Animal Care and Diet

動物飼養(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。建議在無特定病原體（SPF級別）條件下飼養(yǎng)動物，因為細菌感染會降低宿主的免疫應(yīng)答，從而減輕關(guān)節(jié)炎癥。腸道菌群會影響宿主免疫系統(tǒng)對LPS 的易感性。LPS甚至?xí)鹉承PS超級易感品系的小鼠致死。Chondrex公司建議在開展實驗前先測試實驗用小鼠對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)和補體激活的小鼠品系都應(yīng)該對CAIA敏感。

圖一為目前為止測試過的小鼠品系。

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

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

T細胞缺陷的C.B-17 scid/scid 小鼠可用于CAIA，因為在炎癥發(fā)生的過程中，不需要T細胞識別抗原來產(chǎn)生抗體（17）。這中品系小鼠相比正常T細胞的小鼠，會產(chǎn)生更嚴(yán)重的關(guān)節(jié)炎癥， T細胞在愈合過程中有調(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 CAIA 具 有 抗 性（12,19,27）.

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

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LPS不敏感，但CAIA聯(lián)合LPS 會使這些小鼠產(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。一些雜交小鼠會對LPS有應(yīng)答，所以1.5mg每只小鼠的劑量一會產(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：

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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