6、肿瘤坏死因子-α及细胞间粘附分子-1和P-选择素在稳定存活的肝移植受者血清中的表达程度。方法： ELISA 法检测22 例原位肝移植患者和12例正常人血清白细胞介素-6、肿瘤坏死因子-α及细胞间粘附分子-1和P-选择素表达情况。流式细胞术分析外周血T细胞表型。同时对近期6例临床原位肝移植病例进行动态观察。 结果： 肝移植病人外周血CD3+、CD4+、CD8+ T细胞百分比以及CD4+/CD8+比值与正常人组间比较无明显差异（P>0.05），但肝移植组 CD3+CD25+ T 细胞百分比明显高于正常人组（P<0.05）。白细胞介素-6、细胞间粘附分子-1和P-选择素水平肝移植组显著高于正常人组（P<0.05）。近期6例临床原位肝移植病例血清白细胞介素-6、肿瘤坏死因子-α及细胞间粘附分子-1和P-选择素水平的动态观察，未发现规律性变化。结论：细胞间粘附分子-1和P-选择素参与了肝移植后效应T细胞的活化过程，肿瘤坏死因子-α和白细胞介素-6则可能分别介导了对移植物的免疫损伤和修复。     

Clinical observation of immunity in patients with secondary infection from severe acute pancreatitis

Objectives

To observe immune system changes in patients with secondary infection from severe acute pancreatitis (SAP).

Methods

Seventy-nine patients were recruited. The percentages of CD4+, CD8+, natural killer (NK), HLA-DR+ cells and B lymphocytes, and the CD4+/CD8+ ratio, were determined. In addition, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-4 (IL-4) serum levels were determined on days 1, 7, 14, and 28.

Results

Fifteen patients had a secondary infection. The immune response of the infected group was quite different from the non-infected group, with a higher percentage of CD4+ and HLA-DR+ cells on days 1, 7, 14 and 28, a higher percentage of CD8+ and NK cells on days 14 and 28, a reduced CD4+/CD8+ ratio, and a reduction in B lymphocytes. The cytokine levels in the infected group were different from the non-infected group, with a rise in TNF-α and IL-6 through the first 2?weeks, but dropping at 1?month. IL-10 and IL-4 increased initially, but then dropped over the next 3?weeks.

Conclusions

An early excessive immune response followed by a subsequent immune deficiency is closely related to secondary SAP infection.     

晚期肺癌患者调节性T细胞和IL-10表达的变化

探讨晚期肺癌患者的CD4＋CD25＋调节性T细胞（Treg）、IL-10及其他T细胞亚群表达的临床意义。检测晚期肺癌患者92例和正常对照者64名的IL-10（ELISA法）、CD4＋CD25＋调节性T细胞及其他T细胞亚群（流式细胞仪法）。结果显示：肺癌组血清IL-10明显大于对照组（278±34ng/L vs 122±65ng/L,P〈0.01）、CD4＋CD25＋Treg细胞明显大于对照组[（17.8±5.2）%vs（7.1±0.4）%,P〈0.01]、CD3＋、CD4＋、CD8＋CD28＋和NK细胞明显小于对照组[（58.4±7.8）%vs（78.2±6.4）%,P≤0.05;（34.4±7.6）%vs（44.9±8.4）%,P〈0.01;（9.4±3.6）%vs（16.5±2.7）%,P〈0.01;（9.4±3.6）%vs（18.5±7.2）%,P〈0.01]。CD8＋T细胞明显高于对照组[（37.8±6.5）%vs（31.8±5.1）%,P〈0.01]。CD4＋CD25＋Treg细胞、IL-10与CD8＋CD28＋细胞、NK细胞呈明显负相关。这些结果表明,CD4＋CD25＋Treg细胞与IL-10增多为晚期肺癌患者免疫功能受损的表现。     

T cell activation and disease severity in HIV infection.

In vitro studies have indicated that T lymphocyte activation may be of importance in the pathogenesis of HIV infection. In order to define the role of immune activation in vivo, we assessed the expression of the T cell activation markers HLA-DR and CD25 by flow cytometry in peripheral blood in relation to disease severity and the surrogate markers CD4 and beta 2-microglobulin in 157 patients with HIV infection and 53 healthy seronegative blood donors. Percentage levels of CD3+HLA-DR+ T lymphocytes were significantly higher (P < 0.0001) and percentage levels of CD3+CD25+ T lymphocytes significantly lower (P < 0.0001) in all HIV+ patients compared with controls. A significant correlation was observed between increasing percentage levels of CD3+HLA-DR+ T lymphocytes and both declining CD4 counts (r = 0.52; P < 0.001) and increasing beta 2-microglobulin levels (r = 0.56; P < 0.001). Percentage levels of CD4+HLA-DR+ and CD4+ CD25+ lymphocytes were significantly higher in all HIV+ patients compared with controls (P < 0.001). Levels of activated (HLA-DR+ and CD25+) CD4+ lymphocytes showed a significant step-wise linear increase with increasing disease severity (P < 0.001). High levels of CD3+HLA-DR+ T lymphocytes were found in a greater proportion (81.8%) of asymptomatic HIV+ patients (Centres for Disease Control (CDC) group II) than low CD4 counts (51.5%) (P < 0.001). Compared with controls, HIV+ patients had higher percentage levels of CD8+HLA-DR+ lymphocytes (P < 0.001), but similar levels of CD8+CD25+ lymphocytes. These results indicate that T cell activation is not only a consistent but also an early feature in HIV infection. Monitoring levels of activated T cells and their subsets is of value in assessing progression of HIV-related disease.     

子痫前期患者外周血CD8+ CD25+ FoxP3+ 调节T 细胞的变化及意义

目的:探讨外周血CD8+ CD25+ FoxP3+调节T 细胞(Treg)在子痫前期(PE)疾病过程中的作用。方法:研究对象包括子痫前期孕晚期患者46 例,其中轻度子痫24 例(MPE 组),重度子痫22 例(SPE 组),并选择24 例与患者孕龄匹配的健康孕妇作为对照组(HP 组)。流式细胞仪检测分析外周血CD8+ CD25+ FoxP3+ Treg 比例;Luminex200 检测血清IL-6、IL-17A、IL-10、IL-1β、IL-33 和TGF-β1 浓度。分离10 例健康对照者单个核细胞(PBMCs),IL-33 刺激培养后检测CD8+ CD25+ FoxP3+Treg 比例的变化。结果:MPE 和SPE 组CD8+ CD25+ FoxP3+ Treg 比例分别为[0.32(0.19-0.63)%]、[0.13(0.02-0.41)%],两组均低于HP 组[0.48(0.21-0.96)%],三组差异有统计学意义(P<0.05)。与HP 组相比,MPE 组和SPE 组血清IL-6、IL-17A浓度均升高,且SPE 组患者血清IL-6、IL-17A 浓度升高更明显,差异有统计学意义(P<0.05);IL-10、IL-1β和IL-33 浓度在HP、MPE 及SPE 三组间差异无统计学意义(P>0.05);与HP 对照者相比,MPE 和SPE 组血清TGF-β浓度均升高,差异有统计学意义(P<0.05);但MPE 和SPE 组之间差异无统计学意义(P>0.05)。PE 患者CD8+ CD25+ FoxP3+ Treg 比例与血清IL-17A 负相关(r =-0.338,P =0.021),与IL-33 正相关(r =0.548,P =0.001)。PBMCs 用IL-33 刺激5 d 后,与空白对照组相比,CD8+ CD25+FoxP3+ Treg 比例显著增高(P<0.05)。结论:子痫前期患者外周血CD8+ CD25+ FoxP3+ Treg 细胞比例降低可能在其疾病过程中发挥重要作用。     

重度烧伤患者血糖与炎性因子及免疫功能的相关性研究

目的研究重度烧伤患者血糖与炎性因子及免疫功能的相关性。 方法选取2012年1月至2017年12月在成都医学院第一附属医院烧伤整形外科治疗的重度烧伤患者96例作为试验组,另选取成都医学院第一附属医院经体检证明健康的志愿者60名作为对照组,检测两组空腹血糖(FPG)、炎性因子及T淋巴细胞水平,比较两组间FPG、炎性因子以及T淋巴细胞水平之间的差异;再将试验组患者根据FPG浓度分为血糖正常组(17例,FPG <7.0 mmol/L)和血糖升高组(79例,FPG≥7.0 mmol/L),比较两组炎性因子以及T淋巴细胞水平之间的差异,并对重度烧伤患者的FPG与炎性因子及T淋巴细胞水平进行Pearson相关性分析。指标比较采用t检验。 结果试验组患者FPG(11.39±2.28) mmol/L、C反应蛋白(CRP)(39.67±4.15) mg/L、肿瘤坏死因子-α (TNF-α) (108.12±6.49) ng/L、白细胞介素(IL)-6 (139.77±7.43)ng/L、IL-8(91.56±4.91)ng/L,均高于对照组[FPG(5.07±1.15) mmol/L、CRP(5.91±0.76) mg/L、TNF-α(12.94±1.69) ng/L、IL-6(14.11±3.93) ng/L、IL-8(17.22±2.89) ng/L],差异均有统计学意义(t＝5.365、4.365、11.912、9.834、6.139,P值均小于0.01);试验组患者免疫指标CD4^＋ T淋巴细胞水平为(29.77±4.12)%、CD4^＋/CD8^＋为0.91±0.24,分别低于对照组(41.89±5.36)%、1.59±0.37,而试验组的CD8^＋ T淋巴细胞水平为(32.69±4.73)%,高于对照组(25.83±3.52)%,差异均有统计学意义(t＝3.931、2.433、2.696,P值均小于0.05)。试验组患者中,血糖升高组的血浆CRP(45.19±5.45) mg/L、TNF-α(121.81±5.43) ng/L、IL-6(153.31±5.57) ng/L、IL-8(106.56±5.65) ng/L、CD8^＋T淋巴细胞水平(35.46±4.11)%,均高于血糖正常组[CPR(36.67±3.45) mg/L、TNF-α(91.58±4.93) ng/L、IL-6(114.65±6.33) ng/L、IL-8(79.62±3.74) ng/L、CD8^＋ T淋巴细胞水平(28.62±4.03)%],差异均有统计学意义(t＝2.341、2.894、4.167、3.018、2.763,P值均小于0.05);CD4^＋ T淋巴细胞水平(28.89±3.79)%、CD4^＋/CD8^＋(0.82±0.37)均低于血糖正常组[(33.47±4.98)%、(1.17±0.52)],差异均有统计学意义(t＝2.158、2.247,P值均小于0.05);FPG与CRP、TNF-α、IL-6、IL-8以及CD8^＋T淋巴细胞呈正相关(r＝0.651、0.571、0.781、0.425、0.543,P值均小于0.05),与CD4^＋T淋巴细胞、CD4^＋/CD8^＋呈负相关(r＝－0.636、－0.519,P值均小于0.05)。 结论重度烧伤患者机体会发生应激反应,出现应激性高血糖且机体内炎性因子及免疫功能会发生明显的改变,血糖与炎性因子及免疫水平有显著的相关性,血糖越高者,机体炎症反应越明显,免疫功能越低下。     

原发性胆汁性肝硬化患者共刺激分子B7-H4的检测及临床意义

目的 探讨原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者共刺激因子B7-1t4的表达及其与疾病发病机制的关系.方法 分别采用荧光实时定量PCR法(real-time PCR)、酶联免疫吸附试验(ELISA)及流式细胞术(FCM)检测65名PBC患者外周血单个核细胞(PBMC)B7-H4mRNA表达水平、血清IL-2水平以及CD4+、CD8+ T细胞亚群和T细胞表面B7-H4表达百分率,同时监测抗线粒体抗体(anti-mitochondrial antibody,AMA)及临床各项生化指标的关系.结果 (1)PBC患者PBMC B7-144 mRNA水平及T细胞表面B7-H4表达百分率显著低于非PBC肝硬化组及健康对照组(P<0.01).(2)活化72 h后各实验组及对照组IL-2含量及CD4+、CD8+、CD4+ CD8+T淋巴细胞表达水平均低于活化前,以非PBC肝硬化组与健康对照组降低显著(P<0.05);PBC组IL-2含量及CD4+、CD4+ CD8+ T淋巴细胞表达水平高于非PBC肝硬化组与健康对照组(P<0.01).(3)AMA-M2阳性患者血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)及γ-谷氨酰转肽酶(GGT)水平升高,其中ALP及GGT升高显著(P<0.05);AMA-M2阳性患者与阴性患者T细胞表面B7-H4表达百分率差异无统计学意义(P>0.05).结论 共刺激因子B7-H4对PBC患者体内T细胞活化增殖及细胞因子分泌的抑制作用减弱,为研究PBC的发生发展和阐明PBC的发病机制提供了试验资料.     

Dynamics of T cell activation accompanying CD4 recovery in antiretroviral treated HIV-infected Ugandan children

Africans have elevated T cell activation compared to residents of Europe or the USA. Levels of T cell activation also correlate with disease progression in HIV-infected individuals. We sought to determine if treatment with antiretroviral therapy (ART) would reduce levels of T cell activation (CD38 and HLADR co-expression) in HIV-infected Ugandan children. The median CD8+ T cell activation level among 199 ART-treated children (30%) was lower than in 57 ART-naïve children (45%, p < 0.001), but remained higher than in 30 HIV-uninfected children (18%, p < 0.001). Among ART-treated children, CD4% was inversely correlated with both CD8− (ρ = − 0.61, p < 0.001) and CD8+ (ρ = − 0.38, p < 0.001) T cell activation. Prospectively, CD4 recovery correlated with post-treatment CD8+ T cell activation level (p = 0.008). Our data suggest that significant decreases in T cell activation accompany CD4 recovery in ART-treated HIV-infected African children, to levels that approach but do not reach those of uninfected children.     

The immunoregulatory role of IL-35 in patients with interstitial lung disease

Pulmonary fibrosis involves various types of immune cells and soluble mediators, including TGF-β and IL-35, a recently identified heterodimeric cytokine that belongs to the IL-12 cytokine family. However, the effect of regulatory IL-35 may play an important role in fibrotic diseases. The aim of this paper is to explore the immunoregulatory role of IL-35 in the development of fibrosis in interstitial lung disease (ILD). To gain a better understanding of this issue, the concentrations of IL-35 and different profibrotic cytokines in fibrotic (F-ILD) and non-fibrotic (NF-ILD) patients by ELISA were compared to that of intracellular IL-35 and IL-17 on CD4+ T cells stimulated in the presence of BAL or with different ratios of recombinant IL-35 (rIL-35) and TGF-β (rTGF-β), which were evaluated by flow cytometry. We observed that BAL concentration of IL-35 was lower in F patients (p < 0.001) and was negatively correlated with concentrations of TGF-β (p < 0.001) and IL-17 (p < 0.001). In supplemented cell cultures, BAL from NF but not F patients enhanced the percentage of IL-35 + CD4+ T (p < 0.001) cells and decreased the percentage of IL-17 + CD4+ T cells (p < 0.001). The percentage of IL-35 + CD4+ T cells correlated positively with BAL concentration of IL-35 (p = 0.02), but correlated negatively with BAL concentrations of IL-17 (p = 0.007) and TGF-β (p = 0.01). After adjusting the concentrations of recombinant cytokines to establish a TGF-β: IL-35 ratio of 1:4, an enhanced percentage of IL-35 + CD4+ T cells (p < 0.001) but a decreased percentage of IL-17 + CD4+ T cells (p < 0.001) was observed. After adding recombinant IL-35 to the BAL from F patients until a 1:4 ratio of TGF-β: IL-35 was reached, a significantly increased percentage of IL-35 + CD4+ T cells (p < 0.001) and a decreased percentage of IL-17 + CD4+ T cells (p = 0.003) was found. These results suggest that IL-35 may induce an anti-fibrotic response, regulating the effect of TGF-β and the inflammatory response on CD4+ T cells. In addition, the TGF-β: IL-35 ratio in BAL has been shown to be a potential biomarker to predict the outcome of F patients with ILD.     

Changes in intracellular cytokine levels in newborn and adult lymphocytes induced by HSV-1

Changes in the expression of intracellular interleukin-2 (IL-2), interleukin-4 (IL-4), interferon (IFN)-γ, and tumor necrosis factor (TNF)-α in newborn and adult lymphocytes induced by herpes simplex virus (HSV)-1 were examined. Cord blood mononuclear cells (CBMC) or adult peripheral blood mononuclear cells (PBMC) were infected with HSV-1 and cultured with phorbol 2-myristate 13-acetate (PMA) plus ionomycin in the presence of monensin for 4 hr. Surface antigen and intracellular cytokines were stained simultaneously and analyzed by flow cytometry. The percentage of cells that expressed IL-2, IFN-γ, and TNF-α was significantly increased in HSV-1-infected CD3⁺, CD4⁺, CD8⁺, CD45RA⁺, and CD45R0⁺ lymphocytes compared with uninfected lymphocytes from adult PBMC. The percentage of cells that expressed IL-2 and TNF-α was increased significantly in HSV-1-infected CD3⁺, CD4⁺, CD8⁺, and CD45RA⁺ lymphocytes compared with uninfected lymphocytes from CBMC. IFN-γ was under the detectable level in HSV-1-infected and uninfected lymphocytes from CBMC. Intracellular IL-4 was not detected in HSV-1 or in uninfected lymphocytes from PBMC and CBMC. These results demonstrate that HSV-1 enhances intracellular levels of IL-2, IFN-γ, and TNF-α in adult lymphocytes and defective IFN-γ production in cord blood. J. Med. Virol. 56:145–150, 1998. © 1998 Wiley-Liss, Inc.     

Immunomodulatory Effects of Lingzhi and San-Miao-San Supplementation on Patients with Rheumatoid Arthritis

Rheumatoid arthritis (RA) is an autoimmune joint disease. We evaluated a standard preparation of Lingzhi (Ganoderma lucidum) and San-Miao-San (Rhizoma atractylodis, Cortex phellodendri, Radix achyranthes bidentatae) capsules (TCM group) for its supplementary treatment efficacy for RA. There was no significant difference in the absolute count, percentage, and ratios of CD4+/CD8+/natural killer/B lymphocytes between the TCM and placebo groups after taking the capsules (all p > 0.05). There was no significant change in concentrations of plasma cytokines of interferon-γ-induced protein-10 (IP-10), monocyte chemoattractant protein-1, monokine induced by IFN-γ, regulated upon activation normal T-cell expressed and secreted, interleukin (IL)-8, and IL-18 after taking the capsules for 8 and 24 weeks (all p > 0.05). The percentage change in ex vivo-induced level of inflammatory cytokine IL-18 was significantly lower in the TCM group than in the placebo group after taking the capsules for 24 weeks (p < 0.05). Therefore, Lingzhi and San-Miao-San capsules might exert a beneficial immunomodulatory effect in patients with rheumatoid arthritis.     

Quantitative intracellular cytokine measurement: age-related changes in proinflammatory cytokine production

The proinflammatory cytokines play a central role in mediating cellular and physiological responses, and levels may reflect immune system effectiveness. In this study, the effect of ageing on the inflammatory response was examined using a novel method to detect production of the proinflammatory cytokines, i.e. tumour necrosis factor-alpha (TNF-α), IL-6 and IL-1β. Peripheral blood mononuclear cells (PBMC) obtained from healthy donors of different ages were incubated for 0, 24, 48 and 72 h with or without phorbol 12-myristate 13-acetate (PMA) stimulation. At each time point these cells were permeabilized and incubated with secondary conjugated FITC MoAbs specific for each cytokine. A flow cytometric system was developed to quantify specific intracellular fluorescence in T cells (CD3⁺) and monocytes (CD14⁺). TNF-α, IL-6 and IL-1β production in cell culture supernatants was also measured using ELISAs. In older subjects, flow cytometry detected significant increases in intracellular T cell TNF-α and IL-6 (P < 0.05). IL-1β was not detected in any of the T cell samples. Likewise, the monocytes of older subjects demonstrated increased intracellular levels of all three cytokines, but these increases were not significant (P > 0.05). These changes in intracellular proinflammatory cytokine levels may explain some of the exaggerated inflammatory responses seen in elderly patients.     

Reduction in interleukin-2-producing cells but not Th1 to Th2 shift in moderate and advanced stages of human immunodeficiency virus type-1-infection: direct analysis of intracellular cytokine concentrations in CD4+ CD8- T cells

Previous studies have suggested that CD4+ T lymphocytes shift from the Th1 type to the Th2 type during disease progression in patients with the human immunodeficiency virus type-1 (HIV-1). In the present study, we used a modified method that allowed a direct measurement of intracellular cytokines in CD4+ CD8- T cells. A total of 48 HIV-1-infected (HIV+) and 16 HIV-1-uninfected (HIV-) individuals were studied. The percentages of CD4+ CD8- T cells producing interleukin-2 (IL-2), interferon-gamma (IFN-gamma), interleukin-4 (IL-4), or interleukin-5 (IL-5) in HIV+ and HIV- subjects were 23.6% versus 34.9% (P < 0.01), 13.7% versus 13.2%, 1.3% versus 1.0%, and 1. 2% versus 0.9%, respectively. The population of IL-2-producing cells decreased proportionately with reductions in CD4 counts (< 200/mm3, 200-500/mm3, and > 500/mm3 to 18.0%, 23.5%, and 30.5%, P < 0.05, respectively). There was an inverse correlation between the percentage of IL-2-producing cells and plasma viral load (r = - 0. 446, P < 0.05). However, the percentages of CD4+ CD8- T cells producing other cytokines were not different between HIV+ and HIV-. Our cross-sectional study demonstrated a decrease in IL-2-producing cells but not the Th1 to the Th2 shift in the CD4+ CD8- T cell population in the moderate and advanced stages of HIV-1-infection.     

Gammadelta T cells and interleukin-6 levels could provide information regarding the progression of human renal allograft

We have determined the percentage of alphabeta and gammadelta T cells by flow cytometry as well as serum interleukin-6 (IL-6) and soluble interleukin-6 receptor (sIL-6R) levels by enzyme-linked immunosorbent assay in kidney allograft recipients with acute, chronic or stable graft evolution. The percentage of CD4 and CD8 T cells in transplanted patients was lower than in the control group (P < 0.001) with the exception of CD8 gammadelta T cells from patients with stable evolution (P > 0.05). The serum levels of IL-6 and sIL-6R in acute and chronic rejection were higher than in the controls (P < 0.05). No differences in IL-6 levels were observed between the stable evolution and the control groups (P > 0.05). The levels of sIL-6R were higher in stable evolution patients than in the controls (P < 0.05) and no differences were observed between the chronic and stable evolution patients (P > 0.05). IL-6 decreased in patients with a favourable evolution, increased in those with an increased renal dysfunction and was maintained when the renal dysfunction was not modified. These results suggest that gammadelta T cells could participate in renal allograft maintenance and that IL-6 but not sIL-6R serum levels may provide a prognostic marker for measuring the evolution of kidney allograft.     

TLR4 基因敲除对小鼠内脏脂肪组织免疫细胞及脂肪因子的影响

目的:探讨TLR4 基因敲除对小鼠免疫细胞及脂肪因子的影响。方法:取20 周龄的雄性野生型C57BL/6 小鼠和TLR4-/ -小鼠的脾脏和附睾脂肪组织,分离细胞,用流式检测F4/80、CD11b、CD11c、CD3、CD4、CD8 分子的表达;qPCR 检测附睾脂肪组织内IL-6、HMGB1、TNF-α、脂联素和抵抗素的表达。结果:与野生型C57BL/6 小鼠相比,TLR4-/ - 小鼠脾脏和附睾脂肪组织中M1 型(F4/80+ CD11b+ CD11c+ )巨噬细胞比例上升(P<0.05),M2 型(F4/80+ CD11b+ CD11c- )巨噬细胞比例下降(P<0.05),这种趋势在附睾脂肪组织中表现更为显著。同时发现附睾脂肪组织中CD4+ T 细胞比例下降(P<0.05),CD8+ T细胞比例上升(P<0.05);IL-6、HMGB1、抵抗素表达升高(P<0.05);TNF-α和脂联素表达降低(P<0.05)。结论:TLR4 基因敲除可导致内脏脂肪组织脂肪因子和免疫细胞的紊乱。