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。     

胆囊癌组织P62蛋白的表达及其与血管生成关系

目的 :检测胆囊癌组织中P6 2蛋白、碱性成纤维细胞生长因子(bFGF)的表达和微血管密度 (MVD) ,探讨它们的相互关系 ,以及与胆囊癌临床病理特征的关系。方法 :4 1例胆囊癌和 2 2例慢性胆囊炎组织中 ,P6 2蛋白和bFGF的表达用SP免疫组化染色法进行检测。胆囊癌组织中MVD用抗CD34单抗 (mAb)做SP免疫组化染色进行检测。结果 :4 1例胆囊癌组织中 ,P6 2和bFGF表达阳性率分别为 6 3.4 %和 75 .6 % ,均高于 2 2例慢性胆囊炎组织 (P <0 .0 1)。P6 2的表达与胆囊癌淋巴结转移有关 (P <0 .0 5 ) ,与癌组织的分级、临床分期无显著关系。bFGF的表达与胆囊癌临床病理分期及组织学分级有关 (P <0 .0 5 ) ,而与淋巴结转移无显著关系。P6 2表达率与bFGF表达率呈正相关关系 (r=0 .5 2 1;P <0 .0 1)。两者的表达均与患者的年龄、性别、肿瘤的种类、是否伴有胆囊结石无关。胆囊癌组织中MVD明显高于慢性胆囊炎组织 (P <0 .0 1)。P6 2及bFGF表达阳性组织MVD高于表达阴性组织 (分别P <0 .0 5和P <0 .0 1)。MVD与胆囊癌的Nevin分期及淋巴结转移有关 (P<0 .0 1) ,与患者的年龄、性别、肿瘤种类、分化程度以及是否伴有胆囊结石无关。结论 :P6 2蛋白及bFGF的表达与胆囊癌组织中血管的生成相关 ,在胆囊癌的发生和发展过程中可能具有重     

Reconstitution of chemokine-induced actin polymerization in undifferentiated human leukemia cells (HL-60) by heterologous expression of interleukin-8 receptors

The chemokines interleukin-8 (IL-8) and GRO62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> bind in neutrophils to the interleukin-8 receptor 62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> and 62w13757845637/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0"> (IL-8R62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> and 62w13757845637/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0">) triggering reorganization of the actin cytoskeleton and activation of phospholipase C (PLC). Reconstitution of chemokine-induced activation of PLC indicated coupling of IL-8R62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> and 62w13757845637/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0"> to pertussis toxin-insensitive 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE1.gif" alt=" $$G\alpha _{i2^ - } $$ " align="middle" border="0"> or 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE2.gif" alt=" $$G\alpha _{i3^ - } $$ " align="middle" border="0"> . To identify the signal transduction mechanisms of chemokine-induced actin response, undifferentiated human leukemia cells (HL-60 cells) constitutively expressing 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE3.gif" alt=" $$G\alpha _{16^ - } $$ " align="middle" border="0"> , 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE4.gif" alt=" $$G\alpha _{i2^ - } $$ " align="middle" border="0"> and 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE5.gif" alt=" $$G\alpha _{i3^ - } proteins$$ " align="middle" border="0"> were chosen for reconstitution studies. Expression of recombinant receptors after transfection of the cells with the cDNA of IL-8R62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> and 62w13757845637/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0"> was confirmed by binding studies with radiolabeled ligands. IL-8R62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> bound IL-8 with high affinity (Kd62w13757845637/xxlarge8764.gif" alt="sim" align="MIDDLE" BORDER="0">1 nM) and GRO62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> with low affinity (Kd62w13757845637/xxlarge8764.gif" alt="sim" align="MIDDLE" BORDER="0"> 1 62w13757845637/xxlarge956.gif" alt="mgr" align="MIDDLE" BORDER="0">M), whereas IL-8R62w13757845637/xxlarge946.gif" alt="beta" align="MIDDLE" BORDER="0"> bound both IL-8 and GRO62w13757845637/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0"> with high affinity (Kd62w13757845637/xxlarge8764.gif" alt="sim" align="MIDDLE" BORDER="0">1 nM). Flow cytometric actin measurements indicated that high affinity ligand-receptor interactions in both receptor transfectants displayed inducible responses. Pretreatment of transfectants with pertussis toxin caused ADP-ribosylation of G-proteins and blocked chemokine-induced polymerization, indicating involvement of 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE6.gif" alt=" $$G\alpha _{i2^ - } $$ " align="middle" border="0"> or 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE7.gif" alt=" $$G\alpha _{i3^ - } proteins$$ " align="middle" border="0"> , but not 62w13757845637/11_2005_Article_BF02265165_TeX2GIFIE8.gif" alt=" $$G\alpha _{16^ - } proteins$$ " align="middle" border="0"> in this response.accepted by M.J. Parnham     

Effect of activated antigen-specific B cells on ES-62-mediated modulation of effector function of heterologous antigen-specific T cells in vivo

There is currently great interest in the idea of using helminth-derived molecules for therapeutic purposes and indeed we have shown that ES-62, a filarial nematode-derived phosphorylcholine-containing glycoprotein, significantly reduces the severity of arthritis in a murine model. Clearly, knowledge of mechanism of action is important when considering molecules for use in treating disease and although much is known regarding how ES-62 interacts with the immune system, gaps in our understanding remain. A feature of filarial nematode infection is a defective, T helper 2 (Th2)-polarized antigen-specific T-cell response and in relation to this we have recently shown that ES-62 inhibits clonal expansion and modulates effector function towards a Th2 phenotype, of antigen-specific T cells in vivo. ES-62 is also known to directly modulate B-cell behaviour and hence to determine whether it was mediating these effects on T cells by disrupting B-T-cell co-operation, we have investigated antigen-specific responses using an adoptive transfer system in which traceable numbers of tg ovalbumin (OVA)-specific T cells and hen egg lysozyme (HEL)-specific B cells respond to a chemically coupled form of OVA-HEL that contains linked epitopes that promote cognate T- and B-cell interactions. Surprisingly, these studies indicate that activated B cells restore T-cell expansion and prevent Th2-like polarization. However, ES-62-treated double cell transfer mice demonstrate a more generalized immunosuppression with reduced levels of Th1 and -2 type cytokines and antibody subclasses. Collectively, these results suggest that whilst ES-62 can target B-T-cell co-operation, this does not promote polarizing of T-cell responses towards a Th2-type phenotype.     

Autophagy in immunity and cell-autonomous defense against intracellular microbes

Autophagy was viewed until very recently primarily as a metabolic and intracellular biomass and organelle quality and quantity control pathway. It has now been recognized that autophagy represents a bona fide immunologic process with a wide array of roles in immunity. The immunologic functions of autophagy, as we understand them now, span both innate and adaptive immunity. They range from unique and sometimes highly specialized immunologic effectors and regulatory functions (referred to here as type I immunophagy) to generic homeostatic influence on immune cells (type II immunophagy), akin to the effects on survival and homeostasis of other cell types in the body. As a concept-building tool for understanding why and how autophagy is intertwined with immunity, it is useful to consider that the presently complex picture has emerged in increments, starting in part from the realization that autophagy acts as an evolutionarily ancient microbial clearance mechanism defending eukaryotic cells against intracellular pathogens. In this review, we build a stepwise model of how the core axis of autophagy as a cell-autonomous immune defense against microbes evolved into a complex but orderly web of intersections with innate and adaptive immunity processes. The connections between autophagy and conventional immunity systems include Toll-like receptors, Nod-like receptors, RIG-I-like receptors, damage-associated molecular patterns such as HMGB1, other known innate and adaptive immunity receptors and cytokines, sequestasome (p62)-like receptors that act as autophagy adapters, immunity-related GTPase IRGM, innate and adaptive functions of macrophages and dendritic cells, and differential effects on development and homeostasis of T- and B-lymphocyte subsets. The disease contexts covered here include tuberculosis, infections with human immunodeficiency virus and other viruses, Salmonella, Listeria, Shigella, Toxoplasma, and inflammatory disorders such as Crohn's disease and multiple sclerosis.     

酸洗脱血小板HLA-Ⅰ类抗原的研究

目的: 建立一种有效酸洗脱血小板表面HLA-Ⅰ类抗原的方法,评估最适的酸洗脱条件以及这一技术临床应用的可行性。方法: 血小板用不同pH值(pH=2、3、5、7)的柠檬酸缓冲液在0 ℃进行酸洗脱,多色流式细胞技术检测血小板表面HLA-Ⅰ类抗原及P-选择素(CD62P)的表达,Annexin V染色检测血小板的早期凋亡,电阻法检测血小板的最大聚集率。结果: 随着柠檬酸缓冲液pH值的降低(从pH=7到pH=2),血小板表面HLA-Ⅰ类抗原的表达显著减少,但同时血小板的活化率(CD62P表达)和早期凋亡率(Annexin V表达)均显著增加。相对PBS处理,利用pH=3的柠檬酸缓冲液处理血小板可显著减少血小板表面HLA-Ⅰ类抗原的表达(P<0.05),尽管血小板活化率和早期凋亡率均显著增加(均P<0.05),但其聚集功能未受显著影响(P>0.05)。结论: 利用pH=3的柠檬酸缓冲液0 ℃洗脱血小板HLA-I类抗原可作为预防血小板输注无效的方法进行尝试。这项酸处理技术在临床应用前尚需进一步改进和标准化。     

Expression of CD62L on Donor CD4<Superscript>+</Superscript> T Cells in Allografts: Correlation with Graft-Versus-Host Disease after Unmanipulated Allogeneic Blood and Marrow Transplantation

Introduction  The aim of this study was to investigate the association of donor CD4⁺ T cells expressing CD62L with transplant outcomes. Materials and Methods  We report a prospective analysis of 31 patients who were treated with a Bu/Cy regimen, followed by unmanipulated blood and marrow transplantation. Results  Median number (range) of CD4⁺CD62L⁺, CD4⁺CD45RA⁺CD62L⁺, and CD4⁺CD45RO⁺CD62L⁺ cells infused were 0.31(0.05–1.10)×10⁸/kg, 0.22(0.03–0.95)× 10⁸/kg, and 0.17(0.01–0.81)×10⁸/kg, respectively. The incidence of grades II to IV aGVHD was 36%. In a multivariate analysis, infusion of >0.22 × 10⁸ CD4⁺CD45RA⁺CD62L⁺ cells infused/kg increased the risk of grades II to IV aGVHD (HR = 4.741, 95% CI = 1.037–21.662, P = 0.045). Thirteen of 31 patients experienced cGVHD, the risk of cGVHD was increased in patients receiving >0.45 × 10⁸ CD4⁺CD45RA⁺ cells infused/kg (HR = 4.614, 95% CI = 1.265–16.829, P = 0.021). Conclusion  Our results suggest that a high cell dose of CD4⁺CD45RA⁺CD62L⁺ cells increase the incidence of grades II–IV aGVHD. A high number of CD4⁺CD45RA⁺ cells infused were associated with increased risk of cGVHD in our transplant settings. Ying-Jun Chang: performed research, analysis and interpretation of data, and drafting of the article, and gave final approval of the version to be published; Xiang-Yu Zhao: performed research, analysis and interpretation of data, and drafting of the article and gave final approval of the version to be published; Ming-Rui Huo: performed research, analysis and interpretation of data, and drafting of the article and gave final approval of the version to be published; Xiao-Jun Huang: involved in conception and design, revising the article critically, and final approval of the version to be published.     

Dissecting the complexity of the memory T cell response

Memory immune responses are classically attributed to the reactivation of long-lived, antigen-specific T lymphocytes that persist in a quiescent state. Determining mechanisms for the generation of memory T cells and dissecting the functional nature of the memory T cell pool has been encumbered by an inability to distinguish recently activated effector T cells from memory T cells. We have established new activation and biochemical criteria that distinguish effector and memory T cells and have applied these criteria to follow memory generation from activated cells in vivo. We found that the resultant memory T cell pool is heterogeneous and consists of effector-like and resting memory-like subsets that differ in expression of the homing receptor, CD62L. We discuss these findings in the context of memory T cell heterogeneity identified in human and mouse systems. These results suggest that more than one type of previously activated T cell can mediate recall or memory immune responses and that elucidating the fundamental phenotypic and functional features of memory T cell subsets is therefore critical to deciphering the complex nature of the memory immune response.     

Beclin 1、LC3-II及P62蛋白在上皮性卵巢癌中的表达及意义

目的探讨上皮性卵巢癌中Beclin、LC3-Ⅱ及P62蛋白的表达及临床意义。方法选取2011年6月—2013年3月我院手术切除并经病理学检查确诊的上皮性卵巢癌标本50例为实验组,同期手术切除正常卵巢组织20例作为对照,通过免疫组织化学sP法,检测标本中Beclin1、LC3．Ⅱ及P62蛋白的表达情况,并结合临床病理资料进行分析。结果在上皮性卵巢癌组织中,Beclin 1、LC3-Ⅱ蛋白的阳性表达率分别为46．00％（23／50）和42．00％（21／50）,低于正常卵巢组织的95．00％（19／20）和85．00％（17／20）;而P62蛋白的阳性表达率为72．00％（36／50）,高于正常卵巢组织的10．00％（2／20）,差异均具有统计学意义（P值均〈0．05）。在上皮性卵巢癌组织中,Beclin1、LC3-Ⅱ蛋白的表达与FIGO分期、病理分级具有相关性（P〈0．05）,而P62蛋白的表达与FIGO分期具有相关性（P〈0．05）。Beclinl蛋白和LC3-Ⅱ蛋白在上皮性卵巢癌组织中的表达呈正相关（P〈0．01）,P62蛋白的表达与Beclin1、LC3-Ⅱ蛋白均呈负相关（P〈0．01）。结论在上皮性卵巢癌组织中,Beclin1、LC3-Ⅱ和P62蛋白的异常表达,可导致自噬体形成减少和自噬功能减弱;可能与卵巢癌的发生、发展和预后不良有关。