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

KD的病因尚不清楚。有报道指出KD与外周血淋巴细胞(PBL)中Vbeta2^+和Vbeta8.1^+T细胞的选择性增殖有关。为了更好地了解KD的遗传学，我们研究了新鲜分离的PBL和T细胞克隆(TCC)的T细胞受体(TCR)Vbeta^&lt；2+&Gt；和Vbeta8.1^+的增殖情况。血样来自KD急性期(n=20)和恢复期(n=20)、年龄匹配的非感染性疾病儿童(n=18)和健康成人(n=20)。在这四组中，其Vbeta2+和Vbeta8.1+T细胞的百分比没有显著差异。CD4^+或CD8^+T细胞亚群也是如此。从6例KD患者的受累皮肤、淋巴结或外周血中建立的105例TCC也呈Vbeta2或Vbeta8.1TCR阴性。相比之下，只有11个或7个分别产生了可检测到的IL-2或IL-6水平。植物血凝素和佛波酯刺激大多数移行细胞产生较高数量的肿瘤坏死因子-α、白介素2和白介素6。这些结果表明，表达TCR-β而不是Vbeta2或Vbeta8.1受体的CD4^+T辅助细胞主要通过产生肿瘤坏死因子-α参与了KD的免疫病理。在其他关于KD与Vbeta2和Vbeta8.1选择性扩增有关的报道中，使用了体外激活的PBL。因此，他们的方法可能允许在体外修改TCR的用法。此外，我们检测了急性期和恢复期KD患者血清中TSST-1浓度和抗TSST-1抗体。TSST-1最近被报道为KD的可能病因。但我们没有找到任何证据表明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.