宫颈癌患者外周血T淋巴细胞及NK细胞的检测及临床意义

目的 探讨宫颈癌患者外周血T淋巴细胞和NK细胞的活性及临床意义.方法 采用流式细胞仪测定45乳腺癌患者外周血T淋巴细胞和NK细胞的活性,以20例正常人同期监测结果作对照.结果 宫颈癌患者外周血CD+3,CD+4,NK细胞数量及CD+4/CD+8比值较正常对照组均明显下降(P＜0.05),而CD+8细胞水平显著升高.外周血T淋巴细胞亚群和NK细胞数量的改变与宫颈癌临床病理指标有关,分期越晚,CD+3细胞、CD+4细胞、CD+4/CD+8细胞比值及NK细胞数量越低,CD+8细胞水平越高;I、II期宫颈癌患者与III、IV期患者之间有显著差异(P＜0.05).结论 宫颈癌患者细胞免疫功能低下,且临床病理分期越晚,其免疫功能越低,检测T淋巴细胞亚群、NK细胞可用于宫颈癌患者的免疫监测.     

妇科恶性肿瘤患者细胞免疫功能状态分析

目的探讨妇科恶性肿瘤患者细胞免疫功能状况及其临床意义。方法采用流式细胞术技术检测108例妇科恶性肿瘤患者及17例健康正常人的外周血淋巴细胞亚群,并作比较分析。结果与健康正常人比较,妇科恶性肿瘤患者CD4+T淋巴细胞百分率、NK细胞百分率、CD4+/CD8+比值明显升高(P<0.05),CD3+、CD8+T淋巴细胞百分率明显降低(P<0.05),CD19+(B淋巴细胞)在两组间比较的差异无统计学意义(P>0.05)。结论妇科恶性肿瘤患者存在免疫功能紊乱,外周血T淋巴细胞亚群和NK细胞检测可作为细胞免疫功能的指标之一。     

胃癌患者围手术期外周血T细胞亚群及NK细胞水平变化的比较分析

目的探讨胃癌患者围手术期外周血T淋巴细胞亚群及NK细胞表达的临床意义。方法采用流式细胞仪检测69例胃癌患者术前及术后7天及35例健康者外周血T淋巴细胞亚群及NK细胞表达水平。结果健康者与胃癌患者对比、胃癌患者术后7天与术前对比、Ⅰ~Ⅱ期与Ⅲ~Ⅳ期胃癌患者对比,其外周血CD3+、CD4+、CD4+/CD8+及NK细胞表达水平较高,CD8+较低,差异均有统计学意义(P<0.05)。中等分化癌组与低等分化癌组比较无显著变化(P>0.05)。结论胃癌患者免疫功能较低,术后1周有所提高;病理分期越晚免疫功能越差,肿瘤分化程度与机体免疫功能关系不明显。     

生津增效汤对食管癌放疗患者免疫功能及外周血细胞凋亡影响的研究

目的观察中药生津增效汤对食管癌患者放疗前后细胞免疫功能及外周血细胞凋亡的影响。方法将154例食管鳞癌患者随机分为放疗组和治疗组,治疗前后分别检测各组外周血自然杀伤(NK)细胞活性、B淋巴细胞、CD3+、CD4+/CD8+水平和细胞凋亡情况。结果放疗同时口服生津增效汤的患者CD3+、NK细胞、B淋巴细胞、CD4+/CD8+值明显高于治疗前的水平(P<0.05),细胞凋亡率明显下降(P<0.05)。结论生津增效汤可以明显改善食管癌患者放疗后的细胞免疫功能状态,减少外周血细胞凋亡。     

宫颈癌患者外周血T淋巴细胞及NK细胞的检测及临床意义

目的 探讨宫颈癌患者外周血T淋巴细胞和NK细胞活性的临床意义.方法 采用流式细胞仪测定宫颈癌患者外周血T淋巴细胞亚群和NK细胞的活性,并与正常人作对照.结果 宫颈癌患者外周血CD+3,CD+3+,NK细胞数量及CD+4/CD+8比值较正常对照组均明显下降(P<0.05),而CD+8细胞水平显著升高.外周血T淋巴细胞亚群和NK细胞数量的改变与宫颈癌临床病理指标有关,分期越晚,CD+3细胞、CD+3细胞、CD+3/CD+8细胞比值及NK细胞数量越低,CD+8细胞水平越高.结论 宫颈癌患者细胞免疫功能降低,导致肿瘤的发生、发展,应用流式细胞仪检测外周血T淋巴细胞和NK细胞等指标可用于宫颈癌患者的免疫状态监测,对宫颈癌患者的免疫辅助治疗提供依据.     

宫颈癌患者外周血T淋巴细胞及NK细胞检测的临床意义

目的探讨宫颈癌患者外周血T淋巴细胞及NK细胞的检测的临床意义。方法 2009年1月至2011年1月收治的宫颈癌患者214例,选取同期进行健康体检200例(体检组),进行外周血T淋巴细胞及NK细胞检测并比较。结果宫颈癌组与体检组比较P<0.01差异有统计学意义。宫颈癌组Ⅰ期、Ⅲ期与Ⅲ期、IV期比较P<0.05差异有统计学意义。结论外周血T淋巴细胞及NK细胞的检测对宫颈癌患者恶性程度、患者的预后有着良好的指导作用,外周血细胞免疫功能越低下,肿瘤分级越严重。     

急性乙型肝炎外周血淋巴细胞亚群动态分析

目的 探讨急性乙型肝炎(AHB)患者外周血免疫细胞包括T淋巴细胞亚群、B淋巴细胞和自然杀伤(NK)细胞在疾病发展中的动态变化及其意义.方法 流式细胞技术检测17例AHB患者(AHB组)外周血淋巴细胞亚群动态变化和36例慢性乙型肝炎(CHB组)患者及32例正常对照组淋巴细胞亚群分布特点;动态检测AHB患者CD4~+/CD8~+变化,并探讨其与ALT改变及与HBVDNA清除的相关性.结果 AHB组人院4周内外周血CD3~+、CD4~+和CD8~+T细胞频率显著高于CHB组和对照组,NK细胞4周内均低于其余2组(P<0.05).AHB疾病早期ALT高水平异常时CD4~+/CD8~+明显低下;随着ALT恢复正常,CD4~+/CD8~+比值呈逐渐升高趋势.结论 AHB患者外周血免疫细胞的分布与对照组和CHB患者不同,淋巴细胞亚群的动态变化与疾病发展可能有一定的相关性.     

骨髓增生异常综合征患者T淋巴细胞亚群的特征分析

目的:检测和分析骨髓增生异常综合征(M DS)患者T淋巴细胞亚群的分布,评估M DS患者免疫功能状态。方法:采用FC 500流式细胞仪对35例M DS患者和10例正常对照组外周血标本进行T淋巴细胞亚群检测。结果:M DS组的辅助性细胞T细胞(T h),辅助性细胞T细胞/抑制性T细胞(T h/T s)和自然杀伤细胞(NK)的表达均低于正常对照组,T s细胞的表达高于正常对照组;随着M DS疾病的进展,抑制性T细胞(T s)的表达是逐渐增加的,其中只有RA/RA S组与RAEB组之间差异有显著性(P=0.027);M DS患者各组间T h,T h/T s比值和NK细胞的表达均无差异性。结论:T h,T s和NK细胞参与人体重要免疫功能,T淋巴细胞亚群免疫失调,则导致细胞免疫功能紊乱,T h,T s和T h/T s比值成为监测人体免疫功能、反映机体免疫状态的重要指标。     

系统性红斑狼疮患者外周血CD40-CD40L测定的临床意义

目的探讨系统性红斑狼疮(SLE)患者在疾病的不同阶段外周血单个核细胞(PBMC)CD40-CD40L、活化淋巴细胞亚群的表达水平。方法采用流式细胞术检测SLE患者不同时期淋巴细胞表达的CD40-CD40L的百分率,同时对淋巴细胞的表型及HLA-DR进行测定。结果活动期SLE患者淋巴细胞表达的CD40、CD40L均明显高于正常对照组(P<0.05);活动期SLE患者表达的HLA-DR、CD3+HLA-DR+细胞、CD4+HLA-DR+细胞、CD8+HLA-DR+细胞、CD8+细胞、CD19+细胞均显著高于正常对照组(P<0.05),而缓解期与正常对照组之间,差异均无统计学意义(P>0.05);活动期SLE患者表达NK细胞明显比正常对照组低(P<0.05);活动期SLE患者表达的CD3+、CD4+与正常对照组相比,虽有降低,但差异无统计学意义(P>0.05)。相关分析表明,SLE患者(活动期和缓解期)淋巴细胞表达CD40-CD40L的水平与淋巴细胞活化程度呈正相关。结论活动期SLE患者外周血PBMC存在CD40与CD40L的异常表达,以及活动期SLE患者淋巴细胞的异常活化,这些免疫细胞异常在SLE的发病中起到重要的作用,并且异常程度与疾病活动度相关。     

急性髓性白血病患者外周血淋巴细胞亚群的检测及其临床意义

目的探讨急性髓系白血病(AML)患者外周血T淋巴细胞亚群、DNT细胞、NK细胞的表达水平及其临床意义。方法采用流式细胞术对40例初发AML患者、32例AML完全缓解(CR)患者及20例正常人外周血T淋巴细胞亚群、DNT细胞及NK细胞水平进行检测。结果初发AML组与正常对照组比较,CD4+细胞、CD4+/CD8+比值、DNT细胞及NK细胞明显降低(P<0.05);CD3+细胞、CD8+细胞变化不显著(P>0.05)。复治CR组与对照组比较,NK细胞明显降低(P<0.05),其他各值差异无统计学意义。结论初发AML患者的T淋巴细胞亚群、DNT细胞、NK细胞变化明显,而治疗缓解后淋巴亚群基本恢复正常,说明T淋巴细胞亚群、DNT细胞、NK细胞水平检测在评价AML疗效及判断预后方面具有一定的临床价值。     

布鲁氏菌病患者外周血树突状细胞、Th1／Th2细胞因子含量的检测及意义

目的：观察布鲁氏菌病患者外周血树突状细胞表型、Th1／Th2细胞含量的检测及意义。方法选取诊治的布鲁氏菌病患者50例为病例组,另选取同期进行健康体检的正常人50例作为正常组。采用Real time-PCR测定2组Th1相关转录因子T细胞表达的T盒（T-bet）、GATA连接蛋白3（GATA-3）、维A酸相关核孤儿受体γt（RORγt）、叉头蛋白3（Foxp3）及Th1／Th2细胞因子肿瘤坏死因子α（TNF-α）、干扰素γ（IFN-γ）、白介素6（IL-6）、白介素10（IL-10）mRNA含量。酶联免疫吸附实验（ELISA）测定TNF-α、IFN-γ、IL-6、IL-10蛋白表达及补体C3、C4含量。流式细胞术测定树突状细胞表型、Th1、Th2及T淋巴细胞亚群（ CD4＋T细胞、CD8＋T细胞、NK细胞）含量。结果病例组外周血Th1细胞相关转录因子T-bet、RORγt、Foxp3及Th1细胞、Th1／Th2、TNF-α、IFN-γ含量较对照组显著降低,Th2细胞及IL-6、IL-10含量较对照组显著升高,差异有统计学意义（ P ＜0．05）;病例组CD8＋3、CD8＋0、CD8＋6阳性的树突状细胞比例、CD4＋T细胞、NK细胞及补体C3、C4含量较对照组显著降低,CD8＋T细胞含量较对照组显著升高,差异有统计学意义（ P ＜0．05）;TNF-α、IFN-γ含量与CD4＋T细胞、NK细胞、C3、C4含量呈正相关性（ r值分别为2．879、3．214、3．255和2．978, P ＜0．05）,与CD8＋T细胞含量呈负相关性（ r值分别为－3．146和－3．011, P ＜0．05）。 IL-6、IL-10含量与CD4＋T细胞、NK细胞、C3、C4含量呈负相关性（ r值分别为－2．124、－2．343、－3．423、－2．789、－2．993、－2．566、－3．758, P ＜0．05）,与CD8＋T细胞含量呈正相关性（ r值分别为3．465、3．129, P ＜0．05）。结论布鲁氏菌病患者外周血成熟树突状细胞数目减少,同时Th1／Th2细胞及相关细胞因子失衡,且与机体天然免疫和细胞免疫功能降低有关,这可能在布鲁氏菌病发生发展过程中发挥重要作用。     

Natural killer cell activity and serum immunoglobulins in epileptic patients

Serum immunoglobulins and the activity of natural killer (NK) cells of 50 epileptic patients (eight with idiopathic generalized epilepsy and 42 with cryptogenic partial epilepsy) and 28 controls have been studied. The values of IgA, IgG and IgM were the same-in patients and controls. The NK activity in controls was linearly related to the effector-to-target ratio, but this linear relationship was not observed in epileptic patients. The cytotoxic activity of NK cells at the lowest effector-to-target ratio was significantly greater in patients than in controls. This increase was observed in each therapy group. Our results seem to confirm a disturbance of the immune system in epileptic patients and suggest that this modification of cellular immunity is not a drug effect but is related to the illness itself.     

Safrole-modulated immune response is mediated through enhancing the CD11b surface marker and stimulating the phagocytosis by macrophages in BALB/c mice

Safrole, a component of piper betle inflorescence, is a documented rodent hepatocarcinogen and inhibits bactericidal activity and the release of superoxide anion (O(2-)) by polymorphonuclear leukocytes (PMNs). In the present study, we investigated the effects of safrole on immune responses, including natural killer (NK) cell cytotoxicity, phagocytic activity and population distribution of leukocytes from normal BALB/c mice. The cells population (cell surface markers) and phagocytosis by macrophages and monocytes from the peripheral blood mononuclear cells (PBMCs) were determined, and NK cell cytotoxicity from splenocytes of mice after oral treatment with safrole was performed using flow cytometric assay. Results indicated that safrole did not affect the weights of body, spleen and liver when compared with the normal mice group. Safrole also promoted the levels of CD11b (monocytes) and Mac-3 (macrophages) that might be the reason for promoting the activity of phagocytosis. However, safrole reduced the cell population such as CD3 (T cells) and CD19 (B cells) of safrole-treated normal mice by oral administration. Furthermore, safrole elevated the uptake of Escherichia coli-labelled fluorescein isothiocyanate (FITC) by macrophages from blood and significantly stimulated the NK cell cytotoxicity in normal mice in vivo. In conclusions, alterations of the cell population (the increase in monocytes and macrophages, respectively) in safrole-treated normal BALB/c mice might indirectly influence the immune responses in vivo.     

Adenosine kinase, epilepsy and stroke: mechanisms and therapies

Adenosine is an inhibitory modulator of brain activity with neuroprotective and anticonvulsant properties. Adenosine levels are regulated mainly by adenosine kinase (ADK), an enzyme that is responsible for the removal of adenosine via phosphorylation to AMP. Recent evidence indicates that expression of ADK undergoes rapid coordinated changes during brain development and following brain injury, such as after epileptic seizures and stroke. Thus, transient downregulation of ADK after acute brain injury protects the brain from seizures and cell death. Conversely, chronic overexpression of ADK causes seizures in epilepsy and promotes cell death in epilepsy and stroke. These findings have direct implications for the rational definition of ADK as a therapeutic target. In recent years, novel treatment strategies have been developed that make use of the intracerebral transplantation of cells that are ADK deficient and, thus, release adenosine. A new era of cell-based delivery of adenosine has begun, which holds great promise for novel therapies for epilepsy and stroke.     

儿童系统性红斑狼疮患者外周血淋巴细胞亚群与疾病活动的研究

目的探讨儿童系统性红斑狼疮(SLE)患儿外周血中淋巴细胞亚群的变化及临床意义。方法采用流式细胞术检测59例SLE患儿及52名健康对照组患儿的外周血淋巴细胞亚群T细胞、B细胞、Th细胞、Tc细胞、NK细胞和总淋巴细胞数,并对结果进行比较,同时分析SLE患儿外周血淋巴细胞亚群与SLE疾病活动指数(SLEDAI)的相关性及其与狼疮肾炎、补体的相关关系。结果与健康对照组相比,SLE患儿Th细胞和Th百分比、NK细胞和NK细胞百分比及Th/Tc比值明显降低,而B细胞百分比、Tc细胞百分比增高,差异均有统计学意义(P<0.05);SLE患儿B细胞与SLEDAI评分呈正相关,NK细胞百分比与SLE评分呈负相关(P<0.05);狼疮肾炎组患儿的T细胞、Tc细胞计数和Tc细胞百分比均明显高于无狼疮肾炎的患儿,而狼疮肾炎组患儿Th细胞百分比、Th/Tc比值和NK细胞、NK细胞百分比明显低于无狼疮肾炎的患儿(P<0.05);补体降低组患儿的B细胞和B细胞百分比及SLEDAI评分均明显高于补体正常组(P<0.05)。结论 SLE患儿外周血淋巴细胞亚群存在异常,淋巴细胞亚群与SLEDAI评分、狼疮肾炎和补体之间存在显著的相关性,表明淋巴细胞亚群可能反映SLE患儿病情的严重程度。     

Differential effects of female sex hormones on cellular recruitment and tracheal reactivity after formaldehyde exposure

Female sex hormones (FSHs) exert profound regulatory effects on the course of lung inflammation due to allergic and non-allergic immune responses. As pollution is one of the pivotal factors to induce lung dysfunction, in this study we investigated the modulatory role of FSHs on lung inflammation after a formaldehyde (FA) exposure. For this purpose, lung and systemic inflammatory responses were evaluated in terms of leukocytes countings in bronchoalveolar lavage (BAL), peripheral blood and bone marrow lavage from 7-day ovariectomized (OVx) and Sham-OVx rats subjected to FA inhalation for 3 consecutive days. The hypothesized link between effects of FSHs on expression of adhesion molecules and mast cells degranulation was also studied. Once exposed to FA, Sham-OVx rats increased the number of total cells recovered in BAL and of leukocytes in peripheral blood, and decreased the counts in bone marrow. By contrast, in OVx rats upon FA exposure there was a reduction of the total cells counts in BAL and of blood leukocytes; lung expressions of ICAM-1 and Mac-1 were depressed, but the number of bone marrow cells did not vary. Estradiol treatment of OVx rats increased the total cells in BAL and decreased the number of blood leukocytes, whereas the number of bone marrow cell remained unaltered. Progesterone treatment, in turn increased the total cells in BAL and blood leukocytes, but decreased the number of bone marrow cells. OVx rats exposed to FA developed tracheal hyperresponsiveness to methacholine (MCh). A similarly altered response was found between the tracheal segments of Sham-OVx rats after FA exposure and that found in tracheae of naïve rats. Estradiol treatment prevented FA-induced tracheal hyperresponsiveness to MCh whereas progesterone was ineffective in this regard. In addition, OVx rats upon FA exposure significantly increased both, the ability of mast cell degranulation and serum corticosterone levels. In conclusion, it was found that FSHs act by distinct control mechanisms on FA-induced lung inflammation and tracheal hyperresponsiveness, since at low circulating levels of FSHs (such as those after OVx) there is some resistance to the development of a lung inflammatory response, but the cholinergic tracheal responsiveness is exacerbated. Our data also help to understand the involvement of FSHs on mast cells activity after pollutants exposure and add information regarding the role of FSHs on the mechanisms related to endothelium-leukocyte interactions.     

Neuronal MDR-1 gene expression and persistent low levels of anticonvulsants in a child with refractory epilepsy

SUMMARY: It is estimated that 20-25% of epileptic patients fail to achieve good control with antiepileptic drug (AED) treatment; thus, refractory epilepsy (RE) has been described in patients who have adequate therapeutic levels of AEDs without control of seizures. Multidrug resistance genes have been reported to be highly expressed in brain of patients with RE. Persistent low plasma levels of AEDs and high brain expression of the multidrug resistance product P-glycoprotein (P-gp) have been previously communicated in a case report of RE secondary to tuberous sclerosis. Here, the authors report a case of an 8-year-old boy diagnosed with partial RE with focal seizures who was admitted to hospital for a severe episode of subintrant crisis. The patient received polytherapy with carbamazepine (CBZ), phenytoin (PHT), and valproic acid (VA); however, habitual doses of these AEDs failed to control the patient's symptoms. AED blood levels were monitored for 25 consecutive days and showed low values in 8/25 (33%) for CBZ, 10/25 (40%) for PHT, and 25/25 (100%) for VA of samples studied. Because the patient developed focal status epilepticus, surgical treatment by callosotomy was done, resulting in a significant improvement in epileptic symptoms. The immunostaining of brain specimens showed significantly increased expression of P-gp not only in vascular endothelial cells and related astrocytes but also in neurons. Overexpression of P-gp in the brain does not explain the low blood levels of AEDs described in these cases. Different mechanisms such as drug-drug interactions and drug transporters can be involved in the results observed. The P-gp overexpression and/or its pharmacologic induction should be considered as a potential mechanism responsible for drug resistance to epilepsy treatment and highly suspected in patients with persistent subtherapeutic AEDs plasma levels.     

Contribution of plasma cortisol to corticosteroid-suppressed peripheral blood natural killer cell activity in Crohn's disease

We recently showed that patients with active ileocecal Crohn's disease (CD) have a temporarily suppressed peripheral blood natural killer (NK) cell activity during treatment with oral budesonide or prednisolone. This suppression was caused by a decrease in the number of CD16⁺ NK cells in the circulation. In the present study we evaluated the contribution of cortisol in plasma to this suppressed NK cell activity. The CD patients took part in a controlled study where they received either oral budesonide or prednisolone for 10 weeks. Before treatment, and at 4 and 10 weeks of treatment, peripheral blood NK cell activity, numbers of circulating CD 16⁺ NK cells, and plasma cortisol levels were analysed. These parameters were determined both before and 30 min after administration of adrenocorticotropic hormone (ACTH). The ACTH-induced plasma cortisol increase was accompanied by a stimulated NK cell activity, when both are suppressed by corticosteroid treatment, without changing the number of CD16⁺ NK cells. Therefore, a low plasma cortisol level contributes to the corticosteroid mediated NK cell suppression in active ileocecal CD.     

Multidrug transport in human autoimmune T line cells and peripheral blood lymphocytes.

We studied multidrug transport in human autoimmune T line cells and peripheral blood leukocytes, because multidrug transport is pleiotropically limiting the intracellular accumulation of immunosuppressive and chemotherapeutic agents. We observed that a subpopulation of peripheral blood leukocytes containing CD4+, CD8+ and Ig+ lymphocytes expressed a multidrug transport system. Lymphocytic multidrug transport was seen with all peripheral blood samples analyzed, but showed considerable variations between individuals. In further studies with human lymphocytic cell lines multidrug transport was seen with 18/23 human CD4+ T cell lines. Interestingly, expression of multidrug transport was independent of T cell activation. No significant transport was observed with EBV-transformed human B lymphocytes, rat T line cells or rat, mouse, or guinea pig leukocytes.