Watabe D, Kanno H, Yoshida A, Kurose A, Akasaka T, Sawai T
Department of Pathology, Iwate Medical University School of Medicine, 19-1 Uchimaru, Morioka 020-8505, Japan.
Br J Dermatol. 2007 Aug;157(2):259-65. doi: 10.1111/j.1365-2133.2007.08039.x. Epub 2007 Jun 26.
The adhesion of CD4+ T cells to endothelial cells and their subsequent migration to skin tissue are essential to develop the psoriatic skin lesion. However, few studies have examined the role of adhesion molecules in the binding of T cells from patients with chronic plaque psoriasis to endothelial cells in vitro; thus, the adhesion molecules responsible for the development of skin lesions are still unclear.
To identify the responsible adhesion molecules in the interaction between CD4+ T cells in patients with chronic plaque psoriasis and cytokine-stimulated endothelial cells.
An in vitro adhesion assay between Calcein-labelled peripheral blood mononuclear cells (PBMC) and cytokine-stimulated human endothelial cultures, which exhibit a higher adhesion capacity to PBMC, was established, and the adhesion-inhibitory effects of a panel of antiadhesion molecule antibodies on the adhesion of PBMC from patients with psoriasis to endothelial cells were examined. Then, the inhibitory effects of selected antibodies acting on the interaction between CD4+ T cells from patients with psoriasis (purified by negative magnetic cell sorting) and cultured endothelial cells were examined.
A significant increase (P < 0.01) in the adhesion of psoriatic PBMC to both endothelial cultures, human skin microvascular endothelial cells from adults (HMVEC-Ad) and human coronary arterial endothelial cells (HCAEC), compared with healthy PBMC, was demonstrated in our in vitro cell adhesion assay. Pretreatment of both endothelial cultures with tumour necrosis factor (TNF)-alpha (1000 U mL(-1)) induced the most frequent adhesion of PBMC from patients with psoriasis among the three inflammatory cytokines examined, i.e. TNF-alpha, interleukin-1beta and interferon-gamma [TNF-alpha-treated vs. nontreated: P < 0.001 (in both HMVEC-Ad and HCAEC)]. In both endothelial cultures treated with TNF-alpha, PBMC from patients with psoriasis exhibited significantly more frequent adhesion compared with those from healthy individuals (P < 0.001). The TNF-alpha-stimulated HMVEC-Ad, which exhibited the most frequent adhesion of PBMC, were selected for adhesion-inhibition experiments using monoclonal antibodies (mAbs) to adhesion molecules that are upregulated in psoriatic lesions, and the combination of antilymphocyte function-associated antigen type 1 (LFA-1) and anti-intercellular adhesion molecule 1 (ICAM-1) mAbs gave the greatest reduction of adhesion of PBMC from patients with psoriasis (approximately 69% reduction; P < 0.01). This combination of mAbs significantly reduced also the adhesion of CD4+ T cells from patients with psoriasis to TNF-alpha-stimulated HMVEC-Ad (approximately 62% reduction), compared with pretreatment with isotype control mAbs (P < 0.01).
These findings indicate that the LFA-1/ICAM-1 interaction plays a major role in the adhesion of CD4+ T cells to endothelial cells and that TNF-alpha might play an important role for the induction of adhesion molecules on endothelial cells at psoriatic skin lesions.
CD4 + T细胞与内皮细胞的黏附及其随后向皮肤组织的迁移对于银屑病皮损的形成至关重要。然而，很少有研究在体外检测黏附分子在慢性斑块状银屑病患者T细胞与内皮细胞结合中的作用；因此，导致皮肤病变的黏附分子仍不清楚。
确定慢性斑块状银屑病患者CD4 + T细胞与细胞因子刺激的内皮细胞相互作用中起作用的黏附分子。
建立了用钙黄绿素标记的外周血单个核细胞（PBMC）与细胞因子刺激的人内皮细胞培养物之间的体外黏附试验，该培养物对PBMC具有更高的黏附能力，并检测了一组抗黏附分子抗体对银屑病患者PBMC与内皮细胞黏附的抑制作用。然后，检测了作用于银屑病患者（通过阴性磁珠细胞分选纯化）CD4 + T细胞与培养的内皮细胞之间相互作用的选定抗体的抑制作用。
在我们的体外细胞黏附试验中，与健康PBMC相比，银屑病PBMC与两种内皮细胞培养物，即成人皮肤微血管内皮细胞（HMVEC-Ad）和人冠状动脉内皮细胞（HCAEC）的黏附均显著增加（P < 0.01）。在检测的三种炎性细胞因子，即肿瘤坏死因子（TNF）-α、白细胞介素-1β和干扰素-γ中，用TNF-α（1000 U mL(-1)）预处理两种内皮细胞培养物诱导银屑病患者PBMC的黏附最为频繁[TNF-α处理组与未处理组相比：P < 0.001（在HMVEC-Ad和HCAEC中均如此）]。在用TNF-α处理的两种内皮细胞培养物中，银屑病患者的PBMC黏附均显著高于健康个体（P < 0.001）。选择对银屑病皮损中上调的黏附分子使用单克隆抗体（mAb）进行黏附抑制实验，其中抗淋巴细胞功能相关抗原1（LFA-1）和抗细胞间黏附分子1（ICAM-1）mAb组合对银屑病患者PBMC的黏附减少作用最大（减少约69%；P < 0.01）。与用同型对照mAb预处理相比，该mAb组合也显著降低了银屑病患者CD4 + T细胞与TNF-α刺激的HMVEC-Ad的黏附（减少约62%）（P < 0.01）。
这些发现表明LFA-1/ICAM-1相互作用在CD4 + T细胞与内皮细胞的黏附中起主要作用，并且TNF-α可能在银屑病皮损处内皮细胞黏附分子的诱导中起重要作用。
