Bondeson J, Browne K A, Brennan F M, Foxwell B M, Feldmann M
Kennedy Institute of Rheumatology, Hammersmith, London, United Kingdom.
J Immunol. 1999 Mar 1;162(5):2939-45.
Macrophages are the major cytokine producers in chronic inflammatory diseases, but the biochemical pathways regulating cytokine production are poorly understood. This is because genetic tools to dissect signaling pathways cannot be used in macrophages because of difficulties in transfection. We have developed an adenoviral technique to achieve high efficiency gene delivery into macrophages and recently showed that spontaneous TNF-alpha production in rheumatoid arthritis joint cells, chiefly from macrophages, is 75% blocked by adenoviral transfer of IkappaBalpha. In this report we use the same adenovirus to investigate whether the production of a number of proinflammatory cytokines (e.g., TNF-alpha, IL-1beta, IL-6, and IL-8) from human macrophages depends on NF-kappaB. While the cytokine response to certain inducers, such as LPS, PMA, and UV light, is blocked by overexpression of IkappaBalpha, the response to zymosan is not. In contrast, anti-inflammatory mediators (IL-10 and IL-1 receptor antagonist) induced by LPS are only marginally inhibited by IkappaBalpha excess. These studies demonstrate several new points about macrophage cytokine production. First, there is heterogeneity of mechanisms regulating both the proinflammatory and anti-inflammatory cytokines within populations of a single cell type. In addition, the results confirm the utility of the adenoviral technique for functional analysis of cytokine induction. The results also confirm that there are autocrine and paracrine interactions regulating cytokine synthesis within a single cell type. The selectivity of NF-kappaB blockade for proinflammatory but not anti-inflammatory mediators indicates that in macrophages, NF-kappaB may be a good target for the treatment of chronic inflammatory diseases.
巨噬细胞是慢性炎症性疾病中主要的细胞因子产生者,但调节细胞因子产生的生化途径却知之甚少。这是因为由于转染困难,用于剖析信号通路的遗传工具无法在巨噬细胞中使用。我们开发了一种腺病毒技术,以实现高效的基因传递到巨噬细胞中,并且最近表明类风湿性关节炎关节细胞中自发产生的肿瘤坏死因子-α(TNF-α),主要来自巨噬细胞,通过腺病毒转导IκBα可被75%阻断。在本报告中,我们使用相同的腺病毒来研究人类巨噬细胞中多种促炎细胞因子(如TNF-α、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-8(IL-8))的产生是否依赖于核因子-κB(NF-κB)。虽然细胞因子对某些诱导剂(如脂多糖(LPS)、佛波酯(PMA)和紫外线)的反应被IκBα的过表达所阻断,但对酵母聚糖的反应却没有。相反,LPS诱导的抗炎介质(IL-10和IL-1受体拮抗剂)仅被IκBα过量略微抑制。这些研究揭示了关于巨噬细胞细胞因子产生的几个新要点。首先,在单一细胞类型群体中,调节促炎和抗炎细胞因子的机制存在异质性。此外,结果证实了腺病毒技术在细胞因子诱导功能分析中的实用性。结果还证实了在单一细胞类型内存在调节细胞因子合成的自分泌和旁分泌相互作用。NF-κB阻断对促炎介质而非抗炎介质的选择性表明,在巨噬细胞中,NF-κB可能是治疗慢性炎症性疾病的一个良好靶点。