Shibuya Asuka, Ando Minoru, Tsuchiya Ken, Akiba Takashi, Nihei Hiroshi
Department of Medicine IV, Tokyo Women's Medical University, Japan.
Nihon Jinzo Gakkai Shi. 2004 Oct;46(7):700-8.
Dialysis patients are weak in immune host defense, which is associated with their high morbidity of infection. Polymorphonuclear leukocytes (PMNLs) and mononuclear cells play a key role in innate host defense. PMNLs and monocytes have bactericidal activity through the process of phagocytosis. Monocytes and lymphocytes contribute to the development of innate immunity by their cytokine actions. We studied the intracellular cytokine syntheses in response to ex-vivo stimuli, which may reflect the potential reactivity of immune cells in cytokine syntheses when pathogens invade humans. Furthermore, phagocytic activity was assessed in granulocytes and monocytes. Twenty HD, 15 CAPD, and 10 age-matched controls were enrolled in this study. One milliliter of whole blood from each subject was incubated with lipopolysaccharides (LPS) or mitogens for 4 hours at 37 degrees C. Monoclonal antibodies to CD14+ and CD4+ were used for identifying monocytes and helper T cells, respectively. Intracellular cytokines were stained using FASTIMMUNE staining kits. Interleukin-1beta and TNF-alpha syntheses were examined in monocytes, which are the most important early-response cytokines in innate immunity. IFN-gamma and IL-4 syntheses were examined in helper T cells to observe their polarization into Th1 and Th2 cells. IFN-gamma is a key factor in establishing innate immunity. The percentage of cells that stained positive for each cytokine was analyzed using a flow cytometer. The following results were obtained: 1) In CAPD patients, IL-1beta and TNF-alpha response to LPS in monocytes were significantly reduced, as compared to other subjects. Polarization of helper T cells was reduced, resulting in a significant decrease in Th1 cells. 2) In HD patients, monokine responses were not altered, but polarization of helper T cells was skewed toward a Th1 type. Phagocytic activities were not impaired in both dialysis groups. In conclusion, mononuclear cells from CAPD patients have the potential to exhibit failure of a cytokine response to ex-vivo stimuli in terms of innate immunity.
透析患者的免疫宿主防御功能较弱，这与他们感染的高发病率相关。多形核白细胞（PMNLs）和单核细胞在先天性宿主防御中起关键作用。PMNLs和单核细胞通过吞噬过程具有杀菌活性。单核细胞和淋巴细胞通过其细胞因子作用促进先天性免疫的发展。我们研究了体外刺激后细胞内细胞因子的合成，这可能反映病原体侵入人体时免疫细胞在细胞因子合成中的潜在反应性。此外，还评估了粒细胞和单核细胞的吞噬活性。本研究纳入了20名血液透析（HD）患者、15名持续性非卧床腹膜透析（CAPD）患者和10名年龄匹配的对照者。将每个受试者的1毫升全血与脂多糖（LPS）或有丝分裂原在37℃孵育4小时。分别使用抗CD14 +和抗CD4 +单克隆抗体来识别单核细胞和辅助性T细胞。使用FASTIMMUNE染色试剂盒对细胞内细胞因子进行染色。检测了单核细胞中白细胞介素-1β和肿瘤坏死因子-α的合成，它们是先天性免疫中最重要的早期反应细胞因子。检测了辅助性T细胞中干扰素-γ和白细胞介素-4的合成，以观察它们向Th1和Th2细胞的极化情况。干扰素-γ是建立先天性免疫的关键因素。使用流式细胞仪分析每种细胞因子染色阳性的细胞百分比。获得了以下结果：1）与其他受试者相比，CAPD患者单核细胞中白细胞介素-1β和肿瘤坏死因子-α对LPS的反应显著降低。辅助性T细胞的极化减少，导致Th1细胞显著减少。2）HD患者中，单核因子反应未改变，但辅助性T细胞的极化偏向Th1型。两个透析组的吞噬活性均未受损。总之，就先天性免疫而言，CAPD患者的单核细胞有可能表现出对体外刺激的细胞因子反应失败。
