Fine Jay S, Rojas-Triana Alberto, Jackson James V, Engstrom Laura W, Deno Gregory S, Lundell Daniel J, Bober Loretta A
Department of Immunology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.
Inflammation. 2003 Aug;27(4):161-74. doi: 10.1023/a:1025076111950.
We have characterized leukocyte migration to the pleural cavity in a methylated-BSA (mBSA)-induced model of murine delayed-type hypersensitivity and evaluated the ability of IL-4 and IL-10 to modulate this response. Neutrophils, macrophages, T cells, and dendritic cells migrated to the pleural cavity in a time-dependent fashion following direct intrapleural antigen challenge, with neutrophils comprising the majority of exudate leukocytes in the cavity within the first 24 h and the number of mononuclear cells increasing at later times. Real-time quantitative PCR analysis of infiltrating leukocytes revealed a marked elevation of steady-state mRNA levels of IL-1beta and TNFalpha and the chemokines KC, MIP-2, CXCL9, CXCL10, CXCL11, CCL2, CCL3, and CCL4 at 6 h postchallenge, which diminished over time. In contrast, gammaIFN mRNA levels were maximal at 24 h and CCL5 expression was sustained throughout 72 h. ELISA analysis of pleural exudate fluid revealed significant elevations of KC and CCL2 protein levels at 6 h postantigen challenge and a peak increase in gammaIFN protein at 24 h, confirming our mRNA observations. Administration of recombinant murine IL-4 or IL-10 prior to challenge significantly blocked cell trafficking to the pleural cavity as well as peak levels of exudate gammaIFN, with IL-4 being more potent in impairing these responses. IL-4 administration also increased the proportion of naive T cells in the pleural cavity, as judged by CD62L and CD45RB expression. These results indicate that this in vivo model demonstrates a pattern of events associated with Th1-mediated leukocyte trafficking and underscore the potential utility of this in vivo model for evaluating therapeutic inhibitors of leukocyte trafficking.
我们已在甲基化牛血清白蛋白（mBSA）诱导的小鼠迟发型超敏反应模型中，对白细胞向胸腔的迁移进行了特征描述，并评估了白细胞介素-4（IL-4）和白细胞介素-10（IL-10）调节这种反应的能力。在胸腔内直接进行抗原攻击后，中性粒细胞、巨噬细胞、T细胞和树突状细胞以时间依赖性方式迁移至胸腔，中性粒细胞在前24小时内构成胸腔渗出白细胞的大部分，而单核细胞数量在随后的时间增加。对浸润白细胞的实时定量PCR分析显示，攻击后6小时，白细胞介素-1β（IL-1β）、肿瘤坏死因子α（TNFα）以及趋化因子KC、MIP-2、CXCL9、CXCL10、CXCL11、CCL2、CCL3和CCL4的稳态mRNA水平显著升高，随后随时间下降。相比之下，γ干扰素（γIFN）mRNA水平在24小时达到峰值，CCL5表达在整个72小时内持续存在。胸腔渗出液的酶联免疫吸附测定（ELISA）分析显示，抗原攻击后6小时，KC和CCL2蛋白水平显著升高，γIFN蛋白在24小时达到峰值增加，证实了我们对mRNA的观察结果。在攻击前给予重组小鼠IL-4或IL-10可显著阻断细胞向胸腔的迁移以及渗出液γIFN的峰值水平，其中IL-4在损害这些反应方面更有效。通过CD62L和CD45RB表达判断，给予IL-4还增加了胸腔中初始T细胞的比例。这些结果表明，该体内模型展示了与Th1介导的白细胞迁移相关的一系列事件模式，并强调了该体内模型在评估白细胞迁移治疗性抑制剂方面的潜在用途。
