image
Home Journals Acta Academiae Medicinae Sinicae
Acta Academiae Medicinae Sinicae

Abbreviation (ISO4): Acta Academiae Medicinae Sinicae      Editor in chief: Xuetao CAO

About  /  Aim & scope  /  Editorial board  /  Indexed  /  Contact  / 
Online first  /  Current issue  /  All issues  /  Top access  /  RSS

Acta Academiae Medicinae Sinicae >

2017 , Vol. 39 >Issue 1: 156 - 161

DOI: https://doi.org/10.3881/j.issn.1000-503X.2017.01.026

Orginal Article

Advances in Mouse Models of Sjögren’s Syndrome

  • YUAN Siyuan 1 ,
  • WANG Xinxiang 2 ,
  • HAO Weixin , 3
Expand
  • 1Department of Neurology,the Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China
  • 2Experimental Center,Dongfang Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100078,China
  • 3Department of Traditional Chinese Medicine,PUMC Hospital,CAMS and PUMC,Beijing 100730,China
Corresponding author:HAO Weixin Tel:010-69151700,E-mail:

Received date: 2016-06-03

  Online published: 2017-02-20

Supported by

Supported by the Beijing Municipal Natural Science Foundation(7132198)

Copyright

《中国医学科学院学报》编辑部 所有
Fold

Abstract

Sjögren’s syndrome (SS) is a common chronic systemic autoimmune disease;however,its pathogenic mechanisms remain unclear. Proper mouse models of SS are essential for experiments. This article summarizes the recent advances in spontaneous mouse models of SS,genetically engineered mouse models of SS,and experimentally induced mouse models of SS.

Key words： Sjö; gren’; s syndrome; mouse models

Cite this article

YUAN Siyuan , WANG Xinxiang , HAO Weixin . Advances in Mouse Models of Sjögren’s Syndrome[J]. Acta Academiae Medicinae Sinicae, 2017 , 39(1) : 156 -161 . DOI: 10.3881/j.issn.1000-503X.2017.01.026

干燥综合征(Sjögren’s syndrome,SS)是一种常见的慢性系统性自身免疫性疾病,主要临床表现为口干、眼干[1-3]。随着病情进展,SS还可累及肺脏、肾脏、肝脏、甲状腺等内脏器官,并出现血液、关节、皮肤等系统受损的表现。小鼠与人的遗传相似度较高,并且很适合做基因克隆与转基因修改,因此近年来被广泛用于人类SS等多种疾病发生机制的研究。本文总结了SS自发小鼠模型、基因工程SS小鼠模型和实验诱导型SS小鼠模型的研究进展。

自发小鼠模型

SS自发小鼠模型是指不经人工诱发,在自然条件下自然产生的SS小鼠模型,或者由于基因突变的异常表现通过遗传育种保留下来的小鼠模型。遗传易感性是自发小鼠模型最大的特征。通过SS自发模型小鼠的研究,一方面可以很好地了解SS疾病发生与发展过程中不同时间点SS发病的特性,另一方面可以对SS易感基因位点以及耐药基因位点进行鉴定。在SS基础研究中,自发小鼠模型运用最广。
NOD小鼠 20世纪90年代,日本科学家对远交系ICR小鼠作近交培育至第6代时,从白内障易感亚型中分离出NOD小鼠[4]。大约4周龄时,NOD小鼠胰腺便开始遭到淋巴细胞的浸润而导致胰腺细胞破坏,造成胰岛素分泌减少。因此,NOD小鼠常被用于制造1型糖尿病自发模型小鼠。
除胰腺外,在NOD小鼠外分泌腺中亦可发现炎性细胞浸润而导致的泪腺炎与涎腺炎[5]。大约从12周龄开始,NOD小鼠外分泌腺便可检测到炎性细胞,至16周左右将会出现明显的外分泌功能障碍。除泪腺炎与涎腺炎炎性细胞浸润的病理与人类所发生的SS表现相似外,NOD小鼠血清中亦能产生多种与SS发生相关的自身抗体,如:抗核抗体(antinuclear antibody,ANA)、干燥综合征抗原A抗体(Sjögren’s syndrome A/Ro,SSA/Ro)、干燥综合征抗原B抗体(Sjögren’s syndrome B/La,SSB/La)、抗相对分子质量为120 000α-胞衬蛋白抗体、抗毒蕈碱受体3抗体及胰岛细胞自身抗体等。此外,据报道NOD小鼠血清及唾液腺中亦能找到一些与SS发生相关的细胞因子,如:白细胞介素(interleukin,IL)-10、干扰素-γ(interferon-γ,INF-γ)、IL-7、膜结合型干细胞因子(membrane bound stem cell factor,MSCF)、IL-17,IL-11、IL-5及IL-8[6]。由于NOD小鼠以上发病机制与人类高度相似,因此被作为主要模型应用于SS研究[5,7-9]。
NOD.B 10-H2b 小鼠:NOD.B 10-H2b 小鼠主要用于原发性SS研究[10],其最大特点便是不再有1型糖尿病发生,仅表现出SS[10-11]。与人体SS发生相类似,NOD.B 10-H2b 小鼠SS的发生,主要表现为B淋巴细胞异常活跃而产生过多的自身抗体[10]。有学者通过应用中华眼镜蛇蛇毒因子干预NOD.B 10-H2b 小鼠,以探究补体在NOD.B 10-H2b 小鼠SS的发生中所起作用,结果发现中华眼镜蛇蛇毒因子由于能够特异性阻断C3分子,降低小鼠外分泌腺腺体淋巴细胞的浸润及ANA表达水平[12]。亦有学者运用NOD.B 10-H2b 小鼠探究性激素在SS发病过程中所起作用,结果显示卵巢切除的NOD.B 10-H2b 小鼠患SS的几率明显上升且症状表现突出,经激素替代疗法后可得到明显改善[13]。
NOD其他衍生杂交小鼠:许多其他NOD衍生杂交小鼠被用来检测某些特定的基因与蛋白在原发性SS发病机制的作用。研究发现,NOD.IFN-γ-/-与NOD. IFN-γ R-/-小鼠外分泌腺并没有外分泌功能的损伤,提示IFN-γ是引发上皮细胞损伤的关键因素[14]。此外,NOD Igμ-/-小鼠被用来研究B淋巴细胞与自身抗体引起SS外分泌腺外分泌功能下降的机制[15]。NOD.IL4-/-小鼠被用来探究T辅助细胞(T helper cell,Th)1与Th2引起SS唾液腺炎性细胞浸润与唾液腺外分泌功能降低所起到的作用,结果发现,虽然在NOD小鼠中可见到明显的口腔干燥表现及唾液腺中明显的T淋巴细胞浸润,但在NOD.IL-4-/-小鼠并没有出现口腔干燥的症状,表明Th1与Th2在引起SS小鼠口腔干燥起到决定作用[16]。
NFS/sld小鼠 基因突变NFS/sld小鼠具有抑制舌下腺分化的常染色体隐性基因,能够影响腺体中腺泡细胞分化成黏液分泌细胞[17]。NFS/sld小鼠模型α-胞衬蛋白的异常水解是引起小鼠唾液腺损伤的最主要原因[18]。胸腺切除的NFS/sld小鼠出现α-胞衬蛋白活化,在出生后3 d即可出现SS样表现,并且其淋巴细胞浸润仅局限于外分泌腺[19]。浸润的淋巴细胞主要为CD4+T淋巴细胞,可见少量的CD8+T淋巴细胞与B220+B淋巴细胞。活化的CD4+T淋巴细胞则导致唾液腺中IFN-γ、IL-2、IL-10、IL-12p40mRNA表达升高。此外,NFS/sld小鼠外分泌腺中亦能检测到导管上皮细胞抗体。NFS/sld小鼠体内检测到的相对分子质量为120 000的器官特异性抗原与人体内α-胞衬蛋白序列相一致[20]。但是当利用免疫荧光标记法检测SS患者血清时,抗α-胞衬蛋白自身抗体与抗SSA/Ro自身抗体和抗SSB/La自身抗体相比灵敏度与特异性相差较大[21]。
IQI/Jic小鼠 与NOD小鼠一样,IQI/Jic小鼠也是从ICR小鼠培育而来。6月龄的IQI/Jic小鼠唾液腺及泪腺中能够见到明显的淋巴细胞浸润灶,并且在雌性IQI/Jic小鼠胸腺中B细胞数量明显增多[22]。值得注意的是,在IQI/Jic小鼠血清中仅能检测到结核抗体表达水平升高而自身抗SSA/Ro与SSB/La并无异常[22]。涎腺炎可发生在雌性IQI/Jic小鼠的各年龄段,且随着小鼠年龄的增长症状逐渐加重;雄性IQI/Jic小鼠涎腺炎发病较轻,且与年龄无关。与人体发生SS特征相似,CD4+T细胞所造成的外分泌腺体破坏较轻,而B220+B则能引起外分泌腺大范围的炎性细胞浸润。研究表明,IQI/Jic小鼠体内T淋巴细胞活性增强能够增加器官内激肽释放酶-13(kallikrein-13,Klk-13)的产生,因此Klk-13的过多产生可能是IQI/Jic小鼠出现SS样表现的重要原因之一[23-24]。
Aly/aly小鼠 Aly/aly小鼠主要是由于常染色体纯合子隐性基因突变而导致淋巴组织发育不全,表现为淋巴结和派伊尔淋巴结缺乏,脾脏与胸腺结构紊乱。Aly/aly小鼠在14周龄时开始自发出现唾液腺及涎腺炎症,并随年龄增长逐渐加重。除引起唾液腺及涎腺炎症外,Aly/aly小鼠还可出现肾炎、肺炎和胰腺炎等疾病。因此Aly/aly小鼠模型亦常用于皮炎、胰腺炎和骨质疏松症等疾病的研究[25-27]。组织病理学观察发现,其炎性细胞浸润主要以CD4+T细胞为主,浸润特征为由导管周围区域向腺泡小叶逐渐扩散。Aly/aly小鼠的发病特点是泪腺浸润损伤情况比唾液腺更为严重,唾液腺中淋巴细胞浸润损害较轻,有时甚至无淋巴细胞浸润[28]。

基因工程小鼠模型

基因工程小鼠模型包括转基因小鼠模型和基因敲除小鼠模型,转基因小鼠模型是指利用转基因技术将目的基因转移到小鼠胚胎细胞内,使之稳定在特定的组织与器官表达或在胚胎发育过程中特定时间上表达的一类动物模型;基因敲除小鼠模型是指使用生物学技术使动物体内某一特定基因灭活或破坏造成缺陷的动物模型。SS基因工程小鼠模型是对小鼠SS相关基因进行有目的地修饰,使小鼠出现SS样改变,该小鼠模型主要用于SS发病机制及筛选防治SS药物的研究。
分化抑制因子 分化抑制因子(inhibitor of differentiation 3,Id3)能够抑制碱性螺旋-环-螺旋结构域转录因子与DNA的结合,对免疫细胞及非免疫细胞的增殖及分化发挥调节作用[29]。Id3同时也参与T淋巴细胞的选择与分化的信号转导。Id3 KO小鼠的基本特征为免疫系统改变,结外边缘区B细胞增殖,前B细胞残存及出现主要组织相容性复合体限制性[30]。并且这一品系的小鼠将出现与人类相类似的SS[31]。Id3 KO 小鼠在8周龄时,唾液腺与泪腺导管和血管周围将出现淋巴细胞浸润,且随年龄增长浸润情况加重。6至12月龄时泪腺与唾液腺将只表现为CD4+T淋巴细胞浸润,12月龄时可在血清中检测到抗SSA/Ro与SSB/La抗体阳性。值得注意的是,此系列小鼠早在6至18周龄时便会出现SS样表现,但此时小鼠的外分泌腺并未遭到破坏且此时小鼠血清检查也未发现异常自身抗体,表明SS发病还有其他因素起作用。
芳香化酶基因 芳香化酶基因主要是调控雌性激素的产生,芳香化酶基因敲除(aromatase knockout,ArKO)小鼠在12周龄时会表现出T淋巴细胞异常增生且出现SS相类似表现[32]。这一类小鼠发病特征以B220+细胞浸润为主,导致腺泡细胞结构破坏及唾液腺功能损伤。ArKO小鼠血清中抗α-胞衬蛋白抗体及α-胞衬蛋白水解片段与SS患者组织损伤相类似。ArKO小鼠体内未能检测到抗核抗体。此外,在ArKO小鼠体内可发现轻度脾肿大,骨髓中见有异常B淋巴细胞增生。对雌激素缺乏引起的SS小鼠发病机制的研究表明,使用雌激素、大豆异黄酮等对这一类型的SS进行治疗可能有效[33-34]。然而最近有一项报道对此持反对意见。Rahimi Darabad等[31]研究表明,雌激素缺乏的ArKO年轻小鼠虽然会影响其与雌激素相关泪腺特异性基因表达,但其并不会表现出SS相关症状。以上研究提示,ArKO小鼠,尤其是年轻小鼠泪腺功能与结构的变化除了受雌性激素不足影响外,可能亦受其他因素的影响。

实验诱导型小鼠模型

实验诱导型小鼠模型是指通过对正常小鼠进行相应干预以诱导其出现SS表现。目前主要使用的诱导方法为注射同种小鼠或者异种小鼠自身抗原或组织匀浆进行免疫,或用佐剂进行免疫;亦有通过接种病毒进行诱导。诱导型小鼠模型在功能代谢形态结构等方面有所改变,可出现明显的淋巴细胞浸润,导管、血管扩张,腺泡萎缩。但实验诱导的的外分泌腺淋巴细胞浸润是否与自身免疫反应过程有关,目前尚未明确,这类动物模型仅是从局部模拟了人类SS的表现。目前这类模型主要用于疾病的发生和治疗研究。
Ro抗原免疫诱导反复对BALB/c小鼠腹腔进行Ro60 KD自身抗原注射能够引起表位扩展,出现口腔干燥等一些与SS相类似的临床表现[35]。其唾液腺中所浸润的淋巴细胞主要包括45% CD4+T细胞、18%CD8+T细胞和35% CD19+B细胞等。此外,38周龄时在小鼠体内可检测到抗SSA/Ro与抗SSB/La抗体的产生。到目前为止,Ro自身抗体引起小鼠SS的发病机制尚有争论[36]。此类SS小鼠模型最大的缺点是BALB/c小鼠需反复进行腹腔注射Ro60 KD自身抗原5个月以上才能成模。
最近一项研究显示,使用Ro60 KD自身抗原干预不同品系的小鼠,所得结果不尽相同[37]。SJL/J小鼠对此并不产生免疫反应;C57BL/6小鼠经Ro60 KD自身抗原刺激后虽有自身抗体产生,但并不能观察到表位扩展现象。并且SJL/J、C57BL/6与PL/J小鼠外分泌腺均未出现淋巴细胞浸润情况及外分泌腺功能丧失。此外,DBA-2 与BALB/c小鼠外分泌腺均能出现淋巴细胞浸润,但只有BALB/c小鼠外分泌功能下降。
毒蕈碱受体3肽段免疫诱导 多项研究证实,SS患者体内产生的抗毒蕈碱受体3抗体由于能够与唾液腺腺泡细胞上毒蕈碱受体3特异性结合,导致唾液腺外分泌功能丧失[38]。实验显示,毒蕈碱受体3基因敲除小鼠唾液腺外分泌功能与正常小鼠相比下降20%,而毒蕈碱1受体基因敲除小鼠唾液腺外分泌功能并未出现任何变化,同时将小鼠毒蕈碱受体1与受体3基因敲除后小鼠唾液腺则将毫无外分泌功能,可见在SS小鼠唾液腺外分泌功能下降过程中。毒蕈碱受体3发挥了更为关键的作用[39]。
最近的一项研究报道了毒蕈碱受体3细胞外第2环功能表位肽段(208-227)的自身抗体诱发幼年雌性NOD/LtJ小鼠免疫反应的情况,结果显示NOD/LtJ小鼠体内Th-1、Th-2、Th-17等细胞因子表达下降,泪腺与唾液腺中淋巴细胞浸润情况得到改善[40]。这一结果显示毒蕈碱3受体肽段对此类SS可能具有一定治疗作用。邹新乐等[41]利用毒蕈碱受体3肽段N49免疫Balb/c小鼠成功建立了SS小鼠模型。
碳酸酐酶免疫诱导 碳酸酐酶(carbonic anhydraseⅡ,CAⅡ)是一种锌酶,主要功能是催化二氧化碳可逆的水合作用。尽管机体内CAⅡ抗体是自身免疫性胰腺炎的典型表现,但有研究显示许多SS患者体内亦能检测到CAⅡ抗体表达[39,42-43]。对PL/J(H-2u)小鼠使用CAⅡ进行免疫诱导能够产生自身免疫性涎腺炎,在涎腺腺泡周围能够观察到明显的淋巴细胞浸润灶[44]。但到目前为止,CAⅡ参与SS的发病机制尚不明确[45]。

小 结

小鼠模型由于具有价廉、实用、饲养条件要求较简单、寿命短、基因可塑性强等优势,作为一种模式生物被广泛用于研究人类疾病的发病与治疗。通过对SS小鼠模型的研究,能够发现SS发病机制的诸多信息。需要指出的是,尽管近年来在对SS小鼠模型的研究上取得了重大突破,但到目前为止尚没有任何一种模型能够完全模拟人类SS发病各阶段的特征。人类与啮齿类动物的免疫系统始终存在着很大差别。因此,在科研工作中,科研人员想要选择一种适用SS小鼠模型,必须熟练掌握各种SS小鼠模型的发病背景,参照多种SS小鼠模型的发病特征,以对SS的发病进行一个全方位的掌握。同时,研发出更贴切的SS动物模型也非常重要。

The authors have declared that no competing interests exist.

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
Barone F,Colafrancesco S.Sjögren’s syndrome:from pathogenesis to novel therapeutic targets[J]. Clin Exp Rheumatol,2016,34(4 Suppl 98):58-62.Primary Sjgren's syndrome (pSS) is a chronic inflammatory autoimmune disease, characterised by a chronic infiltration of exocrine glands, mainly salivary glands, with the histological features of focal lymphocytic sialoadenitis. Disease spectrum is broad and the occurrence of several extra-glandular manifestations, and in rare cases lymphoma development, is well known. A specific approved treatment for pSS is still lacking and the detection of novel therapeutic biologic target is ongoing. The identification of biological fingerprints seems essential in order to stratify patients both in clinical trials and in real life. Discovery of new components of the inflammatory response will be the key in the future for the identification of novel additional therapeutic options.

PMID

[2]
Rischmueller M,Tieu J,Lester S.Primary Sjögren’s syndrome[J]. Best Pract Res Clin Rheumatol,2016,30(1):189-220.

[3]
Pers JO,Saraux A.Future treatments for Sjögren’s syndrome[J]. Presse Med,2016,45(6 Pt 2):e193-e200.

[4]
Kikutani H,Makino S.The murine autoimmune diabetes model:NOD and related strains[J]. Adv Immunol,1992,51:285-322.This chapter summarizes the experimental findings and observations based on the nonobese diabetic (NOD) mouse model and discusses the pathogenesis and the etiology of autoimmune diabetes. To elucidate the etiology of insulin-dependent diabetes mellitus (IDDM), several animal models have been developed, including experimentally induced and spontaneously developing models. One is low-dose streptozocin-induced diabetes. A high dose of streptozocin destroys completely pancreatic cells but not and cells, resulting in acute diabetes. Multiple injections of subdiabetogenic doses of streptozocin can also induce diabetes a few weeks after treatment. In the latter case, mononuclear infiltration in and around pancreatic islets is observed. IDDM in NOD mice is also immunologically mediated. IDDM in BB rats and NOD mice is very similar to the disease in humans. The availability of a large number of data and materials in mouse genetics and immunology has accelerated the use of NOD mice by diabetologists and immunologists who are interested in the pathogenesis of IDDM and the mechanism of autoimmunity. Several important findings obtained from studies on NOD mice have provided new insights into the immunological and genetic control of IDDM. The development of IDDM in NOD mice is divided roughly into two phases: initiation of autoimmune insulitis and promotion of islet destruction and overt diabetes.

DOI PMID

[5]
Inoue H,Kishimoto A,Ushikoshi-Nakayama R,et al.Resveratrol improves salivary dysfunction in a non-obese diabetic (NOD) mouse model of Sjögren’s syndrome[J]. J Clin Biochem Nutr,2016,59(2):107-112.

[6]
Delaleu N,Immervoll H,Cornelius J,et al.Biomarker profiles in serum and saliva of experimental Sjögren’s syndrome:associations with specific autoimmune manifestations[J]. Arthritis Res Ther,2008,10(1):R22. doi:10.1186/ar2375.

[7]
Li CL,He J,Li ZG,et al.Effects of multi-glycosides of tripterygium wilfordiion in the treatment of Sjögren’s syndrome in the non-obese diabetic mouse model[J]. Chin J Dent Res,2015,18(2):95-101.To investigate the effects of the multi-glycosides of Tripterygium wilfordii (GTW) on Sjgren's syndrome (SS) in the non-obese diabetic (NOD) mouse model.Twenty-seven 8-week-old, female NOD mice were divided into the GTW group, the hydroxychloroquine (HCQ) group, and control (normal saline) group, and received corresponding treatment for 16 weeks. The treatment-induced changes in stimulated total saliva flow rate (STFR), level of serum anti-SSA/SSB, ratio of regulatory T (Treg) cells, histology of the submandibular gland (SMG) and the gene expression profile that is related to inflammation and autoimmunization were evaluated.Compared to the untreated (control) mice, STRF, SMG index and Treg/CD4+ cell ratio were significantly higher, whereas anti-SSA, anti-SSB and lymphoid foci were remarkably lower in GTW-treated mice. HCQ-treated mice showed similar results except SMG index was not different from the untreated mice. NOD mice showed 19.03% altered gene expression with maturation from the age of 8 weeks to 24 weeks. Treatment with HCQ and GTW reduced the change in gene expression to 13.09% and 7.14%, respectively.GTW is as effective as HCQ in the treatment of Sjgren's syndrome in the NOD mouse model.

PMID

[8]
Humphreys-Beher MG,Hu Y,Nakagawa Y,et al.Utilization of the nonobese diabetic (NOD) mouse as an animal model for the study of secondary Sjögren’s syndrome[J]. Adv Exp Med Biol,1994,350:631-636.Sj02gren’s syndrome (S.S.) in the human patient population is an autoimmune inflammatory disease presenting clinical symptoms of xerophthalmia and xerostomia 1 . This condition predominantly affects women. Most diagnoses of S.S. is made in association with autoimmune connective tissue diseases such as rheumatoid arthritis or systemic lupus erythematosus. 2 However, it can also be obseved as an isolated phenomenon, described as primary S.S.

DOI PMID

[9]
Gervais EM,Desantis KA,Pagendarm N,et al.Changes in the submandibular salivary gland epithelial cell subpopulations during progression of Sjögren’s syndrome-like disease in the NOD/ShiLtJ mouse model[J]. Anat Rec (Hoboken),2015,298(9):1622-1634.

[10]
Robinson CP,Yamachika S,Bounous DI,et al.A novel NOD-derived murine model of primary Sjögren’s syndrome[J]. Arthritis Rheum,1998,41(1):150-156.

[11]
Kong L,Robinson CP,Peck AB,et al.Inappropriate apoptosis of salivary and lacrimal gland epithelium of immunodeficient NOD-scid mice[J]. Clin Exp Rheumatol,1998,16(6):675-681.

[12]
Fearon DT,Carroll MC.Regulation of B lymphocyte responses to foreign and self-antigens by the CD19/CD21 complex[J]. Annu Rev Immunol,2000,18:393-422.The membrane protein complex CD19/CD21 couples the innate immune recognition of microbial antigens by the complement system to the activation of B cells. CD21 binds the C3d fragment of activated C3 that becomes covalently attached to targets of complement activation, and CD19 co-stimulates signaling through the antigen receptor, membrane immunoglobulin. CD21 is also expressed by follicular dendritic cells and mediates the long-term retention of antigen that is required for the maintenance of memory B cells. Understanding of the biology of this receptor complex has been enriched by analyses of genetically modified mice; these analyses have uncovered roles not only in positive responses to foreign antigens, but also in the development of tolerance to self-antigens. Studies of signal transduction have begun to determine the basis for the coreceptor activities of CD19. The integration of innate and adaptive immune recognition at this molecular site on the B cell guides the appropriate selection of antigen by adaptive immunity and emphasizes the importance of this coreceptor complex.

DOI PMID

[13]
Mostafa S,Seamon V,Azzarolo AM.Influence of sex hormones and genetic predisposition in Sjögren’s syndrome:a new clue to the immunopathogenesis of dry eye disease[J]. Exp Eye Res,2012,96(1):88-97.Sjgren's syndrome (SS) is a chronic autoimmune disease characterized by lymphocytic infiltration, destruction of lacrimal and salivary glands and the presence of serum autoantibodies. Most women that suffer from SS are post-menopausal however, not all post-menopausal women develop SS, suggesting that other factors, in addition to the decrease in ovarian hormones, are necessary for the development of SS. The purposes of this study were to investigate a) the time course of lymphocytic infiltration and apoptosis in the lacrimal gland after ovariectomy, b) if a predisposed genetic background for SS aggravates the effects of decreasing levels of sex hormones in the lacrimal glands and c) if physiological doses of estrogen or androgen prevent the effects observed after ovariectomy. Six weeks old mice that are genetically predisposed to SS (NOD.B10.H2(b)) and control (C57BL/10) mice were either sham operated, ovariectomized (OVX), OVX+17 estradiol (E(2)) or OVX+Dihydrotestosterone (DHT). Lacrimal glands were collected at 3, 7, 21 or 30 days after surgery and processed for immunohistochemistry to measure CD4(+), CD8(+) T cells, B220(+) B cells, nuclear DNA degradation and cleaved caspase-3 activity. Quantification of the staining was done by light microscopy and Image Pro Plus software. The results of our study show that lymphocytic infiltration preceded lacrimal gland apoptosis after ovariectomy. Moreover, removal of ovarian sex hormones accelerated these effects in the genetically predisposed animal and these effects were more severe and persistent compared to control animals. In addition, sex hormone replacement at physiological levels prevented these symptoms. The mechanisms by which decreased levels of sex hormones caused lymphocytic infiltration and apoptosis and the interaction of lack of sex hormones with the genetic elements remain to be elucidated.

DOI PMID

[14]
Cha S,Brayer J,Gao J,et al.A dual role for interferon-gamma in the pathogenesis of Sjögren’s syndrome-like autoimmune exocrinopathy in the nonobese diabetic mouse[J]. Scand J Immunol,2004,60(6):552-565.Abstract Sjogren's syndrome-like autoimmune exocrinopathy (AEC) in the nonobese diabetic (NOD) mouse progresses from a preimmune phase to an immune phase, resulting in dry mouth and/or dry eyes. In the present study, the impact of the prototypical T-helper type 1 cytokine, interferon-gamma (IFN-gamma), on the onset of AEC was investigated using both the IFN-gamma and the IFN-gamma receptor gene knockout mice, NOD.IFN-gamma(-/-) and NOD.IFN-gammaR(-/-), respectively. Neither the NOD.IFN-gamma(-/-) nor the NOD.IFN-gammaR(-/-) mice exhibited increased acinar cell apoptosis and abnormal salivary protein expression, typically observed in parental NOD mice prior to disease. Without these preimmune phase abnormalities, NOD.IFN-gamma(-/-) and NOD.IFN-gammaR(-/-) mice showed no subsequent autoimmune responses against the salivary glands at 20 weeks. Interestingly, real-time polymerase chain reaction and electrophoretic gel mobility shift assays suggested that IFN-gamma and STAT1, as well as the transcriptional activity of STAT1 in NOD glands, were increased at birth. Unlike the neonatal submandibular glands of NOD or NOD-scid mice that show abnormal glandular morphogenesis at birth, the submandibular glands of the newly constructed congenic strain, NOD-scid.IFN-gamma(-/-), were found to be normal. Taken together, IFN-gamma appears to play a critical role not only during the later immune phase of AEC, but also the early preimmune phase, independent of effector functions of immune cells. How exactly IFN-gamma functions during this period remains speculative.

DOI PMID

[15]
Robinson CP,Brayer J,Yamachika S,et al.Transfer of human serum IgG to nonobese diabetic Igmu null mice reveals a role for autoantibodies in the loss of secretory function of exocrine tissues in Sjögren’s syndrome[J]. Proc Natl Acad Sci USA,1998,95(13):7538-7543.

[16]
Brayer JB,Cha S,Nagashima H,et al.IL-4-dependent effector phase in autoimmune exocrinopathy as defined by the NOD. IL-4-gene knockout mouse model of Sjögren’s syndrome[J]. Scand J Immunol,2001,54(1-2):133-140.

[17]
Hayashi Y,Kojima A,Hata M,et al.A new mutation involving the sublingual gland in NFS/N mice. Partially arrested mucous cell differentiation[J]. Am J Pathol,1988,132(2):187-191.A new mutation in mice affecting the mucous cell differentiation of the sublingual glands is described. The normal mouse sublingual glands are mucus-secreting and virtually all the acinar cells differentiate to mucus-rich cells by the day of birth. In contrast, all endpieces of newborn mutant mice consisted of acini of immature cuboidal cells. However, normal mucous cells, staining intensively with mucin-specific stains such as Alcian blue at pH 2.5 or mucicarmine, appeared in the mutant mice from an early age singly or in groups in a small number of acini, and their number apparently increased with age to occupy over 30% of the total acinar cells. Ultrastructurally, irregular secretion granules of varying electron-density, distinct from ordinary sublingual mucin granules, were frequently observed in the cytoplasm of the immature acinar cells in the mutant phenotype. The genetic analysis showed that a single autosomal recessive gene determined the observed abnormality. This is the first salivary gland mutation and will provide a critical model for the study of salivary mucous cell differentiation.

DOI PMID

[18]
Haneji N,Hamano H,Yanagi K,et al.A new animal model for primary Sjögren’s syndrome in NFS/sld mutant mice[J]. J Immunol,1994,153(6):2769-2777.

[19]
Hayashi Y,Haneji N,Hamano H,et al.Effector mechanism of experimental autoimmune sialadenitis in the mouse model for primary Sjögren’s syndrome[J]. Cell Immunol,1996,171(2):217-225.We have recently established a new animal model for primary Sj枚gren's syndrome in NFS/sld mutant mice thymectomized 3 days after birth (3dTX) bearing an autosomal recessive gene with sublingual gland differentiation arrest. In this study, we analyze developing mechanisms of experimental autoimmune sialadenitis (EAS) in the mouse model, focusing on local expressions of cytokine and cell adhesion molecule genes by reverse transcriptase-polymeric chain reaction (RT-PCR) and immunohistochemistry, kinetic analysis of splenic lymphocytes expressing activation markers, and I-Aq class-II molecules by flow cytometry (FACS). We found up-regulation of local cytokine genes (IL-1 beta, TNF-alpha, IL-2, IFN-gamma, IL-6, IL-10, IL-12p40) and cell adhesion molecule genes (ICAM-1, LFA-1, CD44, Mel-14) in the salivary glands from mice with EAS by RT-PCR, which were supported by immunohistochemistry. FACS analysis demonstrated that a significant proportion of splenic CD4+ T cells express activation markers (CD44, LFA-1, Mel-14low, CD45RB(low)) at a high level and an increase in expression of B220+ B cells bearing I-Aq class-II molecules. These data suggest that spontaneous EAS in 3dTX NFS/sld mutant mice may be triggered by an in situ activation of autoreactive CD4+ T cells comprising unique cytokine profile (high levels of IL-2, IFN-gamma, IL-10, and IL-12p40 mRNA) in the salivary glands.

DOI PMID

[20]
Ishimaru N,Yoneda T,Saegusa K,et al.Severe destructive autoimmune lesions with aging in murine Sjögren’s syndrome through Fas-mediated apoptosis[J]. Am J Pathol,2000,156(5):1557-1564.When we evaluated the age-associated changes in autoimmune exocrinopathy in a NFS/ sld murine model for primary Sjgren's syndrome (SS), severe destructive autoimmune lesions developed in the salivary and lacrimal glands in the aged mice, compared with those observed in the younger model. We detected a decreased secretion of saliva and tear flow in the aged group. A significant increase of TUNEL + -apoptotic epithelial duct cells in the salivary glands was detected in the aged SS animal model. A higher proportion of mouse salivary gland cells bearing Fas was found in the aged group, whereas no significant changes were seen on tissue-infiltrating CD4 + T cells bearing FasL in the salivary glands from young and aged mice. We detected an increased cleavage product of organ-specific autoantigen, 120-kd -fodrin, in the aged salivary gland tissues on immunoblotting, and an increase in serum autoantibody production against 120-kd -fodrin by enzyme-linked immunosorbent assay. An increase in the proliferative response of splenic T cells against organ-specific autoantigen was observed, whereas nonspecific concanavalin A responsiveness was decreased in the aged mice. In addition, a decrease in Fas expression was found on splenic CD4 + T cells in the aged mice, and anti-Fas mAb-stimulated apoptosis was down-regulated on CD4 + T cells. These results indicate that age-associated dysregulation of CD4 + T cells may play a crucial role on acceleration of organ-specific autoimmune lesions in a murine model for primary SS through Fas-mediated apoptosis.

DOI PMID

[21]
Zandbelt MM,Vogelzangs J,Van De Putte LB,et al. Antialpha-fodrin antibodies do not add much to the diagnosis of Sjögren’s syndrome[J]. Arthritis Res Ther,2004,6(1):33-38.

[22]
Saegusa J,Kubota H.Sialadenitis in IQI/Jic mice:a new animal model of Sjögren’s syndrome[J]. J Vet Med Sci,1997,59(10):897-903.

[23]
Takada K,Takiguchi M,Konno A,et al.Autoimmunity against a tissue kallikrein in IQI/Jic mice:a model for Sjögren’s syndrome[J]. J Biol Chem,2005,280(5):3982-3988.

[24]
Kaczor-Urbanowicz KE,Martin Carreras-Presas C,Aro K,et al. Saliva diagnostics-current views and directions [J]. Exp Biol Med (Maywood), 2016.[2016-12-08].

rnals.sagepub.com/doi/pdf/10.1177/1535370216681550.

[25]
Seo Y,Fukushima H,Maruyama T,et al.Accumulation of p100,a precursor of NF-kappaB2,enhances osteoblastic differentiation in vitro and bone formation in vivo in aly/aly mice[J]. Mol Endocrinol,2012,26(3):414-422.We previously reported that alymphoplasia (aly/aly) mice, which have a natural loss-of-function mutation in the Nik gene, which encodes a kinase essential for the processing of p100 to p52 in the alternative nuclear factor-κB (NF-κB) pathway, show mild osteopetrosis with an increase in several parameters of bone formation: bone formation rate, mineral apposition rate, and osteoblast number. We therefore investigated the molecular mechanisms triggered by the alternative NF-κB pathway in the regulation of osteoblast differentiation using primary osteoblasts (POB) prepared from aly/aly mice. Alkaline phosphatase (ALP) activity and mineralization induced by the presence of β-glycerophosphate and ascorbic acid were enhanced in POB from aly/aly compared with wild-type (WT) mice. Furthermore, osteoblastic differentiation induced by bone morphogenetic protein 2 (BMP2), as shown by ALP activity, mRNA expression of osteocalcin, Id1, Osterix and Runx2, and Sma- and Mad-related protein (Smad)1/5/8 phosphorylation, was also enhanced in POB from aly/aly mice. The ectopic bone formation in vivo that was induced by BMP2 was enhanced in aly/aly mice compared with controls. Transfection of a mutant form of p100, p100ΔGRR, which cannot be processed to p52, stimulated ALP activity and Smad phosphorylation. In contrast to p100ΔGRR, overexpression of p52 inhibited these events. Both BMP2-induced ALP activity and Smad phosphorylation were reduced in POB from p100-deficient mice, which carry a homozygous deletion of the COOH-terminal ankyrin repeats of p100 but still express functional p52 protein. p52 and p100ΔGRR interacted with a BMP receptor, ALK2, in overexpressed COS7 cells and changed the ALK2 protein levels in opposite directions: p52 reduced ALK2 and p100 increased it. Thus, the alternative the NF-κB pathway via the processing of p52 from p100 negatively regulates osteoblastic differentiation and bone formation by modifying BMP activity.

DOI PMID

[26]
Wang HX,Yi SQ,Li J,et al.Effects of splenectomy on spontaneously chronic pancreatitis in aly/aly mice[J]. Clin Dev Immunol,2010,2010:614890. doi:10.1155/2010/614890.Background and Aim. Mice with alymphoplasia (aly/aly) mutation characterized by a lack of lymph nodes, Peyer's patches, and well-defined lymphoid follicles in the spleen were found. In this study, we used splenectomized aly/aly mice to elucidate the effects of secondary lymphoid organs in the development of aly/aly autoimmune pancreatitis. Methods. Forty-eight 10-week-old aly/aly mice were divided into two groups for splenectomy and sham operation. Histological and immunohistochemical analyses of the pancreas were performed at the ages of 20, 30, and 40 weeks old after operation, respectively. Results. Our results showed that mononuclear cell infiltration was restricted to the interlobular connective tissues at the age of 20 weeks, and not increase obviously at the age of 30 and 40 weeks in splenectomized aly/aly mice. Furthermore, an apparent decrease in the expressions of CD4+ T, CD8+ T, and B cells was detected in the pancreatic tissues compared with sham aly/aly mice, however, no significant difference in macrophage expression between mice with and without a splenectomy. Conclusions. Inflammation infiltration and development of the pancreatitis in aly/aly mice were suppressed effectively after splenectomy, which was, at least partly, correlated to inhibition of the infiltration of T and B cells in pancreatic tissues but not to macrophages.

DOI PMID

[27]
Kobayashi S,Ueda A,Ueda M,et al.Pathology of spontaneous dermatitis in CBy.ALY-aly mice[J]. Exp Anim,2008,57(2):159-163.CBy.ALY-aly/aly (C-aly) mice develop progressive dermatitis in areas of the face and dorsal neck from around three months of age. Staphylococcus aureus was detected in the skin lesions of C-aly mice. However, even when the mice were raised under S. aureus free conditions, a similar, though less severe, dermatitis was still observed. The mice showed a marked increase in the number of eosinophils in the peripheral blood and skin lesions, with no changes in plasma IgE levels. These findings suggest that the dermatitis of these mice may be an atypical allergic condition with little or no involvement of IgE. C-aly mice may be a useful animal model of chronic dermatitis or pruritus without elevated IgE levels.

DOI PMID

[28]
Tsubata R,Tsubata T,Hiai H,et al.Autoimmune disease of exocrine organs in immunodeficient alymphoplasia mice:a spontaneous model for Sjögren’s syndrome[J]. Eur J Immunol,1996,26(11):2742-2748.Mice homozygous for an autosomal recessive mutation aly (alymphoplasia) lack both lymph nodes and Peyer's patches, and show defects in both humoral and cellular immunity. Histopathological analysis revealed chronic inflammatory changes in exocrine organs such as the salivary gland, lacrimal gland, and pancreas of the homozygotes (aly/aly), but not the heterozygotes (aly/+). In these exocrine organs, mononuclear cells consisting mainly of CD4+ T cells infiltrate periductal areas, and, in some cases, the cell infiltration extended to lobules. The inflammatory changes in exocrine organs were transferred by a T cell-enriched fraction of spleen cells from homozygous animals. These results suggest that autoimmune mechanisms mediated by self-reactive T cells may be involved in the inflammatory lesions of various exocrine organs in the homozygous mice, although these mice show immunodeficiency. Inflammatory changes were also observed in the lung of the homozygotes. Since Sj枚gren's syndrome is characterized by diffuse lymphocyte infiltration in the periductal areas of the lacrimal and salivary glands and is occasionally associated with pulmonary disease, aly/aly mice may serve as a unique spontaneous model of Sjgren's syndrome.

DOI PMID

[29]
Pan L,Sato S,Frederick JP,et al.Impaired immune responses and B-cell proliferation in mice lacking the Id3 gene[J]. Mol Cell Biol,1999,19(9):5969-5980.

[30]
Quong MW,Romanow WJ,Murre C.E protein function in lymphocyte development[J]. Annu Rev Immunol,2002,20:301-322.

[31]
Rahimi Darabad R,Suzuki T,Richards SM,et al.Does estrogen deficiency cause lacrimal gland inflammation and aqueous-deficient dry eye in mice[J]. Exp Eye Res,2014,127:153-160.Abstract Researchers have proposed that estrogen deficiency will lead to a Sjgren's syndrome (SjS)-like lacrimal gland inflammation, aqueous tear deficiency and dry eye. The purpose of this study was to determine whether this proposal is correct. Lacrimal glands were obtained from adult, age-matched wild type (WT) and aromatase knockout (ArKO) mice, in which estrogen synthesis is completely eliminated. Tissues were also obtained from autoimmune MRL/Mp-lpr/lpr (MRL/lpr) mice as inflammation controls. Tear volumes in WT and ArKO mice were measured and glands were processed for molecular biological and histological evaluation. Our results demonstrate that estrogen absence does not lead to a SjS-like inflammation in lacrimal tissue or to an aqueous-deficient dry eye. There was no upregulation of genes associated with inflammatory pathways in lacrimal glands of male or female ArKO mice. Such inflammatory activity was prominent in autoimmune MRL/lpr tissues. We also found no evidence of inflammation in lacrimal gland tissue sections of estrogen-deficient mice, and tear volumes of ArKO males were actually increased as compared to those WT controls. Interestingly, our study did show that estrogen absence influences the expression of thousands of lacrimal gland genes, and that this impact is sex- and genotype-specific. Our findings demonstrate that estrogen absence is not a risk factor for the development of SjS-like lacrimal gland inflammation or for aqueous-deficient dry eye in mice. Copyright © 2014 Elsevier Ltd. All rights reserved.

DOI PMID

[32]
Shim GJ,Warner M,Kim HJ,et al.Aromatase-deficient mice spontaneously develop a lymphoproliferative autoimmune disease resembling Sjögren’s syndrome[J]. Proc Natl Acad Sci USA,2004,101(34):12628-12633.Sjgren's syndrome (SS) is an incurable, autoimmune exocrinopathy that predominantly affects females and whose pathogenesis remains unknown. Like rheumatoid arthritis, its severity increases after menopause, and estrogen deficiency has been implicated. We have reported that estrogen receptor-alpha and -beta-knockout mice develop autoimmune nephritis and myeloid leukemia, respectively, but neither develops SS. One model of estrogen deficiency in rodents is the aromatase-knockout (ArKO) mouse. In these animals, there is elevated B lymphopoiesis in bone marrow. We now report that ArKO mice develop severe autoimmune exocrinopathy resembling SS. By 1 year of age, there is B cell hyperplasia in the bone marrow, spleen, and blood of ArKO mice and spontaneous autoimmune manifestations such as proteinuria and severe leukocyte infiltration in the salivary glands and kidney. Also, as is typically found in human SS, there were proteolytic fragments of alpha-fodrin in the salivary glands and anti-alpha-fodrin antibodies in the serum of both female and male ArKO mice. When mice were raised on a phytoestrogen-free diet, there was a mild but significant incidence of infiltration of B lymphocytes in WT mice and severe destructive autoimmune lesions in ArKO mice. In age-matched WT mice fed a diet containing normal levels of phytoestrogen, there were no autoimmune lesions. These results reveal that estrogen deficiency results in a lymphoproliferative autoimmune disease resembling SS and suggest that estrogen might have clinical value in the prevention or treatment of this disease.

DOI PMID

[33]
Li Y,Liu C,Hou W,et al.Retrograde ductal administration of the adenovirus-mediated NDRG2 gene leads to improved sialaden hypofunction in estrogen-deficient rats[J]. Mol Ther,2014,22(5):908-918.One of the most common oral manifestations of menopause is xerostomia. Oral dryness can profoundly affect quality of life and interfere with basic daily functions, such as chewing, deglutition, and speaking. Although the feeling of oral dryness can be ameliorated after estrogen supplementation, the side effects of estrogen greatly restrict its application. We previously found that N-myc downstream-regulated gene 2 (NDRG2) is involved in estrogen-mediated ion and fluid transport in a cell-based model. In the present study, we used an ovariectomized rat model to mimic xerostomia in menopausal women and constructed two adenovirus vectors bearing NDRG2 to validate their therapeutic potential. Ovariectomized rats exhibited severe sialaden hypofunction, including decreased saliva secretion and ion reabsorption as well as increased water intake. Immunohistochemistry revealed that the expression of NDRG2 and Na + reabsorption-related Na + /K + -ATPase and epithelial sodium channels (EnaC) decreased in ovariectomized rat salivary glands. We further showed that the localized delivery of NDRG2 improved the dysfunction of Na + and Cl reabsorption. In addition, the saliva flow rate and water drinking recovered to normal. This study elucidates the mechanism of estrogen deficiency-mediated xerostomia or sialaden hypofunction and provides a promising strategy for therapeutic intervention.

DOI PMID

[34]
Carvalho VD,Silveira VA,do Prado RF,et al. Effect of estrogen therapy,soy isoflavones,and the combination therapy on the submandibular gland of ovariectomized rats[J]. Pathol Res Pract,2011,207(5):300-305.

[35]
Scofield RH,Asfa S,Obeso D,et al.Immunization with short peptides from the 60-kDa Ro antigen recapitulates the serological and pathological findings as well as the salivary gland dysfunction of Sjögren’s syndrome[J].J Immunol,2005,175(12):8409-8414.Sjgren's syndrome is a poorly understood autoimmune inflammatory illness that affects the salivary and lacrimal glands as well as other organ systems. We undertook the present study to determine whether mice immunized with short peptides from the 60-kDa Ro (or SSA) Ag, which is a common target of the autoimmunity of Sjgren's syndrome, develop an illness similar to Sjgren's syndrome. BALB/c mice were immunized with one of two short peptides from 60-kDa Ro that are know to induce epitope spreading. The animals were analyzed for the presence of anti-Ro and anti-La (or SSB) in the sera by immunoblot and ELISA. Salivary glands were collected and examined by histology after H&E staining. Salivary lymphocytes were purified and studied for cell surface makers by fluorescence-activated cell sorting. Timed stimulated salivary flow was measured. As reported previously, BALB/c mice immunized with 60-kDa Ro peptides developed an immune response directed against the entire Ro/La ribonucleoprotein particle that was similar to that found in humans with lupus or Sjgren's syndrome. Functional studies showed a statistical decrease in salivary flow in immunized mice compared with controls. Furthermore, there were lymphocytic infiltrates in the salivary glands of immunized animals that were not present in controls. The infiltrates consisted of both CD4- and CD8+ T lymphocytes as well as B lymphocytes. BALB/c mice immunized with 60-kDa Ro peptides develop anti-Ro, salivary gland lymphocyte infiltrates, and salivary dysfunction that is highly reminiscent of human Sjgren's syndrome.

DOI PMID

[36]
Chiorini JA,Cihakova D,Ouellette CE,et al.Sjögren syndrome:advances in the pathogenesis from animal models[J]. J Autoimmun,2009,33(3-4):190-196.

[37]
Kurien BT,Dsouza A,Igoe A,et al.Immunization with 60 kD Ro peptide produces different stages of preclinical autoimmunity in a Sjögren’s syndrome model among multiple strains of inbred mice[J]. Clin Exp Immunol,2013,173(1):67-75.

[38]
Iizuka M,Wakamatsu E,Tsuboi H,et al.Pathogenic role of immune response to M3 muscarinic acetylcholine receptor in Sjögren’s syndrome-like sialoadenitis[J]. J Autoimmun,2010,35(4):383-389.Abstract The aim of this study was to clarify the role of the immune response to muscarinic type 3 receptor (M3R) in the pathogenesis of Sj02gren's syndrome (SS). M3R(-/-) mice were immunized with murine M3R peptides and their splenocytes were inoculated into Rag1(-/-) (M3R(-/-)→Rag1(-/-)) mice. M3R(-/-)→Rag1(-/-) mice had high serum levels of anti-M3R antibodies and low saliva volume. Histological examination showed marked infiltration of mononuclear cells in the salivary glands and immunohistochemistry demonstrated that the majority of these cells were CD4(+) T cells with a few B cells and several IFN-γ- and IL-17-producing cells. Apoptotic cells were present in the salivary glands of M3R(-/-)→Rag1(-/-) mice. Moreover, transfer of only CD3(+) T cells from M3R(-/-) immunized with M3R peptides into Rag1(-/-) mice resulted in cell infiltration and destruction of epithelial cells in the salivary glands, suggesting that M3R reactive CD3(+) T cells play a pathogenic role in the development of autoimmune sialoadenitis. Our findings support the notion that the immune response to M3R plays a crucial role in the pathogenesis of SS-like autoimmune sialoadenitis. Copyright 08 2010 Elsevier Ltd. All rights reserved.

DOI PMID

[39]
Inagaki Y,Jinno-Yoshida Y,Hamasaki Y,et al.A novel autoantibody reactive with carbonic anhydrase in sera from patients with systemic lupus erythematosus and Sjögren’s syndrome[J]. J Dermatol Sci,1991,2(3):147-154.Carbonic anhydrase (CA) is an extremely basic zinc metalloenzyme with a wide phyletic distribution, and the enzyme is important for the regulation of acid-base status. A novel autoantibody reactive with carbonic anhydrase was demonstrated. Several different classes of CA are known in mammals. Using the immuno blotting method and immun-dot analysis, we found this autoantibody to be reactive with CA in the sera from patients with Sjgren's syndrome (20.8%), including a patient with Sjgren's syndrome and renal tubular acidosis, and in patients with systemic lupus erythematosus (31.6%). The autoantibody varied in the extent of its cross-reactivity among human CA I (or B), human CA II (or C), bovine CA I, bovine CA II, rabbit CA, and dog CA. The titers continued to float and tended to parallel disease activity. Positive reactivity of autoantibody was observed on eccrine sweat glands and the distal tubules of the kidney by the indirect immunofluorescent method.

DOI PMID

[40]
Yang L,Ju J,Zhang W,et al.Effects of muscarinic acetylcholine 3 receptor (208-227) peptide immunization on autoimmune response in nonobese diabetic mice[J]. Clin Dev Immunol,2013,2013:485213 doi:10.1155/2013/485213.The second extracellular loop (LFWQYFVGKRTVPPGECFIQFLSEPTITFGTAI, aa 205–237) of muscarinic acetylcholine 3 receptor (M3R) has been reported to be an epitope for autoantibodies generated during certain autoimmune disorders, including Sj02gren’s syndrome (SS). Autoantibodies against M3R228–237 have been shown to interfere with the function of M3R. However, few studies have been performed on the M3R205–227 peptide of the second extracellular loop. In the current study, we sought to investigate the effect of M3R208–227 peptide immunization on autoimmune response in NOD/LtJ mice. We synthesized the M3R208–227 peptide and immunized NOD/LtJ mice to investigate whether peptide-specific antibodies could be generated and whether immunization would lead to changes in autoimmune response in NOD/LtJ mice. Our results demonstrate that the secretions of Th-1, Th-2, and Th-17 cytokines are downregulated and lymphocytic infiltration is improved in the salivary glands and lacrimal glands following immunization with M3R208–227 peptide in NOD/LtJ mice, suggesting that peptide immunotherapy using the M3R208–227 peptide may represent a potential therapeutic alternative.

DOI PMID

[41]
邹新乐,李想,徐凯彪,等.毒蕈碱受体3抗原表位多肽诱导干燥综合征动物模型的研究[J]. 现代免疫学,2009,29(1):25-28.利用毒蕈碱受体3(M3R)抗原表位多肽N49免疫BALB/c小鼠,以生理盐水注射小鼠作为对照组,检测各组小鼠血清抗N49多肽抗体和颌下腺淋巴细胞浸润情况。结果显示,多肽免疫组小鼠血清抗N49多肽抗体阳性率达100%(13/13),抗体滴度普遍从免疫后第21天开始显著上升,其中有5只(5/13)小鼠出现颌下腺淋巴细胞浸润;对照组小鼠血清抗N49多肽抗体均为阴性,仅有1只(1/14)出现轻度颌下腺淋巴细胞浸润。进一步以M3R分子不同抗原表位多肽N251和N412为包被抗原,检测以上实验小鼠血清中的抗体情况,显示N49多肽免疫组小鼠血清中抗N251和N412多肽抗体阳性率分别为100%(13/13)和85%(11/13),且两者抗体滴度变化趋势与抗N49多肽抗体一致,而对照组小鼠血清中抗N251和N412多肽抗体均为阴性。表明N49多肽较成功地建立了BALB/c小鼠的干燥综合征(SS)模型,诱导出了典型的自身抗体、颌下腺淋巴细胞浸润和自身抗原M3R分子内表位扩展等特征,为进一步研究M3R等自身抗原在SS发生中的作用机制奠定了基础。

[42]
Dite P,Novotny I,Trna J,et al.Autoimmune pancreatitis[J]. Best Pract Res Clin Gastroenterol,2008,22(1):131-143.

[43]
Caccavo D,Afeltra A,Rigon A,et al.Antibodies to carbonic anhydrase in patients with connective tissue diseases:relationship with lung involvement[J]. Int J Immunopathol Pharmacol,2008,21(3):659-667.Abstract The aim of this study is to evaluate the presence of antibodies to carbonic anhydrase I and/or II (ACAI and ACAII) in patients affected by connective tissue diseases (CTD) and to investigate their association with lung involvement evaluated by High resolution CT scan (HRCT). Ninety-six patients affected by CTD were studied, i.e. 33 rheumatoid arthritis (RA), 8 psoriatic arthritis (PA), 8 ankylosing spondilitis (AS), 23 Systemic Lupus Erythematosus (SLE), 10 Sjogren Syndrome (SS), and 14 Systemic Sclerosis (SSc). ACA were detected by ELISA. The lung involvement was evaluated by means of a previously described HRCT score. According to a receiver operator characteristic curve, patients were divided into those with HRCT score > or = 10 and those with HRCT score or = 10 was predictive of interstitial lung disease. ACAI and/or ACAII were detected in 30/96 patients (31.2%) (P or = 10 and both their CRP and ACAI levels were significantly higher when compared with patients showing a HRCT score less than 10 (P or = 10 than in those with HRCT score less than 10 (P = 0.014 and P = 0.007, respectively). Due to the lower levels of complement fractions detected in patients with HRCT score > or = 10, a possible immune-complex-mediated pathogenic mechanism of lung involvement could be suggested.

DOI PMID

[44]
Nishimori I,Bratanova T,Toshkov I,et al.Induction of experimental autoimmune sialoadenitis by immunization of PL/J mice with carbonic anhydrase Ⅱ[J]. J Immunol,1995,154(9):4865-4873.Experimental autoimmune sialoadenitis was induced in PL/J (H-2u) mice by intradermal immunization with human carbonic anhydrase II (CAII) and adjuvant containing monophosphoryl lipid A and trehalose diorynomycolate. Mice immunized with CAII showed a significant increase in the number and size of foci with lymphocytic infiltration in the salivary gland compared with mice immunized with adjuvant alone and untreated mice. In mice immunized with CAII, lymphocytic foci were observed around intercalated and intralobular ducts in the salivary glands, resulting in atrophy and replacement of acinar units. The epithelial cells of salivary ducts adjacent to the lymphocytic foci showed both degenerative and regenerative changes. Similar lymphocytic infiltrations were observed in the pancreas and kidney of a few mice immunized with CAII. Among several mouse strains with different H-2 haplotypes (p, q, r, s, and u), strains bearing H-2s and H-2u were susceptible to CAII-induced sialoadenitis. These results indicate that sialoadenitis induced by the immunization of CAII in mice may serve as a disease model of Sj枚gren's syndrome and that CAII or its derived peptides in association with the MHC may be one Ag recognized by an autoimmune response in this syndrome.

DOI PMID

[45]
Nishimori I,Miyaji E,Morimoto K,et al.Diminished cellular immune response to carbonic anhydrase Ⅱ in patients with Sjögren’s syndrome and idiopathic chronic pancreatitis[J]. Jop,2004,5(4):186-192.Abstract CONTEXT: A serum antibody to carbonic anhydrase II has been reported in patients with Sj02gren's syndrome and idiopathic chronic pancreatitis. OBJECTIVE: To evaluate cellular immune response to carbonic anhydrase II in patients with Sj02gren's syndrome and idiopathic chronic pancreatitis. PATIENTS: Idiopathic chronic pancreatitis (n=23), Sj02gren's syndrome (n=12), alcoholic chronic pancreatitis (n=3) and normal controls (n=13). MAIN OUTCOME MEASURES: Proliferation assay of peripheral blood mononuclear cells. RESULTS: Notable increased proliferation of the mononuclear cells upon stimulation with carbonic anhydrase II was observed in 2 patients with idiopathic chronic pancreatitis (9%) and 2 patients with Sj02gren's syndrome (17%) but not in patients with alcoholic chronic pancreatitis nor in normal controls. Among the four study groups, there was no significant difference in the prevalence rate of the positive proliferative responses (P=0.444). CONCLUSION: Carbonic anhydrase II may not be a major target antigen for the immunological process in the pathogenesis of Sj02gren's syndrome and idiopathic chronic pancreatitis. Serum antibody to carbonic anhydrase II may be detected in these patients as a consequence of the immune reaction against other antigens which mimic carbonic anhydrase II.

PMID

Options
Outlines

/

〈 〉