Van Montfrans Catherine, Rodriguez Pena Maria Sol, Pronk Inge, Ten Kate Fiebo J W, Te Velde Anje A, Van Deventer Sander J H
Department of Experimental Internal Medicine, Academic Medical Center, Amsterdam, The Netherlands.
Gastroenterology. 2002 Dec;123(6):1865-76. doi: 10.1053/gast.2002.37067.
BACKGROUND & AIMS: Regulatory CD4(+) cells secreting the anti-inflammatory cytokine interleukin (IL)-10 play a key role in maintaining the immune balance in the intestinal mucosa. In this study we engineered primary CD4(+) cells to express IL-10 and investigated the efficacy of this approach in offering protection against experimental colitis.
Spleen-derived CD4(+) cells were transduced by using a retroviral vector to simultaneously express IL-10 and green fluorescent protein (GFP). The therapeutic benefit of CD4(+) cells transduced with IL-10 GFP was studied in experimental colitis, induced by transfer of CD45RB(high) CD4(+) cells to severe combined immunodeficient mice, and in acute trinitrobenzene sulfonic acid (TNBS)-induced colitis.
Transferred engineered GFP fluorescent cells were detected for at least 15 weeks in peripheral blood, spleens, colon, and lymph nodes draining the intestine of recipient SCID mice. IL-10-GFP CD4(+) cells prevented CD45RB(high)-induced transfer colitis effectively, whereas no effect was observed after transfer of nontransduced CD4(+) cells. IL-10-GFP CD45RB(high) CD4(+) cells lost the capacity to induce colitis. By contrast, no therapeutic benefit was observed in TNBS-induced colitis.
Primary murine CD4(+) cells that were engineered to express IL-10 by retroviral transduction act as regulatory cells in CD45RB(high)-induced transfer colitis. This approach may induce long-term maintenance of mucosal immune homeostasis in Crohn's disease.
分泌抗炎细胞因子白细胞介素（IL）-10的调节性CD4(+)细胞在维持肠道黏膜免疫平衡中起关键作用。在本研究中，我们对原代CD4(+)细胞进行基因工程改造使其表达IL-10，并研究该方法对实验性结肠炎的保护效果。
利用逆转录病毒载体转导脾源性CD4(+)细胞，使其同时表达IL-10和绿色荧光蛋白（GFP）。在将CD45RB(high) CD4(+)细胞转移至严重联合免疫缺陷小鼠诱导的实验性结肠炎以及急性三硝基苯磺酸（TNBS）诱导的结肠炎中，研究转导了IL-10 GFP的CD4(+)细胞的治疗益处。
在受体SCID小鼠的外周血、脾脏、结肠及引流肠道的淋巴结中，至少15周都能检测到转导的基因工程改造的GFP荧光细胞。IL-10-GFP CD4(+)细胞有效预防了CD45RB(high)诱导的转移型结肠炎，而未转导的CD4(+)细胞转移后未观察到效果。IL-10-GFP CD45RB(high) CD4(+)细胞失去了诱导结肠炎的能力。相比之下，在TNBS诱导的结肠炎中未观察到治疗益处。
通过逆转录病毒转导改造使其表达IL-10的原代小鼠CD4(+)细胞，在CD45RB(high)诱导的转移型结肠炎中起调节细胞的作用。该方法可能会诱导克罗恩病黏膜免疫稳态的长期维持。
