Knedla A, Riepl B, Lefèvre S, Kistella S, Grifka J, Straub R H, Gay S, Schölmerich J, Müller-Ladner U, Neumann E
Department of Internal Medicine and Rheumatology, Justus-Liebig-University of Giessen, Department of Rheumatology and Clinical Immunology, Kerckhoff-Klinik Bad Nauheim, Germany.
Ann Rheum Dis. 2009 Jan;68(1):124-9. doi: 10.1136/ard.2007.086116. Epub 2008 Mar 13.
The viral gene transfer of interleukin 1 receptor antagonist (IL1ra) and interleukin 10 (IL10) into rheumatoid arthritis (RA) synovial fibroblasts (RASFs) has shown protective effects on cartilage destruction in the severe combined immunodeficiency (SCID) mouse model of RA. Nevertheless, side effects of viral transduction are possible and a number of cytokines or cytokine inhibitors are not available encoded in viral vehicles. As the production of viruses coding for bioactive proteins is cost and time intensive, we established an in vivo long-term release model using osmotic minipumps in the SCID mouse model for RA.
Isolated RASFs were cultured for four passages and coimplanted together with human cartilage and an Alzet osmotic miniature pump model 2004, containing 200 microl of IL10 and IL1ra for 40 days in SCID mice. Implants were removed after 40 days and evaluated histologically. The actual rates of IL10 and IL1ra in murine serum were measured by ELISA.
Release of IL10 and IL1ra by the pumps was effective as both could be measured in significant amounts in the serum of the mice. IL10 and IL1ra release showed protective effects towards the coimplanted cartilage, similar to the adenovirally IL10/IL1ra-transduced RASFs. The mean (SD) invasion scores for the implants with the osmotic pumps were: invasion 0.7 (0.5), degradation 0.5 (0.3) (all parameters significant vs controls, p<0.05).
The results demonstrate that the combination of osmotic pumps with the SCID mouse model for RA can be used as approach for application and evaluation of cartilage-protective molecules. Furthermore, the effect of cartilage-protective cytokines is independent of the type of application.
在类风湿关节炎（RA）的重症联合免疫缺陷（SCID）小鼠模型中，将白细胞介素1受体拮抗剂（IL1ra）和白细胞介素10（IL10）进行病毒基因转移至RA滑膜成纤维细胞（RASFs）已显示出对软骨破坏具有保护作用。然而，病毒转导可能存在副作用，并且许多细胞因子或细胞因子抑制剂无法通过病毒载体编码获得。由于编码生物活性蛋白的病毒生产成本高且耗时，我们在RA的SCID小鼠模型中建立了一种使用渗透微型泵的体内长期释放模型。
将分离的RASFs培养四代，然后与人类软骨以及含有200微升IL10和IL1ra的Alzet渗透微型泵模型2004一起在SCID小鼠中共植入40天。40天后取出植入物并进行组织学评估。通过酶联免疫吸附测定法（ELISA）测量小鼠血清中IL10和IL1ra的实际含量。
泵对IL10和IL1ra的释放是有效的，因为在小鼠血清中均可检测到大量的这两种物质。IL10和IL1ra的释放对共植入的软骨显示出保护作用，类似于腺病毒介导的IL10/IL1ra转导的RASFs。带有渗透泵的植入物的平均（标准差）侵袭评分如下：侵袭0.7（0.5），降解0.5（0.3）（所有参数与对照组相比均有显著差异，p<0.05）。
结果表明，渗透泵与RA的SCID小鼠模型相结合可作为应用和评估软骨保护分子的一种方法。此外，软骨保护细胞因子的作用与应用类型无关。
