不同复苏溶液对失血性休克大鼠中性粒细胞CD11a、CD11b表达的影响

目的观察不同复苏溶液对失血性休克大鼠PMN表面CD11a、CD11b表达的影响.方法成年Wistar大鼠随机分为0.9%NaCl(NS)、7.5%NaCl(HS)、5%NaCl-3.5%NaAc(HSA)三组,每组6只.动物麻醉后,自股动脉放血,于10min内使MAP下降至5.07-5.47kPa,维持90min.按4mL/kg体重分别注入NS、HS、HSA,5min内输完.随后输注三倍最大失血量的复方氯化钠溶液,40min内输完.分别于休克前、后、给液后1.5h、3h、6h取血0.2mL,流式细胞仪测定PMN表面CD11a、CD11b表达的变化.结果休克后,各组PMN表面CD11a表达下降,但与休克前比较无显著差别.给液后1.5h,各组CD11a表达继续下降;给液后3h,各组CD11a表达稍有回升;给液后6h,各组CD11a表达又呈下降趋势,HSA组CD11a表达显著低于休克前水平;各组间于各时相点均无显著差别.休克后,各组PMN表面CD11b表达增加,与休克前比较有非常显著差别.给液后3h内,各组CD11b表达随观察时间的延长呈进行性增加;给液后6h,NS组CD11b表达继续增加,HS组和HSA组CD11b表达有下降趋势.各组CD11b表达于给液后各时相点均较休克前和休克时非常显著增加,HS组和HSA组CD11b表达于给液后各时相点均低于NS组,但无显著差别.结论液体复苏后,失血性休克大鼠PMN表面CD11a表达呈下降趋势,CD11b表达呈上升趋势,HS和HSA有减弱这种趋势的作用.     

严重烧伤病人PMNCD11b/CD18受体表达及特异性iRNA对其调节作用

本文探讨了严重烧伤对病人外周血中性粒细胞 (PMN)CD11b/CD18受体表达的影响及特异性免疫核糖核酸 (iRNA)对其调节作用。结果发现 :(1)严重烧伤病人PMNCD11b/CD18受体表达率明显下降 ,至伤后第 10天时 ,分别只有正常的6 7 1%和 6 8 9% ,且其下降程度与烧伤面积成正比 ;(2 )伤后早期应用特异性iRNA可明显提高烧伤病人PMNCD11b/CD18受体表达率 ;(3)临床观察发现 ,治疗组伤后创面细菌培养阳性率 ,创面脓毒症及菌血症的发生率明显低于对照组 (P均 <0 0 5 ) ,创面愈合时间明显短于对照组 (P <0 0 1)。该结果为临床应用特异性iRNA防治烧伤后感染提供了实验依据。     

CD11c/CD18研究进展

ＣＤ１１ｃ／ＣＤ１８是主要分布于髓样细胞表面的一种跨膜糖蛋白，属于β２整合素家族。它既是纤维蛋白原、脂多糖和ｉＣ３ｂ的受体，又可介导髓样细胞与其它细胞、底物的粘附，参与机体的免疫炎性反应及细胞的分化增殖。     

乳腺癌中CD66b阳性肿瘤相关中性粒细胞的意义

目的 探讨CD66b阳性肿瘤相关中性粒细胞在乳腺癌中的浸润及意义。方法 收集安徽医科大学第一附属医院病理科存档的80例乳腺癌手术切除组织,采用免疫组化SP法观察中性粒细胞的浸润情况,Graphpad软件分析其临床病理特征;下载TCGA数据库,R软件分析乳腺癌中肿瘤相关中性粒细胞的浸润及其与预后的相关性。结果 80例乳腺癌组织中,肿瘤相关中性粒细胞的浸润密度与无瘤生存期呈负相关(P=0.007 7),与肿瘤病理分期(P=0.005 0)、淋巴结转移(P=0.023 7)、远处转移呈正相关(P=0.001 0),与三阴型乳腺癌相关(P=0.012 0);TCGA数据库显示,肿瘤相关中性粒细胞的数量与淋巴结侵犯(P=0.037 0)、远处转移相关(P=0.013 0)。结论 乳腺癌组织中CD66b阳性肿瘤相关中性粒细胞可作为潜在的转移预测指标。     

槲皮素对LPS诱导中性粒细胞活性化效应的抑制作用

目的研究槲皮素(Quercetin,Que)对细菌脂多糖(Lipopolysaccharide,LPS)诱导的中性粒细胞(Polymorphonuclear,PMN)活化效应的影响。方法运用免疫荧光法和流式细胞术,对接受1h LPS刺激的PMN表面黏附分子(CD62L,CD11b／CD18)的表达进行测定,同时应用MTT法对不同状态下的PMN活性进行测定。结果Que对LPS诱导的中性粒细胞活化效应有明显抑制作用,表现为抑制细胞表面黏附分子CD62L的表达和促进CD11b／CD18的表达,同时Que对LPS增加细胞活性的效应有抑制作用。结论槲皮素通过对抗LPS对PMN黏附分子CD62L,CD11b／CD18的表达的影响,抑制LPS诱导的中性粒细胞活化效应,从而阻止PMN对血管内皮细胞的黏附,减少炎症细胞向炎症局灶的浸润,这可能是槲皮素发挥抗炎作用的一个重要机制。     

猪苓多糖协同卡介苗对巨噬细胞表达CD11b、CD18以及协同刺激分子的影响

目的：了解猪苓多糖协同对巨噬细胞J774 A.1CD11b、CD18及协同刺激分子表达的影响.方法：应用流式细胞术检测猪苓多糖协同卡介苗刺激J774 A.10.5 h、1 h、3 h、6 h、12 h、24 h及48 h后,其CD11b、CD18及协同刺激分子CD86、CD40表达的变化.结果：BCG（50 mg/L）组作用1 h后,J774 A.1 CD11b、CD18的表达高于空白组;联合应用PPS（50 mg/L）组刺激12 h,其CD11b、CD18的表达明显高于BCG组（P＜0.05）.BCG组0.5 h、3 h、12 h、24 h、48 h,其协同刺激分子的表达增高;联合应用PPS协同刺激J774 A.10.5 h、1 h、3 h、6 h,协同刺激分子的表达高于BCG组.结论：卡介苗能提高巨噬细胞CD11b、CD18及协同刺激分子的表达,联合应用猪苓多糖可使其作用增强.     

地奥司明对肾缺血再灌注大鼠肾组织MPO和中性粒细胞CD11b、CD54及CD62L水平的影响

 目的：观察地奥司明（DOSM）对肾缺血/再灌注（I/R）大鼠肾组织髓过氧化物酶（MPO）和中性粒细胞CD11b、CD54及CD62L表达水平的影响。方法: 180只SD大鼠随机分成3组:假手术组(SO)、肾缺血/再灌注模型组(I/R)和地奥司明+肾缺血/再灌注模型组(DOSM+I/R)。采用ELISA方法测定肾脏组织中MPO的水平,流式细胞分析法检测中性粒细胞表面CD11b、CD54和CD62L的表达水平。结果: 在1h、3h、6h、12h、24h和48h不同时间点的肾组织中,I/R和DOSM＋I/R两组MPO活性均随时间逐渐升高,于6h达最高,后逐渐下降,两组相比有显著性差异（P＜0.05或P＜0.01）。中性粒细胞CD11b、CD54及CD62L的表达在相同时段I/R组显著高于SO组（P＜0.05或P＜0.01);而在DOSM+I/R组显著低于I/R组（P＜0.05或P＜0.01)。结论：DOSM可显著降低肾I/R病理过程中MPO活性,显著降低中性粒细胞CD11b、CD54及CD62L的表达,有助于减轻I/R时炎性细胞的浸润。     

粘附分子CD11a、CD11b、CD62L在恶性淋巴增殖性疾病的表达

目的：观察恶性淋巴增殖性疾病肿瘤细胞表面β２整合素（ＣＤ１１ａ、ＣＤ１１ｂ）及Ｌ－选择素（ＣＤ６２１）的表达变化及其临床意义。方法：用流式细胞仪检测３５例初诊或复发急性淋巴白血病（ＡＬＬ）、４例慢性淋巴细胞白血病（ＣＬＬ）、３０例多发性骨髓瘤（ＭＭ）、１４例淋巴肉瘤白血症及２５例正常人骨髓单个核细胞粘附分子ＣＤ１１ａ、ＣＤ１１ｂ、ＣＤ６２Ｌ的表达。结果：（１）与正常造血细胞比较，ＣＤ１１ｂ、ＣＤ１     

Anti-CD11b/CD18 antibodies reduce inflammation in acute colitis in rats.

The influx of monocytes and neutrophils into the inflamed tissue could be an important aspect in the pathogenesis of inflammatory bowel disease (IBD). A membrane protein involved in the monocyte/neutrophil adherence to endothelium is CD11b/CD18 or alpha M beta 2 (complement receptor type 3 = CR3). In the present study the role of CD11b/CD18 in experimental IBD was studied by treatment with ED7 and OX42, two MoAbs against CD11b/CD18. Colitis was induced in rats by a single, rectal administration of 30 mg 2,4,6-trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol 30%. Two hours before and 3 days after induction of colitis, the animals were given an i.v. dose of 0.5 mg of either ED7 or OX42 in 1 ml PBS. Controls received PBS or an irrelevant MoAb. Four days after the last treatment with the antibodies, the rats were killed, and macroscopic damage scores of the colon were determined. Macrophages and granulocytes were studied by immunohistochemistry and quantified by Interaktives Bild Analysen System (IBAS), and myeloperoxidase (MPO) activity in colonic tissue was measured. After treatment with ED7 and OX42 the mean damage score of the colon was reduced from 4.2 in IBD animals to 1.0 and 1.3, respectively. Smaller areas of ulcerations and a decrease in the number of ulcerations were observed compared with PBS-treated rats. Furthermore, the amount of infiltrating monocytes and leucocytes in the submucosa was enormously reduced, as well as MPO activity in the colonic tissue. These results show that treatment with MoAbs against CD11b/CD18 reduces clinical signs of experimental IBD in rats by a partial blockade of infiltrating macrophages and granulocytes.     

高渗盐水预处理可减轻中性粒细胞介导的肝脏缺血再灌注损伤

目的：探讨高渗盐水预处理对肝脏缺血再灌注损伤的拮抗作用及其机制。方法：建立大鼠局部肝脏缺血再灌注模型。设假手术组（sham组）、缺血再灌注组（IR组）和高渗盐水预处理组（HTS组），分别于再灌注后1 h、3 h、6 h、12 h和24 h处死大鼠，测定谷丙转氨酶（ALT）；抗凝血流式细胞仪测定中性粒细胞CD11b/CD18(Mac-1)的阳性率；RT-PCR和Western blotting分别测定肝内细胞间黏附分子1（ICAM-1）的mRNA和蛋白表达；比色法测定肝脏内髓过氧化物酶（MPO）活性；常规病理及电镜观察肝脏的病理学改变及肝脏内中性粒细胞的浸润情况。结果： ① HTS组在3 h、6 h和12 h血清ALT水平明显低于IR组（P<0.05）。②HTS组在6 h和12 h中性粒细胞Mac-1表达显著弱于IR组（P<0.05）。③HTS组肝脏MPO活性在再灌注后6 h、12 h和24 h明显低于IR组（P<0.05）。④HTS组大鼠肝脏内ICAM-1mRNA及蛋白表达明显低于IR组。⑤HTS组肝内中性粒细胞浸润、肝细胞浊肿和肝窦狭窄程度轻于IR组。结论： HTS预处理能够通过抑制中性粒细胞Mac-1和肝内ICAM-1的表达，明显抑制肝内中性粒细胞的黏附和聚集，起到拮抗肝脏缺血再灌注损伤的作用。     

Increased CD11/CD18 expression on peripheral blood leucocytes of patients with sarcoidosis.

Sarcoidosis is a multisystem disease of unknown etiology characterized by non-caseating granulomata, formed mainly from macrophages surrounded by lymphocytes and plasma cells. Using a novel method for the preparation of blood leucocytes for flow cytometry, we report increased expression of LeuCAMs (CD11/CD18) on peripheral blood leucocytes of 11 Caucasian and 10 Afro-Caribbean patients with sarcoidosis compared with age-, sex- and race-matched controls. Whilst the percentages of the cells expressing CD11/CD18 were no different, the density, expressed as mean fluorescence intensity (MFI), was greater for all leucocytes in sarcoids than in normal individuals. The expression of intercellular adhesion molecule-1 (ICAM-1), a ligand for LFA-1 which is expressed on all leucocytes, was not significantly different from normal, whereas HLA-DR was expressed more intensely on sarcoid monocytes (P less than 0.01) and blood lymphocytes (P less than 0.005) than control cells. Our findings are consistent with leucocyte activation although we were unable to confirm reports of elevated tumour necrosis factor-alpha (TNF-alpha) in the patients' plasma using an ELISA. Increased expression of adhesion molecules on peripheral blood leucocytes may play a role in the cellular extravasation, aggregation, and granuloma formation seen in sarcoidosis.     

Altered expression of CD11/CD18 on the peripheral blood phagocytes of patients with tuberculosis.

Tuberculosis (TB), caused by Mycobacterium tuberculosis, is characterized by granulomatous lesions made up of epithelioid cells, giant cells and mononuclear leucocytes. Cell-cell adhesion is important in granuloma formation and in the leucocyte migration which accompanies it. We have recently shown increased expression of the adhesion molecules CD11/CD18 (LeuCAMs, beta 2 integrins) on peripheral blood leucocytes from patients with sarcoidosis (Shakoor & Hamblin, 1992). Here we have studied the expression of CD11/CD18 and CD29 (VLA beta 1 integrin) on the peripheral blood leucocytes of 10 TB patients by flow cytometry. The density (expressed as mean fluorescence intensity) of CD11b on monocytes and polymorphs was increased (P < 0.005), as was CD11c (P < 0.005) and CD18 (P < 0.05) on polymorphs. CD11a expression was significantly reduced on polymorphs (P < 0.05). No differences were found in the expression of CD29, the percentages of cells expressing any molecule and, in contrast to sarcoidosis, the density of any molecule on lymphocytes. Although the cytokine tumour necrosis factor (TNF) has been implicated in the process of up-regulation, an ELISA for TNF failed to detect significant levels in plasma. The results suggest increased peripheral phagocyte CD11/CD18 expression is a feature of TB, which may contribute to the pathological processes involved.     

Hemin,a heme oxygenase substrate analog,inhibits the cell surface expression of CD11b and CD66b on human neutrophils

BACKGROUND: Neutrophils are signaled to sites of infection and inflammation by different chemotactic stimuli. In order to reach the airways they have to adhere to, and then migrate through, the endothelium of pulmonary vessels. Carbon monoxide (CO) is a gaseous mediator, endogenously produced in the human airways. Increased CO production has been demonstrated during airway inflammation and CO as well as hemin, a substrate for CO producing enzymes, has been shown to affect neutrophil migration. Our objective was to investigate if the neutrophil cell surface expression of CD11b, CD66b and CD63 was changed during intermittent allergic rhinitis and to establish whether CO could affect the expression of these markers of cellular activation. METHODS: Blood from 10 healthy volunteers was drawn and incubated with different concentrations of hemin. Blood from 12 other healthy volunteers and from 12 patients with intermittent allergic rhinitis was also drawn during grass pollen season. Neutrophils were then isolated from all these three sets, and their expression of CD antigens measured using flow cytometry. RESULTS: Patients with symptomatic intermittent allergic rhinitis exhibited lower levels of CD11b and CD66b on the neutrophil cell surface. Incubation with hemin decreased the expression of CD11b and CD66b. CD63 was generally weakly expressed and not significantly affected by hemin incubation. CONCLUSION: Our results demonstrate that expressions of neutrophil cell surface glycoproteins are changed during the season in patents with intermittent allergic rhinitis and that hemin, a substrate for CO production, may act as an inhibitor of neutrophil activation. This indicates a possible role for CO in the immune defense system.     

A novel point mutation in CD18 causing the expression of dysfunctional CD11/CD18 leucocyte integrins in a patient with leucocyte adhesion deficiency (LAD)

Leucocyte adhesion deficiency type 1 (LAD-1) is characterized by the incapacity of leucocytes to carry out their adhesion functions via their CD11/CD18 antigens, which are also referred to as the leucocyte integrins. The patients generally suffer from poor wound healing and recurrent bacterial and fungal infections. In severe cases, the infections are often systemic and life-threatening. A LAD patient (AW) of moderate phenotype has been identified but, unlike most other cases, the level of CD11/CD18 antigens on her leucocytes are uncharacteristically high for a LAD patient. Molecular analysis revealed that she is a compound heterozygote for CD18 mutations. She has inherited a D231H mutation from her father and a G284S mutation from her mother. By transfection studies, it was established that the G284S mutation does not support CD11/CD18 antigen expression on the cell surface. In contrast, the D231H mutation does not affect CD18 forming integrin heterodimers with the CD11 antigens on the cell surface. However, the expressed integrins with the D231H mutation are not adhesive to ligands.     

Mouse CD23 regulates monocyte activation through an interaction with the adhesion molecule CD11b/CD18

CD23 is expressed on a variety of hemopoietic cells. Recently, we have reported that blocking CD23 interactions in a murine model of arthritis resulted in a marked improvement of disease severity. Here, we demonstrate that CD11b, the α chain of the β₂ integrin adhesion molecule complex CD11b/CD18 expressed on monocytes interacts with CD23. Using a recombinant fusion protein (ZZ-CD23), murine CD23 was shown to bind to peritoneal macrophages and peripheral blood cells isolated from mice as well as the murine macrophage cell line, RAW. The interactions between mouse ZZ-CD23 and CD11b/CD 18-expressing cells were significantly inhibited by anti-CD11b monoclonal antibodies. A functional consequence was then demonstrated by inducing an up-regulation of interleukin-6 (IL-6) production following ZZ-CD23 incubation with monocytes. The addition of Fab fragments generated from the monoclonal antibody CD11b impaired this cytokine production by 50%. Interestingly, a positive autocrine loop was identified as IL-6 was shown to increase CD23 binding to macrophages. These results demonstrate that similar to findings using human cells, murine CD23 binds to the surface adhesion molecule, CD11b, and these interactions regulate biological activites of murine myeloid cells.     

Canine leukocyte cell adhesion molecules (LeuCAMs): Characterization of the CD11/CD18 family

The canine LeuCAM (CD11/CD18) family is characterized using a panel of newly developed anti-canine LeuCAM monoclonal antibodies (mAb) as well as a cross-reactive anti-human CD11b mAb. The new anti-canine LeuCAM mAb were developed by immunizing mice with purified canine CD11/CD18 antigen derived from an anti-canine CD18 affinity column. Six mAb with reactivity to the canine LeuCAM family were cloned and characterized. These 6 included a new anti-canine CD18 mAb, 2 anti-canine CD11a mAb, 2 anti-canine CD11c mAb, and a mAb which recognizes a novel canine CD11/CD18-related macrophage antigen, designated LeuCAMmac, which is described in a separate report. The cell and tissue distribution of canine LeuCAMs was found to be similar to that reported in other species with a few notable exceptions. One prominent exception was the finding that canine CD11c, in contrast to human CD11c, was much more widely distributed on dendritic cells than it was on tissue macrophages. The molecular organization of the canine LeuCAM family was also found to be similar to that reported in other species, with the possible exception of the canine CD11b alpha chain, which may exist as several different species. It is anticipated that these anti-canine LeuCAM mAb will prove useful in immunophenotypic studies of canine hemolymphatic system neoplasia.