关键词： autoimmunity   anti-CD20   B cells   Treg   NONOBESE DIABETIC MICE   ANTIGEN PRESENTATION   INDUCED ARTHRITIS   MOUSE MODEL   THYROIDITIS   THERAPY   EXPRESSION   RITUXIMAB   LYMPHOCYTES   SUPPRESSION  

摘要： B-cell depletion therapy can be effective for treating B-cell lymphomas as well as many human and murine autoimmune diseases. B-cell-deficient mice are normally resistant to spontaneous autoimmune thyroiditis (SAT), but they develop SAT if regulatory T cells are transiently depleted during the first 3-6 weeks after birth. This was also a critical time when B-cell depletion effectively inhibited development of SAT in adult mice. The current study was undertaken to test the hypothesis that transient depletion of B cells using anti-CD20 would be sufficient to suppress SAT if B cells were depleted early in life and that inhibition of SAT would be due to the activity of Treg that functioned most effectively when B cells were absent or low. The results presented here support this hypothesis and indicate that development of autoimmune disease in adults is effectively inhibited when anti-CD20 is administered 1-3 weeks after birth. After 3 weeks, transient B-cell depletion is no longer effective, and B-cell depletion must be maintained to effectively suppress autoimmune disease. B-cell depletion in 1- to 3-week-old mice depletes all B-cell subsets, whereas B-cell depletion initiated in adults spares many marginal zone B cells. Following early B-cell depletion, splenic Treg increase in number, and depletion of Treg reverses the inhibitory effect of anti-CD20 on disease development. Early transient depletion of B cells could be useful for preventing autoimmune disease in individuals at high risk for developing autoimmune diseases as adults.

关键词： Balb/c小鼠   调节性B细胞   B10细胞   流式细胞仪  

摘要： 目的分析年龄、性别和环境因素对Balb/c小鼠脾脏B10细胞的影响。方法以年龄、性别和不同环境对小鼠进行分组,流式细胞仪分析小鼠脾脏CD5+CD1dhiB细胞的比例。体外培养小鼠脾细胞,在不加任何刺激或LPS+PIM刺激5 h后,流式细胞仪分析CD19+IL-10+B细胞比例。结果 6月龄小鼠脾脏中CD5+CD1dhiB细胞比例平均为2.9%,明显高于2月龄小鼠的2.28%(P<0.05);在不加任何刺激的情况下,两组小鼠脾脏中IL-10+B细胞的比例均很低且无统计学差异(P>0.05);加入LPS+PIM刺激5 h后,6月龄小鼠脾脏中IL-10+B细胞的比例平均为1.93%,高于2月龄小鼠的1.25%(P<0.05)。细胞表型和细胞内IL-10染色分析均显示年龄匹配的雌性和雄性小鼠脾脏B10细胞的表达无明显差异(P>0.05)。普通级和SPF级小鼠CD5+CD1dhiB细胞和LPS+PIM诱导的IL-10+B细胞比例无明显差异(P>0.05),而在不加任何刺激的情况下,普通级小鼠脾脏IL-10+B细胞比例平均为0.87%,明显高于SPF小鼠的0.38%(P<0.01)。结论脾脏B10细胞可随Balb/c小鼠年龄增大而扩增,B10细胞的表达与性别差异无明显关系,微生物环境可促进B10细胞的活化,但对B10细胞总数无明显影响。

关键词： 高迁移率族蛋白B1   调节性T细胞   烫伤   乌司他丁   免疫  

摘要： 目的:研究乌司他丁对烫伤大鼠脾脏高迁移率族蛋白B1(HMGB1)的表达以及淋巴细胞免疫功能的影响。方法:96只Wistar大鼠随机分为假烫组、烫伤组和烫伤+乌司他丁治疗组(n=32)。后两组大鼠制作30%TBSAⅢ度烫伤模型,伤后即刻腹腔注射林格液或乌司他丁+林格液抗休克,创面涂碘伏抗炎。假烫组于37℃水浴中12 s行假烫。各组分别于伤后1、3、5、7 d腹主动脉无菌采血,并分离获取大鼠脾脏组织。Westernblot检测大鼠脾组织HMGB1的表达,流式细胞术检测血液中CD4^+CD25^+调节性T细胞(CD4^+CD25^+Treg)的变化。结果:与假烫组比较,烫伤组大鼠脾组织HMGB1水平在伤后1～7 d显著升高(P<0.01),伤后3 d时表达量最高,分别为0.66±0.10、0.22±0.05(t=10.91,P<0.01);乌司他丁治疗后1～7d HMGB1水平有不同程度的降低,其中伤后5 d表达量最低,分别为0.32±0.07、0.63±0.07(t=9.61,P<0.01)。与假烫组比较,烫伤组CD4^+CD25^+Treg表达在伤后逐渐上升,伤后3 d时达到峰值(5.42±0.56)%;乌司他丁治疗后,CD4^+CD25^+Treg于伤后3、5、7 d显著低于烫伤组(P<0.01),伤后5 d时最低(2.23±0.21)%。结论:严重烫伤后大鼠脾脏HMGB1水平的持续升高对其调节性T细胞的异常具有显著的影响;乌司他丁作为一种抗炎药物能够降低伤后HMGB1的表达,改善烫伤大鼠的免疫功能。

关键词： COLLAGEN-INDUCED ARTHRITIS   TGF-BETA   TOLERANCE INDUCTION   MONOCLONAL-ANTIBODY   ULCERATIVE-COLITIS   B10 CELLS   MICE   DEPLETION   RITUXIMAB   EFFECTOR  

摘要： The ability of activated B cells to protect against various experimental autoimmune or allergic diseases makes them attractive for use in cell-based therapies. We describe an efficient way to generate B cells with strong suppressive functions by incubating naive B cells with a relevant Ag conjugated to cholera toxin B subunit (CTB). This allows most B cells, irrespective of BCR, to take up and present Ag and induces their expression of latency-associated polypeptide (LAP)/TGF-beta and after adoptive transfer also their production of IL-10. With OVA as model Ag, when naive T cells were cocultured in vitro with B cells pretreated with OVA conjugated to CTB (OVA/CTB) Ag-specific CD4(+) Foxp3 regulatory T (Treg) cells increased >50-fold. These cells effectively suppressed CD25(-)CD4(+) effector T (Teff) cells in secondary cultures. Adoptive transfer of OVA/CTB-treated B cells to mice subsequently immunized with OVA in CFA induced increase in Foxp3 Treg cells together with suppression and depletion of Teff cells. Likewise, adoptive transfer of B cells pulsed with myelin oligodendrocyte glycoprotein peptide(35-55) (MOGp) conjugated to CTB increased the number of Treg cells, suppressed MOGp-specific T cell proliferation and IL-17 and IFN-gamma production, and prevented the development of experimental autoimmune encephalomyelitis. Similar effects were seen when B cells were given "therapeutically" to mice with early-stage experimental autoimmune encephalomyelitis. Our results suggest that B cells pulsed in vitro with relevant Ag/CTB conjugates may be used in cell therapy to induce Ag-specific suppression of autoimmune disease. The Journal of Immunology, 2012, 188: 1686-1697.

关键词： Graves病   调节性B细胞   白细胞介素10   转化生长因子-β  

摘要： 目的:观察Graves病(GD)小鼠模型脾脏调节性B细胞(regulatory B cells,Bregs)及相关细胞因子表达量的变化,探讨Bregs在GD发生、发展中的免疫调节作用。方法:将6周龄BALB/c雌性小鼠随机分为对照组(肌注空载腺病毒)、GD组(肌注TSHR-A亚单位-腺病毒建立GD小鼠模型)。每3周免疫1次,第3次免疫3周后处死。留取小鼠血清,化学发光法检测FT4水平,细胞培养生物学方法检测TSAb活性。取甲状腺,制备石蜡切片行HE染色。取脾脏,部分制成单个核细胞悬液,行三标记免疫荧光染色、流式细胞分析CD1dhiCD5+CD19+Bregs比例;部分脾脏提取总RNA,实时定量RT-PCR检测Bregs相关细胞因子白细胞介素10(IL-10)mRNA和转化生长因子-β(TGF-β)mRNA的表达水平。结果:发生GD的小鼠甲状腺滤泡细胞增生肥大,且血清FT4及TSAb水平升高。小鼠血清TSAb与FT4水平呈显著正相关(r=0.90,P<0.01);TSHR-A亚单位-腺病毒免疫组发生GD的小鼠脾脏CD19+B细胞较对照组明显升高(P<0.05),B细胞中CD1dhiCD5+CD19+Bregs所占比例和脾组织IL-10 mRNA、TGF-βmRNA表达量较对照组明显降低(P<0.05);且血清TSAb活性与CD1dhiCD5+CD19+Bregs占脾脏B细胞比例、脾组织IL-10 mRNA、TGF-βmRNA表达量均呈显著负相关(P<0.05)。结论:TSHR-A亚单位-腺病毒免疫建立的GD模型中Bregs表达存在缺陷,提示Bregs表达可能在GD的发生及发展中发挥免疫调节作用,且GD疾病严重程度可能与Bregs缺乏程度有关。

关键词： RHEUMATOID-ARTHRITIS  

摘要： A letter to the editor is presented in response to the article "B cell depletion enhances T regulatory cell activity essential in the suppression of arthritis," by K. M. Hamel and colleagues in the 2011 issue.

关键词： Foxp3   TGF-b   LAP   LRRC32   GARP   Tregs   autoimmunity   tolerance   transplantation  

摘要： Since its rediscovery in the mid-1990s,FOXP31 regulatory T cells(Tregs)have climbed the rank to become commander-in-chief of the immune *** possess diverse power and ability to orchestrate the immune system in time of inflammation and infection as well as in time of harmony and *** be the commander-in-chief,they must be equipped with both offensive and defensive *** review will focus on the function of transforming growth factor-b(TGF-b)as the sword,the wand,and the shield of *** as a sword,this review will begin with a discussion of the evidence that supports how Tregs utilize TGFb to paralyze cell activation and differentiation to suppress immune *** will next provide evidence on how TGF-b from Tregs acts as a wand to convert naıve T cells into iTregs and Th17 to aid in their combat against inflammation and ***,the review will present evidence on the role of TGF-b produced by Tregs in providing a shield to protect and maintain Tregs against apoptosis and destabilization when surrounded by inflammation and constant *** triadic function of TGF-b empowers Tregs with the responsibility and burden to maintain homeostasis,promote immune tolerance,and regulate host defense against foreign pathogens.

关键词： 自身免疫甲状腺炎   调节性B细胞   白细胞介素-10   转化生长因子-β   高碘诱导   免疫反应  

摘要： 目的<br>　　 自身免疫甲状腺炎(Auto immune thyroiditis,AIT)是典型的器官特异性自身免疫性疾病,AIT患者体内存在免疫耐受障碍,免疫应答调节受损以致免疫应答和炎症反应过强而造成甲状腺细胞的自身免疫性损伤,发病机制与细胞免疫和体液免疫异常均相关。B细胞主要通过呈递抗原和产生抗体参与体液免疫,其分化为不同亚群参与免疫反应的正向免疫效应与负向免疫调节过程。新近研究发现存在一种新的B细胞亚群——调节性B细胞(regulatory B cell,Breg),其与B细胞在自身免疫病中通常扮演致病性角色不同,Bregs通过分泌白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)等抑制性细胞因子介导免疫耐受,发挥免疫调节作用[3]。本研究通过高碘诱发NOD.H-2h4小鼠发生自身免疫甲状腺炎,观察此动物模型脾脏Bregs及相关细胞因子表达量的变化,探讨Bregs在AIT发生、发展中的免疫调节作用。<br>　　 方法<br>　　 选取4周龄NOD.H-2h4雌性小鼠40只,体重20克左右,随机分为2个对照组(给予无菌双蒸水)和2个高碘饮水组(给予含0.005%碘化钠水,相当于小鼠正常碘摄入量的100倍),每组10只。于实验开始后第10、20周麻醉分别处死1个对照组和1个高碘饮水组。留取各组NOD.H-2h4小鼠甲状腺,HE染色观察淋巴细胞浸润情况并进行甲状腺炎症程度评分;酶联免疫吸附试验(ELISA)测定各组小鼠血清甲状腺球蛋白抗体(TgAb)水平;制备各组NOD.H-2h4小鼠脾脏单个核细胞悬液,免疫荧光染色流式细胞仪分析CD1dhiCD5+CD19+调节性B细胞比例的变化;提取小鼠脾细胞总RNA,实时定量RT-PCR检测调节性B细胞相关细胞因子白细胞介素-10(IL-10)和转化生长因子-β(TGF-β)mRNA的表达水平。应用SPSS17.0软件处理和分析数据。<br>　　 结果<br>　　 NOD.H-2h4小鼠饮高碘水10周及20周后甲状腺组织有不同程度的淋巴细胞浸润,甲状腺炎的发生率、甲状腺炎症程度组织学评分、血清TgAb水平均明显高于饮用无菌双蒸水的对照组(P＜0.05);发生AIT的小鼠脾脏B细胞中CD1dhiCD5+CD19+Bregs所占比例和脾组织及脾细胞中IL-10mRNA、TGF-βmRNA表达量较对照组明显降低(P＜0.05)。随着高碘饮水时间的延长,甲状腺炎的发生率及甲状腺炎症程度组织学评分呈升高趋势,而脾脏Bregs表达则呈降低趋势。<br>　　 结论<br>　　 高碘诱导的发生AIT的NOD.H-2h4小鼠脾脏CD1dhic05+CD19+调节性B细胞及相关细胞因子IL-10和TGF-βmRNA表达较对照小鼠明显降低,且在AIT的发生发展过程中保持较低水平,提示调节性B细胞可能参与AIT的发生、发展。

关键词： CD19+CD24hiCD38hiB细胞   调节性B细胞   系统性红斑狼疮   外周血  

摘要： 研究背景：系统性红斑狼疮(systemiclupuserythematosus，SLE)是一种经典的、慢性、多系统受累的自身免疫病。其病因未明，免疫系统紊乱贯穿整个发病过程，T淋巴细胞的异常、B淋巴细胞的过度活化最终导致免疫系统的异常应答是其主要发病机制。B细胞主要通过呈递抗原和产生抗体发挥免疫调节作用。调节性B细胞(Bregs)是新近发现的一群新的B细胞亚群；是免疫系统中重要的新成员之一，具有免疫抑制作用；在SLE和RA等自身免疫性结缔组织病相关的鼠模型中已经证实Bregs发挥了重要作用。一项新的研究表明SLE患者外周血中的CD19+CD24hiCD38hiB细胞亚群具有调节功能。<br>　　 目的：探讨CD19+CD24hiCD38hiB细胞亚群在SLE患者外周血中的表达及临床意义。<br>　　 方法：病例选取2011年5月至2011年12月在苏州大学附属第一医院风湿科就诊的SLE患者36例，诊断均符合1997年修订的美国风湿病学会(AmericanCollegeofRheumatology,ACR)分类标准（见附表1）。收集SLE患者一般情况和临床资料，根据SLE疾病活动指数(systemiclupuserythematosusdiseaseactivityindex，SLEDAI)积分评估疾病的活动性（见附表2），并结合临床资料将患者分为活动组和稳定组。同时选取正常对照组20名；采用流式细胞术检测SLE患者和正常对照组之间外周血中CD19+CD24hiCD38hi、CD19+CD24intCD38int、CD19+CD24hiCD38-B细胞亚群百分比水平。采用SPSS18.0软件进行统计分析。<br>　　 结果：(1)SLE患者外周血中CD19+CD24hiCD38hiB细胞亚群百分比显著高于对照组(P＜0.05)；且SLE活动组CD19+CD24hiCD38hiB细胞亚群百分比显著高于稳定组(P＜0.05)。(2)SLE患者外周血中CD19+CD24hiCD38-B细胞亚群百分比显著低于对照组(P＜0.05)。(3)SLE患者外周血中CD19+CD24intCD38intB细胞亚群百分比与对照组无统计学差异(P＞0.05)。(4)CD19+CD24hiCD38hiB细胞在SLE患者外周血中表达水平与SLEDAI正相关(P＜0.05)；与病程、年龄、C3、ESR、CRP及SLEDAI等临床指标间无明显相关性(P＞0.05)。(5)CD19+CD24hiCD38-B细胞在SLE患者外周血中表达水平与ESR明显负相关(P＜0.05)；而与病程、年龄、C3、ESR、CRP及SLEDAI等其他临床指标间无明显相关性(P＞0.05)。(6)CD19+CD24intCD38intB细胞在SLE患者外周血中表达水平与疾病的病程、年龄、C3、ESR、CRP及SLEDAI等临床指标间无明显相关性(P＞0.05)。<br>　　 结论：(1)与正常对照组比较，CD19+CD24hiCD38hiB细胞在SLE患者外周血中的表达上调；而SLE患者外周血中CD19+CD24hiCD38-B细胞亚群百分比显著减低；说明CD19+CD24hiCD38hiB细胞亚群对效应T细胞分泌促炎症因子缺乏抑制功能并非由于数量不足所致，而是存在功能缺陷。<br>　　 (2)CD19+CD24hiCD38hiB细胞在SLE患者外周血中的表达水平与SLEDAI正相关；且该群细胞表达水平在SLE活动组显著高于稳定组，提示该群细胞可能与疾病活动相关。