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

我们以前曾报道过结核分枝杆菌对人胚胎肺成纤维细胞系有细胞毒性(J干扰素细胞因子研究，2001，抗菌药)。化学试剂，2002,2005)。据报道，细菌的细胞毒性与它们的毒力有关。为了探讨其细胞毒性、生物医学和免疫学特性的机制，我们用12株卡介苗研究了人肺腺癌细胞株A549对一氧化氮(NO)的诱导活性及其对NO的敏感性。其中，菲普斯、俄罗斯和日本三个菌株对A549产生的NO产量高于其他菌株。这些菌株与遗传背景相近，显示出IL-1和肿瘤坏死因子-α对NO产生的协同作用。因此，这些菌株既能保持卡介苗的原有特性，又能对结核分枝杆菌具有保护作用。另一方面，为了研究对生长型…的敏感性用含10%ADC和0~1 mM NaNO_2的米德布鲁克7H9肉汤培养21天，观察NO对细菌的抑制或毒性作用。巴西、葛兰素史克和俄罗斯三个卡介苗菌株对NO非常敏感，但其他菌株对NO都是耐受的。卡介苗对NO的敏感性与其遗传背景无关。本研究以水苏糖为原料合成了一系列糖衍生物，并研究了它们对结核分枝杆菌、禽型分枝杆菌和金黄色葡萄球菌的抗菌活性，并对其构效关系进行了研究。结果表明，化合物OCT359(烯丙基O-(2，3，4，6-四-O-乙酰基--Dgalactopyranosyl)-(1-&gt；6)-O-(2，3，4-tri-O-acetyl-a-D-galactopyranosyl)-(1-&gt；6)-0-2，3，4-tri-O-acetyl-β-D-glucopyranoside))α)具有体外抗菌活性。甘露三糖的C-1位烯丙基和乙酰氧基是其抗菌活性所必需的。较少

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