系统性红斑狼疮抗核抗体易感基因染色体定位

目的 :确定新西兰小鼠自身抗染色质抗体、抗DNA抗体、抗组蛋白抗体的遗传易感基因染色体定位。方法 :建立新西兰黑色品系 (NZB)和新西兰白色品系 (NZW)子代F1的回交小鼠模型 (NZB×NZW)×NZB和 (NZB×NZW)×NZW ,采用覆盖小鼠 19条染色体的微卫星遗传标记及数量性状位点 (QTL)分析进行基因定位。结果 :确定了自身抗染色质抗体产生的易感基因 ,位于NZW小鼠第 9染色体中段 30厘摩临近 (Lods=3 0 1) ;另外自身抗染色质抗体、抗DNA抗体、抗组蛋白抗体产生的共同易感基因 ,位于NZB及NZW小鼠第 17染色体 ,即H 2复合体及TNF α的基因区域 (Lods值 >4 )。结论 :(NZB×NZW)F1小鼠自身抗核抗体的产生受多基因调控 ,候选易感基因不仅来自于NZB ,也来自于NZW。     

新西兰小鼠CD8+T细胞数量异常的易感基因定位

目的 :定位自发性SLE模型 (NZB×NZW)F1小鼠CD8 T细胞数量异常的遗传易感基因。方法 :建立 (NZB×NZW )F1×NZW回交小鼠模型 ,采用覆盖小鼠 1 9条常染色体的多态性微卫星遗传标记及数量性状位点分析进行基因定位。结果 :CD8 T细胞数量异常的易感基因与小鼠第 1条染色体尾端 92 3cM处的微卫星遗传标记D1Mit36肯定连锁 (Lods值 >3) ,且回交小鼠该遗传标记B W型组的外周血淋巴细胞中CD8 T细胞百分比明显低于W W型组 (P <0 0 0 1 )。结论 :(NZB×NZW )F1小鼠外周血CD8 T细胞数量异常的候选易感基因位于第 1条染色体尾端 92 3cM附近 ,来源于NZB小鼠。     

初步探讨(NZB×NZW)F1×NZW回交鼠和狼疮性肾炎相关的易感基因

目的:探讨与狼疮性肾炎相关的新西兰黑鼠(NZB)易感基因染色体位点.方法:通过建立(NZB×NZW)F1×NZW回交小鼠模型,以尿蛋白为狼疮性肾炎表现型,利用微卫星遗传标记进行基于数量性状位点(QTL)分析的全基因组扫描方法,寻找源于NZB的遗传易感基因位点并确定染色体位置.结果:根据QTL分析发现两个与狼疮鼠蛋白尿发生有关的易感基因位点,分别位于NZB小鼠第4号染色体的D4Mit71区域及第17号染色体D17Mit22区域(LOD＞3.3).同时携有两个NZB位点的小鼠尿蛋白阳性率最高,而只有一个位点的小鼠其尿蛋白阳性率较NZW纯合鼠也明显增高(P＜0.005).结论:NZB第4号染色体的D4Mit71区域及第17号染色体的D17Mit22区域可能为(NZB×NZW)F1×NZW回交鼠狼疮性肾炎蛋白尿易感位点.两个位点之间具有协同增效作用.     

CD5阳性B1细胞增殖易感基因的定位

目的 :定位NZB、NZW及NZB WF1小鼠外周血B1细胞增殖的易感基因。方法 :建立 (NZB×NZW )F1×NZB回交小鼠模型 ,用覆盖小鼠全部常染色体的多态性微卫星遗传标记数量性状位点 (QTL)分析进行基因定位。结果 :B1细胞增殖的易感基因与小鼠的第 2、4、17号染色体上的微卫星遗传标记D2Mit 10 1、D4Mit 11及D17Mit 6 1肯定连锁 (Lods值 >3) ,其中D2Mit10 1、D4Mit11位点来源于NZB ,D17Mit 6 1位点来源于NZW。在D2Mit10 1位点附近存在Ltk、Rag1、2基因 ,在D4Mit 11位点附近存在Lck基因 ,在D17Mit6 1位点附近存在H 2及TNF α基因。结论 :NZB、NZW及NZB WF1小鼠外周血B1细胞增殖受多基因调控 ,易感基因分别位于NZB的Ltk、Rag1、2及Lck基因编码区和位于NZW的H 2、TNF α基因编码区 ,这些易感基因之间有相互促进的叠加效应。     

SLE模型小鼠高IgG血症易感基因-Fcgr2b基因突变及对其表达的影响

目的:测定系统性红斑狼疮(SLE)小鼠模型 (NZB×NZW)F1双亲NZB ,NZW小鼠Fcgr2b基因启动子区核酸序列及其表达的改变,明确Fcgr2b基因的突变性质、对该基因表达的影响及与高IgG血症的关系。方法:采用核酸序列分析测定NZB ,NZW小鼠Fcgr2b基因启动子区突变性质;RT PCR检测Fcgr2b基因表达的改变;ELISA法测定比较(NZB×NZW)F1,NZB和NZW小鼠及(NZB×NZW)F1×NZW回交小鼠Fcgr2b基因B/W型与W /W型组间血清总IgG水平。结果:NZB小鼠Fcgr2b基因启动子区与正常鼠BALB/c相比存在2个部位碱基缺失,分别为13bp及3bp。NZW小鼠除有3个碱基置换外,与BALB/c鼠该基因启动子区序列相同;NZB小鼠Fcgr2b基因mRNA的表达较正常鼠BALB/c降低;NZB小鼠血清总IgG水平明显高于NZW小鼠(P <0 0 5 ) ,回交小鼠Fcgr2b基因B/W型组血清总IgG水平明显高于W/W型组(P <0 0 0 0 1)。结论:NZB小鼠Fcgr2b基因启动子区存在碱基缺失,且该缺失突变可引起其表达的降低而导致高IgG血症     

Fcgr2b基因及H-2复合体对SLE模型小鼠IgG抗体产生的调节作用

目的:分析Fcgr2b基因对SLE小鼠血清总IgG抗体产生的影响;Fcgr2b基因单独及Fcgr2b基因与H-2复合体共同作用对SLE小鼠血清IgG抗DNA抗体产生的调控。方法:建立(NZB×NZW)F1×NZW回交小鼠模型,用特异性荧光抗体染色,流式细胞术检测及PCR技术进行基因分型,以ELISA法测定血清总IgG及抗DNA抗体水平进行分析比较。采用扩增片段长度多态性(AmpFLP)分析NZB,NZW小鼠Fcgr2b基因启动子区是否有多态性。结果:(NZB×NZW)F1×NZW回交小鼠Fc-gr2b基因B/W型组血清总IgG水平明显高于W/W型组(P<0.0001)。Fcgr2b基因独作用时,回交小鼠Fcgr2b基因B/W型组与W/W型组间血清IgG抗DNA抗体水平差异不显著(P>0.05)。Fcgr2b基因与H-2复合体共同作用时,H-2复合体为d/z型时,无论Fcgr2b基因是B/W型或W/W型,其血清IgG抗DNA抗体水平明显高于含H-2复合体Z/Z型组(P<0.01);H-2复合体为d/z型时,含Fcgr2b基因B/W型组血清IgG抗DNA抗体水平明显高于W/W型组(P<0.01)。NZB小鼠Fcgr2b基因启动子区扩增片段长度短于非自身免疫病小鼠NZW,C57BL/6及BALB/c小鼠,提示可能存在碱基缺失。结论:血清总IgG水平由Fcgr2b基因调控;IgG抗DNA抗体产生主要由H-2复合体调控,Fcgr2b基因单独作用对该抗体产生的影响不明显,但与H-2复合体具有协同作用。     

活性染色质诱导抗核抗体生成及肾损伤

系统性红斑狼疮 (SLE )的病因和诱导抗核抗体 (ANA )生成的激发原迄今不明。本实验试图用ConA活化淋巴细胞的染色质免疫同系BALB/c小鼠 ,寻找诱导ANA生成的真正免疫原 ,阐明SLE发生的激发原是自身活化细胞的核成分 ,而且证明它所诱导的ANA具有致病性。从ConA活化的脾细胞中提取染色质 ,然后免疫同系BALB/c小鼠 ,用ELISA方法测定IgG类抗双链DNA (dsDNA )、抗组蛋白抗体 ,用免疫荧光法检测抗核抗体核型和免疫复合物沉积 ,用免疫印迹法测定抗核抗体谱 ,在光镜下检测肾损伤及电镜下检测肾小球沉积物 ,用考马斯亮蓝法检测尿蛋白含量。结果显示 ,活性染色质能诱导IgG类抗dsDNA、抗组蛋白等多种抗核抗体生成 ,且肾小球有显著免疫复合物沉积和蛋白尿形成。该实验表明 ,活化淋巴细胞的染色质是诱导SLE发生的真正自身免疫原。     

狼疮性肾炎发病机理的研究进展

系统性红斑狼疮 (SLE)是一种累及多脏器的自身免疫性炎症性结缔组织病 ,病因复杂 ,受遗传、免疫、神经内分泌、环境等多因素影响。狼疮性肾炎(LN)是SLE最常见而严重的临床表现 ,已成为国内外风湿病学界研究的热点 ,本文将其发病机理的研究近况作一综述。1　遗传因素小鼠狼疮性肾炎 (LN)模型的遗传分析及人类SLE家系研究表明 ,遗传因素确实促进SLE发病 ,具有复杂的多基因遗传的特点 ,LN相关的易感基因不断被揭示。 (SWR×NewZealandBlack(NZB) )F(1)(或SNF(1) )鼠有倾向发展成LN ,虽然NZB鼠狼疮的几个易感遗传位点已被证实 ,S…     

NZB/W F1狼疮模型小鼠泪腺病变的特征

目的:观察狼疮易感NZB/W F1小鼠泪腺随周龄变化特点以及明确是否存在相应眼部体征改变。方法:应用BALB/c小鼠作对照,采用泪液分泌试验进行泪液分泌量测定,ELISA法行血清抗dsDNA抗体检测,采用H-E染色观察小鼠泪腺结构的病理改变及炎症程度并进行炎症评分。结果:NZB/W F1小鼠泪液分泌量低于BALB/c小鼠,29周龄开始其差值逐渐增大,到35周龄NZB/W F1小鼠泪液分泌量(4.6 mm±0.7 mm)显著低于同周龄BALB/c小鼠(8.7mm±0.7mm);NZB/WF1小鼠泪腺显著的灶性炎症细胞浸润出现在35周龄,密集成片浸润的炎症细胞甚至部分取代原有的腺泡结构,其炎症评分(3.3分±0.5分)明显高于同周龄BALB/c小鼠(1.1分±0.6分),小叶内及小叶间导管结构仍保存。结论:随周龄增加,NZB/W F1小鼠自发出现加重类似继发性干燥综合征的泪腺炎症变化,其眼部体征异常出现在35周龄。     

活化淋巴细胞与慢性GVHR诱导的SLE样小鼠模型的比较

目的 :将本室建立的用活化淋巴细胞诱导的系统性红斑狼疮 (SLE)样小鼠模型与国际上公认的用慢性GVHR诱导的SLE样小鼠模型进行比较 ,进一步探索SLE的发病机理。方法 :分别将亲代Balb c小鼠淋巴细胞经静脉和用ConA活化的(Balb c×C5 7BL 6 )F1代小鼠淋巴细胞经皮下途径输入F1代小鼠 ,用ELISA测定IgG类抗dsDNA抗体和抗组蛋白抗体 ,用免疫荧光法检测抗核抗体 (ANA)荧光核型和肾小球内免疫复合物沉积 ,用免疫印迹法检测抗可溶性核抗原 (ENA)抗体。结果 :亲代淋巴细胞免疫F1代小鼠所致的慢性GVHR和活化F1代小鼠淋巴细胞均可诱导F1代小鼠产生高滴度的抗dsDNA抗体、抗组蛋白抗体等ANA ,并且肾脏都有明显的IgG类免疫复合物沉积。但亲代淋巴细胞免疫组ANA核型以颗粒型、核仁型为主 ,ENA多肽谱多在 32、47、6 7kD处显色 ;而活化淋巴细胞免疫组以胞浆型、周边型、均质型为主 ,ENA多肽谱在 2 8、47、6 7kD处显色。结论 :这 2种方法均可诱导出SLE样综合征 ,但其抗核抗体谱有所不同。     

Case 10

An 8-year-old boy presented with a 3-year history of a macrogenitosomia with the subsequent development of adiposogigantism, pronounced acne, and a moustache. Therapy directed toward the treatment of an adrenogenital syndrome had been initiated at the age of 6 years. Enlargement of the right testis was first noted at the age of 8 years. On admission to the Department of Urology, bone age was determined at 16.5 years, whereas psychological development corresponded to that of an 8-year-old child. Both testicles were descended; the right testicle was twice the size of the left one.

Follicle-stimulating hormone and luteinizing hormone were not detectable, plasma Cortisol was normal, α₁ -fetoprotein and human chorionic gonadotropin were negative, testosterone was markedly elevated, and 17-ketosteroids were slightly elevated. Neurologic examination revealed no abnormalities.

A right orchiectomy was performed. On gross inspection the surgical specimen consisted of a tumor in the lower part of the testicle, measuring 2.5 cm in diameter, with a brown, lobulated cut surface. A sharp border against the surrounding testicular tissue was observed.     

Interleukin-10

Interleukin-10 (IL-10) is a potent anti-inflammatory and immunosuppressive cytokine secreted by several cell types. Most anti-inflammatory effects of IL-10 are caused by its ability to deactivate macrophages and monocytes, whereas its immunosuppressive properties are due to functional inhibition of both antigen-presenting cells and T cells. On the other hand, IL-10 also exerts immunostimulatory effects, especially on B cells, CD8+ cytotoxic T cells and natural killer cells. In vivo administration of recombinant IL-10 (rIL-10) efficiently prevents experimental septic shock induced by endotoxin, staphylococcal superantigen or cecal ligation and puncture, as well as experimental autoimmune diseases mediated by T helper type 1 (T(H)1) cells and other inflammatory disorders. rIL-10 exerts paradoxical effects in cancer models, where it promotes tumour rejection, probably due to its stimulatory properties on cytotoxic cells. On the other hand, rIL-10 increases the severity of experimental infections caused by fungi or bacteria, and enhances systemic autoimmune features in mice with spontaneous lupus syndrome. Although the therapeutic potential of rIL-10 in human diseases seems promising, the multiple facets of rIL-10 in experimental immunopathology indicate that the success of clinical trials with rIL-10 will depend both on the appropriate selection of the patient populations to be treated and on the early detection of possible adverse effects.     

Pure trisomy 10p involving an isochromosome 10p

We report a child with trisomy 10p due to a translocation of the long arm of chromosome 10 to the short arm of chromosome 14 and isochromosome formation of 10p [46,XX,i(10)(p10),der(14)t(10;14)(q10;p10)]. Most reported cases of trisomy 10p involve double segmental imbalance. In contrast, the clinical features described in the current case represent pure trisomy 10p and, thus, delineate the 10p trisomy syndrome phenotype. Mechanisms of the chromosomal rearrangements in this case are suggested.     

Family with partial monosomy 10p and trisomy 10p

We report on a family with an abnormality of 10p. The propositus has monosomy for the distal region of 10p and severe psychomotor delay, growth failure, congenital heart defect, multicystic kidney, grade V vesicoureteric reflux, and neurosensory hearing loss. The mother and the elder brother of the propositus carry a balanced reciprocal translocation (5q;10p)(q35.3;p12.3). A retarded and epileptic maternal aunt was found to have dup(10p). Study of the family history led to the successful obstetric management of a subsequent twin pregnancy in which an affected fetus with dup(10p) was identified and selectively terminated, while the other normal twin was delivered at term without problems. © 1995 Wiley-Liss, Inc.     

Interleukin-10 (IL-10) in experimental visceral leishmaniasis and IL-10 receptor blockade as immunotherapy

Interleukin-10 (IL-10) is thought to promote intracellular infection, including human visceral leishmaniasis, by disabling Th1 cell-type responses and/or deactivating parasitized tissue macrophages. To develop a rationale for IL-10 inhibition as treatment in visceral infection, Th1 cytokine-driven responses were characterized in Leishmania donovani-infected BALB/c mice in which IL-10 was absent or overexpressed or its receptor (IL-10R) was blockaded. IL-10 knockout and normal mice treated prophylactically with anti-IL-10R demonstrated accelerated granuloma assembly and rapid parasite killing without untoward tissue inflammation; IL-12 and gamma interferon mRNA expression, inducible nitric oxide synthase reactivity, and responsiveness to antimony chemotherapy were also enhanced in knockout mice. In IL-10 transgenic mice, parasite replication was unrestrained, and except for antimony responsiveness, measured Th1 cell-dependent events were all initially impaired. Despite subsequent granuloma assembly, high-level infection persisted, and antimony-treated transgenic mice also relapsed. In normal mice with established infection, anti-IL-10R treatment was remarkably active, inducing near-cure by itself and synergism with antimony. IL-10's deactivating effects regulate outcome in experimental visceral leishmaniasis, and IL-10R blockade represents a potential immuno- and/or immunochemotherapeutic approach in this infection.     

Structure of interleukin-10/interleukin-10R1 complex

The class II alpha-helical cytokine family consists of eleven members including the interferons, interleukin-10 (IL-10) and several newly discovered IL-10 homologs. The molecules display a vast array of biologic activities including the ability to induce an antiviral state, modulate inflammatory responses, and inhibit cell growth. Biologic activity is dependent on cytokine-dependent aggregation of two different cell-surface receptors. The detailed protein-protein interactions that initiate these biologic responses are amenable to study using X-ray crystallographic methods. In this article, I summarize my laboratory's contributions to understanding these recognition processes using IL-10 as the prototypic class II cytokine.     

Interleukin-10.

Despite the short history of interleukin-10, accumulated evidence indicates that this interleukin plays a major role in suppressing immune and inflammatory responses. Yet interleukin-10 also maintains cell viability and acts as a cofactor to promote the growth of lymphoid and myeloid cells in vitro. Here we review the present knowledge on the structure and function of interleukin-10.