诱发性系统性红斑狼疮小鼠模型研究进展

建立系统性红斑狼疮小鼠模型对于探索人类系统性红斑狼疮的病因、发病机制和治疗有重要意义.目前研究较充分的诱发性狼疮小鼠模型有:慢性移植物抗宿主病模型、烃类化合物诱发的模型、肽诱发的模型、脂多糖诱发的模型、空肠弯曲杆菌与弗氏完全佐剂诱发的模型、药物诱发的模型等.其常用品系、诱导方法、诱导所需时间及诱导后形成的自身免疫病变等有各自的特点.     

姥鲛烷诱导法建立小鼠狼疮肾炎模型

目的:采用姥鲛烷诱导建立C57BL/6小鼠狼疮肾炎模型,并进行生物学鉴定和形成机制的探讨。方法:模型组小鼠单次腹腔注射0. 5 mL姥鲛烷,对照组同方式注射等量生理盐水。注射后监控生存状态;第10天采用流式细胞术分析脾脏巨噬细胞、粒细胞、树突状细胞及B细胞的活化情况;间接免疫荧光法每月检测血清抗核抗体(ANA)及抗双链DNA(dsDNA)抗体的滴度,Albustix试纸法分析尿蛋白含量;注射后8个月处死小鼠并取肾脏,经直接免疫荧光法观测免疫复合物沉积状况,免疫组化法分析干扰素γ(IFN-γ)与白细胞介素4(IL-4)的表达,并进行HE染色及透射电镜病理检查。结果:第10天时脾脏巨噬细胞、粒细胞、树突状细胞及B细胞呈不同程度活化(P<0. 05),B细胞表面标志物B7-1、B7-2及MHC-Ⅱ表达上调(P<0. 05);3～8个月时血清ANA和抗dsDNA抗体水平进行性升高(P<0. 05),3只小鼠出现腹水,1只消瘦并最终于7个月时死亡;4个月时3只小鼠出现蛋白尿,8个月时蛋白尿阳性率100%,尿蛋白含量1～20 g/L;8个月时腹腔内出现大量的异位淋巴组织,肾小球及...     

Pristane诱导小鼠系统性红班狼疮动物模型的研究

目的:建立pristane诱导的系统性红斑狼疮(SLE)小鼠模型,并对该小鼠模型的发病机制进行初步的探讨。方法:6-8周龄雌性BALB/c小鼠单次腹腔注射pristane0.5mL,对照组单次腹腔注射PBS0.5mL,注射前及注射后每2周行流式细胞术(FCM)检测外周血中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例及细胞活化状态B220+Aβ1dhigh),ELISA检测血清中自身抗体(anti-dsDNA,anti-smRNP,anti-ribosomalP0)的含量。至6个月处死动物,FCM检测腹腔细胞中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例和脾脏中细胞的活化(B220,Aβ1d),采用直接免疫荧光法标记小鼠肾脏免疫球蛋白复合物及H&E染色评估小鼠肾脏免疫复合物的沉积及损伤情况。结果:小鼠腹腔注射pristane第2个月开始血清总IgG升高,第3个月起出现自身抗体阳性,到个6月时达到最高,并维持高水平至被处死;Pristane处理6个月后,Pristane处理组小鼠出现关节炎症状,肾脏免疫复合物的大量沉积和明显肾脏损伤。Pristane注射2周起,小鼠外周血中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例明显高于PBS注射组,小鼠腹腔细胞中IFN-α分泌细胞的比例也明显升高;同时外周血和脾细胞中B细胞表面MHCII分子Aβ1d的平均荧光强度(MFI)均高于对照组,表明pristane处理组小鼠中B细胞发生了显著活化。结论:BALB/c小鼠腹腔注射pristane可诱导构建小鼠SLE模型,其SLE的发病可能与IFN-α的持续分泌导致B细胞的异常活化有关。该模型的建立为进一步研究SLE的发病机制提供了良好的动物模型。     

同基因背景小鼠核蛋白诱导狼疮鼠模型

背景：自发性狼疮鼠模型不能对基因以外其他的致病因素进行研究。 目的：以同基因背景Balb/c小鼠核蛋白免疫小鼠后诱导狼疮鼠模型。 方法：选取4~6周SPF级Balb/c小鼠30只,等分为3组。V1组肌肉注射提取的同系Balb/c小鼠核蛋白,间隔3周免疫1次,共免疫4次;V2组注射等体积PBS;V3组为正常对照。检测小鼠末次免疫后3周的24 h尿蛋白、血清抗核抗体、抗双链DNA抗体、小鼠肾脏直接免疫荧光。 结果与结论：V1组小鼠24 h尿蛋白、血清抗双链DNA抗体、抗核抗体均明显高于V2组、V3组,且V1组小鼠肾小球有免疫球蛋白G免疫复合物沉积,可见肾小球轮廓,V2组、V3组未见肾小球轮廓,只见非特异性的微弱荧光。说明以同基因背景Balb/c小鼠核蛋白免疫Balb/c小鼠能够成功诱导狼疮鼠模型。     

系统性红斑狼疮小鼠模型的研究近况

系统性红斑狼疮(SLE)小鼠模型的研究对揭示SLE发病原因、发病机制及探索治疗方法具有重要价值。本文介绍了几种SLE小鼠模型的研究近况,以BXSB小鼠为主,同时介绍(NZBxNZW)F1、MRL/lpr及诱导的SLE样小鼠,从小鼠的品系来源、发病特点、免疫学特征、相关基因以及治疗状况等几个方面进行了综述。     

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。     

恶性疟疾的免疫缺陷小鼠模型研究进展

由于恶性疟原虫的宿主特异性,人们很难在小动物体内进行恶性疟疾红细胞内期的研究,这已成为制约该领域发展的一大难题。免疫缺陷小鼠以其体液免疫或/和细胞免疫缺陷的特点,除了在肿瘤学、免疫学、毒理学、感染性疾病等领域的基础性研究中被广泛应用外,也被用于对疟原虫无性生殖期的研究。本文将针对恶性疟原虫红细胞内期的免疫缺陷小鼠模型的研究进展综述如下。     

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     

Pristane诱导狼疮鼠模型

系统性红斑狼疮(Systemic lupus erythematosus,SLE)是一种累及全身多系统器官组织的自身免疫性疾病,血清中存在以抗核抗体、抗dsDNA抗体为代表的多种自身抗体,目前病因、发病机理尚不完全清楚。合适的动物模型有助于对其进行研究。SLE动物模型分为自发性模型和诱导性模型。自发性狼     

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。     

Silica accelerated systemic autoimmune disease in lupus-prone New Zealand mixed mice

The genetic backgrounds of lupus-prone murine models are a valuable resource for studying the influence of environmental exposure on autoimmune diseases in sensitive populations. Epidemiological studies have shown associations between silica exposure and several autoimmune diseases, including scleroderma and systemic lupus erythematosus. To determine whether silica exposure can exacerbate systemic autoimmunity in genetically predisposed animals, New Zealand mixed mice were intranasally instilled twice with saline or saline suspensions of 1 mg silica or 500 micro g TiO2, a dose equivalent in surface area, and were evaluated with respect to health and immune status. Survival in silica exposed NZM mice was decreased compared to saline and TiO2 exposed mice. Proteinuria levels were elevated in silica exposed mice. Levels of circulating immune complexes, autoantibodies to nuclear antigen (ANA), histone, and double stranded DNA were measured every two weeks by ELISA. Circulating immune complexes showed a trend towards an increased acceleration in levels in the silica exposed mice compared to saline and TiO2 exposed mice. ANA levels were significantly higher in silica exposed animals compared to saline and TiO2 exposed animals (0.237 +/- 0.03 versus 0.140 +/- 0.029 and 0.125 +/- 0.03, P < 0.05) 16 weeks postexposure. Autoantibodies to histone were also significantly elevated after 16 weeks in silica exposed animals compared to saline and TiO2 exposed animals (0.227 +/- 0.03 versus 0.073 +/- 0.015 and 0.05 +/- 0.03, P < 0.05). In contrast, serum IgG levels were decreased in silica exposed NZM mice compared to the saline controls, however, IgM levels were unaffected. Lungs of the silica-exposed mice had increased inflammatory infiltrates as well as fibrotic lesions characterized by excess collagen deposition. Therefore, although NZM mice are susceptible to SLE, silica exposure significantly exacerbated the course of disease.     

Novel spontaneous myelodysplastic syndrome mouse model

BackgroundMyelodysplastic syndrome (MDS) is a group of disorders involving hemopoietic dysfunction leading to leukemia. Although recently progress has been made in identifying underlying genetic mutations, many questions still remain. Animal models of MDS have been produced by introduction of specific mutations. However, there is no spontaneous mouse model of MDS, and an animal model to simulate natural MDS pathogenesis is urgently needed.MethodsIn characterizing the genetically diverse mouse strains of the Collaborative Cross (CC) we observed that one, designated JUN, had abnormal hematological traits. This strain was thus further analyzed for phenotypic and pathological identification, comparing the changes in each cell population in peripheral blood and in bone marrow.ResultsIn a specific‐pathogen free environment, mice of the JUN strain are relatively thin, with healthy appearance. However, in a conventional environment, they become lethargic, develop wrinkled yellow hair, have loose and light stools, and are prone to infections. We found that the mice were cytopenic, which was due to abnormal differentiation of multipotent bone marrow progenitor cells. These are common characteristics of MDS.ConclusionsA mouse strain, JUN, was found displaying spontaneous myelodysplastic syndrome. This strain has the advantage over existing models in that it develops MDS spontaneously and is more similar to human MDS than genetically modified mouse models. JUN mice will be an important tool for pathogenesis research of MDS and for evaluation of new drugs and treatments.     

TACI‐dependent APRIL signaling maintains autoreactive B cells in a mouse model of systemic lupus erythematosus

Autoantibodies contribute to the development of systemic lupus erythematosus (SLE). APRIL (a proliferation‐inducing ligand), a member of the TNF superfamily, regulates plasma‐cell survival and binds to TACI (transmembrane activator CAML interactor) and BCMA (B‐cell maturation antigen). We previously showed that APRIL blockade delayed disease onset in lupus‐prone mice. In order to evaluate the role of APRIL receptors in the development of SLE, APRIL, TACI, BCMA , or double TACI.BCMA null mutations were introduced into the Nba2.Yaa (Y‐linked autoimmune acceleration) spontaneous lupus mouse model. Mortality as a consequence of glomerulonephritis (GN) was reduced in Nba2.APRIL^?/?.Yaa , Nba2.TACI^?/?.Yaa and double‐KO mice compared with Nba2.Yaa mice and correlated with lower levels of circulating antibodies, while splenic populations remained unchanged. In contrast, the appearance of symptoms was accelerated in BCMA‐deficient mice, in which TACI signaling was increased. Finally, lupus‐prone mice deficient for the APRIL‐TACI axis produced less pathogenic antibodies and developed less GN. Disease reduction was attributed to impaired T‐independent type 2 responses when the APRIL‐TACI signaling axis was disrupted. Collectively, our results have identified and confirmed APRIL as a new target involved in B‐cell activation, in the maintenance of plasma cell survival and subsequent increased autoantibody production that sustains lupus development in mice.     

Cerebral leucocyte infiltration in lupus-prone MRL/MpJ-fas lpr mice--roles of intercellular adhesion molecule-1 and P-selectin

The autoimmune disease which affects MRL/MpJ-fas(lpr) mice results in cerebral leucocyte recruitment and cognitive dysfunction. We have previously observed increased leucocyte trafficking in the cerebral microcirculation of these mice; however, the types of leucocytes recruited have not been analysed thoroughly, and the roles of key endothelial adhesion molecules in recruitment of these leucocytes have not been investigated. Therefore the aim of this study was to classify the phenotypes of leucocytes present in inflamed brains of MRL/MpJ-fas(lpr) mice, and dissect the roles of endothelial adhesion molecules in their accumulation in the brain. Immunohistochemical analysis revealed significant leucocyte infiltration in the brains of 16- and 20-week-old MRL/MpJ-fas(lpr) mice, affecting predominantly the choroid plexus. Isolation of brain-infiltrating leucocytes revealed that lymphocytes and neutrophils were the main populations present. The CD3(+) lymphocytes in the brain consisted of similar proportions of CD4(+), CD8(+) and CD4(-)/CD8(-)[double negative (DN)] populations. Assessment of MRL/MpJ-fas(lpr) mice deficient in endothelial adhesion molecules intercellular adhesion molecule-1 (ICAM-1) or P-selectin indicated that cerebral leucocyte recruitment persisted in the absence of these molecules, with only minor changes in the phenotypes of infiltrating cells. Together these data indicate that the brains of MRL/MpJ-fas(lpr) mice are affected by a mixed leucocyte infiltrate, of which the unusual DN lymphocyte phenotype contributes a substantial proportion. In addition, endothelial adhesion molecules ICAM-1 and P-selectin, which modulate survival of MRL/MpJ-fas(lpr) mice, do not markedly inhibit leucocyte entry into the central nervous system.     

狼疮性BXSB小鼠抗DNA抗体水平的动态变化

目的 探讨系统性红斑狼疮性小鼠抗ＤＮＡ抗体水平的动态变化与疾病严重程度之间的关系。方法 测定了不同月龄的狼疮性ＢＸＳＢ小鼠的各种抗ＤＮＡ抗体、血尿素氮和尿蛋白质浓度。结果 ＢＸＳＢ小鼠抗ＤＮＡ抗体量的动态变化无规律性，但血尿素氮和尿蛋白质的浓度随鼠龄的增加而逐步升高。结论抗ＤＮＡ抗体水平并不与ＢＸＳＢ小鼠疾病的严重程度相平行，提示在诊断ＳＬＥ或判断其严重程度时应测定多实验指标为宜。     

Toll-like receptor 8 deletion accelerates autoimmunity in a mouse model of lupus through a Toll-like receptor 7-dependent mechanism

Systemic lupus erythematosus is an autoimmune disorder characterized by increased levels of lymphocyte activation, antigen presentation by dendritic cells, and the formation of autoantibodies. This leads to immune complex-mediated glomerulonephritis. Toll-like receptor 7 (T7) and TLR9 localize to the endosomal compartment and play important roles in the generation of autoantibodies against nuclear components, as they recognize RNA and DNA, respectively. In contrast, very little is known about endogenous TLR8 activation in mice. We therefore tested whether TLR8 could affect autoimmune responses in a murine model of lupus. We introduced a Tlr8 null mutation into C57BL/6 mice congenic for the Nba2 (NZB autoimmunity 2) locus and bearing the Yaa (Y-linked autoimmune acceleration) mutation containing a tlr8 duplicated gene, and monitored disease development, autoantibody production, and glomerulonephritis-associated mortality. Cellular responses were investigated in female Nba2.TLR8^−/− mice bearing no copy of tlr8. The TLR8 deficiency accelerated disease progression and mortality, increased the number of circulating antibodies and activated monocytes, and heightened cellular responses to TLR7 ligation. TLR8-deficient antigen-presenting cells exhibited increased levels of MHC class II expression. The ability of dendritic cells to present antigens to allogeneic T cells after TLR7 ligation was also improved by TLR8 deficiency. TLR8 deletion accelerated autoimmunity in lupus-prone mice in response to TLR7 activation. Antigen-presenting cell function seemed to play a key role in mediating the effects of TLR8 deficiency.     

胰岛素样生长因子1的体内表达对BXSB小鼠发病影响的研究

目的 初步研究人胰岛素样生长因子1(hIGF-1)对系统性红斑狼疮ＢＸＳＢ小鼠发病的影响。方法 用肌肉介导的电脉冲转基因技术,使得含有hIGF-1cDNA的重组真核表达质粒在小鼠体内表达,然后系统检测自身免疫相关性实验指标。结果 虽然hIGF-1促进雄性ＢＸＳＢ小鼠ＩgG类型抗ＤＮＡ抗体的产生,但是明显阻止其尿蛋白浓度的继续升高,抑制巨噬细胞分泌IL-1以及降低脾脏重量。结论 IGF-1具有减缓雄性BXSB小鼠狼疮性肾炎的作用。其作用机制之一可能与巨噬细胞分泌炎性介质的功能下降有关。     

应用蛋白组学与代谢组学技术筛选狼疮生物标志物的研究进展

系统性红斑狼疮（SLE）是一种以自身抗体产生和免疫复合物形成为特点的自身免疫性疾病。其临床表现多种多样,包括肾小球肾炎、皮肤炎、血栓形成,血管炎,癫痫和关节炎等。复杂的发病机理和多样的临床症状给SLE的诊断和监测带来了巨大的挑战。寻找新的生物标志物仍然是SLE研究的主要目标和挑战之一。但值得注意的是,蛋白质组学和代谢组学为生物标志物的发现为此提供了新的思路和平台。越来越多的证据表明,蛋白质和代谢图谱具有高灵敏度和特异性,能更安全有效的诊断SLE和判断病情。通过对该领域的研究可以更好地理解SLE的发病机理,并对其诊断、治疗和预防提供新的思路。