通过携带白细胞介素-1受体拮抗剂的重组腺相关病毒载体进行关节炎的治疗与预防

Therapy and prevention of arthritis by recombinant adeno-associated virus vector with delivery of interleukin-1 receptor antagonist.

作者信息

Pan R Y, Chen S L, Xiao X, Liu D W, Peng H J, Tsao Y P

机构信息

Tri-Service General Hospital and National Defense Medical Center, Taipei, Taiwan, China.

出版信息

Arthritis Rheum. 2000 Feb;43(2):289-97. doi: 10.1002/1529-0131(200002)43:2<289::AID-ANR8>3.0.CO;2-H.

DOI:10.1002/1529-0131(200002)43:2<289::AID-ANR8>3.0.CO;2-H

PMID:10693868

Abstract

OBJECTIVE

To evaluate the recombinant adeno-associated virus vector encoding interleukin-1 receptor antagonist (rAAV-IL-1Ra) complementary DNA for its potential in the treatment and prevention of lipopolysaccharide (LPS)-induced arthritis.

METHODS

The therapeutic effect of rAAV-IL-1Ra on arthritis was studied by injecting knees of Sprague-Dawley rats with LPS and rAAV-IL-1Ra and then evaluating the severity of arthritis by leukocyte counts in synovial fluid, histologic changes of synovium, and uptake of 67Ga citrate in joint tissue. To study the therapeutic effect on recurrent arthritis, we induced recurrent arthritis by a second injection of LPS 80 days after primary LPS and rAAV-IL-1Ra injections and then evaluated the severity of recurrent arthritis. To study the prevention of arthritis, rAAV-IL-1Ra was injected into normal joints. After 100 days, LPS was used to induce arthritis, and the severity of arthritis was evaluated.

RESULTS

The production of the rAAV-IL-1Ra transgene was up-regulated by LPS-induced joint inflammation and proved to be efficacious in the therapeutic and preventative protocols. Not only primary but also recurrent arthritis could be suppressed by a single injection of rAAV-IL-1Ra. We found that the transgene expression of IL-1Ra could be reactivated by a second challenge with LPS delayed for 80 days after rAAV administration. The therapeutic level of IL-1Ra protein reached a mean +/- SD of 5.8+/-0.5 ng/ml in synovial fluid. In addition, the rAAV transgene persisted within normal joints for at least 100 days and could still be induced to express, after LPS insult, a high level of IL-1Ra (mean +/- SD 5.2+/-0.8 ng/ml) that prevented the occurrence of arthritis.

CONCLUSION

This gene therapy, by combining highly efficient and stable rAAV gene delivery, disease-regulated gene expression, and the antiinflammatory effect of IL-1Ra, provides a valuable approach for long-term protection against, and prevention of, arthritis.

摘要

目的

评估编码白细胞介素-1受体拮抗剂（rAAV-IL-1Ra）互补DNA的重组腺相关病毒载体在治疗和预防脂多糖（LPS）诱导的关节炎方面的潜力。

方法

通过向Sprague-Dawley大鼠膝关节注射LPS和rAAV-IL-1Ra来研究rAAV-IL-1Ra对关节炎的治疗效果，然后通过滑膜液中的白细胞计数、滑膜的组织学变化以及关节组织中柠檬酸镓的摄取来评估关节炎的严重程度。为了研究对复发性关节炎的治疗效果，在初次注射LPS和rAAV-IL-1Ra 80天后，通过再次注射LPS诱导复发性关节炎，然后评估复发性关节炎的严重程度。为了研究关节炎的预防，将rAAV-IL-1Ra注射到正常关节中。100天后，用LPS诱导关节炎，并评估关节炎的严重程度。

结果

LPS诱导的关节炎症上调了rAAV-IL-1Ra转基因的产生，并且在治疗和预防方案中证明是有效的。单次注射rAAV-IL-1Ra不仅可以抑制原发性关节炎，还可以抑制复发性关节炎。我们发现，在给予rAAV后80天延迟第二次用LPS刺激可以重新激活IL-1Ra的转基因表达。滑膜液中IL-1Ra蛋白的治疗水平达到平均±标准差5.8±0.5 ng/ml。此外，rAAV转基因在正常关节中持续至少100天，并且在LPS刺激后仍可被诱导表达高水平的IL-1Ra（平均±标准差5.2±0.8 ng/ml），从而预防关节炎的发生。