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膠原誘導(dǎo)性關(guān)節(jié)炎小鼠(CIA)作為人類類風(fēng)濕關(guān)節(jié)炎模型應(yīng)用廣泛,但CIA引起的關(guān)節(jié)炎起病比較緩慢,造模周期較長,一般為6-8周(1-12)。Chondrex公司已開發(fā)出單一種單克隆抗體合劑誘導(dǎo)的小鼠關(guān)節(jié)炎模型(CAIA),明顯縮短了造模周期。與CIA相比,CAIA模型具有以下優(yōu)點(diǎn): (1)可在大多數(shù)小鼠上誘發(fā)關(guān)節(jié)炎,包括CIA不敏感的小鼠;(2)CAIA造模周期短,加速篩選和評(píng)估類風(fēng)濕性關(guān)節(jié)炎治療藥物;(3)應(yīng)用于轉(zhuǎn)基因小鼠來研究基因?qū)﹃P(guān)節(jié)炎發(fā)病的影響;(4)用于研究與人類類風(fēng)濕性關(guān)節(jié)炎相關(guān)的各種炎癥介質(zhì)和因素在疾病中的作用,例如細(xì)菌和病毒毒素(13-17)。
CAIA without LPS
膠原抗體誘導(dǎo)的關(guān)節(jié)炎(不加LPS)
CIA是由于自身抗體形成的免疫復(fù)合物通過激活補(bǔ)體而誘發(fā)關(guān)節(jié)炎癥,因此研究表明多克隆膠原蛋白抗體(8-19) 或單克隆膠原蛋白抗體合劑能誘發(fā)關(guān)節(jié)炎(20-21)。根據(jù)MHC類型,這些自身反應(yīng)性抗體可以識(shí)別位于小鼠II型膠原蛋白的CB11或者CB8片段中的特定抗原定簇(關(guān)節(jié)炎抗原表位)(22-23)。
Chondrex公司生產(chǎn)的能誘發(fā)小鼠關(guān)節(jié)炎的抗體合劑含有 : A2-10 (IgG2a), F10-21 (IgG2a), D8-6 (IgG2a), D1-2G(IgG2b), and D2-112 (IgG2b). 其中兩個(gè)單克隆抗體(F10-21和D8-6)識(shí)別在LysC-2(291-374)的83個(gè)氨基酸肽段,該肽段有LysC內(nèi)切酶作用產(chǎn)生。 另外三個(gè)單克隆抗體(A2-10,D1-2G和D2-112) 能夠識(shí)別二型膠原蛋白CB11片段(124-402)中LysC-1(124-290)的167各氨基酸肽段(13)。這些抗原表位氨基酸序列在各種物種中具有高度保守性,包括雞,小鼠,大鼠,牛,豬,猴子和人 (20-21)。
CAIA with LPS
膠原抗體誘導(dǎo)的關(guān)節(jié)炎(加LPS)
研究表明,聯(lián)合注射低于致炎劑量的單克隆抗體合劑和大腸桿菌多脂(LPS)可誘發(fā)嚴(yán)重的小鼠關(guān)節(jié)炎(13)。通過胃腸道吸收的細(xì)菌毒素(如LPS)與低致炎水平的二型膠原自身抗體對(duì)引發(fā)關(guān)節(jié)炎有發(fā)協(xié)同作用(24)。這種模型相比于傳統(tǒng)CIA 模型有多重優(yōu)勢(shì)。第一,起病時(shí)間短,發(fā)病率高達(dá)100%。第二,可在大多數(shù)小鼠上誘導(dǎo)關(guān)節(jié)炎,包括CIA不敏感的小鼠,T細(xì)胞缺陷的小鼠,特定基因敲除變異的小鼠,詳情請(qǐng)見表二.
圖一顯示CIA模型和CAIA模型特點(diǎn)的比較。CAIA造模周期是CIA建模周期的十分之一。
圖一 CAIA vs. CIA: (a)三角形:100%的小鼠在LPS注射后24-48小時(shí)即可見關(guān)節(jié)炎發(fā)作,在第5-7天病情達(dá)到高峰。(b)圓形:CIA誘發(fā)小鼠關(guān)節(jié)炎模型一般需要4周起病,即使使用CIA易感品系,如DBA/1J和B10.RIII小鼠。
表二 關(guān)節(jié)炎炎癥程度的臨床評(píng)分
得分 | 發(fā)病情況 |
0 | Normal 正常 |
1 | Mild, but definite redness and swelling of the ankle or wrist, or apparent redness and swelling limited to individual digits, regardless of the number of affected digits 輕度的、踝關(guān)節(jié)、腕關(guān)節(jié)發(fā)紅、腫脹 |
2 | Moderate redness and swelling of ankle or wrist 踝關(guān)節(jié)或腕關(guān)節(jié)中度發(fā)紅腫脹 |
3 | Severe redness and swelling of the entire paw including digits 爪子嚴(yán)重發(fā)紅、腫脹,包括指端 |
4 | Maximally inflamed limb involving multiple joints 四肢最-大-程-度發(fā)炎,包括多關(guān)節(jié) |
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