Pasqui A L, Di Renzo M, Auteri A, Federico G, Puccetti L
University of Siena, Italy.
Eur J Clin Invest. 2007 Sep;37(9):742-5. doi: 10.1111/j.1365-2362.2007.01850.x.
Inflammatory and/or immune activation occurs both in animal models (twitcher mice) and in the brain of patients with Globoid cell leukodystrophy (GLD) or Krabbe's disease (KD). In this study we evaluated in vitro the cytokine profile of KD patients and the effect of psychosine, the toxic metabolite which plays a role in the demyelination process in these patients.
We studied cytokine production by peripheral blood mononuclear cells (PBMCs) isolated from four KD patients, diagnosed on the basis of clinical criteria. Cells were cultured and stimulated with appropriate agents and the supernatants collected before and after the addition of psychosine. Tumour necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8) and monocyte chemoattractant factor (MCP)-1) production was evaluated (ELISA method) and compared with a group of 10 normal subjects.
We found a significant increase of TNF-alpha release by PBMCs of KD patients compared with healthy subjects; TNF-alpha production was significantly increased after LPS addition. Psychosine was able to induce a further significant increase (P < 0.05) only in cells obtained from KD patients and not from control subjects. No changes were found in IL-8 and MCP-1 production.
The increased TNF-alpha production permits us to confirm the presence of an inflammatory-immune stimulus in KD patients, which may be induced and potentiated by the pathogenetic metabolite psychosine.
炎症和/或免疫激活在动物模型（震颤小鼠）以及球形细胞脑白质营养不良（GLD）或克拉伯病（KD）患者的大脑中均有发生。在本研究中，我们在体外评估了KD患者的细胞因子谱以及精神鞘氨醇（在这些患者的脱髓鞘过程中起作用的有毒代谢产物）的影响。
我们研究了从4名根据临床标准确诊的KD患者中分离出的外周血单个核细胞（PBMC）产生细胞因子的情况。细胞经培养并用适当的试剂刺激，在添加精神鞘氨醇之前和之后收集上清液。评估肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）和单核细胞趋化因子（MCP）-1的产生（酶联免疫吸附测定法），并与一组10名正常受试者进行比较。
我们发现，与健康受试者相比，KD患者的PBMC释放的TNF-α显著增加；添加脂多糖后，TNF-α的产生显著增加。仅在从KD患者而非对照受试者获得的细胞中，精神鞘氨醇能够诱导进一步的显著增加（P<0.05）。未发现IL-8和MCP-1产生有变化。
TNF-α产生的增加使我们能够证实KD患者中存在炎症-免疫刺激，这可能由致病代谢产物精神鞘氨醇诱导并增强。
