口服耐受预防大鼠复发性类风湿性关系炎的研究

目的：建立实验性类风湿性关节炎（RA）的动物模型,研究口服可溶性鸡Ⅱ型胶原（CCⅡ）诱导免疫耐受对大鼠复发性RA的预防作用,方法：以CCⅡ和完全弗氏佐剂免疫Wistar大鼠,建立RA大鼠模型。在大鼠致炎前和反复造病后,口服CCII,观察其对RA复发的预防作用,并以ELISA法对大鼠血清中的抗CCII抗体进行检测。结果：在Wistar大鼠成功地诱发了RA,发病率为90％,大鼠在致炎前口服CCII可明显延迟RA的发病时间并降低RA的发病率,且大鼠RA的临床症状也明显减轻,而耐受大鼠由CCII引起的迟发性超敏反应（DTH）明显受到抑制,ELISA检测结果显示：口服可溶性CCII对大鼠体内抗CCII抗体的产生有一定的抑制作用。结论：口服CCII可诱导特异性免疫耐受,从而对大鼠RA的发生有着明显的预防作用。     

口服鸡II型胶原治疗大鼠实验性类风湿性关节炎的研究

本文研究口服鸡II型胶原 (CCII)治疗大鼠类风湿性关节炎的作用。以CCII和完全弗氏佐剂免疫Wistar大鼠 ,建立大鼠的类风湿性关节炎 (RA )模型。用CCII进行口服 ,观察实验组和对照组动物关节炎的发病程度。并以ELISA法对发病大鼠血清中的抗CCII抗体的水平进行检测。结果表明 ,口服CCII可以减轻大鼠关节炎的发病程度。其中口服 6μg、 60 μgCCII的实验组大鼠关节炎指数均较对照组明显减低 ,且差异有显著性意义 (P <0 0 1;P <0 0 5 )。而口服 60 0 μgCCII的实验组大鼠关节炎指数虽较对照组大鼠降低 ,但二者相比 ,无显著性差异 (P >0 0 5 )。ELISA检测结果显示 :口服低剂量 (6μg和 60μg )可溶性CCII对大鼠体内抗CCII抗体的产生有一定的抑制作用。口服CCII对关节炎大鼠的发病程度有较明显的抑制作用 ,为应用口服耐受治疗RA等自身免疫性疾病提供了实验基础     

鸡胶原II型诱导大鼠类风关模型的建立

用鸡胶原II型 (chickencollagenII,CCII)和完全弗氏佐剂免疫Wistar大鼠诱导胶原性关节炎 ,建立自身免疫性类风湿性关节炎 (RA)实验动物模型。以ELISA法检测实验组和对照组动物血清中的CCII抗体的水平 ,同时采用组织病理学和X射线摄片的方法显示关节炎大鼠的发病程度和病理学特征 ,并对其血液和滑膜液进行微生物学鉴定。结果显示 ,用CCII和完全弗氏佐剂免疫的实验组大鼠关节炎的发病率为 80 %,同时在免疫后 2 5d ,关节炎指数为最高 (AI=7 8)。ELISA法检测抗CCII抗体结果显示 ,实验组大鼠血清中抗CCII抗体的水平均较各对照组大鼠明显增高 (P <0 0 0 1)。组织病理学和X 射线摄片结果显示 ,关节软骨组织、骨组织和滑膜组织呈典型的关节炎病变。微生物鉴定结果均为阴性。本实验成功建立了胶原诱导的大鼠关节炎模型 ,且其病理特征和人类RA极为相似 ,为进一步深入研究人类RA的发病机制及其临床治疗提供了有价值的实验材料。     

胶原诱导性关节炎大鼠淋巴细胞的过继转移

目的：探讨类风湿性关节炎（RA）发病的免疫学机制以及II型胶原在疾病发生过程中的作用。方法：用鸡II型胶原（CCII）加完全佛氏佐剂（CFA）免疫Wistar大鼠建立胶原诱导性关节炎（CIA）模型，分别用发病大鼠的T淋巴细胞和血清进行过继转移，以ELISA检测外周血中抗CCII抗 体的水平；用病理组织学方法对炎性关节进行分析。结果：成功建立了CIA大鼠模型，发病率为90％，用T淋巴细胞转移的大鼠，可见明显CIA症状，发病率为40％，实验组大鼠外周血中的抗CCII抗体的水平虽较对照组高，但并无显著性意义（P＞0．05），以血清过继转移的大鼠无一发病，但该组外周血中抗CCII抗体的水平明显高于对照组（P＜0．05），组织学鉴定结果可见炎性部位呈典型的CA病变，结论：CIA可以通过淋巴细胞过继转移，机体对I原产生的特异性细胞免疫应答在CIA发生过程中起着重要的作用。为采用T细胞介导的免疫治疗RA提供了实验基础。     

口服鸡Ⅱ型胶原治疗大鼠实验性类风湿性关节炎的研究

本文研究口服鸡Ⅱ型胶原（CCⅡ）治疗大鼠类风湿性关节炎的作用。以CCⅡ和完全弗氏佐剂免疫Wistar大鼠,建立大鼠的类风湿性关节炎（RA）模型,用CCⅡ进行口服,观察实验组和对照组动物关节炎的发病程度。并以ELISA法对发病大鼠血清中的抗CCⅡ抗体的水平进行检测。结果表明,口服CCⅡ可以减轻大鼠关节炎的发病程度。其中口服6μg、60μgCCⅡ的实验组大鼠关节炎指数均对照组明显减低,且差异有显著性意义（P＜0．01;P＜0．05）。而口服600μgCCⅡ的实验组大鼠关节炎指数虽较对照组大鼠降低,胆二者相比,无显著性差异（P＞0．05）。ELISA检测结果显示,口服低剂量（6μg和60μg）可溶性CCⅡ对大鼠体抗CCⅡ抗体的产生有一定的抑制作用。口服CCⅡ对关节炎大鼠的发病程度有较明显的抑制作用。为应用口服耐受治疗RA等自身免疫性疾病提供了实验基础。     

口服MBP诱导免疫耐受预防实验性变态反应性脑脊髓炎的研究

目的：探讨口服耐受对实验性变态反应性脑脊髓炎(EAE)的预防作用。方法：给予健康Lewis大鼠口服MBP抗原诱导免疫耐受，随后以MBP CFA于足底免疫，检测口服耐受对实验大鼠EAE发病的预防作用及对MBP特异性淋转率、细胞因子和抗MBP抗体等免疫功能的变化情况。结果：致病组EAE发病率为91．7％，MBP特异性淋转率、IFN-γ与TNF-α含量升高，IL-10降低；口服耐受组EAE发病率为21．4％，MBP特异性淋转率、IFN-γ、TNF-α回复至正常，IL-10含量低于正常。MBP抗体水平两组均比正常对照组高。结论：口服耐受可降低宿主自身免疫反应性，减低EAE的发病率。     

鸡胶原Ⅱ型诱导大鼠类风关模型的建立

用鸡胶原Ⅱ型（chicken collagen Ⅱ,CCⅡ）和完全弗氏佐剂免疫Wistar大鼠诱导胶原性关节炎,建立自身免疫性类风湿性关节炎（RA）实验动物模型。以ELISA法检测实验组和对照组动物血清中的CCⅡ抗体的水平,同时采用组织病理学和X射线摄片的方法显示关节炎大鼠的发病程度和病理学特征,并对其血液和滑膜液进行微生物学鉴定。结果显示,用CCⅡ和完全弗氏佐剂免疫的实验组大鼠关节炎的发病率为80％,同时在免疫后25d,关节炎指数为最高（AI＝7．8）。ELISA法检测抗CCⅡ抗体结果显示,实验组大鼠血清中抗CCⅡ抗体的水平均较各对照组大鼠明显增高（P＜0．001）。组织病理学和X－射线摄片结果显示,关节软骨组织、骨组织和滑膜组织呈典型的关节炎病变。微生物鉴定结果均为阴性。本实验成功建立了胶原诱导的大鼠关节炎模型,且其病理特征和人类RA极为相似,为进一步深入研究人类RA的发病机制及其临床治疗提供了有价值的实验材料。     

诱导免疫耐受治疗佐剂性关节炎的实验研究

目的：探讨口服Ⅱ型胶原（CⅡ）对大鼠佐剂性关节炎（AA）的治疗作用,研究口服诱导免疫耐受过程中转化生长因子-β（TGF-β）表达的变化.方法：制备大鼠AA模型,观察口服CⅡ前后关节的肿胀度;采用ELISA法检测大鼠血清和关节浸出液中TGF-β的表达水平.结果：口服CⅡ可以明显减轻病变关节的炎症反应,降低发病率,推迟AA的发病并且使病程明显缩短.口服CⅡ诱导免疫耐受的AA大鼠血清和关节浸出液中TGF-β的表达明显高于未口服CⅡ的大鼠（P＜0.05）.结论：口服CⅡ对大鼠佐剂性关节炎有明显的治疗效果;TGF-β是口服耐受治疗AA的重要细胞因子.     

口服免疫耐受大鼠脾淋巴细胞对肾小球系膜细胞中NF-κB p65活性的影响

目的 :探讨口服免疫耐受大鼠脾淋巴细胞对肾小球系膜细胞中NF κBp6 5活性的影响。方法 :将 10只 6～ 8wk龄雌性Wistar大鼠随机分为两组 ,每组 5只。一组用Fx1A抗原灌胃诱导其产生免疫耐受 ,另一组用PBS灌胃作为对照组。 2wk后 ,杀鼠取脾、分离淋巴细胞进行抗原特异性淋巴细胞增殖实验。制备免疫耐受大鼠脾淋巴细胞培养上清 ,用夹心ELISA法测定其中IL 10的水平。将体外培养的系膜细胞用高水平的胰岛素 ,同时加入大鼠的脾淋巴细胞培养上清作用 2 4h。用免疫组化染色及夹心ELISA法检测系膜细胞中NF κBp6 5的活性。结果 :与对照组大鼠相比较 ,口服免疫耐受组大鼠抗原特异性脾淋巴细胞的增殖明显受到抑制 ,其脾淋巴细胞培养上清中IL 10的水平明显升高 (P <0 .0 1)。在体外实验中发现 ,口服免疫耐受大鼠脾淋巴细胞培养上清 ,可抑制胰岛素刺激的系膜细胞内NF κBp6 5的活性 (P <0 .0 5 )。结论 :口服免疫耐受大鼠的脾淋巴细胞 ,可能通过其分泌的IL 10抑制系膜细胞内NF κBp6 5的活性     

体外DCs在抗CD45RB抗体诱导免疫耐受中的作用机制

目的:探讨树突状细胞(dendritic cells,DCs)在抗CD45RB抗体诱导的免疫耐受中所发挥的作用,从而阐明抗CD45RB抗体诱导免疫耐受的机制。方法:采用DCs的常规诱导方法(rmGM-CSF、IL-4和LPS),在诱导过程中加入不同剂量的抗CD45RB抗体,成熟后利用流式细胞仪检测细胞表型、周期和吞噬能力,ELISA法检测IL-12分泌量,混合淋巴细胞培养检测DCs对T细胞增殖能力的影响。结果:DCs经抗CD45RB抗体处理后,CD11C和CD83表达升高,CD86表达下降,自身增殖和吞噬能力增强,但分泌IL-12和刺激T细胞增殖的能力明显下降。结论:耐受性树突状细胞(tolerogenic dendritic cells,tDCs)能显著抑制T细胞的增殖,它的产生是抗CD45RB抗体诱导免疫耐受的主要机制之一。     

口服髓鞘碱性蛋白诱导免疫耐受治疗实验性自身免疫性脑脊髓炎的研究

目的建立实验性自身免疫性脑脊髓炎（ＥＡＥ）动物模型,探讨口服自身抗原诱导免疫耐受对大鼠ＥＡＥ的防治作用。方法在普通Ｗｉｓｔａｒ大鼠经１次足跖真皮内注射完全弗氏佐剂－豚鼠全脊髓匀浆或完全弗氏佐剂－髓鞘碱性蛋白乳剂加百日咳疫苗诱发ＥＡＥ疾病;另在大鼠致炎前及发病后口服髓鞘碱性蛋白（ＭＢＰ）,观察其对ＥＡＥ的防治作用。结果在Ｗｉｓｔａｒ大鼠成功地诱发了ＥＡＥ,发病率将近９０％。大鼠在致炎前口服ＭＢＰ,可明显推迟ＥＡＥ发病时间,降低ＥＡＥ发病率,使神经组织病理改变明显改善。大鼠发生ＥＡＥ后给予ＭＢＰ,可明显控制发病动物的病情,使病程缩短及减轻患鼠神经组织的炎症程度。另外,在耐受鼠由ＭＢＰ引起的迟发型超敏反应（ＤＴＨ）以及体外针对ＭＢＰ的淋巴细胞增殖反应也明显受到抑制。结论口服ＭＢＰ可引起特异性的免疫耐受,从而产生对实验性自身免疫性脑脊髓炎的防治作用     

Experimental animal models for rheumatoid arthritis

Rheumatoid Arthritis (RA) is an autoimmune systemic disorder of unknown etiology and is characterized by chronic inflammation and synovial infiltration of immune cells. RA is associated with decreased life expectancy and quality of life. The research on RA is greatly simplified by animal models that help us to investigate the complex system involving inflammation, immunological tolerance and autoimmunity. The animal models of RA with a proven track record of predictability for efficacy in humans include: collagen type II induced arthritis in rats as well as mice, adjuvant induced arthritis in rats and antigen induced arthritis in several species. The development of novel treatments for RA requires the interplay between clinical observations and studies in animal models. However, each model features a different mechanism driving the disease expression; the benefits of each should be evaluated carefully in making the appropriate choice for the scientific problem to be investigated. In this review article, we focus on animal models of arthritis induced in various species along with the genetic models. The review also discussed the similarity and dissimilarities with respect to human RA.     

血管活性肠肽抑制实验性类风湿性关节炎的研究

类风湿性关节炎是一种自身抗原未明的慢性自身免疫病,以多关节的慢性炎症及渐进性骨和软骨的破坏为主要特征。胶原诱导的关节炎因其临床表现、病理组织学改变以及免疫学表现等方面与类风湿性关节炎的相似性而成为理想的动物模型。在成功建立了胶原诱导的大鼠关节炎模型(CIA)的基础上采用了血管活性肠肽(VIP)注射,研究其干预关节炎发生的效果。结果显示:使用血管活性肠肽组的大鼠CIA发病率和严重程度明显降低,关节肿胀以及骨和软骨的破坏减轻,大鼠血清中抗胶原抗体水平显著降低,大鼠T淋巴细胞对胶原的增殖反应也显著降低。提示VIP可能通过减轻对胶原的免疫应答而抑制关节炎症状,具有可能的临床应用价值。     

Orally administered myelin basic protein in neonates primes for immune responses and enhances experimental autoimmune encephalomyelitis in adult animals

Antigen-driven tolerance is an effective method for suppression of autoimmune diseases. Adult animals can be tolerized against the induction of experimental autoimmune encephalomyelitis (EAE) by both oral and parenteral administration of myelin basic protein (MBP). We have found that in contrast to previous studies of neonatal tolerance in which parenterally administered autoantigens induced tolerance, the oral administration of MBP in neonatal rats did not result in tolerization to MBP, but instead, primed for immunologic responses. Proliferative responses to MBP and its encephalitogenic epitope were present in animals fed with MBP as neonates and co-culture of encephalitogenic T cells with cells from neonatal rats fed with MBP were associated with enhanced MBP responses rather than the suppression observed with cells from adult rats fed with MBP. Furthermore, neonates fed with MBP and immunized 6–8 weeks later with MBP in adjuvant to induce EAE revealed enhancement of disease severity, and were not protected from a second attack upon active reinduction of EAE. Subcutaneous injection of soluble MBP into neonates had no effect on EAE induction as adults, whereas intraperitoneal injection of MBP in neonates was associated with marked suppression of disease in adults. Suppression of EAE began to appear in animals fed with MBP at 4 weeks of age, and was similar to oral tolerance in adult animals when animals were fed at 6 weeks of age. These results suggest that immaturity of the immunoregulatory network associated with oral tolerance and sensitization to autoantigens via the gut in the neonatal period may contribute to the pathogenesis of autoimmune diseases.     

Estrogen altered oral tolerance induction in type II collagen-induced murine arthritis

BACKGROUND: Estrogen plays an important modulatory role in the immune system, and is concerned with the pathophysiology of autoimmune diseases such as rheumatoid arthritis (RA), although the mechanism has not yet been clarified. Oral tolerance, a form of specific peripheral tolerance, which is recognized as a new therapeutic strategy, is related to the function of gut-associated lymphoid tissue. METHODS: In this study, using collagen-induced arthritis as an animal model of RA, the effects of 17beta-estradiol (E2) on oral tolerance induction were investigated. For induction of oral tolerance, mice were fed 60 microg type II collagen (CII) for 10 consecutive days prior to each CII immunization. Mice in the E2 treatment groups were injected with 5 microg (low dose) or 500 microg (high dose) E2 three times during the induction of oral tolerance. RESULTS: Oral tolerance induction suppressed the occurrence of arthritis, the proliferative response of splenocytes to CII and the specific DTH response. However, E2 treatment abrogated the suppression, which might be connected with a change in function of Peyer's patch (PP) lymphocytes. CONCLUSION: These results suggest that oral tolerance induction might be affected by estrogen treatment through alteration of intestinal immune responses.     

Lactobacillus casei potentiates induction of oral tolerance in experimental arthritis

Probiotics have been shown to exert beneficial effects on modulation of diverse diseases. However, no information is available for the effect of probiotics in the induction of oral tolerance in autoimmune diseases. The main purpose of this study was to elucidate whether Lactobacillus casei (L. casei) affect the induction of oral tolerance in experimental rheumatoid arthritis (RA). Type II collagen (CII) alone or together with L. casei was orally administered into collagen-induced arthritis (CIA) rats, and its effects on the clinical and histopathological aspects of RA were investigated. Co-administration of L. casei with CII more effectively suppressed clinical symptoms, paw swelling, lymphocyte infiltration and destruction of cartilage tissues of experimental arthritis than the rats treated with CII alone. The enhanced therapeutic efficacy was associated with an increase in anti-inflammatory cytokines (IL-10 and TGF-beta) while decreasing pro-inflammatory cytokines (IL-1beta, IL-2, IL-6, IL-12, IL-17, IFN-gamma and TNF-alpha). Co-administration of L. casei with CII more effectively suppressed CII-reactive T cell proliferation and the levels of Th1-type IgG isotypes (IgG2a and IgG2b), while up-regulating Foxp3 expression levels and the population of Foxp3(+) CD4(+) T cells. Our study provides evidence that L. casei could potentiate antigen-specific oral tolerance and suppress Th1-type immune responses of arthritic inflammation.     

Role of the Bowel Flora for Development of Immunity to hsp 65 and Arthritis in Three Experimental Models

An infectious aetiology in rheumatoid arthritis (RA) has for long been suggested, although no conclusive evidence for this is at present available. Lately a large interest has been devoted to the involvement of heat shock proteins (hsps) in autoimmune disorders due to their conserved structure and immunogenic properties. Immunity to hsps has been observed both in human autoimmune conditions and in experimental models of autoimmune disease. We have studied the role of the bacterial flora and hsp immunity in the arthritic response in three experimental models of arthritis; type II collagen arthritis (CIA), adjuvant arthritis (AA) and oil induced arthritis (OIA); by using germ free and conventional DA rats.
A high incidence of severe arthritis developed in all the models evaluated irrespectively of whether the animals were in the conventional or germ free state. This confirms earlier results which show a minor effect of the bacterial flora in CIA and AA in high responder strains. These results also show that a severe OIA can develop in germ free animals. Despite the severe arthritic response induced, no serum antibody levels to hsp 65 could be detected in the germ free animals, which was in contrast to the conventional animals where a positive anti-hsp 65 serum response was seen in 35–80% of the animals with CIA, AA or OIA.
These results show that development of a humoral response to hsp 65 in these models of arthritis is dependent on the presence of a bacterial flora. Further, the lack of humoral immunity in germ free animals despite a severe arthritic response indicates that humoral immunity to hsp 65 is not involved in development of disease in these three models of experimental arthritis.     

The in vivo effects of tumour necrosis factor blockade on the early cell mediated immune events and syndrome expression in rat adjuvant arthritis

Anti-TNF therapy is effective in rheumatoid arthritis (RA); however, its mechanisms of action are incompletely understood. T cell-driven mechanisms are thought to play an important role in RA and the effects of TNF blockade on these mechanisms are unclear. Adjuvant arthritis (AA) is a T cell dependent model of inflammatory arthritis. The aims of this study were to investigate the effects of TNF blockade on in vivo T cell cytokine expression and to clarify the role of TNF in the inguinal lymph nodes (ILN) in early arthritis. AA was induced in male DA rats. Rats received either 3 mg/kg or 10 mg/kg PEG sTNF-RI at days 0, 2 and 4 postinduction or 10 mg/kg anti-TNF antibody on day of arthritis induction. Control rats received either saline or normal sheep serum. Paw volume was assessed every 3-4 days. Rats were sacrificed on days 0, 6, 13 and 21 postinduction. Ankles were removed for quantitative radiology and histology. Synovium and ILN were removed for cell culture and to determine mRNA expression of cytokines using semiquantitative RT-PCR. TNF and IFN-gamma protein production was measured using a bioassay and an ELISA. TNF blockade did not suppress mRNA expression of T cell cytokines in the ILN of rats in the early phase of AA, suggesting ongoing T cell activity. TNF protein production by ILN cells in culture was reduced in PEG sTNF-RI treated rats, although mRNA expression was increased in the ILN prior to culture. Early administration of PEG sTNF-RI did not attenuate AA, in contrast to an anti-TNF antibody, which suppressed disease. A shorter half-life for the PEG sTNF-RI compared with the anti-TNF antibody or the development of anti-PEG sTNF-RI antibodies may account for these results.