Analysis of the cytotoxicity of virulent Mycobacterium tuberculosis to human fibroblast cells

我们先前已经报道了毒性结核分枝杆菌对人胚肺成纤维细胞系表现出细胞毒性（J. Interferon Cytokine Res.，2001年，Antimicrob.探员Chemother 2002年，2005年）。细菌的细胞毒性与它们的毒力相关，这是有报道的。为探讨卡介苗的细胞毒作用机制、生物医学和免疫学特性，本研究用12株卡介苗对人肺细胞株A549进行了一氧化氮（NO）诱导活性及对NO敏感性的研究。其中Phipps、Russia和Japan 3个菌株对A549的NO合成有较高的诱导作用。这些菌株与遗传背景接近，且在IL-1和TNF-α的协同作用下产生NO。因此，这些菌株可以保持原始BCG疫苗的原始特性，并且仍然具有对结核分枝杆菌的保护活性。另一方面，为了研究对生长的敏感性， 关于我们 NO对细菌的抑制或毒性作用，将12株菌株用含有10%ADC和（0至1 mM）NaNO_2的Middlebrook 7 H9肉汤培养21天。巴西、葛兰素和俄罗斯3株BCG对NO敏感，其余菌株对NO耐受。卡介苗对NO的敏感性与其遗传背景无关。以水苏糖为原料，合成了一系列糖衍生物，并对其抗结核分枝杆菌、鸟分枝杆菌和金黄色葡萄球菌的活性进行了评价，同时对其构效关系进行了研究。结果表明，化合物OCT 359（烯丙基O-（2，3，4，6-tetra-O-acetyl-α -D-galactopyranosyl）-（1->6）-O-（2，3，4-tri-O-acetyl-a-D-galactopyranosyl）-（1->6）-O-2，3，4-tri-O-acetyl-β-D-glucopyranoside）具有体外抗菌活性。甘露三糖苷的C-1位的烯丙基和乙酰氧基是其抑菌活性所必需的。少

We have previously reported that virulent Mycobacterium tuberculosis exhibits cytotoxicty to human embryonic lung fibroblast cell line (J. Interferon Cytokine Res., 2001, Antimicrob. Agents Chemother., 2002, 2005). The bacterial cytotoxicity correlates their virulence which is reported. In order to investigate the mechanisms of the cytotoxicity, biomedical and immunological character we investigate that the inducing activity of nitric oxide (NO) from human lung cell line A549 and the susceptibility to NO using twelve BCG strains. Among them three strains, Phipps, Russia and Japan, were observed inducing NO production from A549 higher than others. These strains are close to genetic background and exhibited synergistic effect of IL-1 and TNF-α on the NO production. Therefore, these strains may keep original characters of the primarily BCG vaccine and still have protective activity against Mycobacterium tuberculosis. On the other hand, in order to investigate the susceptibility to growth … More inhibitory or toxic effect of NO on bacteria, the twelve strains were cultured with Middlebrook 7H9 broth containing 10 % ADC and (0 to 1 mM) NaNO_2 for 21 days. Three BCG strains, Brazil, Glaxo and Russia, were very sensitive to NO, however, others were tolerant. The character to the susceptibility to NO did not correlate to the genetic background of BCG strains. NO tolerant strains may play a role in the survival in host and these strains could be useful for carrier of recombinant BCG vaccine in future.A series of sugar derivatives were synthesized from stachyose, a sugar compound of Stachys sieboldi Miq, and evaluated for antibacterial activity against Mycobacterium tuberculosis, Mycobacterium avium, and Staphylococcus aureus, and their structure-activity relationships were studied. The results showed that the compound OCT359 (allyl O-(2,3,4,6-tetra-O-acetyl-α -Dgalactopyranosyl)-(1->6)-O-(2,3,4-tri-O-acetyl-a-D-galactopyranosyl)-(1->6)-0-2,3,4-tri-O-acetyl-β-D-glucopyranoside) exhibited in vitro antibacterial activity. The allyl group at C-1 and the acetoxy groups of the manniotrioside were requisite for the antibacterial activity. Less