多发性硬化症发作期外周血和脑脊液淋巴细胞亚群的分析

为分析多发性硬化症 (MS )患者发作期淋巴细胞亚群及给予甲基强的松龙 (MP )治疗后的变化 ,流式细胞仪测定 2 6例处于复发期MS患者外周血 (PB )和脑脊液 (CSF )及 8例MS患者予MP治疗后PB淋巴细胞CD3+ 、CD4 + 、CD8+ 、CD4 5RA+ 、CD4 + /CD4 5RA+ 、CD4 + /CD2 9+ 、CD19+ 、CD5 + /CD19+ 的百分率。结果发现MS患者PB中CD8+ 、CD4 5RA+ 和CD4 + /CD4 5RA+ 百分率降低 ,CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值升高 ;CSF中CD3+ 、CD4 + 、CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值高于PB ;淋巴细胞亚群与临床伤残程度和距此次发作的时间无关 ;MP治疗不影响PB淋巴细胞亚群变化。表明MS患者淋巴细胞通过血脑屏障有选择性 ,淋巴细胞亚群的变化在MS发病机制中起作用     

多发性硬化患者外周血淋巴细胞CD56的表达

目的 探讨CD56^ 淋巴细胞在多发性硬化（MS）的变化。方法 流式细胞仪测定32例复发缓解型MS患者（复发发期23例,缓解期9例）及13例复发期MS患者予糖皮质激素治疗后外周血淋巴细胞CD56表达的阳性百分率。结果 复发期和缓解期MS患者CD56的表达均高于对照组,15例复发期MS患者CD56的阳性百分率与血脑屏障受损呈正相关,复发期MS患者CD56的水平与距发作时间、整个病程、EDSS伤残评分无关。缓解期MS CD56的水平与病程无关,激素对CD56的表达我影响。结论 CD56^ 淋巴细胞涉及MS的发病机制。     

多发性硬化患者外周血白细胞粘附分子的表达

目的 :探讨多发性硬化 (MS)患者外周血白细胞粘附分子表达的水平及予甲基强的松龙 (MP)治疗后的变化。方法 :流式细胞仪测定 2 8例缓解复发型MS患者及 12例复发期MS予静脉MP治疗后外围血淋巴细胞和单核细胞细胞间粘附分子 3(CD5 0 )、细胞间粘附分子 1(CD5 4 )、整合素LFA 1β亚单位 (CD18)、非常晚抗原 4 (VLA 4 )α、β亚单位 (CD4 9d、CD2 9)和 (- )选择素 (CD6 2L)的阳性百分率。结果 :复发期MSCD4 9d和CD2 9在淋巴细胞和单核细胞、CD5 4和CD6 2L在单核细胞的阳性百分率高于缓解期MS和对照组 ,复发和缓解期MSCD5 4在淋巴细胞的阳性百分率高于对照组 ;MP治疗后 ,CD5 4和CD4 9d在淋巴细胞和单核细胞、CD6 2L在单核细胞的阳性百分率下降。结论 :MS患者外周血白细胞粘附分子的表达升高并可作为MS活动期的指标     

CD28在多发性硬化患者CD8^＋淋巴细胞的表达

目的：探讨CD28在多发性硬化（MS）患者CD8^ 淋巴细胞的表达水平。方法：流式细胞仪测定16例复发期MS患者和20例对照组外周血淋巴细胞CD28^ 、 CD8^ 、CD28^-和CD8^ CD28^ 的百分率.结果：复发期MS患者淋巴细胞CD8^ CD28^－百分率低于对照组，CD28^ 和CD8^ CD28^ 的百分率与对照组无明显差异；甲基强的松龙治疗对CD28^ 、CD8^ 、CD28^ 和CD8^ CD28^ 的百分率无影响。 结论：参与MS的发病的CD8细胞是CD8^ CD28^-细胞。     

CD28在多发性硬化患者CD8+淋巴细胞的表达

目的探讨CD28在多发性硬化(MS)患者CD8+淋巴细胞的表达水平.方法流式细胞仪测定16例复发期MS患者和20例对照组外周血淋巴细胞CD28+、CD8+CD28-和CD8+CD28+的百分率.结果复发期MS患者淋巴细胞CD8+CD28-百分率低于对照组,CD28+和CD8+CD28+的百分率与对照组无明显差异;甲基强的松龙治疗对CD28+、CD8+CD28-和CD8+CD28+的百分率无影响.结论参与MS的发病的CD8细胞是CD8+CD28-细胞.     

多发性硬化患者淋巴细胞亚群的研究

目的 观察多发性硬化患者外周血及脑脊液中淋巴细胞亚群的水平。方法 用碱性磷酸酶抗磷酸酶法检测了56例多发性硬化（MS）活动期患者外周血和脑脊液（CSF）的淋巴细胞亚群。结果：活动期MS患者外周血CD4^ 、CD8^ 细胞较对照组减少，CD25^ 细胞、CD4^ /CD8^ 比值较对照组升高（P＜0．05）。CSF中CD4^ 、CD25^ 细胞、CD4^ ／CD8^ 比值较对照组升高，CD8^ 细胞降低（P＜0．05＜0．05），且CSF中淋巴细胞亚群均高于自身外周血中的相应细胞（P＜0．05）。经肾上腺皮质类固醇激素治疗后，随病情缓解，外周血、CSF中的淋巴细胞亚群均有不同程度的改善。结论 淋巴细胞亚群可能参与MS的发病，并与MS的缓解－复发有关。     

多发性硬化患者淋巴细胞亚群的研究

目的观察多发性硬化患者外周血及脑脊液中淋巴细胞亚群的水平. 方法用碱性磷酸酶抗磷酸酶法检测了56例多发性硬化(MS)活动期患者外周血和脑脊液(CSF)的淋巴细胞亚群.结果:活动期MS患者外周血CD4+、CD8+细胞较对照组减少,CD25+细胞、CD4+/CD8+比值较对照组升高(p<0.05).CSF中CD4+、CD25+细胞、CD4+/CD8+比值较对照组升高,CD8+细胞降低(p<0.05<0.05),且CSF中淋巴细胞亚群均高于自身外周血中的相应细胞(p<0.05).经肾上腺皮质类固醇激素治疗后,随病情缓解,外周血、CSF中的淋巴细胞亚群均有不同程度的改善. 结论淋巴细胞亚群可能参与MS的发病,并与MS的缓解-复发有关.     

狼疮性肾炎患者外周血淋巴细胞亚群及活化状态的研究

目的探讨狼疮性肾炎(LN)患者淋巴细胞亚群的变化及所处的不同状态在LN发病机制中的作用。方法采用双标染色技术结合流式细胞术,对32例LN进行研究。结果①LN患者CD3＋CD4＋细胞的百分率明显下降,CD3＋CD8＋细胞的百分率明显增高,CD3＋CD4＋/CD3＋CD8＋的比例∨1(与对照组相比较,P∨0.05)。②LN患者CD45RA＋细胞、CD4＋CD45RA＋细胞、CD4＋CD45RO＋细胞和CD4＋HLA-DR＋细胞的百分率下降(与对照组相比较,P∨0.05或P∨0.001), HLA-DR＋细胞、CD3＋HLA-DR＋细胞、CD19＋HLA-DR＋细胞和CD19＋CD43＋细胞的百分率明显增高(与对照组相比较,P∨0.05或P∨0.01)。结论LN患者免疫功能的紊乱与淋巴细胞活化状态及其亚群之间的平衡失调密切相关。     

老年哮喘患者外周血淋巴细胞内IFNγ与IL4的变化

目的了解老年哮喘患者在不同病期外周血淋巴细胞表达IFN-γ、IL-4的变化.方法应用流式细胞仪,测定哮喘急性发作期、缓解期和健康老年人外周血淋巴细胞表达IFN-γ、IL-4的百分率.结果 CD3+CD8+淋巴细胞IFN-γ表达率,急性发作期组(10.5±4.6%)低于缓解期组(17.5±3.8%,p<0.05)和对照组(18.4±5.9%,p<0.05);IL-4表达率急性发作期组(2.6±2.2%)高于缓解期组(1.3±0.9%,p<0.05)和对照组(1.5±0.8%,p<0.05);CD3+、CD8+淋巴细胞IFN-γ表达率,急性发作期组(31.4±10.3%)显著高于缓解期组(20.2±12.3%,p<0.05)和对照组(21.3±10.4%,p<0.05).结论向Th2偏移的Th1/Th2失衡,可能与老年哮喘患者病情发展有关.     

类风湿关节炎患者关节滑膜液淋巴细胞活化标志的表达及意义

目的:研究类风湿关节炎(rheumatoidarthritis,RA)患者关节滑膜液淋巴细胞活化标志CD69、CD25、CD71、HLA-DR的表达,探讨其在RA致病中的作用。方法:采用流式细胞仪检测RA患者关节滑膜液淋巴细胞活化标志CD69、CD25、CD71、HLA-DR的表达,并设立正常对照组。结果:RA患者关节滑膜液表达CD25和CD71的淋巴细胞与正常对照组相比明显增多(P<0.01);而表达CD69和HLA-DR的淋巴细胞,与正常对照组相比差异无显著性(P>0.05)。结论:关节滑膜液淋巴细胞活化标志的表达增高,在RA发病机制中起着重要的作用。     

CD4+, CD8+, and CD4− CD8− T Cells in CSF and Blood of Patients with Multiple Sclerosis and Tension Headache

Two-colour flow cytometric analysis was performed on paired samples of peripheral blood (PB) and cerebrospinal fluid (CSF) of patients with untreated multiple sclerosis (MS) and, for reference, subjects with muscular tension headache (TH) using anti-CD3, anti-CD4, anti-CD8, and anti-HLA-DR monoclonal antibodies in different combinations. CD4+/CD8+ T-cell ratio was increased in CSF compared to PB in both MS patients and TH subjects to a similar extent. This was mainly due to higher CD4+ T-cell levels in the CSF compartment. The proportion of HLA-DR+ T cells was higher in CSF than PB in both MS and TH; this increase of DR+ T cells in CSF was more prominent in MS. The level of CD4+ CD8+ T cells, which represent a subset of activated T cells, was not different between CSF and PB, either in MS or in TH. The proportion of CD4- CD8- T cells, which were found generally not to be blast cells, was lower in CSF compared to PB in both patient groups. However, their CSF level was higher and their PB level lower in MS compared to TH. Results point to an accumulation of activated T-helper cells in the CSF of both MS patients and healthy subjects. Fetal-type CD4- CD8- T cells bearing the unusual T-cell receptor gamma/delta seem to be selectively recruited to the CSF of MS patients.     

Lymphocyte subsets and activation markers in patients with acute episodes of idiopathic anaphylaxis.

BACKGROUND: Idiopathic anaphylaxis (IA), a type of anaphylaxis in which no external allergen can be identified, is a corticosteroid-responsive disease, that suggests that it may have an immunologic pathogenesis. OBJECTIVE: The objective of this study is to compare patients with acute episodes of IA with normals, patients with chronic idiopathic urticaria, and patients with IA in remission relative to lymphocyte subsets and activation markers. METHODS: This is a prospective cohort study of 38 adults: 5 normals, 4 idiopathic urticaria, 11 IA patients in remission, 9 IA patients with acute attacks who had not yet received prednisone, and 9 IA patients who had received prednisone. The main outcome measures were lymphocyte subset and activation markers determined by two and three color flow cytometry (CD2, CD3, CD4, CD5, CD8, CD16, CD19, CD23, CD25, CD56, and HLA-DR). RESULTS: Comparing patients with acute IA with those in remission, the only significant difference was that the acute IA patients had a significantly higher percentage of CD3+HLA-DR+ cells. Normals had a significantly lower percentage of CD3+ HLA-DR+ cells than all other groups. Patients with acute IA on prednisone as well as IA patients in remission had a significantly higher percentage of CD 19+ CD23+ cells than normals. CONCLUSIONS: These results suggest that there are more activated T cells in patients with acute episodes of IA than in patients in remission. Perhaps, these activated T cells have a role in the pathogenesis of IA.     

Glucocorticoid treatment reduces T-bet and pSTAT1 expression in mononuclear cells from relapsing remitting multiple sclerosis patients

High dose glucocorticoid (GC) treatment has been demonstrated to have a short-term beneficial effect on functional recovery in relapsing multiple sclerosis (MS) patients but the exact mechanism of action of GCs in MS is unclear. We found that high dose intravenous GCs strongly reduced T-bet and pSTAT1 expression in CD4+, CD8+, CD14+ circulating cells in RRMS patients in relapse. pSTAT1and T-bet reduction was associated with the decline of IFNgamma production by PBMCs. A significant increase of AV-positive CD4+ and CD8+ T cells was detectable after GC treatment without any variation in the percentage of annexin V-positive monocytes. By in vitro analysis, patients during relapse, either before or after GC treatment, exhibited a lower proportion of apoptotic lymphocytes than remitting patients and controls. Our study suggests that GCs can modulate T-bet and STAT1 expression and that IFNgamma signalling inhibition contributes to anti-inflammatory action of GCs in the treatment of relapses of MS patients.     

Co-expression of the CD45RA and CD45RO antigens on T lymphocytes in chronic arthritis

The site of T lymphocyte activation in chronic arthritis is unknown. Peripheral blood (PB) lymphocytes from chronic arthritis patients are in a ‘naïve’ or non-activated state, as defined by expression of the CD45RA antigen and lack of HLA class II expression. In contrast, most synovial fluid (SF) T lymphocytes express a ‘memory’ or activated phenotype, as defined by the CD45RO antigen and high HLA class II expression. Following stimulation, naive cells lose CD45RA and gain CD45RO expression to become memory cells with a transitional stage of dual CD45RA, CD45RO antigen expression. To localize where this change in phenotype occurs we used dual colour immunofluorescence labelling to compare the percentage of dual CD45RA, CD45ROpositive T lymphocytes in PB and SF from chronic arthritic patients and from normal PB, assuming this population would be increased at the primary site of T lymphocyte activation. Expression of the intermediate and late activation marker. HLA-DR, was also analysed using dual colour immunofluorescence labelling. The percentage of dual positive T lymphocytes was similar between arthritic PB, SF. and normal PB, as was the density of both CD45RA and CD45RO antigens. Thus, CD45 isoform expression did not indicate where T lymphocytes were activated. However, we identified a previously unreported population of CD45RA⁺ CD45RO⁺ HLA-DR^- T lymphocytes in arthritic and normal PB. In SF, this population was absent, but a substantial number of dual CD45RA, CD45RO-positive HLA-DR⁺ T lymphocytes were identified. This population would not be predicted by the current model of T lymphocyte activation. Division of T lymphocytes into functional groups on the basis of CD45 isoform expression is likely to be more complicated than previously thought. Based on our findings we propose an alternative model of T lymphocyte differentiation.     

Peripheral blood monocyte and T‐lymphocyte activation levels at diagnosis predict long‐term survival in head and neck squamous cell carcinoma patients

This study was performed to determine whether peripheral blood (PB) monocyte and/or lymphocyte activation at diagnosis were associated with long‐term prognosis in patients with head and neck squamous cell carcinoma (HNSCC), and to what extent such prognostic properties relate to human papilloma virus (HPV)‐associated tumor infection of the included patients. This was a long‐term prospective study describing patient survival in relation to PB T lymphocyte and monocyte activation in patients observed for up to 14 years following diagnosis. Sixty‐four patients from a consecutive cohort of newly diagnosed HNSCC patients along with 16 non‐cancer control patients were included over a period of almost 2 years. Monocyte responsiveness was assessed at diagnosis (N = 56 HNSCC/16 non‐cancer controls) by measuring net levels of spontaneous vs lipopolysaccharide‐induced monocyte chemotactic protein (MCP)‐1 secretion in vitro. PB T lymphocyte activation was determined (N = 58 HNSCC/16 controls) by measuring the percentage of T cells expressing CD69 by flow cytometry. Whether HPV infection or not was determined by PCR analysis on formalin fixed paraffin‐embedded tumor tissue. Tumor HPV‐positive patients had better prognosis than HPV‐negative patients. A low net MCP‐1 response in monocytes predicted increased survival (Relative risk (RR) = 2.1; Confidence interval (CI): 1.1–4.0; p < 0.05). A low percentage of CD69 positive T lymphocytes also predicted better prognosis (RR = 2.6; CI: 1.3–5.0; p = 0.005). The predictive power of MCP‐1 monocyte and CD69 T lymphocyte measures were retained when adjusted for age and gender of the patients and shown to be independent of each other (N = 50 HNSCC/16 controls). The results were similar in HPV tumor‐positive and ‐negative patients. Patients with high monocyte‐ and/or T lymphocyte activation status had low survival with 8% 5 year overall survival (OS) compared to 65% 5 year OS for patients with dual low activation levels (RR = 0.27; CI: 0.14–0.56; p < 0.001), mostly secondary to disease‐specific survival. Both tumor HPV‐positive and ‐negative HNSCC patients with high percentage of CD69 positive T lymphocytes and/or high monocyte MCP‐1 secretion had low long‐term survival. The data suggest that the general inflammatory and adaptive immune systems are independently linked to the clinical aggressiveness of both tumor HPV‐negative and ‐positive HNSCC patients.     

Two systemic lupus erythematosus (SLE) global disease activity indexes--the SLE Disease Activity Index and the Systemic Lupus Activity Measure--demonstrate different correlations with activation of peripheral blood CD4+ T cells

Global disease activity measurement in systemic lupus erythematosus (SLE) patients is important for the clinical estimation and adjustment of therapy. By contrast, immune system activation plays a significant role in disease pathogenesis, with CD4+ lymphocytes acting as central cells in the immune response. We investigated which scale better correlates with immunologic changes in the blood of SLE patients, the SLE Disease Activity Index (SLEDAI) or the Systemic Lupus Activity Measure (SLAM) scale. Samples of peripheral blood were obtained from 45 SLE patients with different disease activity as assessed by the SLEDAI and the SLAM scales on the same day. We assessed the percentage of CD4+ T cells with activation-associated receptors: CD69, CD25int, CD95, HLA-DR, and CD4+ T cells with killing properties containing perforin and granzyme B. Our results indicated that the percentage of CD4+CD69+ and CD4+CD25(int) cells did not correlate with either the SLEDAI or the SLAM scale. Significant and positive correlations were observed between percentages of CD4+CD95+ and CD4+HLA-DR+ lymphocytes and SLE activity, but only when activity was measured using the SLAM scale, not with the SLEDAI scale. The percentage of CD4+perforin+ and CD4+granzyme B+ cells also strongly correlated with disease activity measured only with the SLAM scale. We conclude that the SLAM scale better reflects changes of immune system activity in SLE patients compared with the SLEDAI scale.     

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.     

Activated CD4+ and CD8+ T Cell Proportions in Multiple Sclerosis Patients

The goal of this study was to trace the course of multiple sclerosis (MS) by evaluating the lymphocyte subpopulation counts and the levels of CD4+ and CD8+ T cell activation using flow cytometry. Samples obtained from healthy subjects (N?=?40) and patients with MS (N?=?290) were analyzed. Lymphocytes were labeled for the surface markers CD4+, CD8+, CD3+, CD16+, CD19+, CD45+, and CD53+ and the activation marker HLA-DR+. Cell counts were then determined using flow cytometry. A high degree of inter-individual variability was observed in the counts of all lymphocyte subtypes in the MS group. A significantly lower proportion of CD3+ T cells (69?±?14 % in healthy subjects and 60?±?17 % as a percent of total lymphocytes in MS patients), CD4+ T cells (41?±?11 and 28?±?18 %, respectively), and a significantly higher proportion of NK T cells (12?±?5 and 25?±?21 %, respectively) were observed in patients with MS than in healthy subjects. These differences led to a lowered CD4+/CD8+ T cell ratio. Furthermore, a significantly lower proportion of activated CD4+ T cells (HLA-DR+ CD4+; from 48?±?10 to 38?±?15 % as a percent of CD4+ cells) was observed in patients with MS than in healthy subjects. The high level of inter-individual variability in lymphocyte cell counts and the counts of activated T cells suggest that MS is a complex and heterogeneous disease.