膠原抗體誘導(dǎo)小鼠關(guān)節(jié)炎模型之動(dòng)物飼養(yǎng)條件和飲食

Animal Care and Diet

動(dòng)物飼養(yǎng)條件和飲食

建立CAIA模型需要選擇7-8周齡小鼠。老齡鼠的發(fā)病率和病情嚴(yán)重程度較低。建議在無(wú)特定病原體（SPF級(jí)別）條件下飼養(yǎng)動(dòng)物，因?yàn)榧?xì)菌感染會(huì)降低宿主的免疫應(yīng)答，從而減輕關(guān)節(jié)炎癥。腸道菌群會(huì)影響宿主免疫系統(tǒng)對(duì)LPS 的易感性。LPS甚至?xí)鹉承?duì)LPS超級(jí)易感品系的小鼠致死。Chondrex公司建議在開(kāi)展實(shí)驗(yàn)前先測(cè)試實(shí)驗(yàn)用小鼠對(duì)LPS的易感性。盡管飲食影響可以不用考慮，但是Chondrex公司建議飼喂高脂肪食物，研究表明高脂肪食物更容易引發(fā)炎癥 （4）。

Mouse Strains

小鼠品系

注射抗二型膠原單克隆抗體合劑可以繞過(guò)宿主生成抗二型膠原自身抗體的需求，所以可以誘發(fā)MHC （例如H-2q和H-2r）缺陷的小鼠產(chǎn)生關(guān)節(jié)炎.所有能產(chǎn)生正常炎癥反應(yīng)和補(bǔ)體激活的小鼠品系都應(yīng)該對(duì)CAIA敏感。

圖一為目前為止測(cè)試過(guò)的小鼠品系。

DBA/1 (H-2q) and B10.RIII (H-2r) 小鼠對(duì)CIA和CAIA均有較高的敏感性（13）。

BALB/c (H-2d) 小鼠對(duì)CIA具有抗性，但對(duì)CAIA有較高敏感性，是目前最-常-用的品系（25-27）。

T細(xì)胞缺陷的C.B-17 scid/scid 小鼠可用于CAIA，因?yàn)樵谘装Y發(fā)生的過(guò)程中，不需要T細(xì)胞識(shí)別抗原來(lái)產(chǎn)生抗體（17）。這中品系小鼠相比正常T細(xì)胞的小鼠，會(huì)產(chǎn)生更嚴(yán)重的關(guān)節(jié)炎癥， T細(xì)胞在愈合過(guò)程中有調(diào)節(jié)炎癥的作用。

SWR（H-2q）小鼠具有CIA易感基因型，但由于C5缺陷，對(duì)CIA 和CAIA具有抗性。 其他C5缺陷小鼠，例如B10.D2/oSn 和 NOD/LtSz scid/scid 對(duì) CAIA 具 有 抗 性（12,19,27）.

盡管裸大鼠對(duì)CAIA具有高易感性， 裸小鼠對(duì)CAIA具有抗性（29）。裸小鼠可能缺乏特定的促炎細(xì)胞因子的表達(dá)。

圖一-用于膠原 CIA 及 膠原蛋白抗體誘導(dǎo)（CAIA ）的關(guān)節(jié)炎動(dòng)物模型研究的常用小鼠品系

*需要含結(jié)核桿菌的CFA佐劑誘導(dǎo)關(guān)節(jié)炎

C57BL/6小鼠是轉(zhuǎn)基因小鼠模型中最-常-用的品系。盡管C57BL/6小鼠對(duì)LPS不敏感，但CAIA聯(lián)合LPS 會(huì)使這些小鼠產(chǎn)生嚴(yán)重的關(guān)節(jié)炎癥。在這種品系的小鼠中建模需要使用高劑量單克隆抗體合劑，每只小鼠需要5mg(21,30).C57BL/6 與129/Sy小鼠雜交用于建立基因敲除小鼠。一些雜交小鼠會(huì)對(duì)LPS有應(yīng)答，所以1.5mg每只小鼠的劑量一會(huì)產(chǎn)生嚴(yán)重的關(guān)節(jié)炎癥（30-31） 。

以下基因敲除小鼠用于基因?qū)AIA模的影響 (29-39).

1) NOS2 knockout mice (30)

2) Osteopontin knockout mice (31)

3) COX-1 and COX-2 knockout mice (32)

4) MMP-2 (gelatinase A) and MMP-3 (gelatinase B) knockout

mice (33)

5) P2X7 receptor knockout mice (34)

6) c-Jun N-Terminal Kinase knockout mice (35)

7) Prostaglandin E2 receptor knockout mice (36)

8) CD69 null mice (37)

相關(guān)文獻(xiàn)：

4. P. Wooley, Collagen-induced arthritis in the mouse.Methods Enzymol 162, 361-373 (1988).

12. R. Reife, N. Loutis, W. Watson, K. Hasty, J. Stuart, SWR Mice Are Resistant to Collagen-Induced Arthritis but Produce Potentially Arthritogenic Antibodies. Arthritis Rheum. 34, 776-81(1991).

13. K. Terato, D. Harper, M. Griffiths, D. Hasty, X. Ye, et al.,Collagen-induced Arthritis in Mice: Synergistic Effect of E. Coli Lipopolysaccharide Bypasses Epitope Specificity in the Induction of Arthritis With Monoclonal Antibodies to Type II Collagen.Autoimmunity 22, 137-47 (1995).

17. T. Kagari, H. Doi and T. Shimozato. The importance of IL-1β and TNF-α, and the noninvolvement of IL-6, in the development of monoclonal antibody-induced arthritis. J Immunol 169,1459-1466 (2002).

19. W. Watson, P. Brown, J. Pitcock, A. Townes, Passive Transfer Studies With Type II Collagen Antibody in B10.D2/old and New Line and C57Bl/6 Normal and Beige (Chediak-Higashi) Strains: Evidence of Important Roles for C5 and Multiple Inflammatory Cell Types in the Development of Erosive Arthritis.Arthritis Rheum. 30, 460-5 (1987).

25. P. M. Wallace, J. F. MacMaster, K. A. Rouleau, T. J. Brown,J. K. Loy, et al., Regulation of Inflammatory Responses by Oncostatin M. J Immunol. 162, 5547-55 (1999).

26. R. de Fougerolles, A. G. Sprague, C. L. Nickerson-Nutter,G. Chi-Rosso, P. D. Rennert, et al., Regulation of Inflammation by Collagen-Binding Integrins alpha1beta1 and alpha2beta1 in

Models of Hypersensitivity and Arthritis. J Clin Invest. 105, 721-9(2000).

27. S Larox, J. Fuseler, D. Merril, L. Gray, R. Reife, K. Terato,et al. #301 A novel model of polyarthritis induced in mice using monoclonal antibodies to type II collagen. Characterization and

effects of chemically modified tetracycline. Arthritis Rheum 42:s121 (supplement).

29. K. Takagishi, N. Kaibara, T. Hotokebuchi, C. Arita, M.Morinaga, K. Arai, et al., Serum Transfer of Collagen Arthritis in Congenitally Athymic Nude Rats. J Immunol. 134, 3864-7 (1985).

30. H. Kato, K. Nishida, A. Yoshida, I. Takada, C. McCown, et al., Effect of NOS2 Gene Deficiency on the Development of Autoantibody Mediated Arthritis and Subsequent Articular Cartilage Degeneration. J Rheumatol. 30, 247-55 (2003).

31. K. Yumoto, M. Ishijima, S. Rittling, K. Tsuji, Y. Tsuchiya, et al., Osteopontin Deficiency Protects Joints Against Destruction in Anti-Type II Collagen Antibody-Induced Arthritis in Mice. Proc Natl Acad Sci U S A 99, 4556-61 (2002).

32. L. Myers, A. Kang, A. Postlethwaite, E. Rosloniec, S. Morham, et al., The Genetic Ablation of Cyclooxygenase 2 Prevents the Development of Autoimmune Arthritis. Arthritis Rheum 43, 2687-93 (2000).

33. T. Itoh, H. Matsuda, M. Tanioka, K. Kuwabara, S. Itohara, R. Suzuki, et al., The Role of Matrix metalloproteinase-2 and Matrix metalloproteinase-9 in Antibody-Induced Arthritis. J Immunol 169, 2643-7 (2002).

34. J. Labasi, N. Petrushova, C. Donovan, S. McCurdy, P. Lira,et al., Absence of the P2X7 Receptor Alters Leukocyte Function and Attenuates an Inflammatory Response. J Immunol 168,6436-45 (2002).

35. Z. Han, L. Chang, Y. Yamanishi, M. Karin, G. Firestein, Joint Damage and Inflammation in c-Jun N-terminal Kinase 2 Knockout Mice With Passive Murine Collagen-Induced Arthritis. Arthritis Rheum 46, 818-23 (2002).

36. J. McCoy, J. Wicks, L. Audoly, The Role of Prostaglandin E2 Receptors in the Pathogenesis of Rheumatoid Arthritis. J Clin Invest 110, 651-8 (2002).

37. R. Newton, K. Solomon, M. Covington, C. Decicco, P. Haley, et al., Biology of TACE Inhibition. Ann Rheum Dis 60 Suppl 3, 25-32 (2001).

38. R. Holmdahl, L. Jansson, M. Andersson, E. Larsson,Immunogenetics of Type II Collagen Autoimmunity and

Susceptibility to Collagen Arthritis. Immunology. 65, 305-10(1988).

39. S. Thornton, G. Boivin, K. Kim, F. Finkelman, R. Hirsch, Heterogeneous Effects of IL-2 on Collagen-Induced Arthritis. J Immunol 165, 1557-63 (2000).

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