恶性淋巴瘤患儿细胞黏附分子检测分析研究

目的探讨恶性淋巴瘤患儿血中细胞黏附分子CD11a、CD62L、CD54表达水平及化疗和中药联合治疗对其表达水平的影响。方法采用流式细胞术动态检测50例恶性淋巴瘤患儿细胞黏附分子在各种治疗中的表达水平。结果化疗加中药舒肝溃坚汤治疗组与单纯化疗组相比,外周血中CD11a（t=3．58）、CD62L（t=3．02）、CD54（t=2．78）水平均明显增高,差异具有统计学意义（P〈0．01）,中药联合治疗可明显提高患儿细胞黏附分子表达,增加机体免疫功能。结论细胞黏附分子与肿瘤免疫密切相关,在恶性淋巴瘤发生、发展以及药物治疗中发挥了重要作用。舒肝溃坚汤对其具有明显疗效。     

不同透析膜对维持性血液透析患者B细胞活化的影响

目的　观察尿毒症患者反映B细胞活化的血清sCD2 3 浓度水平的变化及不同种类透析膜对维持性血液透析患者血清sCD2 3 水平的影响。方法　采用双抗体夹心ELISA法检测尿毒症非透析患者 ,及使用不同种类透析膜透析器初用及复用 5次时分别观察透析过程中患者血清sCD2 3 水平的变化。结果　尿毒症血液透析患者透析前及尿毒症非透析患者血清sCD2 3 较正常对照组显著增高。血透过程中尿毒症血透患者血清sCD2 3 分子浓度水平规律性的变化。不同类型透析膜中 ,醋酸纤维素膜 (CA)对血清sCD2 3 的影响显著大于血仿膜 (HE)和聚砜膜 (PS) ;透析器复用对血透患者血清sCD2 3 水平无明显影响。结论　血透过程中血清sCD2 3 分子浓度水平规律性的变化 ,是导致机体免疫力下降的可能原因之一。HE膜和PS膜生物相容性优于CA膜 ,透析器复用可改善透析膜的生物相容性     

不同类型透析膜对维持性血液透析患者血清C-反应蛋白水平变化的影响

 目的观察尿毒症维持性血液透析患者血清C-反应蛋白(C reactive protein,CRP)水平的变化以及不同类型透析膜初次与复用对其的影响,从机体免疫功能的角度对透析膜的生物相容性进行评价.方法采用免疫速率散射比浊法测定尿毒症维持性血液透析患者初用和复用醋酸纤维素膜(CA)、血仿膜(HE)和聚砜膜(PS)透析器进行透析时患者血清CRP水平的变化,并加以比较.同时检测健康查体和尿毒症非透析患者的CRP水平作为对照.结果尿毒症血液透析治疗患者透析前及尿毒症非透析患者血清CRP水平均较健康查体对照组显著增高.尿毒症患者透析开始后4.5 h血清CRP水平与透析前相比明显升高(P＜0.05),HD后1.5 h后差异更为明显.使用醋酸纤维素膜(CA)透析器患者的血清CRP水平更明显高于使用血仿膜(HE)和聚砜膜(PS)透析器的患者(P＜0.05).结论血液透析可导致尿毒症血透患者血清CRP水平的增高,这种变化与透析膜的生物相容性有关,而透析器复用可在一定程度上减轻这种变化.     

人参皂苷Re对肿瘤坏死因子-α诱导的白细胞活化的抑制作用及机制

目的研究人参皂苷Re对肿瘤坏死因子-α诱导的白细胞活化的作用及其机制。方法提取并培养人外周血中性白细胞,用酶标仪和流式细胞仪体外观察肿瘤坏死因子-α(20 ng/ml)对中性白细胞黏附相关指标MPO的分泌以及白细胞表面黏附分子CD11b和CD18的表达的诱导作用,同时观察肿瘤坏死因子-α(20 ng/ml)对中性白细胞跨内皮细胞迁移的诱导作用,并测定不同浓度人参皂苷Re对以上变化的影响。结果人参皂苷Re可以剂量依赖性地明显抑制肿瘤坏死因子-α诱导的黏附于内皮细胞表面白细胞MPO的分泌和黏附分子CD11b、CD18的表达,明显抑制肿瘤坏死因子-α诱导的中性白细胞跨内皮细胞的迁移。结论人参皂苷Re可以抑制由肿瘤坏死因子-α诱导的白细胞的活化,包括白细胞与内皮细胞的黏附和游出,该作用与抑制白细胞表面黏附分子CD11b和CD18的表达相关。     

人血白细胞黏附分子表达及黏附能力与辐射剂量的关系

目的观察人离体外周血白细胞^60Coγ射线照射后24h内黏附分子的表达及单个核细胞黏附功能与照射剂量间的关系。方法人外周静脉血经0、2、4和6Gy γ射线照射后，用流式细胞术检测照后0、6、12和24h不同黏附分子（CD11a、CD11b、CD18、CD29、CD49d、CD54）的表达。用细胞黏附方法测定照后6、12和24h单个核细胞对不同底物的黏附情况。结果粒细胞、单核细胞CD29的表达在照后6h，2和4Gy组升高，在其他时间点各剂量照射组表现为下降；单个核细胞对底物IgG的黏附能力在照后6、12和24h的各个剂量组均表现为增强。结论外周血细胞黏附分子表达和黏附能力与照射剂量具有较好的量效关系。     

高压氧干预下急性脑梗死患者细胞黏附分子变化及对预后的影响

目的 对急性脑梗死(acute cerebral infarction,ACI)患者CD11b、CD11c和CD54作动态观察,找出其变化规律及高压氧(hyperbaric oxygen,HBO)对该规律的影响,并进一步阐明HBO对ACI患者脑血管的保护作用;同时对ACI患者治疗前CD11b、CD11c和CD54与近、远期神经功能恢复程度相关性进行探讨.方法 64例ACI患者分2组:HBO组31例,常规治疗联合HBO治疗;治疗对照组33例,仅行常规治疗.2组分别在治疗前(发病≤72 h)及治疗第7、10、12、20天晨抽取外周静脉血测定CD11b、CD11c和CD54,并在相同治疗时间进行神经功能缺损程度评分(neural functional damage scores,NDS).另外选取正常对照组25人,仅抽取周围静脉血测定CD11b、CD11c和CD54.结果 HBO组和治疗对照组治疗前CD11b、CD11c和CD54表达明显升高,达到高峰,2组各指标差异无统计学意义(P＞0.05),第7天均开始下降.HBO组CD11b、CD11c高峰持续时间为7d,治疗对照组持续10 d;HBO组CD54高峰持续时间为10 d,治疗对照组持续12d.经相关性和线性回归分析发现,发病第20天时NDS水平(近期疗效)和发病半年、1年时NDS的水平(远期疗效)分别为97.2％、96.7％及97.6％,可用CD11b和/或CD11c和/或CD54的上调水平以及其他相关因素解释.结论 HBO治疗后可缩短ACI患者CD11b、CD11c和CD54高峰持续时间.说明HBO可干预细胞黏附分子的变化过程,缩短其异常时限,减少它们的表达,对ACI患者有保护作用,并减少白细胞的黏附.同时治疗前CD11b、CD11c和CD54的表达可预测病情的轻重,影响近、远期疗效,为研究如何调控或抑制CD11b、CD11c和CD54的表达、减少对血管的损伤及早期制定更全面的治疗方案提供依据.     

高压氧对脑缺血再灌注小鼠白细胞粘附分子的影响

目的观察脑缺血再灌注小鼠白细胞粘附分子CD11a／CD18、CD11b／CD18、CD62L表达的变化及高压氧（hyperbaric oxygen，HBO）对其表达的影响。方法昆明小鼠63只随机分为缺血再灌注1、3、5d组，HBO1、3、5d组，手术1、3、5d组，共9组，每组7只。应用流式细胞仪分别检测各组白细胞粘附分子CD11a／CD18、CD11b／CD18、CD62L的表达。结果缺血再灌注1d组CD11a／CD18表达水平显著高于同期手术组（P〈0．05）；缺血再灌注3d组CD11b／CD18表达水平显著高于同期手术组（P〈0．05）；缺血再灌注1、3、5d组CD62L表达水平均显著低于同期手术组（P〈0．05）。经HBO处理后，HBO1、3、5d组CD11a／CD18表达水平与同期缺血再灌注组相比，差异均无统计学意义（P〉0．05）；HBO 3d组CD11b／CD18表达水平显著低于同期缺血再灌注组（P〈0．05），但是仍显著高于同期手术组（P〈0．05）；HBO 5d组CD62L表达水平显著高于同期缺血再灌注组（P〈0．05），与同期手术组相比差异无统计学意义（P〉0．05）。结论脑缺血再灌注后，白细胞粘附分子CD11a／CD18、CD11b／CD18表达升高，CD62L表达降低；HBO治疗对缺血再灌注后CD11a／CD18表达差异无统计学意义，但可抑制缺血再灌注后CD11b／CD18的表达，缩短CD62L表达降低的时间。     

人外周血白细胞黏附分子的辐射效应研究

目的 研究外周血白细胞表面黏附分子的表达及其介导的细胞黏附能力变化与辐射剂量的关系。方法 人外周血细胞经不同剂量照射后不同时间用流式细胞仪双色标记分析不同白细胞表面不同黏附分子表达,特异底物包被微孔板法和结晶紫染色分析检测单核细胞对不同底物的黏附能力。结果 单核细胞表面CD11b和粒细胞表面CD29表达下降与辐射剂量间存在良好量效关系,照射后单核细胞对底物β1-整合素和胶原蛋白I的黏附能力改变与辐射剂量存在一定量效关系。结论 外周血白细胞表面黏附分子表达及功能改变展现出的与辐射剂量间的良好关系,为进一步深入研究将其作为评估生物受照剂量指标的可能打下基础。     

低分子肝素和尿激酶对大鼠肾小球炎症保护作用的比较

目的比较低分子量肝素和尿激酶对肾小球炎症反应的保护作用。方法3月龄雌性Wistar大鼠40只,随机分为对照组(NC组)、脂多糖(LPS)组(L组)、LPS 氨甲环酸组(LT组)、LPS 氨甲环酸 低分子量肝素组(LTH组)及LPS 氨甲环酸 尿激酶组(LTU组),每组8只。采用直接免疫荧光检测肾小球纤维蛋白沉积和CD11b阳性细胞分布情况,Western blotting检测内皮细胞细胞间黏附分子-1(ICAM-1)蛋白质表达。结果NC组大鼠肾小球内无纤维蛋白沉积和CD11b阳性细胞;ICAM-1有少量表达。与NC组比较,L组纤维蛋白沉积(9.1%±1.6%)和CD11b阳性细胞(11.2±2.1)增多(P<0.05);ICAM-1表达(0.23±0.09)明显上调(P<0.05)。与L组比较,LT组纤维蛋白沉积(23.4%±3.2%)和CD11b阳性细胞(20.4±3.5)进一步增多;ICAM-1表达(0.44±0.16)进一步上调(P<0.05)。与LT组比较,LTH组和LTU组纤维蛋白沉积(分别为11.3%±1.4%,10.6%±1.5%)和CD11b阳性细胞(分别为7.3±1.8,8.4±1.6)明显减少(P<0.05或0.01);ICAM-1表达(分别为0.19±0.10,0.18±0.09)明显下调(P<0.05),LTH、LTU两组间无统计学差异。结论低分子肝素与尿激酶均能有效减少纤维蛋白沉积,起到减轻肾小球炎症反应的作用。     

维持性血液透析患者外周血单个核细胞早期凋亡

目的　研究维持性血液透析 (MHD)患者外周血单个核细胞 (PBMC)早期凋亡 (Annexin V)。　方法　应用流式细胞仪 (FCM)定量检测终末期肾衰竭未透析患者 (ND)、醋酸纤维膜透析患者 (CA)、聚砜膜透析患者 (PS)和健康自愿者 (C)各10例的 PBMC体外无刺激培养 2 4 h后 Annexin V的表达。　结果　 ND组 Annexin V表达显著高于 C组和 PS组 ,CA组显著高于 C组和 PS组 (P<0 .0 5 ) ,而 PS组与 C组、CA组与 ND组差异不显著。 Annexin V与 Ccr间呈显著负相关 (P<0 .0 5 )。　结论　透析和尿毒症本身可诱导外周血单个核细胞凋亡增加 ,生物相容性较好的透析膜可减轻细胞凋亡     

Effect of intense wrestling exercise on leucocytes and adhesion molecules in adolescent boys

BACKGROUND: In adults, exercise is a powerful and natural stimulator of immune cells and adhesion molecules. Far less is known about exercise responses during childhood and adolescence and whether or not exercise in "real life" activities of healthy adolescents influences immune responses. OBJECTIVE: To determine if strenuous exercise leads to significant changes in leucocyte number and adhesion molecule expression in adolescent boys. METHODS: Eleven healthy, high school boys, aged 14-18.5 years, performed a single, typical, 1.5 hour wrestling practice session. Blood was sampled before and after the session. Flow cytometry was used to evaluate changes in immune responses. RESULTS: The exercise led to significant (p<0.05) and robust increases in granulocytes, monocytes, and all lymphocyte subpopulations. The most significant changes were observed for natural killer cells (p<0.0005). The number of T cytotoxic and T helper cells expressing CD62L increased significantly (p<0.002 and p<0.0005 respectively), as did the number of T cytotoxic and T helper cells not expressing CD62L (p<0.003 and p<0.009 respectively). The density of CD62L on lymphocytes decreased significantly with exercise (p<0.0005), whereas CD11a (p<0.01) and CD54 (p<0.01) increased. CONCLUSIONS: The data show that an intense wrestling bout in adolescent boys leads to profound stimulation of the immune system. The role of these common changes in overall immune status and the development of the immune and haemopoietic systems has yet to be determined.     

Labelling of leucocytes with technetium-99m exametazime causes in vitro upregulation of granulocyte CD11b without correlation to tissue uptake in vivo

The aims of this study were to investigate whether labelling with technetium-99m exametazime alters the expression of adhesion molecule CD11b on granulocytes and monocytes, and to study whether the expression of CD11b on unlabelled or labelled cells correlates with uptake of the labelled cells in the inflamed bowel, in the lungs or in the reticuloendothelial system. Leucocytes were obtained from 25 patients with inflammatory bowel disease who underwent leucocyte scan. The cellular expression of CD11b was analysed using flow cytometry. Labelling with^99mTc-exametazime induced an increased surface expression of CD11b on granulocytes (P<0.01), but not on monocytes. The increase in CD11b expression on granulocytes was lower than the spontaneous mobilization that occurred at 37° C and correlated neither with this, nor withN-formyl-methionyl-phenylalanine induced expression of the same receptor. Basal expression of CD11b on unlabelled granulocytes, but not on monocytes, correlated with bowel and lung uptake 45 min after reinjection of labelled cells, but not with uptake on later images. No correlation was found between the CD11b expression on labelled granulocytes or monocytes and scintigraphic uptake. Our findings show that labelling with^99mTc-exametazime increases the expression of adhesion protein CD11b on granulocytes. The increase in surface expression of CD11b does not correlate with the scintigraphic uptake of labelled cells in the bowel, in the lungs or in the reticuloendothelial system.     

LeuCAM and reactive oxygen species during long-term exercise combined with sleep and energy deficiency

INTRODUCTION: During the Norwegian military ranger-training course, cadets are exposed to prolonged physical exercise combined with sleep-, energy-, and food deficiency. The open-window postexercise hypothesis indicates that after hard physical activity, there is an increased risk of contracting infectious diseases. PURPOSE: The purpose of the present study was to determine leukocyte reactive oxygen species (ROS) levels, total antioxidant status (TAS), leukocyte expression of the cell adhesion molecules CD62L and CD11b, and plasma levels of soluble adhesion molecule L-selectin before, during, and in the recovery phase of a military ranger-training course. METHODS: Ten cadets from the Norwegian Military Academy were recruited to the study. Flow cytometry was used to study the intracellular levels of ROS in leukocytes (basally, as well as after in vitro stimulation with phorbol myristate acetate (PMA)), applying the probes dihydroethidium (DHE) and dihydrorhodamine 123 (DHR) and the leukocyte expression of adhesion molecules CD62L and CD11b. ELISA was used to assess the plasma levels of soluble L-selectin, and TAS in plasma was measured using the ABTS+ reduction assay kit. RESULTS: The basal levels of ROS as well as PMA-stimulated ROS in leukocytes declined gradually during the ranger-training course, being lowest on the last day (P < 0.05). The level of TAS increased (P < 0.01) during the course. A striking decrease (P < 0.001) was observed in leukocyte CD62L expression and was sustained even after 3 d of recovery. The leukocyte expression of CD11b remained unchanged. CONCLUSION: The ranger-training course leads to a partial exhaustion of the leukocyte ROS-generating machinery and to a nearly total extinguishing of leukocyte CD62L expression. These changes may support the open-window hypothesis indicating reduced ability to combat microbial invasions before total restitution.     

Expression of lymphocyte subsets after exercise and dexamethasone in high and low stress responders

PURPOSE: Recent work indicates that among the normal population, persons can be classified as low (LR) or high (HR) stress responders based on hypothalamic-pituitary-adrenal (HPA) axis responses to high-intensity exercise. We studied whether differential activation of the HPA axis affected cytokine production and expression of selected lymphocyte subsets in HR and LR in response to high-intensity exercise after placebo and dexamethasone (DEX; 4 mg). METHODS: Healthy HR (N = 12) and LR (N = 10) underwent two exercise tests at 90% of VO2max, 8 h after placebo or DEX. Expression of lymphocyte surface markers (CD3+, CD4+, CD8+, CD56+), adhesion molecule markers (intercellular adhesion molecule-1/ICAM-1: CD54+ and L-selectin: CD62L+), and concentrations of plasma interleukin 6 (IL-6) were examined before and after exercise. RESULTS: Baseline percentages of CD8+ and CD56+ cells were significantly higher, and concentrations of IL-6 and percentages of CD4+ cells were significantly lower in HR as compared with LR. The percentage of CD54+ and CD62L+ cells was not significantly different in HR and LR. DEX significantly reduced the percentage of CD3+ and CD4+ and increased the percentage of CD8+ and CD56+ subsets; the percent of cells expressing CD54+ increased, whereas CD62L+ decreased. Exercise-induced changes in the percentage of lymphocyte subsets were similar to those induced by DEX. CONCLUSION: In summary, HR and LR have different baseline patterns of IL-6 and lymphocyte subsets, which may reflect differential sensitivity to endogenous glucocorticoids. However, exogenous glucocorticoids induced similar patterns of lymphocyte expression in HR and LR.     

UVB-irradiated T-cells undergoing apoptosis lose L-selectin by metalloprotese-mediated shedding

PURPOSE: L-selectin (CD62L) is a prerequisite for leucocyte adhesion to endothelial cells of blood vessels and consequently for transmigration. Its expression on the cell surface therefore regulates the ability of lymphocytes to enter lymph nodes, to re-enter blood vessels or to invade tissues at sites of inflammation. The aim of this study was to determine the expression of CD62L on apoptotic lymphocytes after UVB irradiation. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from peripheral blood of normal healthy volunteers. Cells were stimulated with phorbol myristate acetate (PMA) and ionomycin for activation. Apoptosis in peripheral T-cells and Jurkat cells was induced by irradiation with UVB (120 mJ/cm2). In addition, T-cells or Jurkat cells were cultured for the indicated time with anti-Fas antibody CH11. The CH11-induced apoptosis was inhibited by the pan-caspase inhibitor zVAD-fmk. For detection of apoptosis, cells were analysed by cytofluorometry for morphological changes typical for apoptosis. The reliability of the apoptotic cell gate was confirmed by staining with FITC-labelled annexin-V in the presence ofpropidium iodide (PI). For FACS analysis of CD62L expression on the cell-surface immunofluorescence was performed using FITC-conjugated anti-CD62L and PE-conjugated anti-CD3 antibodies. Soluble CD62L (sCD62L) in the cell supernatants was measured by standard ELISA technique. Assays were performed in the presence and absence of metalloprotease inhibitor KB8301. RESULTS: PBMC from healthy volunteers undergoing apoptosis following UVB irradiation selectively shed CD62L, whereas the expression of the lineage-specific marker CD3 showed only minor changes. Shedding was blocked by the hydroxamic acid-based metalloprotease inhibitor KB8301. When Jurkat cells were treated with the caspase inhibitor zVAD-fmk, anti-CD95 antibodies did not induce apoptosis, and the expression of CD62L remained unaltered. CONCLUSION: UVB or ionizing radiation induce apoptosis in lymphocytes. The loss of CD62L is associated with apoptosis and will influence lymphocyte trafficking and, by excluding them from CD62L-mediated adhesion and tissue invasion, might contribute to the regulation of inflammation.     

UVB-irradiated T-cells undergoing apoptosis lose L-selectin by metalloprotease-mediated shedding

Abstract. Purpose : L-selectin (CD62L) is a prerequisite for leucocyte adhesion to endothelial cells of blood vessels and consequently for transmigration. Its expression on the cell surface therefore regulates the ability of lymphocytes to enter lymph nodes, to re-enter blood vessels or to invade tissues at sites of inflammation. The aim of this study was to determine the expression of CD62L on apoptotic lymphocytes after UVB irradiation. Materials and methods : Peripheral blood mononuclear cells (PBMC) were isolated from peripheral blood of normal healthy volunteers. Cells were stimulated with phorbol myristate acetate (PMA) and ionomycin for activation. Apoptosis in peripheral T-cells and Jurkat cells was induced by irradiation with UVB (120 mJ/cm2). In addition, T-cells or Jurkat cells were cultured for the indicated time with anti-Fas antibody CH11. The CH11-induced apoptosis was inhibited by the pan-caspase inhibitor zVAD-fmk. For detection of apoptosis, cells were analysed by cytofluorometry for morphological changes typical for apoptosis. The reliability of the apoptotic cell gate was confirmed by staining with FITClabelled annexin-V in the presence of propidium iodide (PI). For FACS analysis of CD62L expression on the cell-surface immunofluorescence was performed using FITC-conjugated anti-CD62L and PE-conjugated anti-CD3 antibodies. Soluble CD62L (sCD62L) in the cell supernatants was measured by standard ELISA technique. Assays were performed in the presence and absence of metalloprotease inhibitor KB8301. Results : PBMC from healthy volunteers undergoing apoptosis following UVB irradiation selectively shed CD62L, whereas the expression of the lineage-specific marker CD3 showed only minor changes. Shedding was blocked by the hydroxamic acid-based metalloprotease inhibitor KB8301. When Jurkat cells were treated with the caspase inhibitor zVAD-fmk, anti-CD95 antibodies did not induce apoptosis, and the expression of CD62L remained unaltered. Conclusion : UVB or ionizing radiation induce apoptosis in lymphocytes. The loss of CD62L is associated with apoptosis and will influence lymphocyte tra Ýcking and, by excluding them from CD62L-mediated adhesion and tissue invasion, might contribute to the regulation of inflammation.