体外CDl34L单抗或CTLA4Ig对狼疮样BXSB小鼠脾细胞分泌IL-6、IFN-γ及自身抗体的影响

目的：探讨体外CD134L单抗或CTLA4Ig对狼疮样BXSB小鼠脾细胞分泌IL-6、IFN-γ及自身抗体的影响。方法：采用未经治疗的狼疮样BXSB小鼠模型，应用CDl34L单抗和／或CTLA4Ig体外特异性阻断CDl34-CDl34L或B7-CD28通路后，用MTT法测定分裂原刀豆蛋白A（ConA）诱导的脾淋巴细胞增殖反应和用ELISA方法测定ConA诱导的脾细胞培养上清液中IL-6、IFN-γ和抗ds-DNA抗体的表达水平，并与经中药狼疮方或强的松治疗的狼疮样BXSB小鼠模型进行比较。结果：（1）在单纯培养或经ConA刺激后培养，相比于正常对照鼠，狼疮样小鼠脾淋巴细胞都表现有增殖反应性的显著增高，IFN-γ、IL-6蛋白量的增高和抗ds-DNA抗体的过度分泌。（2）经强的松或中药狼疮方体内治疗后，狼疮样小鼠脾淋巴细胞体外培养的增殖反应性及IFN-γ、IL-6的分泌都受到明显抑制，抗ds-DNA抗体的产生也明显减少。（3）体外单独应用CD134L单抗或CTLA41g特异性阻断CD134-CD134L或B7-CD28共刺激信号通路，同样可以显著抑制体外培养的狼疮样小鼠脾淋巴细胞的增殖反应及IFN-γ、IL-6的分泌，并可明显减少抗ds-DNA抗体的产生，但它们的抑制作用比强的松、中药狼疮方体内治疗狼疮样小鼠时所表现出的类似效应要弱。（4）联合CD134L单抗和CTLA4Ig，则治疗作用显著提高，其抑制脾淋巴细胞的增殖反应及IFN-γ、IL-6的分泌，抗ds-DNA抗体产生的作用都优于中药狼疮方的体内治疗效果，而与强的松的体内治疗效应相当。结论：CD134-CD134L可以提供独立的、非B7-CD28依赖的，另一种驱动抗原特异性T细胞增殖的协同刺激通路。同时阻断B7与CD28、CD134L与CD134间的相互作用，使自身反应性T淋巴细胞的活化和增殖受到快速而最大限度的抑制，对治疗SLE等自身免疫性疾病可能是一种较理想的免疫干预模式。     

中药狼疮方对狼疮样BXSB小鼠肺组织CD134/CD134L和RANTES表达的影响

目的探讨中药狼疮方对狼疮样BXSB小鼠肺组织CD134／CD134L和RANTES表达的影响。方法采用BXSB小鼠模型,随机分为3组：狼疮方治疗组、强的松治疗组、未治疗组,每组6只,疗程10周。另设与BXSB小鼠同基因的正常C57BL／6小鼠6只为正常对照组。分别取小鼠肺组织,应用逆转录-荧光定量-聚合酶链反应（RT-FQ-PCR）技术定量测定小鼠肺组织CD134、CD134L和趋化因子RANTES的mRNA表达水平。结果①未治疗组小鼠肺组织CD134、CD134L mRNA和RANTKS mRNA的表达水平都显著高于正常对照组（P〈0．01,P〈0．05）;经强的松或中药狼疮方治疗后,BXSB小鼠肺组织CD134、CD134L及RANTES的mRNA表达都受到明显抑制,显著低于未治疗组（P〈0．01,P〈0．05）;且接近正常水平,与正常对照组无显著性差异（P〉0．05）。②BXSB小鼠肺组织RANTES的mRNA表达水平与CD134L的mRNA表达水平呈显著的正相关关系（r=0．793,P〈0．05）,而与CD134的mRNA表达水平无显著的相关关系（r=0．412,P〉0．05）。结论中药狼疮方具有与强的松类似的免疫抑制作用,可显著抑制狼疮样小鼠肺组织CD134／CD134L共刺激信号表达;并下调肺组织RANTKS mRNA表达水平,具有一定的肺脏保护作用。     

狼疮样BXSB小鼠脾细胞CD134/CD134L的表达及治疗影响

为了探讨CD134/CD134L共刺激信号在SLE发病机制中的可能作用及治疗的影响。采用狼疮样BXSB小鼠模型,随机分为三组:狼疮方治疗组、强的松治疗组、未治疗组,每组6只,疗程10周。另设与BXSB小鼠同基因的正常C57BL/6小鼠6只为正常对照组。分别采集上述各组小鼠外周血和脾组织进行检测。结果:(1)与正常对照组比较,未治疗组BXSB小鼠的血清IgG和抗dsDNA抗体水平和脾组织CD4+CD134+、CD8+CD134+、CD19+CD134L+、CD23+CD134L+的百分比及CD134、CD134L的mRNA拷贝数都显著增高(P<0.05或P<0.01)。且血IgG和抗dsDNA抗体水平与脾细胞CD19+CD134L、CD23+CD134L的水平增加呈正相关;(2)狼疮方治疗组或强的松治疗组的血清IgG、抗dsDNA抗体水平和脾组织CD4+CD134+、CD8+CD134+、CD19+CD134L+、CD23+CD134L+的百分比及CD134、CD134L的mRNA拷贝数都显著低于未治疗组(P<0.05或P<0.01),而与正常对照组无显著差异(P均>0.05)。狼疮样BXSB小鼠存在CD134/CD134L的异常表达。狼疮方可减轻狼疮样小鼠高丙种球蛋白血症,减少体内自身抗体产生,对CD134/CD134L的下调作用与强的松相当。     

体外CD134L单抗或CTLA4Ig对狼疮样BXSB小鼠脾细胞分泌IL-6、IFN-γ及自身抗体的影响

目的:探讨体外CD134L单抗或CTLA4Ig对狼疮样BXSB小鼠脾细胞分泌IL-6、IFN-γ及自身抗体的影响。方法:采用未经治疗的狼疮样BXSB小鼠模型,应用CD134L单抗和/或CTLA4Ig体外特异性阻断CD134-CD134L或B7-CD28通路后,用MTT法测定分裂原刀豆蛋白A(ConA)诱导的脾淋巴细胞增殖反应和用ELISA方法测定ConA诱导的脾细胞培养上清液中IL-6、IFN-γ和抗ds-DNA抗体的表达水平,并与经中药狼疮方或强的松治疗的狼疮样BXSB小鼠模型进行比较。结果: (1)在单纯培养或经ConA刺激后培养,相比于正常对照鼠,狼疮样小鼠脾淋巴细胞都表现有增殖反应性的显著增高,IFN-γ、 IL-6蛋白量的增高和抗ds-DNA抗体的过度分泌。(2)经强的松或中药狼疮方体内治疗后,狼疮样小鼠脾淋巴细胞体外培养的增殖反应性及IFN-γ、IL-6的分泌都受到明显抑制,抗ds-DNA抗体的产生也明显减少。(3)体外单独应用CD134L单抗或 CTLA4Ig特异性阻断CD134-CD134L或B7-CD28共刺激信号通路,同样可以显著抑制体外培养的狼疮样小鼠脾淋巴细胞的增殖反应及IFN-γ、IL-6的分泌,并可明显减少抗ds-DNA抗体的产生,但它们的抑制作用比强的松、中药狼疮方体内治疗狼疮样小鼠时所表现出的类似效应要弱。(4)联合CD134L单抗和CTLA4Ig,则治疗作用显著提高,其抑制脾淋巴细胞的增殖反应及 IFN-γ、IL-6的分泌,抗ds-DNA抗体产生的作用都优于中药狼疮方的体内治疗效果,而与强的松的体内治疗效应相当。结论: CD134-CD134L可以提供独立的、非B7-CD28依赖的,另一种驱动抗原特异性T细胞增殖的协同刺激通路。同时阻断B7与 CD28、CD134L与CD134间的相互作用,使自身反应性T淋巴细胞的活化和增殖受到快速而最大限度的抑制,对治疗SLE等自身免疫性疾病可能是一种较理想的免疫干预模式。     

糖皮质激素对BXSB狼疮小鼠肾组织fractalkine/CX3CR1表达的影响

目的：研究BXSB狼疮小鼠肾组织趋化因子fractalkine及其受体CX3CR1的表达以及给予泼尼松治疗后的改变,探讨两者在狼疮肾炎发病机制中的可能作用。方法：12 周龄雄性BXSB狼疮小鼠随机分成泼尼松治疗组（n=6）和实验对照组（n=6）;另取同周龄雄性C57BL/6J小鼠6只作为正常对照组。正常对照组和实验对照组小鼠每天给予0.5 mL生理盐水灌胃;泼尼松治疗组小鼠每天给予0.18 mg/20 g BW的泼尼松溶于0.5 mL生理盐水灌胃。持续10周结束实验。应用逆转录-聚合酶链反应（RT-PCR）及Western印迹检测小鼠肾组织fractalkine和CX3CR1 mRNA和蛋白的表达,并检测小鼠实验室指标以及肾脏组织病理学的变化。结果： BXSB狼疮小鼠肾组织fractalkine以及CX3CR1 mRNA和蛋白表达均较C57BL/6J小鼠明显增高,而经过泼尼松治疗后的BXSB小鼠两者的表达均较未治疗组（实验对照组）明显下降,同时伴有血清免疫球蛋白G（IgG）、IgM、血清抗双链脱氧核糖核酸（dsDNA）抗体水平以及血尿素氮（BUN）、血肌酐（SCr）水平的明显改善,尿蛋白减少;肾小球内免疫复合物沉积和肾脏组织病理学改变亦显著减轻。结论： 实验结果提示fractalkine/CX3CR1可能参与了小鼠狼疮肾炎的发病机制,且糖皮质激素可能通过抑制肾脏fractalkine的表达而发挥其治疗效应。     

三氧化二砷对MRL/lpr狼疮鼠抗ds-DNA抗体和淋巴细胞亚群的影响

目的研究三氧化二砷(ATO)对MRL/lpr狼疮鼠抗ds-DNA抗体和淋巴细胞亚群的影响。方法将MRL/lpr狼疮鼠随机分为:ATO组、生理盐水(NS)组和环磷酰胺(CTX)组。另设正常C57/BL小鼠,随机分为ATO治疗组(0.4 mg/kg.d)和对照组(0.25 mL)。以上每组各6只,隔日腹腔注射,给药2个月后处死,称体重和脾脏重量算出脾脏指数;ELISA法测各组小鼠血清抗ds-DNA抗体水平;流式细胞术测脾脏细胞亚群百分比。结果(1)MRL/lpr小鼠ATO治疗组脾脏指数、血清抗ds-DNA抗体水平和CD19 、CD3 、CD3 CD4 细胞百分比均低于NS对照组(P<0.01或P<0.05),NK细胞百分比明显高于NS对照组(P<0.05);(2)ATO治疗组小鼠的脾脏指数和CD3CD4双阳性细胞百分比显著低于CTX组(P<0.01),CTX治疗组小鼠血清抗ds-DNA抗体水平均较对照组明显降低(P<0.01),但ATO治疗组的抗体水平较CTX治疗组高(P<0.05);(3)在C57/BL小鼠中,ATO治疗组和NS治疗组之间以上指标均无差异。结论ATO抑制MRL/lpr狼疮鼠T、B淋巴细胞活化同时上调NK细胞功能,减少体内自身抗体产生;但是对正常小鼠的抗体水平和细胞亚群并无明显影响。     

狼疮方对狼疮样小鼠肾脏RANTES表达的影响

目的 :观察慢性移植物抗宿主病 (GAHD)狼疮样肾炎小鼠模型肾组织中趋化因子RANTES的表达并探讨中药狼疮方对其的影响。方法 :以强的松为阳性对照 ,观察狼疮方对模型小鼠肾脏RANTES表达的影响 ,并进一步分析其对肾脏病理和功能损害的影响。结果 :狼疮方具有与强的松相似的防止和减轻狼疮样肾炎小鼠肾功能恶化、肾组织病理改变的作用(P <0 0 0 1)。这作用与减轻狼疮小鼠肾组织中RANTES表达显著相关。结论 :狼疮方明显下调狼疮肾炎肾脏RANTES表达 ,这与其减轻病理改变相一致     

Pristane诱导狼疮模型鼠共刺激分子的表达变化

探讨共刺激分子在Pristane诱导狼疮鼠体内的表达变化及意义。采用6~8周龄雌性BALB/c鼠单次腹腔注射0.5ml Pristane,对照组注射0.5ml PBS。通过尿蛋白试纸法检测尿蛋白含量;ELISA法检测外周血自身抗体(抗ds-DNA、抗Sm/RNP)含量;HE染色评估病理学损伤;流式细胞术检测小鼠外周血、脾脏淋巴细胞表面共刺激分子CD28、CD40L/CD40、CD137/CD137L的表达;免疫组织化学法检测小鼠组织中共刺激分子CD28、CD40L、CD137的表达。实验结果显示:与对照组小鼠相比,模型组小鼠出现多种SLE样临床症状和体征;模型组小鼠外周血和脾脏淋巴细胞表面共刺激分子CD28,一个月时表达增高,后逐渐下降至低于正常对照组(P0.05);CD40三个月时表达升高,六个月时表达降低至低于正常对照组(P0.05),而CD40L的表达变化无统计学意义(P0.05);CD137/CD137L三个月时表达增高并维持高表达状态至处死(P0.05);狼疮鼠肺组织中共刺激分子CD28、CD40L和CD137呈现高表达状态,其他组织中变化并不明显。以上结果提示,Pristane诱导狼疮鼠体内共刺激分子表达发生异常变化,可能与其疾病的发生发展有密切关系,为临床治疗系统性红斑狼疮提供了研究基础。     

CTLA-4Ig对B6.MRL-Faslpr/J狼疮小鼠脾脏Th17和Treg细胞表达的影响与其对小鼠狼疮样病征干预作用的相关性研究

目的:初步探讨B7阻断剂CD28与细胞毒T淋巴细胞相关抗原4免疫球蛋白(CTLA-4Ig)对B6.MRL-Faslpr/J狼疮小鼠脾脏Th17和调节性T细胞(Treg细胞)表达的影响与其对小鼠狼疮样病征干预作用之间的相关性。方法:将4个月龄大小雌性B6.MRL-Faslpr/J狼疮小鼠16只,随机分为观察组(Ⅰ组)和对照组(Ⅱ组),分别静脉注射CTLA-4Ig及等量PBS,检测小鼠干预前后24 h尿蛋白、ANA抗体、ds-DNA抗体及干预结束2周后血清IL-17A、脾脏中Th17细胞和Treg细胞百分比。结果:末次干预2周后Ⅰ组的24 h尿蛋白、血清ANA及ds-DNA较Ⅱ组下降均有统计学意义(均P0.05)。末次干预2周后Ⅰ组血清中IL-17A、脾脏Th17细胞比例均较Ⅱ组低,而脾脏的Treg细胞占CD4+T淋巴细胞的比例高于Ⅱ组,差异均有统计学意义(均P0.05)。结论:CTLA4-Ig具有减轻B6.MRL-Faslpr/J狼疮鼠狼疮样病征的作用;上调Treg细胞、下调Th17细胞可能是CTLA-4Ig减轻B6.MRL-Faslpr/J狼疮鼠狼疮样病征的重要机制之一。     

氟达拉宾对系统性红斑狼疮BXSB小鼠狼疮活动的影响

目的 :探讨氟达拉宾对系统性红斑狼疮BXSB小鼠狼疮活动的影响 ,氟达拉宾治疗重型系统性红斑狼疮的可能性、有效性及其可能的机制。方法 :用 30mg (m2 ·d) ,连续 3天氟达拉宾尾静脉注入BXSB小鼠体内 ,用血液分析仪分析用氟达拉宾前后不同时间小鼠外周血白细胞的变化 ,用ELISA方法测定BXSB小鼠血清抗ds DNA抗体、抗核抗体的变化 ,免疫荧光检查肾组织的病理改变 ,尿蛋白试纸检测用氟达拉宾前后BXSB小鼠的蛋白尿 ,流式细胞仪分析T淋巴细胞表面CD4 + Fas+ 、CD8+ Fas+ 、、CD4 5RO+ Fas+ 表达的变化。结果 :用氟达拉宾后BXSB小鼠外周血白细胞数从第 3天开始下降 ,至第 7天时白细胞下降至最低值〔(0 5± 0 2 )× 10 9L- 1 〕 ,白细胞上升至 1 0× 10 9L- 1 的时间是用药后 19天 ;BXSB小鼠血清抗ds DNA抗体、抗核抗体的水平明显下降 ,分别出现在用氟达拉宾后第 14、2 1天时 ;用药后第 2 8天氟达拉宾组 72 7%的BXSB小鼠肾组织进行免疫荧光病理检查 ,其荧光强度由 +～ ++→± ;用氟达拉宾后第 2 1、2 8天尿蛋白从 ++～ +++转±～ -占 81 8% ;用Flu后BXSB小鼠CD4 + Fas+ 、CD8+ Fas+ 、CD4 5RO+ Fas+ 的表达均明显低于用Flu前。结论 :氟达拉宾可明显减少BXSB小鼠血清抗ds DNA抗体、抗核抗体的水平 ,减少BXSB小     

Effect of anti-CD134L mAb and CTLA4Ig on ConA-induced proliferation, Th cytokine secretion, and anti-dsDNA antibody production in spleen cells from lupus-prone BXSB mice

We sought to evaluate the effects of combined downregulation of CD134 and cytotoxic T lymphocyte-associated antigen 4 (CTLA4) on the autoimmune process of lupus. Concanavalin A (ConA)-induced proliferation, T helper cell cytokine secretion, and anti-double stranded DNA (dsDNA) antibody production were measured in cultures of splenic lymphocytes derived from lupus-prone BXSB mice. Splenocytes from six prednisone-treated and six untreated male lupus-prone BXSB mice, as well as from six syngeneically normal C57BL/6 male mice, were stimulated with ConA. BXSB splenocytes from untreated mice were exposed to anti-CD134L mAb, CTLA4 linked to the Fc portion of IgG1 (CTLA4Ig), or both. The magnitude of splenocyte proliferation and the levels of IFN-gamma, IL-6, and anti-dsDNA antibody were: (1) significantly higher in cultures of ConA-stimulated control and other cells than in unstimulated cells, (2) similar in cultures of normal and BXSB cells treated with anti-CD134 and CTLA4Ig or prednisone and (3) significantly reduced in cultures of ConA-stimulated and unstimulated cells treated with anti-CD134L and CTLA4Ig or prednisone compared with cells treated with CD134L or CTLA4Ig alone. Like corticosteroids, anti-CD134L mAb or CTLA4Ig can inhibit T- and B-cell activation by blocking the CD134-CD134L or CD28/CTLA4-B7 co-stimulatory pathway. The combined immune intervention described herein may prove useful for the treatment of autoimmune diseases such as systemic lupus erythematosus.     

狼疮性BXSB小鼠脾脏淋巴细胞增殖与凋亡的初步分析

为了比较全面准确地了解系统性红斑狼疮 (SLE )BXSB小鼠的发病过程中 ,淋巴细胞增殖与凋亡的动力学变化及其机制。采用细胞双色荧光染色的标记技术 ,检测了脾脏淋巴细胞中的增殖细胞和凋亡细胞的百分率 ,并且测定了巨噬细胞吞噬凋亡细胞的能力。结果发现 ,发病的雄性BXSB小鼠和雌性BXSB小鼠脾脏中增殖的CD4 + T淋巴细胞和B淋巴细胞百分率显著高于对照C5 7小鼠 ,而凋亡的B淋巴细胞的百分率显著低于对照C5 7小鼠 ;但是 ,雌雄BXSB小鼠和对照C5 7小鼠巨噬细胞吞噬凋亡细胞的吞噬指数相同。本研究结果表明 ,在BXSB小鼠的SLE发病过程中 ,淋巴细胞的增殖速度异常升高、而凋亡速度下降 ,可能与其脾脏肿大有关 ;而且淋巴细胞的增殖与凋亡的失衡与巨噬细胞的功能无关 ,可能与淋巴细胞内在的异常有关。     

Isolation and functional analysis of autoreactive T cells from BXSB mice with murine lupus

CD4(+) T helper cells play a pivotal role in the pathogenesis of SLE, although the mechanism is still unclear. The present study was designed to isolate and characterize autoreactive T lymphocytes from BXSB mice, a mouse model for human SLE. Splenocytes from 6-month-old male BXSB mice with murine lupus were repeatedly stimulated in vitro with irradiated syngeneic B cells in the presence of recombinant IL-2, resulting in six autoreactive T-cell lines and two T-cell clones. TCR analysis showed that, one of the T-cell lines, ATL1, was almost clonal, as a Vbeta2.1-Jbeta2, a Valpha5.1-Jalpha15 and a Valpha10.1-Jalpha15 chains were predominantly expressed in this line. The two clones derived from ATL1 turned out to be sister clones, using the TCR Vbeta2.1-Jbeta2 and Valpha10.1-Jalpha15 chains. ATL1 cells proliferated in response to stimulation of syngeneic and H-2-matched allogeneic B cells and secreted IFN-gamma. Monoclonal Ab against CD4 and CD28 inhibited the proliferative response of ATL1 for syngeneic B cells. Interestingly, ATL1 did not respond to BXSB spleen or peritoneal macrophages, suggesting that B cells were able to either express accessory molecules necessary for T-cell triggering or present cryptic epitopes recognized by the autoreactive T cells. Moreover, ATL1 was able to help BXSB, but not C57BL/6, B cells producing IgG and IgM Abs against dsDNA and histone in vitro. Passive transfer of viable ATL1 cells into young female BXSB mice significantly accelerated the production of autoantibodies. Possible mechanisms of interaction between ATL1 and lupus B cells are further discussed.     

Antibody production in autoimmune BXSB mice. I. CD40L-expressing B cells need fewer signals for polyclonal antibody synthesis.

Male, but not female, BXSB mice develop severe lupus associated with multiple immune system defects. It was recently shown that one immunological abnormality found in male BXSB mice encompasses B cell expression of CD40 ligand (CD40L) by an expanded population of large B cells. The present study was undertaken to determine how the CD40L-expressing large B cells in male BXSB mice compared with size-matched B cells from female mice in terms of their ability to secrete antibody. It was shown that the large B cells from female mice, similar to the small B cells from either male or female mice, required CD40 signalling, immunoglobulin cross-linking and cytokines for optimal antibody synthesis. In contrast, large B cells from male BXSB mice produced high levels of antibody when stimulated with only two of the three signals, and made significantly more total IgM and IgG, and anti-ssDNA antibody than size-matched B cells from female mice when stimulated with IL-4/IL-5 alone, IL-4/IL-5 plus low levels of anti-IgD-dextran, or IL-4/IL-5 plus anti-CD40 MoAb. The ability of the large B cells from male mice to produce antibody under suboptimal stimulatory conditions correlated with their expression of CD40L, and was inhibited by CD40-immunoglobulin. Taken together, these findings suggested that large CD40L-expressing B cells from male BXSB mice may be able to bypass a need for CD40 signalling from T cells, thus contributing to autoimmune disease by promoting antibody production in the absence of cognate T cell help.     

B cell apoptosis accelerates the onset of murine lupus

To investigate whether the increased rate of lymphocyte apoptosis in systemic lupus erythematosus is involved in the onset of the disease, apoptotic or necrotic T or B lymphocytes from various cell lines were injected intraperitoneally into pre-autoimmune (NZBxNZW)F1 mice (BW) and non-autoimmune BALB/c mice. The intraperitoneal production of cytokines and chemokines, the specific T cell response in the spleen, and the production of anti-histone and anti-dsDNA Ab were investigated. The onset of the disease was characterized by creatinine levels and evaluation of glomerular IgG deposits. In BW, but not in BALB/c mice, injection of apoptotic and not necrotic cells up-regulated IL-6 and IL-10 in resident macrophages. Administration of apoptotic cells augmented the number of Th2 and B lymphocytes recruited in the peritoneal cavity. Only the treatment with apoptotic B cells promoted a systemic Th2 autoimmune response to H2 histones, associated with earlier occurrence of high levels of anti-dsDNA autoantibodies, higher creatinine levels and more numerous glomerular IgG deposits than in BW controls not injected with apoptotic B cells. In genetically susceptible mice exposure to apoptotic of B, but not T, lymphocytes can elicit a Th2 response to H2 histones that helps B cell production of anti-dsDNA Ab and finally triggers the onset of lupus.     

Long-term anti-CD4 treatment of MRL/lpr mice ameliorates immunopathology and lymphoproliferation but fails to suppress rheumatoid factor production

MRL/lpr mice were treated with anti-CD4 mAb to define the role of CD4+ T cells in the pathogenesis of autoimmune disease and the lymphoproliferation characteristic of the strain. Anti-CD4 treatment was not associated with adverse effects, and survival of treated mice was increased over that of rat IgG-treated controls. Renal function was preserved, and the histologic severity of glomerulonephritis was minimal in treated mice. Lymphoid tissues of mice receiving anti-CD4 were effectively depleted of CD4+ T cells, and lymphoproliferation was markedly reduced. Serum IgG, anti-Sm, and anti-dsDNA levels were reduced significantly, while serum IgM and IgM rheumatoid factor levels were unaffected by anti-CD4 treatment. These data show that in MRL/lpr mice lymphoproliferation, renal disease, anti-Sm and anti-dsDNA antibody production, and elevated IgG levels are all linked to CD4+ T cell function. In contrast, both total IgM and IgM rheumatoid factor production appear to be the result of B-cell activity that is not regulated by CD4+ T cells.     

B cells from autoimmune BXSB mice are hyporesponsive to signals provided by CD4+ T cells

Male BXSB mice, unlike female BXSB mice, develop an early-onset, lupus-like disease characterized by high levels of anti-nuclear antibodies (Abs) and total Ig. It has recently been shown that the male BXSB mice contain an expanded population of large B cells which are hyperresponsive to stimulation by anti-CD40 mAb. The present study was undertaken to determine whether their potential for extra CD40 signaling enabled the B cells from male BXSB mice to hyper-respond to CD40L-expressing CD4+ T cells. In contrast to expectations, large B cells from male BXSB mice did not interact with CD4+ T cells in a positive manner; cultures of B cells from antigen (Ag)-primed male BXSB mice, unlike cultures of B cells from Ag-primed female mice, generated few antibody forming cells (AFC) following interaction with activated CD4+T cells. In addition, B cells from male BXSB mice, unlike B cells from female BXSB mice, failed to upregulate MHC class II molecules following interaction with activated CD4+ T cells. Subsequent experiments revealed that the inability of the B cells from the male mice to upregulate MHC class II molecules in response to T cell-mediated activation resided primarily in the population of large B cells. Large B cells from male BXSB mice were also defective in their ability to proliferate following stimulation with activated CD4+ T cells. Taken together, these findings demonstrated that similar to B cells in lupus patients, large B cells from male BXSB mice could function in a hyporesponsive manner, and that this hyporesponsiveness related to the inability of the B cells to interact in a positive manner with CD4+T cells.     

中药复方对重症肌无力淋巴细胞亚群的影响

目的探讨中药复方对重症肌无力（MG）患者的T淋巴细胞介导的免疫调节机制。方法将60例患者随机分为两组,每组30例,治疗组服用黄芪复方,对照组服用泼尼松片,疗程12周。应用流式细胞仪检测MG患者治疗前后外周血淋巴细胞亚群分布的变化情况。结果经治疗后,治疗组和对照组CD4^＋及CD4^＋／CD8^＋比值与本组治疗前比较均有明显下降,CD8^＋明显增加,差异均有统计学意义（P〈0．05）;治疗组和对照组治疗后组间比较各项指标差异均无统计学意义（P〉0．05）。结论淋巴细胞亚群比例分布等免疫功能的变化可能是中药黄芪复方发挥免疫调节作用机制之一。