Fas、Caspase-3和抗核小体抗体在参与SLE患者淋巴细胞凋亡紊乱中的作用

目的探讨SLE患者免疫功能紊乱与淋巴细胞凋亡信号传导途径异常之间的关系。方法应用流式细胞术测定SLE患者淋巴细胞表面Fas、FasL及细胞质中活化caspase-3的表达率,并测定凋亡细胞百分率（AnnexinV^＋PI^-）和坏死细胞百分率（AnnexinV^＋PI^＋）。应用ELISA方法测定血清中抗核小体抗体浓度。结果与健康对照组相比,稳定期和活动期SLE患者组淋巴细胞中凋亡细胞和坏死细胞百分率均显著增加（P〈0．05）,淋巴细胞表面Fas、FasL及细胞质中活化caspase-3的表达率也显著增加（P〈0．05）。与稳定期SLE患者组相比,活动期SLE患者组淋巴细胞中坏死细胞百分率显著增加（P〈0．05）,凋亡细胞百分率略有增加但无统计学意义（P〉0．05）。活动期患者组淋巴细胞表面Fas、FasL以及细胞质中活化caspase-3的表达率略有增加但无统计学意义（P〉0．05）。活动期SLE患者组抗核小体抗体浓度显著高于健康对照组和稳定期患者组（P〈0．05）。SLE患者凋亡细胞百分率和活化caspase-3的表达率与补体C3浓度水平呈负相关关系（P〈0．05）。结论Fas信号传导通路在SLE患者淋巴细胞凋亡紊乱中发挥了重要作用。caspase-3的活化是早期提示淋巴细胞凋亡的重要信号。SLE患者淋巴细胞凋亡活化程度与疾病活动程度和免疫效应功能紊乱密切相关,而淋巴细胞凋亡异常程度与抗核小体抗体水平的高低密切相关。淋巴细胞凋亡加速在SLE患者免疫病理损伤加重和免疫细胞调控紊乱中扮演了重要角色。     

Structural and functional characteristics of lymphocytes in chronic carriers of tick-borne encephalitis virus

Study of the surface phenotype, structure of plasma membrane, and activity of DNA repair system in peripheral blood lymphocytes from patients with different clinical forms of tick-borne encephalitis associated with long persistence of the virus revealed a pronounced imbalance between lymphocyte subpopulations, increased microviscosity of the plasma membrane lipid phase, and decreased activity of the DNA repair system in lymphocytes.     

Chronic restraint stress induces severe disruption of the T-cell specific response to tetanus toxin vaccine

Chronic stress is known to induce immunological disorders. In the present study we examined the consequences of chronic restraint stress on the immune response to tetanus toxin in mice. We investigated the repartition of subsets of lymphoid cells in blood and spleen, the functional ability of lymphocytes to proliferate and to produce cytokines, and antibody titres against tetanus toxin following stress. We report discordance of the stimulation index of lymphocytes in the restraint group: the proliferating rate severely decreased following stimulation with a relevant antigen, whereas it increased with mitogen. Thus, we report a decrease in cytokine production with relevant antigen (interferon-gamma and interleukin-10), without a T helper type 1 and 2 secretion imbalance. Moreover, we observed an alteration in the humoral response, including a delay in isotype maturation and an immunoglobulin G1/G2a imbalance.     

COPD患者外周血T淋巴细胞亚群的变化及乌体林斯的调节作用

目的:探讨COPD患者外周血T淋巴细胞亚群的变化及乌体林斯治疗COPD的作用机制.方法:100例COPD患者随机分为治疗组和对照组,每组各50例,在我院体检的健康成人50例作为健康体检组.对照组根据病情需要给予抗感染、止咳、祛痰、平喘等综合治疗.治疗组在对照组治疗基础上加用乌体林斯1.72 μg/次,肌肉注射,每周1次...     

Dysregulation of IL-2 and IL-8 production in circulating T lymphocytes from young cystic fibrosis patients

It is well documented that patients with cystic fibrosis (CF) are unable to clear persistent airway infections in spite of strong local inflammation, suggesting a dysregulation of immunity in CF. We and others have reported previously that T lymphocytes may play a prominent role in this immune imbalance. In the present work, we compared the reactivity of CD3+ T cells obtained from young CF patients in stable clinical conditions (n = 10, aged 9-16.5 years) to age-matched healthy subjects (n = 6, aged 9-13.5 years). Intracellular levels of interferon (IFN)-gamma, interleukin (IL)-2, IL-8 and IL-10 were determined by flow cytometry after whole blood culture. The data identified T lymphocyte subsets producing either low levels (M1) or high levels (M2) of cytokine under steady-state conditions. We found that the production of IFN-gamma and IL-10 by T lymphocytes was similar between young CF patients and healthy subjects. In contrast, after 4 h of activation with PMA and ionomycin, the percentage of T cells producing high levels of IL-2 (M2) was greater in CF patients (P = 0.02). Moreover, T cells from CF patients produced lower levels of IL-8, before and after activation (P = 0.007). We conclude that a systemic immune imbalance is present in young CF patients, even when clinically stable. This disorder is characterized by the capability of circulating T lymphocytes to produce low levels of IL-8 and by the emergence of more numerous T cells producing high levels of IL-2. This imbalance may contribute to immune dysregulation in CF.     

Subpopulations of human T lymphocytes. IX. Imbalance of T cell subpopulations in patients with progressive systemic sclerosis.

Peripheral blood lymphocytes from twenty patients with progressive systemic sclerosis (PSS) were analysed for the numbers and proportions of B lymphocytes possessing surface immunoglobulin, cells with Fc receptors, T cella and T cells with receptors for IgM (T mu) or IgG (T gamma). In patients with PSS, B cells and lymphocytes with Fc receptors were comparable in both numbers and proportions to those of the control group. Circulating T lymphocytes were significantly fewer in the patient group. T mu cells were decreased and T gamma cells increased, resulting in lower T mu/T gamma ratios as compared to controls. This study demonstrates a profound imbalance between T mu and T gamma cells (containing a population of helper or suppressor cells, respectively). These results are discussed in relation to immunodeficiencies observed in patients with PSS.     

急性心肌梗死后辅助性T细胞功能失衡的意义

目的：探讨急性心肌梗死(AMI)后辅助性T淋巴(Th)细胞功能失衡的意义。方法： 采用流式细胞分析法对33例AMI病人，22例不稳定心绞痛(UA)病人和35例AMI大鼠进行Th细胞胞内干扰素γ(IFN-γ)和白介素4(IL-4)的动态监测，同时应用RT-PCR方法检测AMI大鼠心肌组织IFN-γ和IL-4以及T细胞表面趋化因子受体CCR3、CCR5和CXCR3 mRNA的表达。结果：AMI和UA病人发病24 h内Th1型胞内IFN-γ的水平明显高于对照组。UA患者IFN-γ高表达持续时间较短，发病1周后恢复；AMI患者IFN-γ高表达持续时间较长，发病后1周甚至1月仍增高。Th2型胞内IL-4未见明显变化。AMI大鼠心梗后Th细胞胞内IFN-γ和IL-4均无明显变化，T细胞表面趋化因子受体CCR3、CCR5和CXCR3 mRNA表达亦无显著差异，但AMI 1周、2和1月末心肌组织局部IFN-γ mRAN的表达明显增加。结论： AMI后机体出现T细胞功能失衡，主要表现为Th1细胞功能亢进，可能参与AMI的发病；AMI后T细胞功能失衡可能作为自身免疫病的发病机制之一，参与了AMI后自身免疫性心肌炎的发病和自身免疫因素引起的心肌损伤和心室重塑过程。     

In vitro production of B cell growth factor and B cell differentiation factor by peripheral blood mononuclear cells and bronchoalveolar lavage T lymphocytes from patients with idiopathic pulmonary fibrosis.

The activation of B lymphocytes and formation of immune complexes have been suggested to play an important role in the pathogenesis of idiopathic pulmonary fibrosis (IPF). To investigate the mechanisms of activation of B lymphocytes, we studied the production of B cell growth factor (BCGF) and B cell differentiation factor (BCDF) in patients with IPF and those with interstitial pneumonia associated with collagen vascular diseases (IP-CVD), in comparison with healthy controls. Culture supernatants of peripheral blood mononuclear cells from patients with IPF induced more IgM and IgA production by B lymphocytes than those from healthy controls, indicating a higher production of BCDF in the patients. Culture supernatants of T lymphocytes obtained from bronchoalveolar lavage fluids (BALF) of patients with IPF induced higher proliferation of B lymphocytes than those from healthy controls, indicating a higher production of BCGF. An increase in production of BCGF and BCDF was not observed in patients with IP-CVD. In the light of these results, it was suggested that there may be an imbalance in T lymphocyte subsets that release lymphokines like BCGF and BCDF in patients with IPF, and that the subsets may differ between blood and BALF. It remains to be elucidated whether the activation of B lymphocytes depending on T lymphocytes determines the development of disease in IPF.     

Functional behaviour and immunological phenotype of circulating B lymphocytes in multiple myeloma. Studies with pokeweed mitogen.

Peripheral blood B lymphocytes, depleted of adherent cells, from 10 patients with multiple myeloma were cultured in the presence of PWM with autologous or donor T lymphocytes. The results show that: (1) co-cultures with allogeneic T lymphocytes produced more plasma cells than those with autologous ones; (2) the kappa/lambda ratio overlapped the values obtained in normal controls, irrespective of the light chain produced by the neoplastic plasma cells and (3) the immunological phenotype of plasma cells obtained from PWM stimulated peripheral B cells (RFA2+, RFA3+, A10+) was clearly different from that one of myelomatous plasma cells (RFA2-, RFA3-, A10+). These data confirm the T cell imbalance already seen in myeloma patients; moreover they show that PWM responsive B cell are functionally normal and phenotypically different from bone marrow myeloma cells. These results support the view that most of the peripheral B lymphocytes, previously identified as monoclonal are in fact normal cells bearing adherent monoclonal Ig molecules.     

SLE患者淋巴细胞凋亡和p53蛋白表达的相关性研究

研究SLE患者外周血淋巴细胞在体外培养下细胞凋亡和p5 3蛋白表达与SLE疾病活动相关性。AnnexinV联合PI染色定量法及免疫荧光染色法 ,分析了 4 4例SLE患者和 30例正常人外周血淋巴细胞在体外培养后凋亡发生率 ,凋亡相关基因p5 3蛋白的表达以及淋巴细胞凋亡发生与疾病活动的相关性。在体外培养作用下 ,活动期SLE患者淋巴细胞凋亡发生率较正常人显著增高 (P <0 0 1 ) ,而静止期则明显升高 (P <0 0 5 ) ,疾病活动指数与体外淋巴细胞凋亡率呈正相关 (P <0 0 1 )。p5 3蛋白表达在活动期SLE患者较正常人显著性下降 (P <0 0 1 ) ,静止期明显降低 (P <0 0 5 )。p5 3蛋白的表达与疾病活动指数SLEDAI,抗dsDNA抗体和C3补体之间有明显的相关性 (P <0 0 1 )。SLE患者外周淋巴细胞在体外培养凋亡率较正常人显著增高 ,表明SLE存在体内凋亡或凋亡相关性免疫机制失调     

Immunological studies in typhoid fever. II. Cell-mediated immune responses and lymphocyte subpopulations in patients with typhoid fever.

The development of the cell-mediated immune response (CMIR) to antigens prepared from Salmonella typhi was investigated in patients suffering from typhoid fever and in normal healthy subjects. The leucocyte migration inhibition test, blast transformation of lymphocytes and active rosette-forming cells were used for detecting CMIR. Peripheral blood lymphocytes were analysed for the numbers and proportions of B lymphocytes, T lymphocytes and their subpopulations with receptors for IgM (T micro) or IgG (T gamma) and cells with Fc receptors for IgG. These parameters were correlated with the duration and the severity of illness. The uncomplicated cases of typhoid fever were found to have an intact CMIR as compared to the complicated cases. The ratio of T lymphocyte subpopulations was grossly imbalanced in typhoid patients, the numbers of T lymphocytes and their subpopulations were further altered in he complicated cases as compared to uncomplicated cases. The present study demonstrates a depressed state of CMIR in complicated patients with typhoid fever. CMIR may thus emerge as the cardinal point for recovery in typhoid fever rather than the specific antibodies. The study further demonstrates that imbalance within the subsets of T lymphocytes may be responsible for the depressed state of CMIR in complicated cases of typhoid fever.     

妊娠子宫微环境中uNK细胞NKG2A和NKG2D及其配体表达的研究

目的：研究妊娠子宫微环境中子宫自然杀伤细胞（uNK细胞）NKG2A和NKG2D及其相应配体的表达，探讨NKG2A与NKG2D的不平衡表达在母胎免疫耐受形成中的作用。方法：选择30例孕6-9周的正常妊娠妇女,分离其新鲜蜕膜组织，除去绒毛,分离蜕膜和外周血单个核细胞，采用流式细胞仪测定NK细胞的数量及NKG2A与NKG2D的表达；采用RT-PCR技术检测滋养层组织NKG2A与NKG2D配体人类白细胞抗原-E(HLA-E)、主要组织相容性复合体-Ⅰ类分子相关蛋白A(MICA)mRNA的表达结果：妊娠子宫蜕膜淋巴细胞中NK细胞约占70%,流式细胞分析的结果显示，子宫自然杀伤细胞NKG2A的表达显著高于外周血NK细胞，分别为97.86%±1.75%与33.35%±10.92%（〖AKx-D〗±s），两者差异显著（P<0.05），在滋养层细胞中检测到其配体HLA-E的表达；而与外周血相比，uNK细胞表面NKG2D的表达与之较为相近，分别为93.21%±4.52%与97.80%±1.72%，但两者仍有显著差异（P<0.05）。在滋养层组织未检测到其相应配体MICA mRNA的表达结论：蜕膜中的淋巴细胞主要为NK细胞，其免疫学表型与外周血NK细胞有较大的区别，妊娠期子宫自然杀伤细胞表面高表达抑制性受体NKG2A，同时滋养层组织表达相应的配体人类白细胞抗原-E，这可能是维持母胎界面免疫耐受的重要因素。     

Sister chromatid exchanges, chromosome aberrations and micronuclei in female lymphocytes: correlations with biological rhythms, miscarriages and contraceptive pill use

Our study looked at the variation in peripheral blood lymphocytes, during the menstrual cycle, of frequencies of sister chromatid exchanges (SCE) and micronuclei (MN) in 819 women and cells with aberrant chromosomes (CA) in a selected sample of 136 volunteers. We observed significant fluctuations in SCE and CA frequencies: SCEs reached a maximum value at the end of menstruation and a low at the time of ovulation, whereas CAs showed a continuous increase from the beginning of the menstrual cycle up to the time of ovulation and a progressive decrease thereafter. MN frequency did not fluctuate in a statistically significant way. No statistically significant differences in SCE, CA and MN frequencies were observed when fertile women were compared with women taking the contraceptive pill or those in menopause and no difference was found between women who had undergone physiological or surgically induced menopause. Moreover, no difference was found between women with a history of miscarriages and matched controls. These data together suggest that the natural variations in sexual hormone levels, but not those due to the contraceptive pill or their reduction at menopause, can contribute in modulating the baseline frequencies of SCEs and CAs. Moreover, these data suggest that the increased risks either of producing a chromosome imbalance in the progeny (eliciting miscarriages) or of occurrence of gynaecological diseases is not predictable by evaluating cytogenetic end-points in peripheral blood lymphocytes.     

Loss of mucosal CD4 lymphocytes is an early feature of HIV infection.

T cell subsets in the gut mucosa are distinct populations and their imbalance in HIV has specific implications in infection. Alterations in T cell subsets in duodenal biopsies were investigated in 17 asymptomatic HIV patients, 24 AIDS patients and 10 controls with non-ulcer dyspepsia. Immunohistochemistry and immunofluorescence using MoAbs to CD3, CD4, CD8, CD68, CD45RA, CD45RO and gp120 were performed on frozen sections. In the lamina propria, there was a significant depletion of CD4+ cells at all stages of HIV, but the density of CD8 lamina propria cells was increased. Intraepithelial lymphocytes were decreased in AIDS patients. There was a significant correlation between cellular density and mucosal CD3+ lymphocytes, and between mucosal CD3+ and CD8+ lymphocytes. Although mucosal CD4,CD45RO+ 'memory' cells were decreased, CD8,CD45RO+ 'memory' cells were increased. Mucosal CD4+ lymphocyte depletion occurred early in HIV, and thus their role in mucosal protection against opportunistic infection should be revised. Mucosal CD8+ lymphocytes initially increased, but decreased when CD4 blood counts were depleted, perhaps contributing to loss of host protection against infection. Intraepithelial lymphocyte depletion may also contribute to opportunistic infection.     

Monoclonal B Lymphocytes in Multiple Myeloma

Individual specific antisera were raised against the monoclonal serum component from six IgG myeloma patients. Idiotypic (Id) determinants were identified in direct or indirect immunofluorescence (IFL) on 0.5-44% B lymphocytes from five patients with active disease. Id determinants were not detected on B lymphocytes from one IgG-k myeloma patient with an imbalance in the expression of upsilon heavy chains and kappa light chains. Nor were Id determinants detected on E-rosetting cells. Id structures were usually present together with upsilon chains on monoclonal B lymphocytes, some of which also carried mu and/or delta chains. Blood cells with lymphoplasmacytoid appearance were abundant in two patients. These cells carried IgG with Id determinants both in the cytoplasm and on the surface. Fc receptors were present on most cells carrying surface Id structures but were rare on cells with cytoplasmic monoclonal characteristics. The results further support the hypothesis of multiple myeloma as a differentiating B-lymphocyte tumour.     

Influence of ageing,heat shock treatment and in vivo total antioxidant status on gene-expression profile and protein synthesis in human peripheral lymphocytes

Ageing results in a progressive, intrinsic and generalised imbalance of the control of regulatory systems. A key manifestation of this complex biological process includes the attenuation of the universal stress response. Here we provide the first global assessment of the ageing process as it affects the heat shock response, utilising human peripheral lymphocytes and cDNA microarray analysis. The genomic approach employed in our preliminary study was supplemented with a proteomic approach. In addition, the current study correlates the in vivo total antioxidant status with the age-related differential gene expression as well as the translational kinetics of heat shock proteins (hsps). Most of the genes encoding stress response proteins on the 4224 element microarray used in this study were significantly elevated after heat shock treatment of lymphocytes obtained from both young and old individuals albeit to a greater extent in the young. Cell signaling and signal transduction genes as well as some oxidoreductases showed varied response. Results from translational kinetics of induction of major hsps, from 0 to 24 h recovery period were broadly consistent with the differential expression of HSC 70 and HSP 40 genes. Total antioxidant levels in plasma from old individuals were found to be significantly lower by comparison with young, in agreement with the widely acknowledged role of oxidant homeostasis in the ageing process.     

FcIgG receptor-bearing lymphocytes and monoclonal antibody-defined T cell subsets in atopic dermatitis: effect of treatment with thymopoietin pentapeptide (TP-5)

Lymphocyte subpopulations were determined in the blood of patients with atopic dermatitis (AD) before and after treatment with the thymopoietin pentapeptide TP-5. The relative and absolute numbers of lymphocytes bearing suppressor/cytotoxic cell markers (FcIgG+E+ and T8+ cells) were significantly decreased in the untreated patients and the T4+/T8+ cell ratio was increased, indicating an imbalance between lymphocyte subpopulations in AD. Patients who had been treated for 6 weeks with TP-5 displayed no significant abnormality of any of the lymphocyte subsets studied and comparison of pre- and posttreatment values revealed that there was a statistically significant increase in T8+ cell numbers, that by contrast did not take place in placebo-treated AD patients. The treatment had no demonstrable effect on IgE serum levels or on the spontaneous in vitro production of IgE by cultured lymphocytes from the patients.     

Impaired production of interleukins in patients with cell-mediated immunodeficiencies.

The poor mitogen response to phytohaemagglutinin (PHA) of lymphocytes from three patients with cell-mediated immunodeficiencies was restored to normal when supernatants containing interleukin 2 (IL-2) were added. One of three children with severe combined immunodeficiency also showed a partial response. There was no improvement in the normal mitogenic response of the lymphocytes from patients with either the X linked or common variable forms of hypogammaglobulinaemia. All three patients with cell-mediated immunodeficiencies showed gross imbalance in the ratio of helper/inducer (OKT4+) to suppressor/cytotoxic (OKT8+) T cells. The PHA stimulated culture supernatant from one of these patients failed to induce proliferation of a cytolytic continuous T cell line. Our data suggests that the underlying defect in these patients may be a failure in production of interleukins but not in the acquisition of IL-2 receptors.     

血小板相关抗体与原发性血小板减少性紫癜关系的研究进展

原发性血小板减少性紫癜 (idiopathicthrombocy topenicpurpura ,ITP)是一种自身免疫性血小板减少症 ,血小板减少的机制是由于自身免疫功能异常导致血液中产生血小板相关抗体 ,使循环中血小板破坏增加。ITP是由血小板自身抗体所引起的综合征 ,为临床常见的出血性疾病 ,其主要病理是血小板抗体的产生并吸附于血小板表面后加速血小板从循环中的清除 ,这些抗体作用的血小板膜靶抗原可多种多样 ,其中血小板膜上糖蛋白 (glycoprotein ,GP)Ⅱb/Ⅲa是一重要靶抗原 ,少数针对GPⅠb/ⅠX复合物。由于抗体直接作用于GPⅡb/Ⅲa、GPⅠb/ⅠX等 ,使…     

Impairments in functional subsets of T-suppressor (CD8) lymphocytes, monocytes, and natural killer cells among asbestos-exposed workers

Peripheral blood leukocytes from asbestos-exposed workers were analyzed by dual color flow cytometry using monoclonal antibodies that identify developmental (HLA-DR) and functional (Leu 8) subsets of T helper, suppressor lymphocytes, and monocytes. An increase in the number of T suppressor cells was closely associated with a decrease in T lymphocyte functions while numerical defects in activated monocytes (Leu M3+Ia+) and natural killer cells (Leu 7+) were correlated with a depressed Th/Ts ratio. Furthermore, among asbestos-exposed workers with depressed T cell functions we have demonstrated a significantly higher number of the effector Ts (Leu 2+ Leu 8-) subset which regulates both the Th/Ts lymphocyte system as well as B cells and NK cell activities. These findings identified changes in the T suppressor feedback regulatory loop as being responsible for the immunoregulatory imbalance among long-term asbestos workers. In double blind analyses of demographic and radiographic data these phenotypic changes were not correlated with age, smoking history, or duration of exposure but were associated with radiographic evidence of asbestos-associated effects. This correlation established a direct link between asbestos exposure and the subsequent development of immune dysfunction.