Investigation of cause of Kawasaki disease, molecular infecto-immunolgical analysis

KD的病因尚不清楚。一些报道表明，KD与外周血淋巴细胞（PBL）中Vbeta2^+和Vbeta8.1^+T细胞的选择性扩增有关。为了更好地了解KD的伦理学，我们研究了T细胞受体（TCR） Vbeta^<2+>和Vbeta8.1^+在新分离的PBL和T细胞克隆（TCC） T细胞上的扩增。血液样本来自急性（n=20）和恢复期（n=20） KD患者，年龄匹配的非传染性疾病儿童（n=18）和健康成人（n=20）。四组间Vbeta^<2+>和Vbeta8.1^+T细胞百分比差异无统计学意义。CD4^+或CD8^+ T细胞亚群也是如此。从6例KD患者的受累皮肤、淋巴结或PBL中建立的105例TCC也呈Vbeta2或Vbeta8.1 TCR阴性。在没有任何刺激的情况下，105个TCC中有68个产生了可检测水平的tnf - α。相比之下，分别只有11个或7个产生可检测水平的IL-2或IL-6。植物血凝素和肉豆蔻酸佛醇刺激诱导大多数TCC产生较高数量的tnf - α、IL-2和IL-6。这些结果表明，CD4^+ T辅助细胞表达tcr - β而不是Vbeta2或Vbeta8.1受体，主要通过产生tnf - α参与KD的免疫病理。在其他报道中，KD与Vbeta2和Vbeta8.1的选择性扩增有关，使用了体外活化的PBL。因此，他们的方法可能允许可能允许体外修改TCR的使用。此外，我们测定了KD患者急性期和恢复期血清中TSST-1的浓度和抗TSST-1抗体。最近有报道称TSST-1可能是KD的病因。但我们没有发现任何证据表明KD是由TSST-1引起的。

The etiology of KD is unknown. Some reports indicated that KD is associated with the selective expansion of Vbeta2^+ and Vbeta8.1^+T cells in peripheral blood lymphocytes (PBL). To understand better the ethiology of KD,we investigated T cell receptor (TCR) Vbeta^<2+> and Vbeta8.1^+ expansion on the T cell of freshly isolated PBL and T cell clones (TCC). Blood samples were obtained from patients with acute (n=20) and convalescent (n=20) KD,age matched children with non-infectious disease (n=18), and healthy adults (n=20). Among these four groups, there were no significant differences in the percentages of their Vbeta^<2+> and Vbeta8.1^+T cells. The same was true for the CD4^+ or CD8^+ T cell subsets. One hundred and five TCC established from the affected skin, lymphnode or PBL of six patients with KD were also negative for either Vbeta2 or Vbeta8.1 TCR.Sixty-eight of 105 TCC produced detectable levels of TNF-alpha, in the absence of any stimuli. In contrast, only 11 or 7 produced detectable levels of IL-2 or IL-6, respectively. Stimulation with phytohaemagglutinin and phorbol myristate induced most TCC to produce higher amounts of TNF-alpha, IL-2 and IL-6. These results suggest that CD4^+ T helper cells expressing TCR-beta other than Vbeta2 or Vbeta8.1 receptor, primarily through TNF-alpha production, are involved in the immunopathology of KD.In other reports that KD is associated with the selective expansion of Vbeta2 and Vbeta8.1, in vitro activated PBL were used. Therfore, their method may allow possibly allow in vitro modification of TCR usage.Fouthermore we measured TSST-1 concentrations and anti-TSST-1 antibody in serum from KD patients in both the acute and the convalescent phase. TSST-1 was reported as a possible cause of KD recently. Bur we failed to find any evidence that KD is caused by TSST-1.