Bio-molecular Approaches to Mycobacterial

从耐酸菌Gordona aurantiaca中分离得到一种含分枝菌酸的糖脂--海藻糖-2，3，6 '-三分枝菌酸酯（TTM），并对其生物活性进行了表征。将TTM脂质体包封制剂皮下或囊内（肾脏）注射到小鼠体内，在注射部位产生了与结核杆菌感染病灶相似的组织病理学变化：在这些病变中观察到脓肿形成伴空泡状坏死、巨噬细胞浸润和脓肿周围成纤维细胞增殖。静脉或腹腔注射TTM脂质体使腹腔巨噬细胞处于活化状态。活化的巨噬细胞通过细胞与细胞接触裂解同源肿瘤细胞，包括肥大细胞瘤、骨髓瘤或肺癌细胞。此外，它们还释放了炎症因子，包括过氧化氢、白细胞介素-1和肿瘤坏死因子α，这表明活化的巨噬细胞在坏死和/或脓肿形成中起着重要作用。此外，这种治疗在体内对荷瘤小鼠有效，以延长其对腹腔内攻击的P815肥大细胞瘤细胞或对淋巴结中转移的刘易斯肺癌细胞的存活时间。我们还发现，在腹腔内注射TTM-脂质体的小鼠的腹腔渗出液细胞中，不仅活化的巨噬细胞，而且γ-δ T淋巴细胞大量产生。对腹膜渗出液细胞的淋巴细胞部分的白细胞介素活化细胞分选仪分析显示，γ-δ T细胞在其表面共表达α-β抗原受体。此外，这些T细胞增强了巨噬细胞中saltrin的细胞内杀伤。

We isolated a mycolic acid-containing glycolipid, trehalose-2,3,6'-trimycolate ( TTM ) from an acid-fast bacterium, Gordona aurantiaca, and characterized its biological activity. Subcutaneous or intra-capsular (kidney) injection of the liposome-encapsulated preparation of TTM into mice generated similar histo-pathological changes at the injected sites as that at the infected foci of tubercle bacilli : Abscess formation with caesation necrosis, infiltration of macrophages and proliferation of fibroblasts around the abscess were observed in these lesions. Intravenous or intra-peritoneal injection of TTM-liposomes rendered peritoneal macrophages to be in an activated state. The activated macrophages lysed syngeneic tumor cells including mastocytoma, myeloma or lung cancer cells by cell-to-cell contact. In addition, they released inflammatory factors including hydrogen peroxide, interleukin-1 and tumor-necrosisfactor alpha, suggesting that the activated macrophages were playing a central role in caesation necrosis and/or abscess formation. Furthermore, this treatment was effective in vivo on tumor-bearing mice to prolong their survival times against intraperitoneal challenge of P815 mastocytoma cells or against metastatic Lewis' lung cancer cells in the lymph nodes.We also found that in the peritoneal exudate cells of the mice, which were injected intraperitoneally with TTM-liposomes, not only activated macrophages but also gamma-delta T lymphocytes were generated in a large number. Fluorescein-activated cell sorter analyses of the lymphocytes fraction of the peritoneal exudate cells revealed the gamma-delta T cells co-expressed alpha-beta antigen receptors on their surfaces. In addition, these T cells augmented the intra-cellular killing of salmonella in macrophages.