Tsuji-Yamada J, Nakazawa M, Minami M, Sasaki T
Department of Dermatology, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan.
J Rheumatol. 2001 Jun;28(6):1252-8.
To-determine the relationship between the frequency of cytokine producing cells and systemic sclerosis (SSc), we examined the frequencies of interleukin 4 (IL-4) or IL-2 producing CD4+ or CD8+ T cells in peripheral blood mononuclear cells (PBMC) from patients with SSc.
PBMC from 23 SSc patients and 14 healthy controls were isolated from heparinized blood by density centrifugation. Purified PBMC were stimulated with immobilized anti-CD3 monoclonal antibody for 6 h in the presence of monensin. Cells were fixed, made permeable, and stained for intracellular cytokines in combination with staining for T cell surface markers, CD4, and CD8.
The frequency of CD4+ T cells was negatively correlated with duration of SSc, whereas the frequency of CD8+ T cells was not correlated with duration. The frequencies of IL-4 producing (IL-4+) cells in CD4+ T cells and in CD8+ T cells from patients with SSc were both significantly higher than those from controls. In contrast, the frequencies of IL-2 producing (IL-2+) cells in CD4+ T cells and in CD8+ T cells from patients with SSc were both significantly lower than that from controls. Further, the ratios of IL-4+/IL-4+ + IL-2+ in CD4+ T cells and in CD8+ T cells were both negatively correlated with disease duration of SSc.
These results suggest that type 2 cytokine producing T cells, not only CD4+ T cells but also CD8+ T cells, have important roles in the pathogenesis of SSc, especially in the early phase of SSc.
为了确定细胞因子产生细胞的频率与系统性硬化症（SSc）之间的关系，我们检测了SSc患者外周血单个核细胞（PBMC）中产生白细胞介素4（IL-4）或IL-2的CD4⁺或CD8⁺T细胞的频率。
通过密度离心从23例SSc患者和14名健康对照者的肝素化血液中分离PBMC。纯化的PBMC在莫能菌素存在的情况下用固定化抗CD3单克隆抗体刺激6小时。细胞固定、通透处理后，进行细胞内细胞因子染色，并结合T细胞表面标志物CD4和CD8染色。
CD4⁺T细胞的频率与SSc的病程呈负相关，而CD8⁺T细胞的频率与病程无关。SSc患者CD4⁺T细胞和CD8⁺T细胞中产生IL-4的细胞（IL-4⁺）频率均显著高于对照组。相反，SSc患者CD4⁺T细胞和CD8⁺T细胞中产生IL-2的细胞（IL-2⁺）频率均显著低于对照组。此外，CD4⁺T细胞和CD8⁺T细胞中IL-4⁺/IL-4⁺ + IL-2⁺的比例均与SSc的病程呈负相关。
这些结果表明，产生2型细胞因子的T细胞，不仅CD4⁺T细胞，而且CD8⁺T细胞，在SSc的发病机制中起重要作用，尤其是在SSc的早期阶段。
