慢性移植物抗宿主病狼疮小鼠模型肾组织细胞凋亡及Th1/Th2细胞因子的研究

目的研究慢性移植物抗宿主病(cGVHD)狼疮样小鼠模型。肾组织细胞凋亡及Th1／Th2细胞因子的改变方法20只B6D2F1代杂交鼠，随机分为模型组(10只)及对照组(10只)，12周后处死采用原位末端标记法(TUNEL)染包观察肾组织细胞凋亡；免疫组织化学、Western blot和反转录-聚合酶链反应(RT-PCR)技术检测Fas、FasL基因转录及蛋白表达情况。结果TUNEL法结果显示模型组肾组织细胞凋亡较对照组增多。免疫组织化学提示：各组均有Fas、FasL表达，正常对照组有极少量Fas表达．表达部位在肾小球系膜细胞；FasL表达部位在近端肾小管上皮细胞。聚合酶链反应(PCR)结果：模型组较正常对照组Fas mRNA表达增高。模型组较正常对照组FasL mRNA表达差异无显著性。正常对照组干扰素(IFN)-γ和白细胞介素(IL)-4阳性细胞极少；模型组肾组织血管周围浸润的炎性细胞中可见IFN-γ和IL-4阳性细胞，且IL-4阳性细胞明显高于IFN-γ，提示Th2细胞表达占优势。模型组比正常对照绀INFγ／和IL-4阳性细胞明显升高，RT-PCR结果显示：模型组IFN-γ与正常对照组比较差异无显著性：模型组IL-γ较正常对照组显著升高。结论cGVHD狼疮样小鼠模样小鼠模织细胞凋亡异常及Th1/Th2细胞因子失衡，在狼疮肾炎(LN)的发病中起重要作用。     

凋亡核小体抗核小体抗体与狼疮肾炎

狼疮肾炎(lupus nephritis,LN)是系统性红斑狼疮(systemic lupus erythematosus,SLE)发生率最高的并发症,是,免疫复合物介导的继发性肾小球肾炎.而近十年的研究显示凋亡产生的核小体介导免疫复合物沉积于肾小球基底膜,引起肾组织病理损伤,在LN的发生发展中起重要作用.本文对凋亡紊乱及凋亡细胞清除缺陷在SLE发生机制中的作用.核小体诱导免疫反应的机制以及在介导LN损伤中的作用综述如下.     

肾小球肾炎肾组织megsin的表达及其意义

目的 :观察megsin在肾小球肾炎患者肾小球内表达及其特点 ,对原发性肾小球肾炎 (IgA肾病 ,IgAN)和继发性肾小球肾炎 (狼疮性肾炎 ,LN)进行研究。　 方法 :应用针对人类megsin的多克隆抗体 ,采用免疫组化法检测肾活检组织中肾小球内megsin的表达 ,其中IgAN 10例、LN 2 0例 ,并采用半定量方法与正常肾组织和轻微病变性肾病进行比较。　 结果 :正常肾组织和轻微病变性肾病均可表达一定量的megsin ,而在IgAN和LN的肾小球内megsin均表达明显增加。megsin仅表达于肾小球系膜区域 ,小管间质未见表达。在IgAN系膜增生性病变中megsin表达量较硬化性病变明显增多 (P <0 0 5 )。而在LN肾小球megsin的表达量虽较正常肾组织和轻微病变型肾病显著增多 ,但与原发性肾小球肾炎IgAN相比较 ,其表达量少 ,即使在LNⅣ型时。　 结论 :megsin在原发性和继发性肾小球肾炎均有一定的表达 ,并在其疾病进程中以及肾小球系膜功能调节中起一定作用 ,但在原发性和继发性肾小球肾炎中megsin的表达调节可能存在不同的机制     

尿足细胞监测活动性狼疮肾炎的意义

目的探讨尿足细胞作为无创监测肾活动性损伤的实用性。方法选31例肾活检病理确诊为狼疮肾炎的患者。用抗足细胞表面标志蛋白podoealyxin抗体免疫化学法检测尿沉渣足细胞,并与肾活检病理、血尿、蛋白尿进行对比分析;用抗podoealyxin和抗增殖细胞核抗原的抗体双重免疫组化法检测各型狼疮肾组织和尿中增殖的足细胞。全身活动指标积分用系统性红斑狼疮疾病活动指数评估。结果31例狼疮肾炎病理显示Ⅲ型占25．8％,Ⅳ型占64．5％,Ⅴ型占9．7％。31例患者尿中足细胞阳性者28例,尿足细胞阳性总检出率为90％。除Ⅴ型狼疮肾炎患者无足细胞尿和血尿外,其余病例均有不同程度的足细胞尿和血尿,且尿足细胞数与血尿程度、肾小球病理活动指数呈正相关（r=0．639,P=0．000;r=0．487,P=0．014）。31例患者均有蛋白尿,其量与肾小球病理活动指数不相关。免疫组化双重染色显示,Ⅳ型狼疮肾炎肾小球和尿沉渣均有增殖的足细胞。8例患者在免疫炎症抑制治疗过程中上述各项指标呈平行下降之势,其中6例尿足细胞消失。结论尿足细胞是监测狼疮肾炎活动和评估药物治疗反应的无创指标。     

Bcl-2、Bax在狼疮肾炎患者肾组织中的表达及意义

目的研究细胞凋亡调节蛋白(Bcl-2及Bax)在狼疮肾炎(LN)患者肾组织的表达情况及与LN肾脏病理改变的关系。方法2003-012003-04对广西医科大学第一附属医院的44例活动期LN患者用免疫组化方法检测肾组织中Bcl-2、Bax的表达,观察它们与LN肾脏病理改变的关系。结果(1)LN肾小球Bcl-2及Bax表达均增高,与肾小球细胞增殖程度及增殖细胞核抗原(PCNA)阳性细胞数呈正相关。(2)LN肾间质Bcl-2表达增高,与间质炎症细胞浸润程度正相关。(3)LN肾小管Bax表达增高,与肾小管间质病变呈正相关。结论LN肾组织存在Bcl-2、Bax蛋白异常表达。Bcl-2高表达在LN肾小球细胞增殖及肾间质炎症细胞浸润过程中可能起重要作用。Bax在肾小管过度表达与肾小管间质病变有关。     

甲基泼尼松龙对狼疮小鼠肾脏细胞凋亡及ICE基因表达的影响

为了探讨肾脏细胞凋亡在糖皮质激素治疗狼疮性肾炎机制中的作用，采用甲基泼尼松龙（ＭＰＳ）和近交系纯合子ＭＲＬ－ｌｐｒ／ｌｐｒ自发狼疮小鼠进行体内实验，利用计算机图像分析系统观察了ＭＰＳ对狼疮小鼠肾脏细胞凋亡的诱导，并应用逆转录－巢式聚合酶链反应和Ｎｏｒｔｈｅｒｎ杂交技术观察了ＭＰＳ对哺乳动物凋亡相关基因白细胞介素－１β转化酶（ＩＣＥ）表达的影响。结果表明，ＭＰＳ冲击治疗组小鼠肾小球系膜区细胞的凋亡数目显著高于对照组（Ｐ＜０．０５），肾小管间质细胞和肾血管周浸润细胞的凋亡数目也明显高于对照组（Ｐ均＜０．０１），治疗组肾脏细胞ＩＣＥ基因的表达明显增强。研究提示，ＭＰＳ能够诱导狼疮小鼠的肾脏细胞发生凋亡，ＩＣＥ基因可能参与了狼疮小鼠肾脏细胞的凋亡机制。     

狼疮肾炎肾间质浸润与肾损害进展及肾功能相关性的研究

目的：观察狼疮肾炎肾间质浸润的白细胞类型与肾功能损害进展的关系。方法：检测40例狼疮肾炎患者24h尿蛋白定量、血肌酐，随即进行肾活检及免疫组织化学检测，21例患者进行二次肾活检。结果：狼疮肾炎患者首次肾活检时显示T淋巴细胞与单核细胞所占比例差异无显著性，而第2次肾活检时以单核细胞为主。首次肾活检时间质浸润的淋巴细胞CD4^ ／CD8^ 较正常对照组增加，第2次肾活检时CD^ ／CD8^ 低于正常对照组。第2次肾活检时显著间质浸润的发生率由首次活检时的47.5％升至100％。浸润白细胞总数与血肌酐水平呈正相关，其中T细胞、中性粒细胞亦与血肌酐呈正相关，单核细胞与血肌酐无显著相关性。结论：重复肾活检可发现更多病例肾间质损害，狼疮肾炎肾间质的主要浸润细胞类型呈疾病进展阶段依赖性，CD4^ T细胞在间质损伤机制中起始发作用，CD8^ T细胞的作用可能是继发的，长期的蛋白尿可能对CD8^ 细胞的浸润有激发作用。     

免疫疗法在小儿肾脏疾病中的应用

小儿原发性肾小球疾病以肾病综合征、急性肾小球肾炎最为多见,继发性肾小球疾病则以过敏性紫癜肾炎、狼疮肾炎、乙肝肾炎多见。以上疾病     

狼疮性肾炎与细胞凋亡

引入细胞凋亡这一新概念阐明狼疮性肾炎的发病机制，从狼疮鼠中已研究发现有淋巴细胞凋亡异常及其调控基因的突变，并已揭示出其关键的发病环节；同时在人类狼疮中也已发现有细胞凋亡异常，由此说明细胞凋亡对狼疮性肾炎的致病作用及其治疗的指导意义。     

RANTES在狼疮肾炎肾组织中的表达及其意义

目的　了解狼疮肾炎 (LN)肾组织中化学趋化因子激活正常T细胞表达和分泌因子(RANTES)的表达并探讨其与LN肾脏病理改变和肾功能损害的关系。方法　采用微波免疫组织化学染色 (SP法 )和原位分子杂交技术检测LN肾脏RANTES的表达 ,并进一步分析其与LN活动指数、肾脏病理和功能损害的关系。结果　狼疮肾炎组织中RANTES表达显著增高 ,以WHOⅣ型为显著。RANTES在肾小球和肾小管均有表达 ,但肾小管表达更显著。LN肾组织RANTES表达与肾组织活动指数、肾脏病理改变和肾功能损害显著相关。结论　RANTES可能参与了LN的发病机制 ,肾组织中RANTES的表达可作为反映狼疮肾组织活动病变和进行性肾损害的参考指标。     

Chimerism occurs twice as often in lupus nephritis as in normal kidneys

OBJECTIVE: Systemic lupus erythematosus (SLE) is an immune-mediated disease that particularly affects the kidneys, causing lupus nephritis. In experimental mouse models, lupus nephritis can be mimicked by inducing a chimeric state through the injection of parental T cells in offspring. In humans, pregnancy-induced chimerism may play a role in the pathogenesis of autoimmune diseases such as SLE, but it is likely that only certain chimeric cells have pathogenic potential. In this study, we investigated whether the distribution of chimeric cells is different in the kidneys of women with SLE from that in normal kidneys, and we examined the phenotype of chimeric cells in women with SLE. METHODS: The presence of chimeric cells was investigated by in situ hybridization targeting the Y chromosome in 57 renal biopsy samples from 49 women with lupus nephritis. Fifty-one kidney autopsy specimens without histomorphologic lesions served as controls. Double-staining for the Y chromosome in combination with CD3 and CD34 markers was performed in 5 kidney specimens with lupus nephritis to identify the phenotype of the chimeric cells. RESULTS: Y chromosome-positive cells were found in 27 of 49 patients with lupus nephritis and in 13 of 51 normal controls (P < 0.01). Both CD3+ and CD34+ chimeric cells were identified in lupus nephritis kidney specimens. CONCLUSION: Chimeric cells are present significantly more often in kidneys with lupus nephritis than in normal kidneys, and some of these chimeric cells are T cells. This finding is interesting in light of experimental models demonstrating that lupus nephritis is initiated by chimeric T cells.     

Apoptose und C1q: Erklärungsansätze für die Pathogenese des SLE

Dysregulation of apoptosis may play a major role in the pathogenesis of systemic lupus erythematosus (SLE). A defective clearance of apoptotic cells or inappropriately high rates of apoptosis may lead to a pathologic accumulation of abnormal cell material with a secondary autoimmune response. Experimental findings in apoptotic keratinocytes and C1q knock-out mice suggest an important role of C1q in the clearance of apoptotic cell material. In addition, there are several links between C1q and SLE: Most the patients with C1q deficiency develop a SLE-like syndrome. SLE itself often causes secondary C1q deficiency and autoantibodies to C1q are detected in almost all patients with active lupus nephritis. These observations suggest a central role of C1q in apoptosis and in the pathogenesis of SLE.     

Pathogenic anti-nucleosome antibodies

There is increasing evidence that in systemic lupus erythematosus, nucleosomes, the basic chromatin component, represent both a driving immunogen and a major in vivo target for antibodies. Either a disturbed apoptosis or a reduced clearance of apoptotic cells by phagocytes may lead to an increased exposure of apoptotic nucleosomes to the immune system. These nucleosomes, which have been cleaved and modified during the process of apoptosis, escape normal clearance and encompass epitopes that normally are not encountered by the immune system. This may then lead to tolerance breaking and autoimmunity by the activation of nucleosome-specific autoreactive T cells (that help B cells) and subsequently to the production of anti-nucleosome, anti-histone and anti-DNA autoantibodies. Some anti-nucleosome antibody subsets are pathogenic and are involved in the nephritogenic process in systemic lupus erythematosus. Accordingly, several studies reported: (i) increased plasma circulating nucleosomes that positively correlated with an active disease, (ii) nucleosomes in typical glomerular deposits as well as in the basement membrane of non-lesional skin of systemic lupus erythematosus patients and (iii) a close correlation between nephritis and the presence of anti-nucleosome antibodies. Recent studies reported anti-nucleosome antibodies also in primary anti-phospholipid syndrome and particularly in patients with associated lupus-like disease.     

慢性移植物抗宿主病狼疮样小鼠肾组织RANTES表达及泼尼松的调控作用

目的 观察趋化因子RANTES（“regulated upon activation normal T cell expressed and secreted”）在慢性移植抗宿主病（VGHD）狼疮样小鼠肾组织中的表达及泼尼松的调控作用。方法 GVHD狼疮样小鼠模型参照有关文献建立。按随机设计原则将模型动物分两组：即模型组（A组）和泼尼松治疗组（B组）,另设正常对照组（C组）,每组均有8只动物。RANTES的蛋白表达采用免疫组织化学方法,mRNA表达采用原位杂技术检测。结果 ①模型组（A组）肾组织中RANKTES蛋白表达显著高于正常对照组（C组）（P＜0．05）,肾小球和肾小管间质中均有表达,尤以肾小管－间质表达更为丰富;而泼尼松治疗组（C组）RANTES在肾小球和肾小管－间质表达均显著减少（P＜0．05）。②A肾组织中RANTESmRNA表达显著高于C组（P＜0．05）,经泼尼松治疗后RANTESmRNA以到明显抑制。③A组中RANTES蛋白和mRNA表达呈正相关（r＝0．612,PO＜0．05）。结论 慢性GVHD狼疮样小鼠肾组织中RNNTES蛋白和mRNA表达均显著增高,泼尼松在改善肾组织病理改变的同时,对RANTES表达亦起抑制作用,提示RANTES可能参与了狼疮肾炎的发病机制,泼尼松抑制RANTES过度表达可能是其在狼疮肾炎中发挥治疗作用的机制之一。     

Glomerular deposition of renal tubular epithelial antigen in patients with systemic lupus erythematosus: its possible role in lupus nephritis

Fifty-three renal specimens from 48 patients with SLE were examined for the presence of RTE in the glomeruli. Glomerular RTE, presumably in immune complex form was detected in 60% of the tissues. The deposition of these complexes was related to the severity of histologic changes and activity of SLE. In addition, glomerular localization of RTE was associated with decreased renal function and increased proteinuria. The association between the presence of glomerular RTE antigen, the severity of renal histologic changes and the decreased renal function suggested a possible role for this antigen in the pathogenesis of lupus nephritis.     

狼疮肾炎肾小球微血栓形成及其与抗凝血相关因子抗体抗磷脂抗体的相关性研究

目的 评估肾小球微血栓(GMT)在狼疮肾炎(LN)中的发生率,并探讨针对某些凝血相关因子的抗体和抗磷脂抗体在LN患者GMT形成中的临床意义.方法 连续收集124例LN患者肾活检组织标本和血浆,观察组织标本中GMT的发生率.并分成LN-GMT组和LN-non-GMT组;比较两组患者的疾病活动度、相关实验室检查指标和肾组织活动,慢性指数;测定患者的狼疮抗凝物(LA)、抗心磷脂抗体(ACL)、抗β2糖蛋白I(抗β2GP I)抗体、抗凝血酶抗体、抗纤溶酶抗体、抗组织型纤溶酶原激活物(t-PA)抗体和抗膜联蛋白AⅡ(Annexin A II)抗体.结果 GMT在LN中的发生率约为20.2%;LN-GMT组系统性红斑狼疮疾病活动指数(SLEDAI)、肾组织病变活动指数、肾组织病变慢性指数、尿蛋白定量(24 h)、血清肌酐、血清尿素氮的水平和高血压的发生率都较LN-non-GMT组高(P＜0.01);LN-GMT组LA、IgG型抗B2GP I抗体和抗凝血酶抗体阳性率均显著高于LN-non-GMT组(P＜0.05);两组IgG型ACL抗体、抗纤溶酶抗体、抗t-PA抗体和抗Annexin A II抗体阳性率差异均无统计学意义(P＞0.05);两组各抗体IgM型差异无统计学意义(P＞0.05).结论 LN中伴有GMT形成的患者肾脏病变重于无GMT者;LA、IgG型抗β2GP I抗体和抗凝血酶抗体与LN患者GMT形成相关.     

凋亡细胞促进狼疮性BXSB小鼠发病的研究

目的 探讨凋亡细胞在系统性红斑狼疮发病中的作用。方法 将体外诱导的凋亡胸腺细胞经腹腔注射给同基因同性别的正常Ｃ５７小鼠和具有狼疮倾向的雌雄ＢＸＳＢ小鼠;检测血清中ＬｇＭ和ＩｇＧ类型的抗ｄｓＤＮＡ抗体和抗ｓｓＤＮＡ抗体的水平,尿蛋白浓度以及肾脏中ＩｇＧ类型免疫复合物的沉积。结果 用凋亡腺细胞免疫后,诱导Ｃ５７小鼠产生ＩｇＧ类型的抗ｄｓＤＮＡ抗体和抗ｓｓＤＮＡ抗体;相比之下,诱导雌性ＢＸＳＢ小鼠产生更高水平的ＩｇＧ类型抗ｄｓＤＮＡ抗体和抗ｓｓＤＮＡ抗体;并且使雄性ＢＸＳＢ小鼠的尿蛋白浓度显著升高。结论 凋亡细胞具有免疫原性,它可能是狼疮性ＢＸＳＢ小鼠体内自身抗原的主要来源;狼疮性遗传因素和Ｙａａ基因能促进凋亡细胞的免疫原性和增强凋亡细胞的致肾炎作用。     

Intracellular antigens released from Balb/c mice kidneys under chemically induced apoptosis and/or necrosis

The present investigation assesses the possible role of apoptosis and necrosis in intracellular antigen exposure of kidneys from Balb/c mice. Renal tissues were cultured and treated with chemicals to induce apoptosis and /or necrosis. The expression of intracellular antigens Sm, RNP, Ro and La were monitored with antibodies against these antigens. Main results confirm that renal intracellular antigens are released and exposed onto the surface of apoptotic and necrotic cells, therefore these antigens become an easy target of autoantibodies. This mechanism may be important in the lupus nephritis pathogenesis.     

Accumulation of apoptotic cells in the epidermis of patients with cutaneous lupus erythematosus after ultraviolet irradiation

OBJECTIVE: To examine whether apoptosis contributes to the pathogenesis of skin lesions in patients with cutaneous lupus erythematosus (CLE) after ultraviolet (UV) irradiation. METHODS: In situ nick translation and TUNEL were performed to detect apoptosis in 85 skin biopsy specimens from patients with various subtypes of CLE. Specimens from normal healthy donors and patients with polymorphous light eruption were used as controls. In addition to assessment of primary lesions, provocative phototesting was carried out to investigate events occurring secondary to UV irradiation during a very early stage of lesion formation. RESULTS: A significant increase in apoptotic nuclei was found in the upper epidermal layer of primary and UV light-induced skin lesions of CLE patients compared with controls. In tissue sections obtained from control subjects at 24 hours after a single exposure to UV light, a slight increase in the count of epidermal apoptotic nuclei was present as compared with skin tissue from CLE patients obtained under the same conditions before lesion formation. In sections obtained from controls at 72 hours after irradiation, a significant decrease in the apoptotic nuclei count was observed, consistent with a proper clearance of apoptotic cells in the period between 24 and 72 hours after irradiation. In striking contrast, the number of apoptotic nuclei increased significantly within this period in tissue sections from patients with CLE. CONCLUSION: These data support the hypothesis that apoptotic cells accumulate in the skin of patients with CLE after UV irradiation, as a result of impaired or delayed clearance. The nonengulfed cells may undergo secondary necrosis and release proinflammatory compounds and potential autoantigens, which may contribute to the inflammatory micromilieu that leads to formation of skin lesions in this disease.     

Methodological update detection of antibodies to dsDNA: current insights into its relevance

Ever since its first discovery in 1957, anti-DNA has taken a special place amongst the other antinuclear antibodies. Even today, it stands out between these, because of its high specificity for one particular disease: systemic lupus erythematosus (SLE). Furthermore, anti-DNA has been shown to actually play a role in a key disease feature of SLE: lupus nephritis. Binding of anti-DNA to the glomerular basement membrane of the kidney has been shown to be mediated by nucleosomes. More recently, it has been shown that patients with SLE also have antibodies specific for nucleosomes in their circulation. It may well be that anti-nucleosome detection in the near future will prove to be of more relevance than anti-DNA detection. Nucleosomes also seem to play a key role in the induction of anti-DNA (and anti-nucleosome) production. Mechanisms involved in this process may include defects in apoptosis and/or clearance of apoptotic material. Studies of these mechanisms will help us to decipher the cause of autoantibody production, or indeed of autoimmune diseases such as SLE.