動物をモデルとした川崎病病因の免疫学的・病理学的解析

研究了干酪乳杆菌细胞壁提取物（LCWE）刺激BALB/C和C3H/Hej小鼠腹腔巨噬细胞（PMC）产生IL-1、TNFalpha和IL-6等炎性细胞因子的能力。通过检测C3H/Hej小鼠胸腺细胞对IL-1的增殖活性和杂交瘤B3BI细胞对IL-6的增殖活性，测定mpc培养上清中细胞因子的含量。采用多克隆和单克隆抗小鼠TNFalpha抗体，采用酶联免疫吸附法测定TNFalpha对L929细胞的细胞毒活性。与LPS相比，LCWE诱导BALB/C小鼠产生更高滴度的IL-1和TNFalpha，并产生相似水平的IL-6。在C3H/flej小鼠中，LCWE诱导三种细胞因子的产生均较低或完全不产生。LCWE刺激MPCs的il -1 β和TNFalpha MRNA表达的研究支持产生的细胞因子水平和时间动力学的结果。采用Lehman等人（1988）报道的改良方法，将LCWE注入腹腔，可诱导更多的心肌损伤。令人惊讶的是，LCWE治疗后仅48小时（0.02 mg/g体重）就发现心脏受累，如血管炎（心内膜炎）、冠状动脉炎和心包炎。在BALB/c和C3H/Hej小鼠中48小时后，BALB/C和C3H/Hej小鼠心脏损伤的总频率分别为69%（9/13）和57%(5/14)，而第四周时，心脏损伤的总频率分别为28%（7/25）和8%（2/25）。he染色的组织病理学图显示心肌组织有明显的细胞浸润，尤其是巨噬细胞和中性粒细胞。此外,。结果表明，ABC法对炎症的心脏大动脉和小动脉进行了IL-1、TNFalpha和IL-6等细胞因子抗体的强烈染色。以上结果表明，LCWE在体内和体外都是巨噬细胞和其他产生炎症细胞因子的细胞的强大激活剂，导致小鼠心脏受累。我们认为LCWE注射BALB/C小鼠是研究人类川崎病热生成的良好动物模型。少

The ability of cell wall extract separated from Lactobacillus casei(LCWE)to stimulate peritoneal macrophages(PMC)of BALB/C and C3H/Hej mice, which produced the inflammatory cytokines such as IL-1, TNFalpha and IL-6 was evaluated. The measurement of cytokines in the MPC-cultured supernatants was assayed by the proliferative activity of thymocytes from C3H/Hej mice for IL-1 and of hybridoma B3BI cells for IL-6. TNFalpha was measured by the cytotoxic activity to L929 cells and by the enzyme-linked immunosorbent assay using polyclonal and monoclonal anti-murine TNFalpha antibodies.In comparison with LPS, LCWE induced the production of IL-1 and TNFalpha in higher titers, and similar level of IL-6 production in BALB/C mice. In C3H/flej mice LCWE induced the three kinds of cytokine production in low level or not at all. The study of IL-1beta and TNFalpha MRNA expression of MPCs stimulated with LCWE supported the results of the level and time-kinetics of produced cytokines.The induction of mur … More ine heart damages was performed by an injection of LCWE into the peritoneal cavity using a modified method reported by Lehman et al(1988). Surprisingly, only 48 hours after the treatment of LCWE(0.02 mg/g of body weight cardiac involvement, such as angitis (endocarditis, coronary arteritis and pericarditis were found. in both BALB/c and C3H/Hej mice. The overall frequency of cardiac damages was 69 %(9/13)and 57 %(5/14)in BALB/C and C3H/Hej mice respectively 48 hours later, whereas it was 28 %(7/25)and 8%(2/25)respectively at the fourth week. The histopathological pictures by HEstaining revealed marked cell infiltration in cardiac tissure, especially by macrophges and neutrophils. Moreover, . it was demonstrated that large and small cardiac arteries with inflammation were strongly stained by antibodies to cytokines such as IL-1, TNFalpha and IL-6 in the, ABC method.The above results suggested that LCWE was a powerful in vivo and in vitro activators of macrophages and other cells which produced inflammatory cytokines, which caused murine cardiac involvement. It is thought that LCWE injecxted BALB/C mice would be good animal models for studying the hathogenesis of Kawasaki disease in man. Less