手足口病患儿外周血T淋巴细胞亚群的临床研究

目的 研究于足口病患儿T淋巴细胞亚群的变化趋势,为临床治疗及预后评估提供依据.方法 采用流式细胞仪检测2008年5月1日至8月31日在北京地坛医院住院的346例手足口病患儿急性期的T淋巴细胞亚群,同时测定其中67例患儿恢复期的T淋巴细胞哑群;对其中99例患儿咽拭子标本进行RT-PCR以确定病原.埘不同样本行独立样本t检验、配对t检验或方差分析.结果 手足口病患儿T淋巴细胞业群均数低于相应年龄段正常参考值水平.重症患儿各年龄组中的T淋巴细胞(TL)/淋巴细胞(L)比值、＞1岁组T辅助细胞(Th)/L比值,以及1～2岁患儿的TL、Th及Th/抑制性T淋巴细胞(Ts)值低于普通病例(P＜0.05).患儿T淋巴细胞亚群随病程进展有逐渐上升趋势,普通患儿的TL/L及Th/L在恢复期增高(TL/L:56.3±8.6比61.1±9.1,t=2.56,P＜0.05;Th/L:30.2±7.2比34.9±7.9,t=2.90,P＜0.05),重症患儿除Ts/L和Th/Ts外,所有指标均在恢复期明显增高(P＜0.01);急性期重症与普通崽儿的TL为(1.738±0.976)×10~6/L比(2.696±1.946)×10~6/L(t=2.17,P＜0.05),Th/L为25.9±7.0比30.2±7.2(t=2.34,P＜0.05),Th为(0.864±0.550)×10~6/L比(1.459±0.879)×10~6/L(t=2.90,P＜0.01),L为(3.352±1.458)×10~6/L比(4.664±2.435)×10~6/L(t=2.32,P＜0.05),恢复期两组比较差异无统计学意义(P＞0.05).肠道病毒(EV)71阳性患儿T淋巴细胞亚群各值均低于阴性患儿,但差异无统计学意义(P＞0.05).结论 T淋巴细胞免疫功能可能与手足口病发病及病情发展有关.     

支原体肺炎患儿外周血淋巴细胞亚群监测及临床意义

目的探讨肺炎支原体感染与儿童细胞免疫功能的关系及临床意义。方法对30例支原体肺炎患儿的T淋巴细胞亚群进行检测,并与健康儿童对照。结果支原体肺炎患儿外周血T淋巴细胞亚群CD3＋、CD4＋,均明显高于健康儿童（P〈0．01）。结论支原体感染患儿存在细胞免疫功能紊乱或失调。     

卵巢早衰患者外周血T淋巴细胞亚群变化及其临床意义

采取流式细胞仪检测12例卵巢早衰（POF）患者（观察组）与20例健康育龄妇女（对照组）外周血T淋巴细胞亚群CD3、CD4、CD8变化；应用磁性匀相酶联免疫法测定两组性激素促卵泡生成素（FSH）、黄体生成素（LH）、雌激素（E2）、孕激素（P）、雄激素（T）、泌乳素（PRL）水平。结果观察组CD4低于对照组，CD8高于对照组，CD4^＋／CD8^＋明显降低；FSH、LH水平明显高于对照组，P均〈0．05；E2、PRL水平低于对照组，P均〈0．05。对外周血T淋巴细胞亚群与雌激素水平进行相关性分析显示，血清E2降低，CD3、CD4／CD8分别与E2呈正相关．E2与CD8^＋呈负相关。提示E2有免疫抑制作用，可抑制T细胞免疫应答。     

疣状胃炎外周血T淋巴细胞亚群检测及其临床意义

目的:检测疣状胃炎患者(n=35)外周血T淋巴细胞亚群CD_3+、CD_4+、CD_8+,并与107例慢性浅表性胃炎中的幽门螺杆菌(Hp)感染的患者(n=16)比较。方法:所有患者均通过胃粘膜活检快速尿素酶试验和~(14)C-尿素呼气试验以确定是否有Hp感染,用流式细胞术方法检测疣状胃炎和Hp感染的浅表性胃炎患者外周血T淋巴细胞亚群CD_3+、CD_4+、CD_8+。结果:疣状胃炎Hp感染率明显高于慢性浅表性胃炎者(P<0.01);与Hp感染的浅表性胃炎比较,疣状胃炎CD_3+、CD_8+降低以及CD_4+/CD_8+比值增加(P<0.01,P<0.05)。疣状胃炎中Hp阳性组与Hp阴性组比较CD_8也明显降低(P<0.05)。结论:疣状胃炎发病可能有免疫因素参与。     

不同临床类型HBV感染者外周血T淋巴细胞亚群的变化

目的研究HBV感染者外周血T淋巴细胞亚群的变化及其临床意义.方法在33例慢性乙型肝炎轻度、50例中度、32例重度、64例肝硬化、51例慢性乙型肝炎肝衰竭和30例健康人,采用特异性荧光抗体标记法和流式细胞仪检测外周血T淋巴细胞亚群.结果慢性乙型肝炎重度、乙型肝炎肝硬化和慢性乙型肝炎肝衰竭患者与正常对照组比,CD3^＋、CD4^＋、CD8^＋细胞数和CD4^＋/CD8^＋比值显著下降（P〈0.05）,以慢性乙型肝炎肝衰竭下降最显著;HBeAg阴性患者外周血CD3^＋、CD4^＋和CD8^＋细胞数较HBeAg阳性患者显著减少（P〈0.05）;HBV DNA载量与患者外周血CD3^＋、CD4^＋、CD8^＋细胞数和CD4^＋/CD8^＋比值均无明显相关性.结论严重的HBV感染者存在不同程度的细胞免疫功能低下及免疫调节紊乱,细胞免疫功能下降程度与病毒复制水平无明显相关.     

慢性HBV感染患者外周血T淋巴细胞免疫应答及其临床意义

目的探讨Th1／Th2、Tc1／Tc2亚群比例在慢性HBV感染的临床分型中的变化,以及在慢性HBV感染免疫病理损伤中的作用。方法用PMA、Ionomycin作为刺激剂,采用流式细胞仪（FACS）胞内细胞因子法对慢性HBV感染者外周血CD3＋CD8＋T细胞和CD3＋CD8-T细胞内IFN～y和IL-4的表达进行分析,比较慢性肝炎、肝炎肝硬化（活动性）、慢性重症肝炎各组Th1／Th2、Tc1／Tc2亚群比例变化。结果慢性肝炎、肝炎肝硬化（活动性）、慢性重症肝炎患者的Th1、Tc1细胞均高于正常对照组。慢性重症肝炎组Th1、Tc1显著高于慢性肝炎、正常对照组（P〈0．01）,慢性重症肝炎组Tc1显著高于活动性肝炎后肝硬化组（P〈0．05）。肝炎肝硬化（活动性）组Tc1显著高于正常对照组（P〈0．05）。Th1、Tc1细胞随着慢性乙型肝炎肝脏炎症活动的加剧而增高。而Th2、Tc2细胞则在各组中均无显著性差异（P〉0．05）。慢性重症肝炎患者恢复期Th1和Tc1细胞显著低于治疗前水平（P〈0．01）。结论在慢性HBV感染中,机体免疫平衡偏向Th1类反应,Th1、Tc1细胞与肝脏炎症活动严重程度呈正相关。提示Th1和Tc1细胞在慢性乙型肝炎肝脏病理发生中可能起了重要作用。     

急性白血病患者外周血T淋巴细胞亚群检测的临床意义

急性白血病患者的细胞免疫功能显著降低,表现为总T细胞(T_3)、T辅助细胞(T_H)和T抑制细胞(T_s)阳性率显著低于正常值,化疗后有效患者的T细胞免疫功能基本恢复正常,化疗无效和复发患者的T淋巴细胞亚群与未治组患者的相似,动态观察有助于判断急性白血病患者的预后。     

重症病毒性脑炎患儿T淋巴细胞亚群变化及其临床意义分析

目的观察重症病毒性脑炎患儿T淋巴细胞亚群变化,探讨其临床意义。方法选取2009年6月—2012年6月山东大学齐鲁儿童医院收治的病毒性脑炎患儿60例,其中轻、重型患儿各30例,分别作为轻型组、重型组。同时选取健康儿童30例作为对照组。轻型组患儿给予常规治疗,重型组患儿在常规治疗基础上给予大剂量甲泼尼龙联合丙种球蛋白治疗。比较3组受试者T淋巴细胞亚群及重型组不同预后患儿治疗前后T淋巴细胞亚群。结果轻型组患儿CD+3、CD+4、CD+8细胞分数及CD+4/CD+8细胞比值与对照组比较,差异均无统计学意义(P0.05);重型组患儿CD+3、CD+4细胞分数均低于对照组、轻型组,CD+8细胞分数、CD+4/CD+8细胞比值均高于对照组、轻型组(P0.05)。重型组预后良好患儿治疗后CD+3、CD+4细胞分数及CD+4/CD+8细胞比值均高于治疗前,CD+8细胞分数低于治疗前(P0.05);重型组预后不良患儿治疗后CD+3、CD+4细胞分数及CD+4/CD+8细胞比值高于治疗前(P0.05),而治疗前后CD+8细胞分数比较,差异无统计学意义(P0.05)。结论重症病毒性脑炎患儿常伴有T淋巴细胞亚群改变,检测重症病毒性脑炎患儿T淋巴细胞亚群有助于判断病情严重程度及预后效果。     

慢性HCV感染患者外周血T淋巴细胞亚群的检测

目的探讨慢性丙型肝炎病毒(HCV)感染患者T细胞亚群水平以及与丙型肝炎病毒复制的关系。方法以间接免疫荧光一流式细胞仪、ELISA法和RT-PCR检测了26例慢性HCV感染患者外周血T细胞亚群(其中HCV-RNA阳性17例,HCV-RNA阴性9例)。结果慢性HCV感染患者外周血CD_4~ 淋巴细胞亚群,CD_4~ /CD_8~ 比值明显低于正常对照组(P<0.01)。血清HCV-RNA阳性患者的CD_4~ /CD_8~ 比值明显低于HCV-RNA阴性患者(P<0.05)。结论提示细胞免疫功能受抑制可能是HCV持续感染的原因。     

慢性HBV感染患者T细胞亚群变化规律及可能的意义

目的通过对不同病理阶段的224例HBV感染患者CD4 、CD8 T细胞亚群、HBV DNA等指标的检测结果进行了回顾性分析,以期明确不同病理阶段T细胞亚群变化的规律及可能的临床意义.方法外周血CD4 和CD8 T细胞亚群检测采用流式细胞仪技术;HBV DNA检测采用荧光定量PCR检测;ELISA检测用于血清HBeAg和抗HBe评价.结果肝硬化患者(499.75±304.46,t=-3.12,P=0.041)和肝癌患者(442.10±241.81,t=4.1,P=0.038)CD4 T细胞,显著低于急性肝炎患者(682.21±299.74);肝硬化患者(303.45±225.84,t=3.75,P=0.033)和肝癌患者(296.59±279.51,t=5.39,P=0.026)CD8 T细胞,显著低于急性肝炎患者(542.83±277.35);59例肝硬化患者的CD4 T细胞(477.42±206.39)低于35例HBsAg阳性患者(512.37±261.87),但统计学差异不显著(t=0.76,P=0.62).但前者CD8 T细胞亚群显著高于后者(369.35±203.14 vs 277.81±212.58;t=3.02,P=0.41);肝硬化和肝癌血清HBV DNA阳性患者,均显著低于慢性重型肝炎患者(t=4.21,P=0.038;t=3.92,P=0.041),但肝癌组与肝硬化组间无差异(t=0.49,P=0.37).结论HBV感染肝硬化和肝癌患者CD4 和CD8 T细胞亚群较急性肝炎和健康对照人群显著降低,对于CD8 T细胞亚群显著降低的HBV感染患者,应该排除原发性肝癌的存在.     

慢性HBV感染者外周血T淋巴细胞亚群和NK细胞活性变化

探讨慢性HBV感染者外周血T淋巴细胞亚群及NK细胞活性变化情况。应用流式细胞法检测所有研T淋巴细胞亚群和NK细胞。慢性乙肝、肝炎肝硬化和慢性重型乙肝组患者CD3 、CD4 百分率及CD4 ／CD8 比值与正常组比较均有所下降,且慢性重型乙肝组患者CD4 百分率和CD4 ／CD8 比值与正常组比较差异有显著性。各临床类型慢性HBV感染者NK细胞百分率均降低,与正常对照组比较有统计学意义。慢性HBV感染者细胞免疫功能低下。检测T淋巴细胞亚群及NK细胞活性变化对判断病变程度、指导临床治疗具有一定参考价值。     

Peripheral T-lymphocyte subpopulations in different clinical stages of chronic HBV infection correlate with HBV load

AIM： To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus （HBV） replication in different dinical stages of chronic HBV infection.
METHODS： A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages： immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time poiymerase chain reaction.
RESULTS： CD8^＋ T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages （36.87 ± 7.58 vs 34.37 ± 9.07, 36.87 ± 7.58 vs 28.09 ± 5.64, P 〈 0.001）. The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8^＋ T-cells than CD4^＋ T-cells （36.87 ± 7.58 vs 30.23 ± 6.35, 34.37 ± 9.07 vs 30.92 ± 7.40, P 〈 0.01）, whereas the peripheral blood in patients at the immune- inactive carrier stage and in normal controls contained less CD8^＋ T-cells than CD4^＋ T-cells （28.09 ± 5.64 vs 36.85 ±6.06, 24.02 ± 4.35 vs 38.94 ± 3.39, P 〈 0.01）. ANOVA linear trend test showed that CD8^＋ T-cells were significantly increased in patients with a high viral load （39.41 ± 7.36, 33.83 ± 7.50, 31.81 ± 5.95 and 26.89 ± 5.71, P 〈 0.001）, while CD4^＋ T-cells were significantly increased in patients with a low HBV DNA load （37.45 ± 6.24, 33.33 ± 5.61, 31.58 ± 6.99 and 27.56 ± 5.49, P 〈 0.001）. Nultiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3^＋, CD4^＋ and CD8^＋ cells and CD4^＋/CD8^＋ ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation.
CONCLUSION： Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.     

Liver cell transformation in chronic HBV infection

Epidemiological studies have provided overwhelming evidence for a causal role of chronic HBV infection in the development of hepatocellular carcinoma (HCC), but the molecular mechanisms underlying virally-induced tumorigenesis remain largely debated. In the absence of a dominant oncogene encoded by the HBV genome, indirect roles have been proposed, including insertional activation of cellular oncogenes by HBV DNA integration, induction of genetic instability by viral integration or by the regulatory protein HBx, and long term effects of viral proteins in enhancing immune-mediated liver disease. In this chapter, we discuss different models of HBV-mediated liver cell transformation based on animal systems of hepadnavirus infection as well as functional studies in hepatocyte and hepatoma cell lines. These studies might help identifying the cellular effectors connecting HBV infection and liver cell transformation.     

Functionally aberrant dendritic cell subsets and expression of DC-SIGN differentiate acute from chronic HBV infection

Background

Dendritic cells (DCs) promote pathogen recognition, uptake and presentation of antigen through DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN) and toll-like receptors (TLRs).

Aims and Objectives

We aimed to study temporal changes in DCs, TLRs and DC-SIGN during acute viral hepatitis B (AVHB) infection and compare them to chronic (CHB) and to investigate the earliest time point of activated pathogen recognition receptors in hepatitis B viral infection.

Methods

We measured the frequencies of circulating myeloid (mDC) and plasmacytoid (pDC) dendritic cells and IFN-α production along with the expression of DC-SIGN and Toll Like Receptors (TLR's) in HBV patients at different time points. Also investigated in healthy volunteers, the dynamic changes in TLRs expression after receiving hepatitis B vaccine.

Results

On follow-up of AVHB patients, we found the mDC population was significantly higher at week 4 and 6 (p < 0.02, 0.01), whereas the pDC population was unchanged at week 6 compared with week 0. Whereas frequencies of mDCs and pDCs were found to be elevated in AVHB and CHB patients than HC (p < 0.00 and 0.01, respectively) but was comparable among AVHB vs CHB. The DCs in CHB patients were functionally impaired with significantly low IFN-α production and low DCSIGN expression (p < 0.04 and 0.00, respectively). Even after stimulation by TLR agonists, no change was found in IFN-α production in CHB patients. MyD88 and IL-6, IFN-α mRNA levels were also found down-regulated. Interestingly, on follow-up after HBV vaccine, TLRs expression was found high at day 3 after vaccination.

Discussion

The initial events of immune activation might be responsible for modulating immune response. These novel observations would pave the way for the development of antiviral strategies for chronic HBV infection.

     

慢性HBV感染者CD8^＋ T细胞免疫反应的研究

目的观察不同慢性HBV感染者外周血CD8^＋ T细胞的细胞内毒性颗粒和细胞因子的表达水平。方法用rHBcAg作为刺激剂,采用免疫荧光染色结合流式细胞术分析慢性乙型肝炎中度、重度患者以及无症状HBV携带者外周血CD8^＋ T细胞的IFN-γ/IL-4和穿孔素/颗粒酶B表达水平。结果慢性乙型肝炎重度患者CD8^＋T细胞的穿孔素及颗粒酶B百分率显著高于正常对照（P〈0.01）和HBV携带者（P〈0.05,P〈0.01）,中度患者显著高于正常对照（P〈0.05）。2组慢性乙型肝炎IFN-γ＋/CD8^＋ T细胞（Tc1）百分率均高于正常对照和HBV携带者（P〈0.05）。HBV携带者CD8^＋T细胞的穿孔素和颗粒酶B表达以及Tc1百分率与健康对照差异无统计学意义。IL-4^＋/CD8^＋ T细（Tc2）表达各组间差异无统计学意义（P〉0.05）。结论 CD8^＋ T细胞中穿孔素/颗粒酶B表达率的增加以及Tc1细胞增加与慢性HBV感染的肝损伤有关。     

gammadelta T cell subsets in patients with arthritis and chronic neutropenia

BACKGROUND: An abnormal distribution of subsets of gammadelta T cells, which are a component of the inflammatory infiltrate in arthritic synovium, has been demonstrated in the peripheral blood (PB) of patients with arthritis and neutropenia. OBJECTIVE: To evaluate whether the clinical manifestations of patients with arthritis and neutropenia are related to the specific gammadelta T cell subset predominant in the PB. METHODS: Flow cytometry of PB lymphocytes in six consecutive patients with chronic neutropenia and arthritis was performed. Variable (V) gamma and delta gene families were analysed by polymerase chain reaction. cDNA was subjected to direct automated sequencing of T cell receptor (TCR) genes. RESULTS: Three patients had non-deforming and non-erosive rheumatoid factor (RF)(+) polyarticular rheumatoid arthritis, RF(+) oligoarticular arthritis, or RF(-) non-deforming oligoarticular psoriatic arthritis with persistent expansions of Vgamma1(+)/Vdelta2(+), Vgamma2(+)/Vdelta2(+), or Vgamma1(+)/Vdelta (undetermined (2- 1-)) T cells, respectively. The other three patients, without persistent expansion of gammadelta T cells, had either non-deforming and non-erosive oligo- or polyarthritis with a balanced distribution of several Vdelta and Vgamma genes, or severe erosive RF(+) arthritis with deficiency of all but Vgamma1(+)/Vdelta1(+) T cells. CONCLUSIONS: gammadelta T cell lymphoproliferations in chronic neutropenia and arthritis use different Vgamma and Vdelta gene families, often forming T cell receptor (TCR) structures that are infrequent in normal adult PB. Arthritis with Vgamma1(+)/Vdelta2(+), Vgamma2(+)/Vdelta2(+), or Vgamma1(+)/Vdelta2(-)/Vdelta1(-) gammadelta T cells in the PB is non-deforming and non-erosive, suggesting a protective effect of these cells, as opposed to a more pathogenic contribution of Vgamma1(+)/Vdelta1(+) cells.     

Immunoregulatory T cell subsets in chronic hepatitis B virus infection: the influence of homosexuality

The purposes of this study were 2-fold: (i) To enumerate peripheral immunoregulatory T cell subsets in untreated patients with chronic hepatitis B virus (HBV) infection and (ii) to examine the relationship between disturbances in the balance of lymphocyte subsets with liver disease and the presence of homosexuality. Circulating T lymphocyte subsets were evaluated by monoclonal antibodies to the following cell antigens: OKT3 (total T cells), OKT4 (helper/inducer T cells), and OKT8 (suppressor/cytotoxic T cells). The following groups of subjects were examined: (i) 16 heterosexuals with HBV-associated chronic active hepatitis (CAHB); (ii) 10 heterosexual, healthy HBsAg carriers, and (iii) 16 male homosexuals with CAHB. Controls included 51 healthy heterosexuals and 12 healthy, noninfected male homosexuals. We were able to demonstrate that heterosexuals with CAHB had T4/T8 ratios which did not differ from those of noninfected heterosexuals. Both healthy carriers and healthy homosexuals, however, exhibited significantly lower T4/T8 ratios than did noninfected heterosexuals (p less than 0.05, p less than 0.01, respectively). In addition, homosexuals with CAHB had lower (1.5 +/- 0.1) T4/T8 ratios than did heterosexuals with CAHB (2.0 +/- 0.2). A possible mechanism for these findings is discussed. The data indicate that the presence of homosexuality may be an important factor to consider when evaluating immunoregulatory subsets in CAHB.