免疫组化分析鼠疫疫苗免疫和鼠疫菌攻毒后恒河猴脾组织中T和B淋巴细胞增殖(英文)

目的在我们先前的研究中发现,鼠疫疫苗免疫的猕猴脾组织中B细胞和T细胞数量明显增加。然而,是否这些细胞是由鼠疫疫苗引起的增殖性B细胞和T细胞还不得而知。方法为回答这个问题,本研究使用免疫组化双标记方法检测了猕猴脾组织中T细胞和B细胞的增殖。应用Ki67抗体以及T细胞和B细胞特异性单克隆抗体的免疫组化双标法,对脾组织T细胞和B细胞增殖进行检测。脾组织分别来自于亚单位疫苗SV1(20μg F1+10μg rV270)免疫的猕猴以及分别通过SV2(200μg F1+100μg rV270)、减毒活疫苗EV和铝佐剂免疫并分别攻毒的猕猴。结果与正常动物相比,受试动物脾组织生发中心有较多的B细胞增殖,边缘区有较多的静止B细胞,在动脉周围淋巴鞘区有较多的静止T细胞,提示原始T细胞可能在免疫早期发生增殖。经SV1、SV2或EV76免疫并攻毒的动物脾组织生发中心较仅用SV1免疫动物的生发中心扩大。此外,铝佐剂免疫并攻毒的猕猴脾组织生发中心不完整,这可能归因于强毒株鼠疫菌感染引起的病理损伤。B细胞增殖、静止B细胞增加、静止T细胞增多及生发中心扩大是诱导特异性体液免疫和保持免疫记忆反应的标志。结论这些结果与我们先前的发现的SV1、SV2或者EV76免疫动物激发较高的抗体和IL-4分泌相一致。     

核因子-κB活性对BXSB狼疮小鼠脾脏自发性生发中心形成的影响

目的探讨核因子(NF)-kB活性对BXSB狼疮小鼠脾脏中自发性生发中心形成的影响及其机制。方法18只BXSB自发性狼疮小鼠随机分成对照组和吡咯二硫氨基甲酸酯(PDTC)干预组,PDTC组：隔日腹腔注射PDTC(120mg/kg体重)；对照组：隔日腹腔注射等量溶媒。持续8周结束实验。以电泳迁移率改变实验检测脾脏组织NF-kB活性,双标记流式细胞术检测脾脏B细胞CD154表达及生发中心B细胞凋亡,组织化学方法染色脾脏生发中心,并以图像处理系统半定量分析。结果PDTC显著抑制BXSB狼疮小鼠脾脏组织NF-kB活性,较对照组下降62．82%；BXSB狼疮小鼠脾脏组织可见自发性生发中心形成,抑制NF-kB活性能下调脾脏B细胞CD154表达、使其自发性生发中心形成受阻、促进生发中心B细胞凋亡。结论NF-KB过度活化可通过上调脾脏B细胞CD154的异常表达而促进自发性生发中心形成、并减少生发中心B细胞凋亡。由此,自发性生发中心形成过程中生成的自身反应性B细胞得以逃逸凋亡,分化成产自身抗体浆细胞。提示NF-kB可成为防治系统性红斑狼疮的一个作用靶点。     

核因子-κB活性对BXSB狼疮小鼠脾脏白发性生发中心形成的影响

目的探讨核因子（NF）-κB活性对BXSB狼疮小鼠脾脏中自发性生发中心形成的影响及其机制。方法18只BXSB自发性狼疮小鼠随机分成对照组和吡咯二硫氨基甲酸酯（PDTC）干预组，PDTC组：隔日腹腔注射PDTC（120mg／kg体重）；对照组：隔日腹腔注射等量溶媒。持续8周结束实验。以电泳迁移经改变实验检测脾脏组织NF-κB活性，双标记流式细胞术检测脾脏B细胞CD154表达及生发中心B细胞凋亡，组织化学方法染色脾脏生发中心，并以图像处理系统半定量分析。结果PDTC显著抑制BXSB狼疮小鼠脾脏组织NF-κB活性，较对照组下降62．82％；BXSB狼疮小鼠脾脏组织可见自发性生发中心形成．抑制NF-κB活性能下调脾脏B细胞CD154表达、使其自发性生发中心形成受阻、促进生发中心B细胞凋亡，结论NF-κB过度活化可通过上调脾脏B细胞CD154的异常表达而促进自发性生发中心形成、并减少生发中心B细胞凋亡。由此，自发性生发中心形成过程中生成的自身反应性B细胞得以逃逸凋亡，分化成产自身抗体浆细胞：提示NF-κB可成为防治系统性红斑狼疮的一个作用靶点。     

低氧诱导因子-1α、Delta样配体4在不同乳腺癌分子亚型组织中的表达及临床意义

目的探讨低氧诱导因子(HIF-)1α、Delta样配体(DLL)4在不同乳腺癌分子亚型组织中的表达及临床意义。方法采用免疫组织化学法检测乳腺癌组织〔基底细胞样型17例、人表皮生长因子受体(HER)-2过表达型21例、Luminal A型54例、Luminal B型23例与普通乳腺样型23例〕与癌旁组织(对照组)HIF-1α、DLL4表达。结果 115例乳腺癌组织HIF-1α、DLL4分别为70.43%、69.57%,明显高于对照组(25.00%、22.00%)(P0.05),其中基底细胞样型、HER-2过表达型、Luminal A型与Luminal B型乳腺癌组织HIF-1α、DLL4的阳性表达率均明显高于对照组(P0.05),基底细胞样型乳腺癌组织HIF-1α、DLL4的阳性表达率明显高于Luminal A型,HER-2过表达型乳腺癌组织HIF-1α的阳性表达率明显高于Luminal A型与Luminal B型(P0.05)。乳腺癌组织HIF-1α、DLL4表达与年龄、组织学分级无明显的关系(P0.05),与腋窝淋巴结转移、临床分期具有明显的关系,其中中青年乳腺癌患者HIF-1α、DLL4阳性表达较高,腋窝淋巴结转移者上述因子阳性表达率更高,Ⅲ/Ⅳ期上述因子阳性表达率更高(P0.05))。结论 HIF-1α、DLL4在基底细胞样型、HER-2过表达型乳腺癌组织中的表达较高,而在Luminal A、B型乳腺癌组织中的表达较低;其可能与临床病理特征及预后状况有一定的关系。     

肝细胞癌变过程中核转录因子-кB表达的基础与临床研究

目的 观察肝细胞癌(HCC)形成过程中核转录因子-кB(NF-кB)动态改变及临床价值.方法 雄性SD大鼠以2-乙酰氨基芴(2-FAA)制备肝癌模型,经病理组织学分析肝细胞形态学变化,定量观察NF-кB动态变化.并以自身配对法收集经手术切除后的肝癌及其痛周组织,定量分析肝癌组织中NF-кB表达及病理学特征.结果 诱癌后肝细胞发生颗粒样变性、不典型增生、到高分化肝细胞癌形成;在此过程中,NF-кB表达呈梯度增加.NF-кB阳性表达呈棕黄色颗粒状染色,癌组织NF-кB点灶状表达,定位于胞浆和细胞核;癌周组织NF-кB主要定位于胞浆,未见细胞核阳性.人肝癌组NF-кB明显高于癌周组织(P<0.01),癌组织NF-кB表达阳性率为100%,癌周组织为68.6%(x2=13.1,P<0.01).其表达与分化程度、肿瘤数目和肿瘤直径无关.结论 NF-кB表达参与肝癌的发生、发展,活性抑制可能是肝癌基因治疗的新靶点.     

胃黏膜相关淋巴样组织淋巴瘤染色体易位t(11;18)与BCL10的表达

目的　检测胃黏膜相关淋巴样组织 (MALT)淋巴瘤的染色体易位t(11;18) (q2 1;q2 1)和BCL10蛋白表达的情况。方法　采用RT PCR检测胃MALT淋巴瘤和滤泡性胃炎 (FG)中API2 MLT融合及免疫组化检测BCL10蛋白、Ki 6 7表达情况 ,并结合临床病理进行分析。结果　 14例胃MALT淋巴瘤中有 3例 (2例低恶性 ,1例低～高恶性 )检测到API2 MLT融合 ,8例FG无此融合。BCL10在FG淋巴滤泡生发中心细胞胞质中弱表达 ,在胃MALT淋巴瘤中表达明显增强 ,且 4 2 .5 %的病例细胞核阳性。胃低～高恶性及弥漫大细胞淋巴瘤 (DLBCL)的Ki 6 7标记率显著强于低恶性MALT淋巴瘤 (P<0 .0 5 )。BCL10核表达与Ki 6 7阳性表达之间差异无显著性 (P >0 .0 5 ) ,但随Ki 6 7表达增强 ,BCL10核表达的概率增加。结论　API2 MLT融合和BCL10核表达可能与胃MALT淋巴瘤从低恶性向高恶性转化有关。RT PCR检测API2 MLT融合是检测t(11;18) (q2 1;q2 1)的一项重要工具     

T1b期肾癌及癌旁组织中PCNA、Bax的表达及意义

目的 探讨T1b期肾癌及癌旁组织中增殖细胞核抗原(PCNA)、Bax蛋白表达及意义,为T1b期肾癌行肾部分切除术中安全边距的选择提供理论依据.方法 选取21例行肾癌根治术的T1b期肾癌患者的癌组织标本,病理诊断均为肾透明细胞癌;分别取其癌组织(A组),癌旁0.2 cm(B组)、0.5 cm(C组)、1.0 cm(D组)处的正常肾组织,采用免疫组织化学染色法检测各组不同组织中的PCNA、Bax蛋白表达情况.结果 A组PCNA阳性表达率最高,明显高于其他各组(P均＜0.05);A组Bax蛋白阳性表达率最低,明显低于其他各组(P均＜0.05).C、D组的PCNA蛋白阳性表达率明显低于B组,Bax蛋白阳性表达率明显高于B组(P均＜0.05).结论 癌旁0.5 cm是T1b期肾癌肾部分切除术的安全切除边距.     

肝细胞癌变过程中核因子-κB及其基因的动态表达与定量分析

目的 观察肝癌形成过程中核因子-κB(NF-κB)及NF-κB mRNA动态表达与作用机制.方法 雄性SD大鼠以2-乙酰氨基芴制备肝癌模型,经病理组织学分析肝细胞形态学变化,定量观察NF-κB动态变化,以巢式PCR分析NF-κB mRNA的表达.并以自身配对法收集经手术切除后的肝癌及其癌周组织,定量分析肝癌组织中NF-κB表达及病理学特征. 结果诱癌后在肝细胞呈颗粒样变性,不典型增生,肝细胞癌形成,NF-κB及基因表达呈梯度增加.NF-κB阳性表达呈棕黄色颗粒状染色,癌组织NF-κB点灶状表达,定位于胞质和细胞核,癌周组织NF-κB主要定位于胞质,未见细胞核阳性.癌变过程中NF-κB mRNA表达明显增强.人肝癌组织NF-κB(69.3±40.2)pg/mg,明显高于癌周组织(21.0±17.2)pg/mg(t=6.54,P＜0.01).癌组织NF-κB表达阳性率为100%,癌周组织为68.6%(X2=13.05,P＜0.01).其表达与肿瘤分化程度,肿瘤数目和肿瘤直径无关. 结论 NF-κB异常表达与肝癌的发生发展有关,表达抑制有助于肝痛治疗.     

GTPBP2通过Wnt/β-catenin信号通路调控结直肠癌干细胞增殖、细胞周期、迁移、侵袭和凋亡

目的 探讨GTP结合蛋白(GTPBP)2对CD133阳性结直肠癌干细胞增殖、细胞周期、迁移、侵袭和凋亡的影响及其分子机制。方法 选取30例结直肠癌患者的结直肠癌组织及相应癌旁组织；利用免疫磁珠法分选出人结直肠癌SW480细胞的CD133阳性肿瘤干细胞，将其分为Con组、si-NC组、si-GTPBP2组；实时荧光定量-聚合酶链反应(RT-qPCR)检测GTPBP2 mRNA表达水平；Western印迹检测GTPBP2表达、干细胞表面标志物表达及增殖、细胞周期、迁移、侵袭、凋亡和Wnt/β-catenin信号通路相关蛋白表达；四甲基偶氮唑蓝(MTT)检测细胞增殖；Transwell实验检测细胞迁移和侵袭；流式细胞术检测细胞凋亡和周期。结果 与癌旁组织相比，结直肠癌组织中GTPBP2表达水平明显升高(P<0.05);沉默GTPBP2可明显抑制结直肠癌干细胞增殖、迁移和侵袭，明显阻滞细胞周期进程，并明显促进凋亡，结直肠癌干细胞存活率、S期细胞比例、迁移、侵袭细胞数及细胞周期蛋白(Cyclin)D1、细胞核增殖抗原(Ki)-67、基质金属蛋白酶(MMP)-2、MMP-9、B细胞淋巴瘤(B...     

滤泡辅助T细胞相关分子与支气管哮喘

滤泡辅助性T细胞(T follicular helper cells,Tfh)是近来发现的CD4+T细胞亚群,它能够迁移至次级淋巴中心的B细胞滤泡,促进生发中心反应,包括调节生发中心的形成、发展和成熟以及辅助B细胞产生具有免疫功能的球蛋白及影响类别转换.Tfh能连续产生趋化因子受体5,在其配体CXCL13的作用下,外周的Tfh被募集至生发中心的B淋巴滤泡,参加生发中心的形成.诱导性共刺激分子和CO40L可以与B细胞表面的配体结合可以调节Tfh分泌IL-21参与B细胞的免疫应答,参与体液免疫.支气管哮喘(简称哮喘)发病机制复杂,T淋巴细胞在哮喘的气道炎症反应中起重要作用.本文回顾了Tfh细胞相关分子在哮喘气道炎症中的作用.     

Distinctive High Expression of Antiretroviral APOBEC3 Protein in Mouse Germinal Center B Cells

Tissue and subcellular localization and its changes upon cell activation of virus-restricting APOBEC3 at protein levels are important to understanding physiological functions of this cytidine deaminase, but have not been thoroughly analyzed in vivo. To precisely follow the possible activation-induced changes in expression levels of APOBEC3 protein in different mouse tissues and cell populations, genome editing was utilized to establish knock-in mice that express APOBEC3 protein with an in-frame FLAG tag. Flow cytometry and immunohistochemical analyses were performed prior to and after an immunological stimulation. Cultured B cells expressed higher levels of APOBEC3 protein than T cells. All differentiation and activation stages of freshly prepared B cells expressed significant levels of APOBEC3 protein, but germinal center cells possessed the highest levels of APOBEC3 protein localized in their cytoplasm. Upon immunological stimulation with sheep red blood cells in vivo, germinal center cells with high levels of APOBEC3 protein expression increased in their number, but FLAG-specific fluorescence intensity in each cell did not change. T cells, even those in germinal centers, did not express significant levels of APOBEC3 protein. Thus, mouse APOBEC3 protein is expressed at distinctively high levels in germinal center B cells. Antigenic stimulation did not affect expression levels of cellular APOBEC3 protein despite increased numbers of germinal center cells.     

T-bet在肺结核组织淋巴细胞中的表达特征及其临床意义

目的探讨肺结核组织中T-bet的表达与机体免疫和病理发展变化的关系。方法采用免疫组织化学双染法检测8例肺结核组织中T-bet和CD4^＋、CD20^＋的表达情况。结果在肺结核结节中,CD4^＋T细胞T-bet阳性率为70.1%,CD20^＋B细胞T-bet阳性率为18.2%。结论 T-bet在肺结核组织中高表达,T-bet在肺结核的机体免疫中可能发挥着重要的作用。     

Impaired T- and B-cell development in Tcl1-deficient mice

TCL1, the overexpression of which may result in T-cell leukemia, is normally expressed in early embryonic tissues, the ovary, and lymphoid lineage cells. Our analysis of mouse B-lineage cells indicates that Tcl1 expression is initiated in pro-B cells and persists in splenic marginal zone and follicular B cells. T-lineage Tcl1 expression begins in thymocyte progenitors, continues in CD4(+)CD8(+) thymocytes, and is extinguished in mature T cells. In Tcl1-deficient mice, we found B lymphopoiesis to be compromised at the pre-B cell stage and T-cell lymphopoiesis to be impaired at the CD4(+)CD8(+) thymocyte stage. A corresponding increase was observed in thymocyte susceptibility to anti-CD3epsilon-induced apoptosis. Reduced numbers of splenic follicular and germinal center B cells were accompanied by impaired production of immunoglobulin G1 (IgG1) and IgG2b antibodies in response to a T-dependent antigen. The marginal zone B cells and T-cell-independent antibody responses were also diminished in Tcl1(-/-) mice. This analysis indicates a significant role for Tcl1, a coactivator of Akt signaling, in normal T- and B-cell development and function.     

Expression of the recombination-activating genes in extrafollicular lymphocytes but no apparent reinduction in germinal center reactions in human tonsils.

V(D)J recombination in lymphocytes is mediated by 2 recombination-activating genes, RAG1 and RAG2, which are expressed during lymphocyte development in bone marrow and thymus. Prompted by studies reporting re-expression of the RAGs in germinal center B cells, the expression of RAGs and terminal deoxynucleotidyl transferase (TdT) in human lymphoid tissues was examined using in situ hybridization and immunohistochemistry, respectively. Here it is shown that RAGs and TdT are not reinduced in germinal center reactions. However, RAG(+)/TdT(+) cells are frequently present in extrafollicular areas of tonsils mainly at the boundary between lymphoid tissue and fibrous scaffold. Phenotypic analyses suggest that these cells are B cells. Finally, it is shown that RAG(+)/TdT(+) cells are found more frequently in tonsils than in other peripheral lymphoid tissues. This may reflect an increased influx of RAG(+)/TdT(+) cells as a result of higher antigenic stimulation at this site. Alternatively, this observation may indicate that the tonsils are an additional site of lymphocyte ontogeny.     

Analysis of B-cell antigens in normal reactive lymphoid tissue using four B-cell monoclonal antibodies

Lymph nodes from 13 cases of reactive hyperplasia were examined with four different monoclonal antibodies to B cells. B-1 recognizes an antigen of 30,000 daltons on B cells. CB-2 was prepared with normal spleen and binds to a glycolipid. BA-1 labels surface immunoglobulin- positive cells, but not T lymphocytes or monocytes. B-532 recognizes an antigenic determinant of 45,000 daltons. Using the immunoperoxidase method on frozen sections of reactive lymph nodes, the staining patterns of these four unique antibodies showed dramatic differences. B- 1 labeled 80%-90% of the germinal center cells and 10%-50% of the mantle region. Few interfollicular cells were positive. CB-2 stained predominantly in the mantle area (50%-90% positive cells), with moderate staining in the germinal center as well and less than 1% positive cells in the diffuse cortex. BA-1 exhibited predominant labeling in the mantle (50%-90%), with little staining in the germinal center. A large number of cells in the interfollicular, subcapsular, and medullary regions expressed the BA-1 antigen. The B-5 antibody demonstrated intense staining in the follicle (50%-95%). This staining often appeared to be polarized within the germinal center. The mantle zone demonstrated staining of 30%-50% of the cells. The different staining patterns of the B-cell antibodies, as demonstrated by the in situ distribution of positive cells within the lymph node, may reflect stages of development or activation of the B-cell population.     

In vivo expression of the B7 costimulatory molecule by subsets of antigen-presenting cells and the malignant cells of Hodgkin's disease

The B-lymphocyte/accessory-cell activation antigen B7 (BB1) has been shown in vitro to stimulate T-lymphocyte proliferation and cytokine production via CD28 present on the latter cells. In this study, benign lymphoid tissues, lymphomas, and extralymphoid inflammatory sites were examined immunohistochemically using anti-B7 and other relevant monoclonal antibodies. B7 was expressed by benign transformed germinal center B cells, as it was by B cells of follicular lymphomas. B7 was also expressed by a subpopulation (a mean of 31% to 65%) of macrophages and dendritic cells in a variety of lymphoid tissues. It was present in abundance on all macrophages constituting sarcoid granulomas in lymph nodes. In extralymphoid inflammation, 17% to 35% of macrophages expressed B7 only weakly. Cases of Hodgkin's disease showed expression of B7 by the majority of Reed-Sternberg cells or malignant mononuclear variants, a phenomenon that potentially contributes to the lymphocytic accumulation that is a feature of this condition. CD28+ T cells were seen in all areas where T cells were present. B7+ and CD28+ cells colocalized in, for example, lymphoid follicles, lymph node paracortex, sarcoid granulomas, and Hodgkin's disease tissue, indicating a potential for cellular interaction via these molecules at these sites.