Effects of laboratory versus field exercise on leukocyte subsets and cell adhesion molecule expression in children

In adults, exercise is a powerful and natural stimulator of immune cells and adhesion molecules. Far less is known about these exercise responses during childhood and whether or not exercise in real-life activities of healthy children might influence immune responses. We compared laboratory exercise (10×2 min periods of heavy, constant intensity, cycle ergometer exercise with 1 min rests between exercise in nine subjects, aged 9–15 years) with field exercise (90 min soccer practice in nine different subjects, aged 9–11 years). Blood was sampled before both protocols, 5 min after the 30 min laboratory protocol, and 10–15 min after the 90 min field protocol. Both field and laboratory exercise protocols led to significant (P<0.05) increases in granulocytes, monocytes, and all lymphocyte subpopulations. The mean (SEM) increases were similar for the two protocols except for the significantly greater increase in laboratory compared with field protocols for natural killer cells [142 (39)% vs 12 (16)%, P<0.001] and monocytes [64 (22)% vs 32 (19)%, P<0.001]. Both protocols significantly influenced adhesion molecules (such as CD54) which have not been previously studied in children. However, the adhesion molecule CD8⁺CD62L^– increased to a significantly (P<0.001) greater extent in the laboratory [101 (25)%] versus field [34 (25)%] protocol. Finally, the density of CD62L on lymphocytes significantly decreased with laboratory exercise but showed no change in the field protocol [–20 (3)% vs –3 (3)%, P<0.001]. The rapid and substantial immune response in both laboratory and field protocols suggests that exercise stimulation of the immune system occurs commonly in the real lives of children and may play a role in their overall immune status. Electronic Publication     

Differential cell adhesion molecule expression and lymphocyte mobilisation during prolonged aerobic exercise

This study was undertaken to determine the cell adhesion molecule profile of CD4⁺, CD8^hi and CD56⁺ lymphocytes, which are mobilised to and from the peripheral blood during and after prolonged aerobic exercise. Ten healthy males (21–35 years old) were tested on two occasions, separated by at least 14 days. On the first occasion, subjects were examined in a rested state but did not exercise. On the second occasion, the same subjects were examined at the same time of day before, during and after 2 h of exercise at 65% of peak oxygen consumption. Blood samples obtained at rest (t ₀), during (at 0.5, 1, 1.5 and 2 h, t _0.5, t ₁, t _1.5 and t ₂, respectively) and after (at 4 and 24 h, t ₄ and t ₂₄, respectively) exercise were analysed by two-colour flow cytometry for CD4⁺, CD8^hi and CD56⁺ cell surface expression, and density of CD62L, CD49d and CD11a. At t ₂, circulating concentrations of CD56⁺, CD8^hi and CD4⁺ lymphocytes had increased (P<0.05) by 330%, 105% and 30%, respectively. The majority of CD4⁺, CD8^hi and CD56⁺ lymphocytes mobilised to the blood at t ₂ were CD62L^– and CD11a^hi, although populations of CD4⁺ and CD56⁺ cells that expressed CD62L⁺ and CD11a^lo were also mobilised. Changes in subset concentrations at t _0.5 were positively associated (r=0.63; P<0.01) with their corresponding mean surface density of CD11a at t ₀. Our findings suggest that the differential mobilisation of lymphocytes during prolonged aerobic exercise is linked to the surface expression of CD11a (i.e. lymphocyte-function-associated antigen-1). However, mechanisms unrelated to CD11a expression also appear to be involved. Electronic Publication     

Dendrite-associated cell adhesion molecule, telencephalin, promotes neurite outgrowth in mouse embryo

Telencephalin (TLCN) is a dendrite-associated cell adhesion molecule expressed by neurons within the telencephalon. It belongs to the intercellular adhesion molecule subgroup of the immunoglobulin superfamily. To examine a neurite outgrowth-promoting activity, neurons dissociated from mouse embryos were cultured on the substrate of recombinant mouse TLCN protein. Hippocampal neurons extended multiple neurites on TLCN. The neurite outgrowth on TLCN was suppressed by an anti-TLCN antibody. Non-telencephalic neurons also extended neurites on TLCN. These results demonstrate a neurite outgrowth-promoting activity of TLCN and suggest that both telencephalic and non-telencephalic neurons express TLCN counter-receptor(s) which is coupled to the neurite outgrowth.     

Murine hepatic microvascular adhesion molecule expression is inducible and has a zonal distribution

The structural and functional heterogeneity of hepatocytes and non-parenchymal cells across the liver lobule or acinus has been well documented. The geographic distribution and potential for induced expression of adhesion molecules on murine hepatic microvascular cells has not been reported, although these molecules are able to influence the metastatic outcome of intravascular cancer cells. We have postulated that the expression of adhesion molecules on these cells is susceptible to regulation by environmental factors and that these molecules have a zonal distribution across the acinus. To test this hypothesis, we injected C57BL/6 mice with bacterial lipopolysaccharide, 1 g/g body weight, i.p. At various time points (0–48 h) after stimulation, liver tissue sections were prepared for immunohistochemistry. Confocal microscopy was used to detect the expression of vascular cell adhesion molecule-1 (VCAM-1), E-selectin, intercellular adhesion molecule-1 (ICAM-1) and v integrin. The expression patterns were quantitatively measured by histomorphometry. Under basal conditions, ICAM-1 was weakly expressed in terminal portal veins while minimal VCAM-1 and no E-selectin were detected. Following stimulation with lipopolysaccharide, VCAM-1 and E-selectin were expressed on the endothelium of terminal portal veins and on sinusoidal lining cells with significantly stronger expression in the periportal zone than midzone. VCAM-1 expression peaked at 4 h and decreased gradually by 48 h. E-selectin peaked at 2 h and disappeared by 12 h after stimulation. ICAM-1 expression showed a much stronger and more uniform expression across the acinus with the peak reached by 4 h and sustained for longer than 48 h after lipopolysaccharide administration. The v integrin was not detected under basal conditions or after lipopolysaccharide stimulation. Expression of all these adhesion molecules (ICAM-1, VCAM-1, E-selectin and v integrin) was induced by growth of B16F1 melanoma cells in the peritoneal cavity of the mouse. These results support the hypotheses that expression of microvascular adhesion molecules in the mouse liver is susceptible to regulation by environmental stimuli and has a zonal heterogeneity across the acinus.     

慢性阻塞性肺病患者细胞粘附分子表达研究

对慢性阻塞性肺病（ＣＯＰＤ）患者外周血单个核细胞（ＰＢＭＣｓ）表面的ＣＤ１１ａ／ＣＤ１８，ＣＤ１１ｂ／ＣＤ１８（ＡＰＡＡＰ法）及ＣＤ４４（流式细胞分析法）粘附分子表达及血浆中可溶怀Ｅ．Ｐ．－选择素水平进行检测。结果：ＣＰＯＤ急性加重期患者ＰＢＭＣｓ表面ＣＤ１１ａ、ＣＤ１１ｂ及ＣＤ４４分子表达明显增高，血浆中可溶性Ｅ．Ｐ－选择素水平亦显著增高，与正常人及缓解期患者相比均有显著性差异。综合治疗可使血浆     

细胞粘附分子在狼疮性肾炎及膜增殖性肾炎肾组织中的表达

用免疫组化方法和计算机图像分析系统，对２５例狼疮性肾炎（ＬＮ）ＩＶ型及１５例膜增殖性肾炎（ＭＰＧＮ）Ｉ型患者肾组织内ＩＣＡＭ－１和ＶＣＡＭ－１的表达进行了定量研究。结果显示：ＭＰＧＮ及ＬＮ患者肾组织中ＩＣＡＭ－１及ＶＣＡＭ－１表达均显著增加，ＬＮ患者ＩｃＡＭ－１在肾小球内皮细胞表达最强，且与内皮细胞增殖程度显著正相关。     

The intercellular adhesion molecule (ICAM) family of proteins

Macromolecular adhesive associations between cells are important for transmitting spatial and temporal information that is critical for immune system function. One such group of proteins, the intercellular adhesion molecules (ICAMs), has grown as newly identified members are revealed. In addition, the functions of the ICAMs, in general, have begun to be better understood, including intracellular signaling events. This information has led to the design of novel therapeutic agents that may prove effective in a variety of disease states.     

Ontogeny and induction of adhesion molecule expression in human fetal intestine.

In this study we examined the distribution of the adhesion molecules ICAM-1, VCAM-1 and E-selectin in human fetal intestine, to determine whether they may have a role in the development of gut-associated lymphoid tissue. Secondly, we studied the tempo of induction of these molecules after T cell activation in explants of human fetal intestine cultured in vitro. In the fetus from 11 to 20 weeks gestation, endothelial expression of ICAM-1 and diffuse staining of VCAM-1 was observed in the lamina propria. In contrast, there was intense expression of ICAM-1 and VCAM-1 in the developing Peyer's patches, suggesting that these molecules may be involved in the accumulation or organization of lymphoid tissue in the gut. After T cell activation in fetal intestinal explants, the expression of ICAM-1 and VCAM-1 was increased on most endothelial cells, leucocytes, and stromal cells in the lamina propria. Expression was maintained for at least 4 days. In contrast, the induction of E-selectin was rapid, and the expression was transient, despite the continuing presence of activated T cells and macrophages. This suggests that other factors are required to prevent the down-regulation of E-selectin to maintain the sustained expression sometimes observed in vivo.     

Immunolocalization of the cellular adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and platelet endothelial cell adhesion molecule (PECAM), in human edometrium throughout the menstrual cycle

In the present study the presence and distribution of cellularadhesion molecules involved in leukocyte binding were investigatedin human endometrium. Endometrial biopsies (n = 45) were collectedfrom women at all stages of normal menstrual cycles. Consecutivecryostat sections of endometrium were immunostained with monoclonalantibodies to intercellular adhesion molecule-1 (ICAM-1) andplatelet endothelial cell adhesion molecule (PECAM) and haematoxylinand eosin. Primary antibody binding was visualized using a streptavidin–biotinsystem. Strong staining for PECAM was observed in endothelialcells of all vessel types and in focal areas of stroma includingsingle cells, small clusters and larger aggregates of cells.At menstruation, however, almost the entire stroma stained forPECAM which was temporally related to a massive influx of leukocytes.ICAM-1 staining, which was consistently less intense than PECAMstaining, was detected in vascular endothelial cells duringthe cycle, reaching a peak at menstruation. Unlike PECAM, ICAM-1staining did not occur consistently across all vessel types.Stromal staining for ICAM-1 was rare except at menstruation,when almost the entire stroma showed positive staining for ICAM-1.No glandular or luminal epithelial staining was detected foreither PECAM or ICAM-1. This study demonstrates that PECAM andICAM-1 are expressed on endothelial cells of veins, arteriolesand capillaries, and stromal cells within human endometrium.     

Fractalkine和CX3CR1━新发现的一对与白细胞趋化和粘附功能密切相关的分子

趋化因子超家族由许多成员组成 ,以往根据趋化因子结构域中的保守半胱氨酸的数目和位置将其分为ＣＸＣ (α)、ＣＣ ( β)和Ｃ (γ) 3型 ,最近又有学者[1 ] 发现了一种新的趋化因子 ,其趋化因子结构域具有ＣＸ3Ｃ基序 ,因此被称为第四型或δ型趋化因子。其首位成员被称为ｆｒａｃｔａｌｋｉｎｅ。Ｆｒａｃｔａｌｋｉｎｅ分子是Ｂａｚａｎ等[1 ] 经Ｂｌａｓｔ分析法从美国国家生物技术信息中心 (ｎａｔｉｏｎａｌｃｅｎｔｅｒｆｏｒｂｉｏｔｅｃｈｎｏｌｏｇｙｉｎｆｏｒｍａｔｉｏｎ ,ＮＣＢＩ)的表达序列标签 (ｅｘ ｐｒｅｓｓｅｄｓｅｑｕ…     

Lack of stimulatory effect of dienogest on the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 by endothelial cell as compared with other synthetic progestins

Objectives: Monocyte adhesion to endothelial cells is an important initial event at the onset of atherosclerosis. It is partially mediated by the expression of adhesion molecules on the endothelial cell surface. While estrogens inhibit the development of atherosclerosis, the effect of co-administered progestin remains controversial. We examined the effect of progestins on cytokine-stimulated human umbilical venous endothelial cell (HUVEC) expression of adhesion molecules. Methods: In HUVECs, mRNA expression of progesterone receptors (PRs) and androgen receptors (AR) was determined by RT-PCR. HUVECs were stimulated by interleukin-1β (IL-1β) for 24 h with or without various steroids, and then the cell-surface expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was semiquantified by ELISA. Results: In all preparations of HUVECs used in this study, RT-PCR confirmed mRNA expression of both isoforms of PR, PR-A and PR-B, as well as AR. Addition of progesterone (10⁻¹⁰–10⁻⁷ M) or dienogest (DNG) (10⁻¹⁰–10⁻⁸ M) did not affect IL-1β-stimulated ICAM-1 or VCAM-1 expression. In contrast, medroxyprogesterone acetate, norethindrone acetate and levonorgestrel (10⁻¹⁰–10⁻⁸ M) dose-dependently increased cell adhesion molecules. The progestin-induced increase was blocked by the concomitant addition of mifepristone, a PR antagonist, but not by hydroxyflutamide, an AR antagonist, indicating that the progestin stimulation was mediated predominantly via PR. Conclusions: These results suggest that DNG, unlike other synthetic progestins, lacks stimulation of cell adhesion molecules. For the prevention of atherosclerosis, estrogen in combination with DNG may be a suitable regimen in hormone replacement therapy in postmenopausal women.     

肿瘤细胞癌胚抗原的细胞粘附活性研究

本实验对肿瘤细胞系的癌胚抗原(CEA)细胞粘附活性进行了研究。CEA 阳性肿瘤细胞系 LoVo及 HeLa 的单细胞悬液在37℃,RPMI1640全培养基中可以相互粘附而各自形成大小不等的细胞凝团。这种细胞间的粘附不仅可以被抗 CEA 多克隆抗体特异性阻断,而且受到外加 CEA 抗原的竞争抑制。细胞粘附试验也获得了同样结果。实验提示,CEA 为一粘附分子。     

Comparative study of adhesion molecule expression in cultured human macro- and microvascular endothelial cells

Culture systems as models for disease are only valid as long as they are comparable to in vivo conditions. The phenotype of cultured endothelial cells (ECs) has only been sporadically compared to the corresponding phenotype in vivo. Thus, we compared by immunolocalization the endothelial expression of ICAM-1, VCAM, and E-selectin in vivo in stimulated/unstimulated human umbilical vein endothelial cells (HUVEC) as a model for macrovascular ECs and stimulated/unstimulated HPMEC (human pulmonary microvessel endothelial cells) as a model for pulmonary microvascular ECs with that in human lungs in vivo (normal and ARDS). Proinflammatory stimuli in vitro were used to stimulate conditions relevant for ARDS. ICAM-1 expression in stimulated HUVEC/HPMEC correlated well with in vivo expression (macro- and microvessels). For E-selectin, the staining pattern in macro/microvessels correlated moderately with unstimulated and well with stimulated HUVEC/HPMEC. For VCAM a good correlation was found for stimulated/unstimulated HUVEC and unstimulated HPMEC. The expression patterns in stimulated HUVEC corresponded well for all three molecules with those in vivo. Thus, the expression patterns in vitro are only partially transferable to in vivo conditions. The study suggests that E-selectin- and VCAM-coated beads could potentially serve in the isolation process of arteriolar and venular ECs.     

Time-course of adhesion molecule expression in rectal mucosa of gluten-sensitive subjects after gluten challenge.

Adhesive interactions between endothelium and circulating cells, such as monocytes, neutrophils and lymphocytes, are crucial for localizing the inflammatory response. We investigated the inflammatory response of rectal mucosa to local gluten challenge as a dynamic model of antigen-induced tissue injury, during which the expression of adhesion molecules on leucocytes and endothelial cells could be sequentially observed. Expression of ELAM-1, ICAM-1 and VCAM-1 was monitored in 10 treated and eight untreated patients with gluten sensitivity (coeliac disease), and in five disease controls for up to 4 h (short challenge), while a further seven treated coeliacs were monitored for up to 24 h (long challenge) following rectal gluten challenge. In the former, the expression of VCAM-1 and ELAM-1 was significantly raised 4 h after gluten challenge compared with controls. VCAM-1 and ELAM-1 expression was also increased in mucosae of treated patients, but to a lesser extent. VCAM-1 expression continued to increase for up to 24 h after gluten, while ELAM-1 had begun to wane by 4 h, reaching basal levels by 24 h. In contrast, the expression of ICAM-1 did not change in any of the disease groups studied. These findings relate to significant increases in lymphocytes (CD3+ cells) after 8 h, and neutrophils (CD15+ cells) after 4 h in the lamina propria. This approach has permitted novel studies of the inflammatory response to a defined antigen in sensitized (gluten-sensitive) human patients.     

雾化吸入灭活草分枝杆菌降低哮喘小鼠核因子κB及细胞间黏附分子1的表达

 目的:研究雾化吸入灭活草分枝杆菌对支气管哮喘小鼠气道炎症,以及哮喘肺组织中核因子κB(NF-κB)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1)的影响,探讨雾化吸入灭活草分枝杆菌防治哮喘的机制。方法:将24只雄性BALB/c小鼠按随机数字表法分为3组,每组8只：正常对照组（A）、哮喘模型组（B）和治疗组（C）。以鸡卵清蛋白致敏制造小鼠支气管哮喘模型。C组在激发后给予雾化吸入灭活草分枝杆菌治疗5 d,每天1次。各组动物处死后提取肺组织和支气管肺泡灌洗液（BALF）。进行病理HE染色及AB-PAS染色观察气道炎症浸润及黏液分泌情况,并行病理半定量分析。对BALF中炎症细胞进行分类计数。实时荧光定量PCR检测肺组织NF-κB、ICAM-1和VCAM-1的mRNA表达水平。结果:治疗组嗜酸性粒细胞比例低于模型组(P<0.05),气道炎症病变及黏液分泌情况较模型组减轻(P<0.05, P<0.01)。哮喘模型组的肺组织中NF-κB mRNA含量与正常组相比显著升高(P<0.01),而治疗组肺组织中的NF-κB mRNA 含量明显低于模型组(P<0.05);模型组的ICAM-1 mRNA 水平比正常组高(P<0.05),但治疗后明显降低(P<0.01);VCAM-1的 mRNA水平在各组间无显著差异。相关性检验发现小鼠肺组织中VCAM-1的mRNA与ICAM-1的mRNA呈明显正相关(r=0.84,P<0.01),但NF-κB的mRNA与ICAM-1的mRNA、VCAM-1的mRNA无明显相关性(均P>0.05)。结论:雾化吸入草分枝杆菌对支气管哮喘小鼠气道炎症及黏液分泌有抑制作用;NF-κB参与哮喘发病过程,雾化吸入灭活草分枝杆菌降低哮喘小鼠的NF-κB水平。同时雾化吸入灭活草分枝杆菌可降低黏附分子尤其是ICAM-1的表达,是其控制炎症的另一个重要机制。     

Bioimmunoassays for proinflammatory cytokines involving cytokine-induced cellular adhesion molecule expression in human glioblastoma cell lines

Twelve human glioblastoma/astrocytoma cell lines were tested for cellular adhesion molecule expression following cytokine induction in order to identify a cell line that would be suitable for functional cytokine bioimmunoassays. Many of the glioblastoma/astrocytoma cell lines were shown to inducibly express intercellular adhesion molecule-1 (ICAM-1, CD54) and vascular cell adhesion molecule-1 (VCAM-1) following stimulation with interleukin-1 (IL-1), interleukin-1β (IL-1β), tumour necrosis factor- (TNF-), tumour necrosis factor-β (TNF-β), and interferon-γ (IFN-γ), but not with any of the several other cytokines tested. The cell line U-138MG, a human glioblastoma-derived line, was the most sensitive one to IL-1/β, TNF-/β and IFN-γ for ICAM-1 expression, comparing well with proinflammatory cytokine-induced ICAM-1 expression in the endothelial cell hybrid EA-hy926 line, and was shown to be useful for the functional assay of the biological potencies of these individual cytokines. Such bioimmunoassays, which are developed by routine ELISA techniques, should provide valuable alternatives to existing bioassays for these cytokines.     

Up-regulation of the granulocyte adhesion molecule Mac-1 by autoantibodies in autoimmune vasculitis

The characteristic finding of autoantibodies in patients with vasculitis has raised the possibility that these antibodies play a role in the pathogenesis of the disease. The expression of adhesion molecules (AM) on leucocytes and endothelial cells is believed to be integral to the development of vasculitis. We therefore investigated the effect of sera, positive for anti-neutrophil cytoplasmic antibodies (ANCA) or anti-nuclear antibodies (ANA) from patients with vasculitis, on granulocyte expression of the adhesion molecule Mac-1 (CD11b). Autoantibody-positive sera from 15 out of 35 patients with vasculitis stimulated an up-regulation of Mac-1 on granulocytes. In most cases this effect was reproduced by the autoantibody-positive purified IgG fraction. Autoantibody-negative samples did not stimulate AM up-regulation. Of interest, preincubation of sera with purified antigens did not inhibit AM up-regulation by the autoantibody samples. Blocking the Fc receptors on granulocytes did result in a decrease of Mac-1 up-regulation, but this trend was not statistically significant. These results suggest that both ANCA and ANA have the capacity to up-regulate granulocyte AM expression, and that while Fc interaction with granulocyte Fc receptors is important, it is not the only mechanism whereby such autoantibodies activate cells.     

Synovial expression of cell adhesion molecules in polymyalgia rheumatica

Polymyalgia rheumatica (PMR) is a common disorder of the elderly: the pathogenesis of the syndrome is still debated, though active synovitis of the shoulder has recently been confirmed. To investigate the pathogenesis of this synovitis we evaluated cell adhesion molecule (CAM) expression in shoulder synovial tissue from patients with PMR, correlated synovial expression with the serum levels of soluble forms, and assessed the changes associated with corticosteroid treatment. Arthroscopic synovial biopsies were obtained from 12 untreated and seven corticosteroid (CS)-treated cases. CAM expression was evaluated by MoAb staining on frozen sections and computerized image analysis. Soluble CAM were quantified by ELISA. Endothelial cells expressed intercellular adhesion molecule-1 (ICAM-l), E- and P-selectins. Infiltrating cells were ICAM-1 and β₁-integrin-positive, while L-selectin expression was limited to intravascular leucocytes. Synovial lining cells strongly expressed vascular cell adhesion molecule-1 (VCAM-1), and less intensely ICAM-1. Only the soluble form of ICAM-1 (sICAM-1) was elevated in untreated patients. CS treatment was associated with a decrease in ICAM-1, VCAM-1 and E- and P-selectin expression. sICAM-1 levels were in the normal range in treated patients. VLA-5 and 6 expression was widely distributed among cell types, and was not CS-sensitive. Active shoulder synovitis is associated with different CAM expression in PMR. ICAM-l expression is widely distributed and correlates with elevated levels of the soluble form; it is significantly lower in CS-treated asymptomatic cases.     

Alterations in blood leucocyte adhesion molecule profiles in HIV-1 infection.

CD4 and CD8 lymphocyte numbers in the gut lamina propria are grossly altered in HIV-1 infection, out of proportion to alterations in the circulation. Such alterations in lymphocyte counts in the tissues may be due to altered leucocyte migration from the blood. One factor affecting leucocyte migration is adhesion molecule expression. Levels of adhesion molecule expression on peripheral CD4 and CD8 lymphocytes, monocytes and neutrophils from HIV-1-infected (AIDS and non-AIDS) and low-risk control individuals were compared. CD11a, CD62L, CD44, CD49d and beta7 integrin expression were examined by FACS analysis of fresh whole blood. Significant alterations in adhesion molecule expression were detected in HIV infection. The most striking alterations were observed in the CD8 lymphocyte population. CD11a expression was increased and CD62L and CD44 decreased. The CD4 lymphocyte population followed a similar, though less striking, pattern of alteration in adhesion molecule expression. Neutrophils displayed significantly reduced expression of both CD11a and CD62L, but only after onset of AIDS. Monocytes from infected individuals without AIDS displayed a different pattern of altered adhesion molecule expression compared with individuals with AIDS. These findings suggest that in HIV infection, leucocyte functions, such as migration, which require adhesion molecules are abnormal.