Cross-dressed CD8α+/CD103+ dendritic cells prime CD8+ T cells following vaccination

Activation of naïve cluster of differentiation (CD)8⁺ cytotoxic T lymphocytes (CTLs) is a tightly regulated process, and specific dendritic cell (DC) subsets are typically required to activate naive CTLs. Potential pathways for antigen presentation leading to CD8⁺ T-cell priming include direct presentation, cross-presentation, and cross-dressing. To distinguish between these pathways, we designed single-chain trimer (SCT) peptide–MHC class I complexes that can be recognized as intact molecules but cannot deliver antigen to MHC through conventional antigen processing. We demonstrate that cross-dressing is a robust pathway of antigen presentation following vaccination, capable of efficiently activating both naïve and memory CD8⁺ T cells and requires CD8α⁺/CD103⁺ DCs. Significantly, immune responses induced exclusively by cross-dressing were as strong as those induced exclusively through cross-presentation. Thus, cross-dressing is an important pathway of antigen presentation, with important implications for the study of CD8⁺ T-cell responses to viral infection, tumors, and vaccines.Professional antigen-presenting cells (APCs) are typically required to activate naïve cluster of differentiation (CD)8⁺ T cells, either by direct priming or cross-priming. In direct priming, infected (viral infection) or directly transfected (DNA vaccination) APCs synthesize the foreign antigen and use endogenous MHC class I pathways of antigen presentation to present antigen and prime CD8⁺ T cells. In cross-priming, APCs are able to capture, process, and present exogenous antigen onto MHC class I molecules through a process known as cross-presentation (1). Cross-priming has been shown to be an essential pathway for immunity to many viral infections and tumors. Although the pathways that lead to cross-presentation remain incompletely understood, increasing evidence suggests that only certain dendritic cell (DC) subsets are efficient in this process.Cross-dressing involves the transfer of intact MHC class I/peptide complexes between cells without the requirement for further processing, representing an alternative pathway of indirect antigen presentation (2, 3). Although cross-dressed DCs can activate memory CD8⁺ T cells following viral infection in vivo (4), it remains unclear whether cross-dressing can prime naïve CD8⁺ T-cell responses, what DC subtypes are required to prime CD8⁺ T cells by cross-dressing, and how robust this pathway is compared with traditional pathways of indirect antigen presentation. These questions must be addressed before the physiologic relevance of cross-dressing can be evaluated in context.To address these questions, we have taken advantage of Batf3-deficient mice and engineered MHC class I single chain trimer (SCT) constructs. Batf3^−/− mice have a selective loss of CD8α⁺ and CD103⁺ DCs, without abnormalities in other hematopoietic cell types or architecture (5). DCs from Batf3^−/− mice are deficient in cross-presentation, and cytotoxic T lymphocyte (CTL) responses to viral infection and syngeneic tumors are impaired in Batf3^−/− mice. Thus, Batf3^−/− mice represent a valuable model system to study cross-presentation, cross-dressing, and the role of CD8α⁺/CD103⁺ DCs following DNA or cellular vaccination. We have previously engineered completely assembled MHC class I SCT whereby all three components of the complex (heavy chain, β₂m, and peptide) are attached by flexible linkers (6). Through progressive molecular engineering, even peptides with low binding affinities can be successfully anchored in the peptide binding groove by a disulfide trap between the first linker and the heavy chain (7–9). Using these experimental tools, we demonstrate that cross-dressing is a robust pathway of antigen presentation following DNA and cellular vaccination, capable of priming naïve and memory CD8⁺ T cells. In addition, we demonstrate that CD8α⁺/CD103⁺ DCs are required to prime CTLs by cross-dressing.     

Circulating Bone Marrow-Derived CD45−/CD34+/CD133+/VEGF+ Endothelial Progenitor Cells in Adults with Crohn’s Disease

Background

Circulating endothelial progenitor cells (EPCs) are bone marrow-derived stem cells able to migrate to sites of damaged endothelium and differentiate into endothelial cells. Altered EPC level and function have been described in various inflammatory diseases and have been shown to augment vasculogenesis in murine models. Previous studies of EPC in the context of Crohn’s disease (CD) have yielded conflicting results.

Aim

To determine whether the circulating levels of EPCs are changed in the context of CD.

Methods

CD patients and healthy controls were recruited. Disease activity was assessed by CDAI. Peripheral blood mononuclear cells were isolated and EPC numbers evaluated by FACS analysis using anti-CD34, anti-VEGF receptor-2, anti-CD133, and anti-CD45 markers.

Results

Eighty-three subjects, including 32 CD patients and 51 controls were recruited, including 19 (59.4 %) and 23 (45 %) males (p = 0.26), aged 34.8 ± 14.9 and 43.3 ± 18.5 years (p = 0.64), in cases and controls, respectively. Mean CDAI was 147 ± 97, disease duration was 12.7 ± 11.1 years, and 28 (87.5 %) were receiving biologics for a mean duration of 21.7 ± 16.8 months. The mean level of peripheral EPCs in CD patients was 0.050 ± 0.086 percent and 0.007 ± 0.013 % in controls (p < 0.01). There was no significant correlation between EPC levels and age (r = ?0.13, p = 0.47), CDAI (r = ?0.26, p = 0.15), disease duration (r = ?0.04, p = 0.84), or duration of treatment with biologics (r = 0.004, p = 0.99).

Conclusion

EPCs are elevated in patients with CD. Further studies are needed to examine the function of EPCs and their possible role as a marker of disease severity or therapeutic response.

     

慢性乙肝患者血清CD4+、CD8+T细胞及CD4+/CD8+水平的变化与干扰素疗效的关系

慢性乙型病毒性肝炎（CHB）的发生机理相当复杂,越来越多的研究证实,机体的免疫功能紊乱,尤其是T淋巴细胞亚群与慢性肝病的发生发展密切相关。而CHB患者细胞免疫状态可影响干扰素的疗效。本文就CHB患者血清CD4^＋、CD8^＋ T细胞及CD4^＋／CD8^＋水平的变化与干扰素的疗效做一综述。     

Increased CD4+CD16+ and CD4+CD56+ T cell subsets in Behçet's disease

Behçet's disease is a systemic vasculitis of unknown etiology. Various immune abnormalities have previously been shown in Behçet's disease. We investigated T lymphocyte subsets associated with cytotoxic activity and natural killer (NK) cells by flow cytometry in 37 patients with Behçet's disease, 38 healthy controls, and 17 diseased control patients. Compared to the healthy controls, CD4⁺CD16⁺ and CD4⁺CD56⁺ subsets were found to be higher in the Behçet's disease group as well as in the disease control group (CD4⁺CD16⁺: BD=5?±?3, DC=14?±?14, HC= 3?±?2, P=0.001; CD4⁺CD56⁺: BD=11?±?5, DC= 18?±?17, HC=8?±?6, P=0.01). CD8⁺CD16⁺ and CD8⁺CD56⁺ T cell subsets were at normal levels in Behçet's disease but found to be elevated in disease controls. Similarly, NK cells (CD16⁺CD56⁺) were high only in the disease control group. Significant increases in CD4⁺CD16⁺ and CD4⁺CD56⁺ cell subsets in Behçet's patients and disease controls suggest that T cell activation patterns of these subsets in Behçet's disease are similar to those in other inflammatory disorders.     

小鼠骨髓间充质干细胞SCA-1~+/CD45~+/CD31~+亚群归巢基因研究

目的:探讨小鼠骨髓间充质干细胞(mBMSCs)亚群SCA-1+/CD45+/CD31+归巢分子基础。方法:以小鼠心脏干细胞表面分化抗原检测mBMSCs后,以CD45、CD31为标准分选得到4个亚群。体外将4个亚群采用transwell小室,检测各组的归巢能力。分别将各亚群细胞注入心肌梗死48h模型。小鼠体内注射后48h、96h、7d处死小鼠取其心脏,完成小动物活体成像并检测其荧光强度。可见SCA-1+/CD45+/CD31+组归巢能力优于其他各组。而后采用基因芯片完成Agilent小鼠全基因4×44K芯片,从分子水平对SCA-1+/CD45+/CD31+亚群归巢能力进行探讨。结果:将SCA-1+/CD45+/CD31+亚群与其他亚群有关归巢基因的聚类分析及Network分析的结果比较可见,Dcx及MADCAM1基因为两结果交集,是有关归巢的关键基因。结论:mBMSCs为一多克隆的细胞群体。SCA-1+/CD45+/CD31+亚群归巢能力方面优于其他亚群;其分子机制在于Dcx及MADCAM1基因表达较其他亚组多见。     

急性脑梗死患者T淋巴细胞亚群CD4+、CD8+、CD4+/CD8+ 和C反应蛋白变化及其意义的研究

目的 探讨急性脑梗死患者T淋巴细胞亚群CD4^＋、CD8^＋、CD4^＋/CD8^＋和C-反应蛋白（CRP）变化及其意义。方法 应用流式细胞仪测定36例发病72 h内急性脑梗死患者及正常对照组血清中CD4^＋、CD8^＋、CD4^＋/CD8^＋及CRP水平,并根据神经功能缺损标准将脑梗死组分为轻、中、重三型,将脑梗死组根据年龄分为老年组和非老年组,分析CD4^＋、CD8^＋和CRP与脑梗死轻重的关系。结果 与正常对照组相比急性脑梗死组中、重型CD4^＋、CD4^＋/CD8^＋明显降低,CD8^＋、CRP明显升高,而轻型脑梗死组变化不明显;中、重型脑梗死随着病情加重CD4＋明显减低。结论 炎症反应参与了急性脑梗死的发病过程,急性脑梗死后患者免疫功能发生紊乱,并与其病情轻重有关。     

依托咪酯对老年外科手术患者CD3~+、CD4~+、CD4~+/CD8~+及炎症状态的影响

目的探讨依托咪酯对老年外科手术患者免疫指标CD3~+、CD4~+、CD4~+/CD8~+及炎症状态的影响。方法全身麻醉(全麻)老年手术患者125例随机单盲法分为观察组63例给予依托咪酯麻醉,对照组62例给予丙泊酚麻醉,观察两组手术一般情况,入室(T0)、诱导后(T1)、手术开始后10 min(T2)、术毕(T3)、术后24 h(T4)时免疫指标(CD3~+、CD4~+、CD8~+、CD4~+/CD8~+比值)及炎症指标肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-8、IL-10水平变化。结果两组手术一般资料无统计学差异(P>0.05),两组T0各项免疫指标差异无统计学意义(P>0.05),观察组T3免疫指标CD3~+、CD4~+均显著高于对照组(P<0.05),T1、T2、T3免疫指标CD3~+、CD4~+、CD8~+均显著高于对照组(P<0.05),两组T0各项炎症指标差异均无统计学意义(P>0.05),观察组T1、T2、T3各项炎症指标均显著高于T0(P<0.05),T4炎症指标IL-6、IL-8、IL-10显著高于T0(P<0.05),T2、T3各项炎症指标均显著高于对照组(P<0.05),T1炎症指标IL-6、IL-8显著高于对照组(P<0.05),T4炎症指标IL-8显著高于对照组(P<0.05),T4炎症指标IL-6、IL-8、IL-10显著高于同组T0(P<0.05)。结论依托咪酯对老年外科手术患者免疫功能影响较小,对炎症状态表现为抗炎状态。     

小鼠骨髓间充质干细胞SCA-1~+/CD45~+/CD31~+亚群抗凋亡能力的研究

目的:探讨小鼠骨髓间充质干细胞(mBMSCs)各亚群在心肌修复中抗凋亡能力。方法:以小鼠心肌干细胞表面分化抗原检测mBMSCs后以CD45、CD31为标准分选得到小鼠骨髓间充质干细胞4个亚群。体外将4个亚群采用无血清低氧(5%)培养,流式细胞仪检测各组的凋亡率。分别将细胞注入小鼠心梗模型体内,注射后48h完成心功能测定。结果:SCA-1+/CD45+/CD31+组抗凋亡能力最强,心脏彩超提示心功能恢复亦优于其他各组。结论:小鼠骨髓间充质干细胞SCA-1+/CD45+/CD31+组抗凋亡能力明显优于其他组,治疗后心功能恢复亦优于其他各组。     

小鼠骨髓间充质干细胞SCA-1~+/CD45~+/CD31~+亚群向心肌样细胞分化的分子基础

目的:探讨小鼠骨髓间充质干细胞(mBMSCs)亚群SCA-1~+/CD45~+/CD31~+向心肌样细胞分化的分子基础。方法:以小鼠心脏干细胞表面分化抗原检测mBMSCs后以CD45、CD31为标准分选得到4个亚群。备亚群和心肌细胞共培养后检测α-actin、Cx43、Desmin、cTnI的表达。并采用基因芯片完成Agilent小鼠全基因4*44K芯片从分子水平对备亚群和心肌细胞共培养结果进行探讨。结果:SCA-1~+/CD45~+/CD31~+亚群和心肌细胞共培养后更易表达出心肌细胞标志性分子。通过Agilent小鼠全基因4*44K芯片将SCA-1~+/CD45~+/CD31~+与其他组的心血管发育基因的聚类分析及Network结果共同比较可见两者无交集,以差异基因均为4倍表达为标准,因而以Network结果为主。可见:SETD2、NCL、EPOR、Rock2为感兴趣基因。结论:mBMSCs为多克隆的细胞群体。SCA-1~+/CD45~+/CD31~+在向心肌定向分化及改善心功能方面优于其他亚群,其分子机制在于SETD2、NCL、EPOR、Rock2基因表达较其他亚组多见。     

Hyperosmotic perfusion of the beating rat heart and the role of the Na+/K+/2CI− co-transporter and the Na+/H+ exchanger

The aim of the present study was to investigate the role of the Na⁺/K⁺/2CI⁻ co-transporter and the Na⁺/H⁺ exchanger on contractile function and electrolyte regulation during hyperosmotic perfusion of the heart. Langendorff perfused rat hearts were subjected to hyperosmolal perfusion in 10-min intervals. Perfusates were made hyperosmotic by adding mannitol to the buffer (370, 450 and 600 mOsmol/kg H₂O). Cardiac contractile function was monitored with a balloon in the left ventricle (LV) coupled to a pressure transducer. Cardiac effluent was sampled repeatedly throughout and after hyperosmotic perfusion and analyzed for content of Na⁺, K⁺ and CI⁻. All three hyperosmotic perfusates initially reduced LV developed pressure (LVDP), but for 370 and 450 mOsmol/kg H₂, LVDP recovered to baseline within 4 min of perfusion. With 600 mOsmol/kg H₂, LVDP recovered slowly and was 50% below baseline after 10 min of hyperosmotic perfusion. Inhibition of the Na⁺/H⁺ exchanger with 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and 3-methyl-sulfonyl-4-piperidinobenzoyl-guanidine methanesulfonate (HOE 694) abolished the recovery of LVDP to the 600 mOsmol/kg H₂ perfusate, whereas inhibition of the Na⁺/K⁺/2CI⁻ co-transporter had no impact on LVDP. Potassium was taken up by the heart during hyperosmotic perfusion and this uptake was significantly reduced with inhibition of the Na⁺/H⁺ exchanger. Intracellular pH was assessed with ³¹P magnetic resonance spectroscopy and hyperosmolality induced a significant alkalosis that was dependent upon the Na⁺/H⁺ exchanger. The rat heart responds to moderate elevations in osmolality with a transient reduction in contractile function, whereas an elevation of 300 mOsmol/kg H₂ persistently reduces contractile function. The Na⁺/H⁺ exchanger, but not the Na⁺/K⁺/2CI⁻ co-transporter, is of importance in contractile recovery and electrolyte regulation during hyperosmotic perfusion in the rat heart. Received: 10 December 1998, Returned for revision: 18 January 1999, Revision received: 1 July 1999, Accepted: 19 July 1999     

重症急性胰腺炎患者CD4+/CD8+细胞比值、IL-16的变化及意义

15例健康人和25例重症急性胰腺炎患者采静脉血,采用流式细胞仪检测CD4^＋、CD8^＋细胞百分比并计算CD4^＋/CD8^＋比值,酶联免疫吸附法检测血清白细胞介素（IL）-16水平.结果显示,与正常健康人相比,患者外周血CD4^＋细胞百分比、CD4^＋/CD8^＋比值明显降低（P＜0.05）,CD8^＋细胞百分比和血清IL-16水平显著增高（P＜0.05）,上述指标变化可能在胰腺炎病理过程中起重要作用.     

What is CD4+CD56+ malignancy and how should it be treated?

CD4+CD56+ malignancy is a rare neoplasm with a typical clinical pattern, an aggressive course and high early relapse rate despite good initial response to chemotherapy. In this review, the impact of different therapeutic approaches on clinical outcome has been studied. We evaluated 91 published cases and our own six patients in terms of clinical features, immunophenotype/cytogenetics and treatment outcome. Treatment was divided into four groups: (A) chemotherapy less intensive than CHOP; (B) CHOP and CHOP-like regimens; (C) therapy for acute leukemia; (D) allogeneic/autologous stem cell transplantation. The median overall survival was only 13 months for all patients. Patients with skin-restricted disease showed no difference in the overall survival from patients with advanced disease (17 and 12 months, respectively). Age >/=60 years was a negative prognostic factor. Age-adjusted analysis revealed improved survival after high-dose chemo/radiotherapy followed by allogeneic stem cell transplantation when performed in first complete remission. This therapeutic approach should be recommended for eligible patients with CD4+CD56+ malignancy. For older patients the best treatment option is still unknown.     

支气管肺泡灌洗液细胞分类及CD4+/CD8+对间质性肺疾病的诊断价值

目的探讨支气管肺泡灌洗液细胞分类及T细胞亚群测定对间质性肺疾病的诊断价值。方法选取我院收治的间质性肺疾病患者59例,行支气管肺泡灌洗及经支气管肺活检,测定灌洗液细胞分类及T细胞亚群CD_4~+/CD_8~+。结果 59例患者最终明确诊断52例,其中结节病16例,特发性肺间质纤维化(IPF) 5例,非特异性间质性肺炎(NSIP) 10例,隐源性机化性肺炎(COP) 11例,外源性过敏性肺炎(HP) 2例,嗜酸性粒细胞肺炎(EP) 2例。肺结核3例,血管炎1例。EP患者BALF嗜酸性粒细胞(EOS)大于25%; HP患者淋巴细胞比例高达70%; NSIP和结节病患者BALF中淋巴细胞比例增高,分别为34. 0±19. 56%和27. 50±18. 53%;结节病患者T细胞亚群CD_4~+/CD_8~+升高为5. 64±3. 90,而COP中降低为0. 95±0. 53。结论 BALF细胞分类及T细胞亚群CD_4~+/CD_8~+测定有助于间质性肺疾病的鉴别诊断。     

Demonstration of Low-Regulatory CD25High+CD4+ and High-Pro-inflammatory CD28−CD4+ T-Cell Subsets in Patients with Ulcerative Colitis: Modified by Selective Granulocyte and Monocyte Adsorption Apheresis

Low-CD25^High+CD4⁺, a subset of regulatory CD25⁺CD4⁺ T cells and high-inflammatory CD28⁻CD4⁺ T cells can exacerbate ulcerative colitis (UC). This study sought to investigate the frequency of CD25^High+CD4⁺ and CD28⁻CD4⁺ T cells in patients with UC and the changes in these cells during Adacolumn granulocyte and monocyte adsorption apheresis (GMA). Subjects were 12 patients with active UC, 11 with quiescent UC, and 14 healthy volunteers (HVs). The mean clinical activity index was 15.7 ± 2.2 in active UC and 4.5 ± 1.1 in quiescent UC. Peripheral blood samples were stained with CD4, CD25, and CD28 antibodies for flow cytometry. Patients with active UC received GMA and blood samples were examined before and after the first GMA session. Patients with active UC (P < 0.04) or quiescent UC (P < 0.02) had a higher percentage of CD28⁻D4⁺T cells compared with HVs, while the percentage of CD28⁺CD4⁺ T cells was lower in both UC groups compared with HVs (P = 0.03 and P < 0.02). Patients with active UC had a lower percentage of CD25^High+CD4⁺T cells compared with quiescent UC patients (P < 0.001). A significant increase in CD25^High+CD4⁺ T cells was associated with GMA (P < 0.03). Low CD25^High+CD4⁺ and high CD28⁻CD4⁺ are prominent features in UC. The increase in CD25^High+CD4⁺ T cells induced by GMA should contribute to improved immune function. Additional studies are warranted, since a low frequency of CD25^High+CD4⁺ ₋ and a high frequency of CD28⁻CD4⁺ ₋ expressing T cells might be a predictor of clinical response to GMA.     

CD95-mediated apoptosis in naïve, central and effector memory subsets of CD4+ and CD8+ T cells in aged humans

Aging is associated with a decrease in naïve (T_N) and central memory (T_CM), and an accumulation of effector memory (T_EM and T_EMRA) T cell subsets. Previously, we have demonstrated an increased sensitivity of T_N and T_CM CD4+ and CD8+ T cells in aging to TNF-α-induced apoptosis. In this investigation, we examined whether similar differential sensitivity is applicable to CD95-mediated apoptosis. We show that T_N and T_CM CD4+ and CD8+ T cells from aged subjects are significantly more sensitive to CD95-mediated apoptosis. Increased apoptosis is associated with increased activation of caspase-8 and caspase-3. Both caspase-8 and caspase-3 inhibitors blocked CD95-mediated apoptosis and activation of caspase-8 and caspase-3 in T_N and T_CM CD4+ and CD8+ T cells. No significant difference was observed in apoptosis or in activation of caspase-8 and caspase-3 in T_EM and T_EMRA CD4+ and CD8+ T cells between young and aged subjects; both populations were relatively and comparably resistant to CD95-mediated apoptosis and caspase activation. No correlation was observed between the sensitivity/resistance of any of the subsets of CD4+ or CD8+T cells to CD95-mediated apoptosis and the expression of CD95. Our data suggest that increased CD95-mediated apoptosis of T_N and T_CM CD8+ and CD4+ T cells may play a role in their decline in human aging.     

Dietary gluten triggers concomitant activation of CD4+ and CD8+ αβ T cells and γδ T cells in celiac disease

Celiac disease is an intestinal autoimmune disease driven by dietary gluten and gluten-specific CD4⁺ T-cell responses. In celiac patients on a gluten-free diet, exposure to gluten induces the appearance of gluten-specific CD4⁺ T cells with gut-homing potential in the peripheral blood. Here we show that gluten exposure also induces the appearance of activated, gut-homing CD8⁺ αβ and γδ T cells in the peripheral blood. Single-cell T-cell receptor sequence analysis indicates that both of these cell populations have highly focused T-cell receptor repertoires, indicating that their induction is antigen-driven. These results reveal a previously unappreciated role of antigen in the induction of CD8⁺ αβ and γδ T cells in celiac disease and demonstrate a coordinated response by all three of the major types of T cells. More broadly, these responses may parallel adaptive immune responses to viral pathogens and other systemic autoimmune diseases.     

系统性红斑狼疮患者外周血CD4+ CD8+和CD22+淋巴细胞活化分子CD69的表达

目的探讨系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL) T细胞(CD4+、CD8+)和B细胞(CD22+)活化分子CD69的表达.方法应用双染色流式细胞术检测CD4、CD8、和CD22细胞亚群CD69分子;在植物凝集素(PHA)刺激后20 h淋巴细胞亚群CD69分子的表达.结果①SLE患者PBMC的CD69分子活动期高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05).②进一步分析CD4+、CD8+和CD22+淋巴细胞亚群的CD69的表达,其中,SLE活动期患者CD4+细胞的CD69表达显著高于静止期(P＜0.001)和正常对照组(P＜0.01)的表达,SLE静止期患者与正常对照组CD69表达差异无显著性(P＞0.05);CD8+细胞活动期高于静止期患者(P＜0.05),其余组间差异无显著性(P＞0.05);CD22+B细胞各组间差异无显著性.③PHA刺激20 h后,CD4+、CD22+B细胞的CD69表达,活动期显著高于静止期患者和正常对照组(P＜0.01).结论 SLE患者外周血CD4+T细胞和CD22+B细胞存在着异常的活化,这种淋巴细胞的异常活化是SLE重要的发病机制之一.     

循环CD133+/KDR+内皮祖细胞水平与急性缺血性卒中患者转归的相关性

目的 探讨急性缺血性卒中患者循环CD133+/KDR+内皮祖细胞(endothelial progenitor cells,EPCs)水平与转归的关系.方法 纳入发病24 h内的首次急性缺血性卒中住院患者以及年龄和性别相匹配的健康体检者.收集患者人口统计学和临床资料.采用流式细胞术检测CD133+/KDR+EPCs水平.在发病后90 d时对所有患者进行随访,采用改良Rankin量表评价临床转归,0~2分定义为转归良好,>2分定义为转归不良.结果 共纳入连续126例缺血性卒中患者以及60例年龄和性别相匹配的健康体检者.在缺血性卒中患者中,大动脉粥样硬化(large artery atherosclerosis,LAA) 33例(26.19%),小动脉闭塞(small artery occlusion,SAO)74例(58.73%),心源性栓塞(cardioembolism,CE)19例(15.08%);82例(65.08%)转归良好,44例(34.92%)转归不良.LAA型(0.071%±0.018%)、CE型(0.068%±0.016%)和SAO型(0.118%±0.012%)患者基线循环EPCs数量均显著低于对照组(0.246%±0.052%;P均<0.05);CE型(P=0.028)和LAA型(P=0.037)均显著低于SAO型;CE型低于LAA型,但差异无统计学意义(P=0.762).转归不良组LAA型(40.91%对18.29%;χ2=7.577,P=0.006)和CE型(29.55%对7.32%;χ2=11.049,P=0.001)和心房颤动(29.55%对10.98%;χ2=6.582,P=0.009)患者的构成比以及年龄[(69.64±9.62)岁对 (61.12±7.31)岁;t=5.570,P<0.001]、基线NIHSS评分[(14.16±4.22)分对 (6.96±2.04)分;t=12.919,P<0.001]、基线收缩压[(176.06±13.42)mmHg对 (164.12±11.69)mmHg,1 mmHg=0.133 kPa;t=5.187,P<0.001]、低密度脂蛋白胆固醇[(2.92±0.52)mmol/L对 (2.49±0.36)mmol/L;t=5.447,P<0.001]、空腹血糖[(8.76±2.88)mmol/L对 (6.82±2.24)mmol/L;t=4.185,P<0.001]、C反应蛋白[(7.62±1.82)mg/L对 (4.57±1.58)mg/L;t=9.790,P<0.001]和D-二聚体[(1.14±0.08)mg/L对 (0.97±0.22)mg/L;t=4.946,P<0.001]水平均显著高于转归良好组,而SAO型患者构成比(29.55%对74.39%;χ2=23.759,P<0.001)以及高密度脂蛋白胆固醇[(0.94±0.68)mmol/L对 (1.16±0.14)mmol/L;t=2.829,P=0.005]和基线EPCs(0.069%±0.018%对0.098%±0.021%;t=7.755,P<0.001)水平显著低于转归良好组.多变量logistic回归分析显示,基线NIHSS评分较高(优势比1.242,95%可信区间1.126~1.372;P<0.001)、CE型(优势比3.460,95%可信区间1.312~5.146;P=0.016)和基线EPCs数量较低(优势比1.632,95%可信区间1.006~3.024;P<0.001)是急性缺血性卒中患者转归不良的独立危险因素.结论 急性缺血性卒中患者循环EPCs水平显著降低,基线EPCs水平较低是缺血性卒中患者90 d时转归不良的独立预测因素.     

癌干细胞CD44~+/ALDH~+在结直肠癌中的表达及其临床病理学意义

目的:观察癌干细胞CD44+/ALDH+在结直肠癌组织中的表达,并探讨其临床病理学意义.方法:利用组织芯片仪制作组织芯片,通过免疫组织化学染色,免疫组织化学双重染色观察结直肠癌组织和不同组织中CD44+,ALDH+,CD44+/ALDH+的表达.结果:CD44,ALAH1在结直肠癌中的表达明显高于正常肠黏膜和伴不典型增生的管状腺瘤(均P<0.05);在结直肠癌中,CD44和ALDH1的表达率与癌组织的病理分级、浸润、淋巴结转移有关(均P<0.05);CD44+/ALDH+在结直肠癌中的表达明显高于正常肠黏膜组织和伴不典型增生的管状腺瘤(均P<0.05),随着癌分化程度的降低,表达率增加.结论:CD44+/ALDH+可作为结直肠癌干细胞有价值的标志物,可作为结直肠癌早期和预防复发转移的可靠指标.