Woods J M, Katschke K J, Tokuhira M, Kurata H, Arai K I, Campbell P L, Koch A E
Department of Medicine, Northwestern University Medical School, Chicago, IL 60611, USA.
J Immunol. 2000 Sep 1;165(5):2755-63. doi: 10.4049/jimmunol.165.5.2755.
The rheumatoid arthritis (RA) joint is characterized by an inflammatory synovial pannus which mediates tissue destruction. IL-13 is a cytokine that inhibits activated monocytes/macrophages from secreting a variety of proinflammatory molecules. The aim of this study was to examine whether gene therapy-delivered IL-13 could reduce the production of key proinflammatory mediators in RA synovial tissue (ST) explants. Adenoviral vectors encoding the genes for human IL-13 (AxCAIL-13) and bacterial beta-galactosidase were generated and examined for protein production. Vectors were used to infect RA ST explants and RA synovial fibroblasts, and conditioned medium (CM) was collected at various times for analysis by ELISA and competitive immunoassay. AxCAIL-13 decreased the production of RA ST explant proinflammatory IL-1beta by 85% after 24 h. Likewise, TNF-alpha levels were decreased by 82 and 75% whereas IL-8 levels were reduced 54 and 82% after 24 and 48 h, respectively, in RA ST explant CM. Monocyte chemotactic protein-1 concentrations were decreased by 88% after 72 h in RA ST explant CM. RA ST explant epithelial neutrophil-activating peptide-78 concentrations were decreased 85 and 94% whereas growth-related gene product-alpha levels were decreased by 77 and 85% at 24 and 48 h, respectively, by AxCAIL-13. Further, IL-13 significantly decreased PGE2 and macrophage inflammatory protein-1alpha production. These results demonstrate that increased expression of IL-13 via gene therapy may decrease RA-associated inflammation by reducing secretion of proinflammatory cytokines and PGE2.
类风湿性关节炎（RA）关节的特征是存在介导组织破坏的炎性滑膜血管翳。白细胞介素-13（IL-13）是一种细胞因子，可抑制活化的单核细胞/巨噬细胞分泌多种促炎分子。本研究的目的是检验基因治疗递送的IL-13是否能减少RA滑膜组织（ST）外植体中关键促炎介质的产生。构建了编码人IL-13基因（AxCAIL-13）和细菌β-半乳糖苷酶的腺病毒载体，并检测其蛋白产生情况。将载体用于感染RA ST外植体和RA滑膜成纤维细胞，在不同时间收集条件培养基（CM），通过酶联免疫吸附测定（ELISA）和竞争性免疫测定进行分析。24小时后，AxCAIL-13使RA ST外植体促炎白细胞介素-1β（IL-1β）的产生减少了85%。同样，在RA ST外植体CM中，24小时和48小时后，肿瘤坏死因子-α（TNF-α）水平分别降低了82%和75%，而IL-8水平分别降低了54%和82%。72小时后，RA ST外植体CM中单核细胞趋化蛋白-1（MCP-1）浓度降低了88%。AxCAIL-13使RA ST外植体上皮中性粒细胞激活肽-78（ENA-78）浓度在24小时和48小时分别降低了85%和94%，而生长相关基因产物-α（GRO-α）水平分别降低了77%和85%。此外，IL-13显著降低了前列腺素E2（PGE2）和巨噬细胞炎性蛋白-1α（MIP-1α）的产生。这些结果表明，通过基因治疗增加IL-13的表达可能通过减少促炎细胞因子和PGE2的分泌来减轻RA相关炎症。
