Nanney L B, Mueller S G, Bueno R, Peiper S C, Richmond A
Department of Plastic Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
Am J Pathol. 1995 Nov;147(5):1248-60.
The alpha-chemokines have been implicated as regulators of proliferation and differentiation of normal keratinocytes and as mediators of keratinocyte maturation and migration in inflammatory processes that involve the skin. Using the cutaneous wound repair model, we examined the sites and temporal sequence of the appearance of melanoma growth stimulatory activity or growth-regulated gene (MGSA/GRO;ligand) and the type B interleukin (IL)-8 receptor (IL-8RB) to which MGSA/GRO binds. Human burn tissues (n = 44) representing days 2 to 12 after injury were obtained during surgical debridement, fixed in 4% paraformaldehyde, and embedded in paraffin. Immunolocalizations were performed with polyclonal antisera for both ligand and receptor, as well as a monoclonal antibody for the IL-8 RB. Western blot analysis confirmed the presence of the IL-8 RB in immunoprecipitates of epidermal keratinocyte lysates. In normal skin, MGSA/GRO protein was restricted to sites populated by differentiated keratinocytes (suprabasal compartments, inner root sheath cells, and dermal sweat ducts). MGSA/GRO protein was barely detectable within epithelial margins and islands of burn wounds where the migrating/proliferating keratinocyte populations reside, but staining intensities increased as cells matured into the outer layers. Weak diffuse staining was detected in areas of neutrophilic infiltration (granulation tissue and overlying exudates). By contrast, in normal skin the IL-8 RB was detected in specific locations within epidermal and dermal compartments of healing wounds. In the dermis, polyvalent antibodies detected receptor immunoreactivity most prominently in dermal sweat ducts and endothelium of capillaries, whereas this immunoreactivity was inconspicuous in sections stained with the monoclonal antibody. Receptor immunostaining was noted in migrating/proliferating keratinocytes in epithelial margins and islands but was in the outer layers or in hypertrophic epidermis adjacent to wounds. This same pattern was observed in epidermal appendages such as hair follicles and eccrine sweat ducts. In granulation tissues, IL-8 RB was noted in numerous fibroblasts and in subpopulations of macrophages and smooth muscle. The presence of both MGSA/GRO and its receptor in human burn wounds implicate this cytokine as an autocrine or paracrine mediator of epidermal regeneration in both the inflammatory and proliferative phases of cutaneous wound repair.
α趋化因子被认为是正常角质形成细胞增殖和分化的调节因子，也是涉及皮肤的炎症过程中角质形成细胞成熟和迁移的介质。利用皮肤伤口修复模型，我们检测了黑色素瘤生长刺激活性或生长调节基因（MGSA/GRO；配体）以及MGSA/GRO所结合的B型白细胞介素（IL）-8受体（IL-8RB）出现的部位和时间顺序。在手术清创过程中获取了代表受伤后第2至12天的人类烧伤组织（n = 44），用4%多聚甲醛固定，石蜡包埋。用针对配体和受体的多克隆抗血清以及针对IL-8RB的单克隆抗体进行免疫定位。蛋白质印迹分析证实表皮角质形成细胞裂解物的免疫沉淀物中存在IL-8RB。在正常皮肤中，MGSA/GRO蛋白局限于由分化的角质形成细胞占据的部位（基底上层、内根鞘细胞和真皮汗腺导管）。在迁移/增殖的角质形成细胞群体所在的烧伤伤口上皮边缘和岛状区域几乎检测不到MGSA/GRO蛋白，但随着细胞成熟进入外层，染色强度增加。在嗜中性粒细胞浸润区域（肉芽组织和覆盖的渗出物）检测到微弱的弥漫性染色。相比之下，在正常皮肤中，在愈合伤口的表皮和真皮区域的特定位置检测到IL-8RB。在真皮中，多价抗体在真皮汗腺导管和毛细血管内皮中最显著地检测到受体免疫反应性，而在用单克隆抗体染色的切片中这种免疫反应性不明显。在上皮边缘和岛状区域的迁移/增殖角质形成细胞中注意到受体免疫染色，但在伤口外层或肥厚表皮中没有。在毛囊和小汗腺等表皮附属器中也观察到相同的模式。在肉芽组织中，在许多成纤维细胞以及巨噬细胞和平滑肌亚群中注意到IL-8RB。人类烧伤伤口中同时存在MGSA/GRO及其受体表明这种细胞因子在皮肤伤口修复的炎症和增殖阶段是表皮再生的自分泌或旁分泌介质。
