Antachopoulos Charalampos, Demchok Joanne P, Roilides Emmanuel, Walsh Thomas J
Immunocompromised Host Section, Pediatric Oncology Branch, National Cancer Institute, Bethesda, MD 20892, USA.
Mycoses. 2010 Jul;53(4):321-8. doi: 10.1111/j.1439-0507.2009.01725.x. Epub 2009 Jul 22.
Previous studies have not systematically assessed the effect of fungal biomass on polymorphonuclear leucocyte (PMN)-induced hyphal damage (HD) of filamentous fungi. We hypothesised that fungal biomass is a significant factor affecting PMN-induced HD. One isolate each consisting of a volume of 2 x 10(4) conidia ml(-1) of Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Rhizopus oryzae, Rhizopus microsporus, Cunninghamella bertholletiae, Scedosporium prolificans and Fusarium solani were incubated for six different time periods yielding biomass values between 0.01 and 0.1 optical density (OD, 405 nm). Polymorphonuclear leucocyte were added at effector-target (E : T) ratios of 5 : 1, 10 : 1, 20 : 1, 50 : 1 and 100 : 1, and HD was assessed by XTT [2,3-bis-(2-methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-carboxanilide] metabolic assay. Hyphal damage decreased with increasing biomass following the sigmoid (E(max)) model (median R(2): 0.87). Hyphal damage at 0.01 OD exceeded HD at 0.1 OD (P < 0.01) by >twofold in 64 out of 80 comparisons. The sigmoid curves were shifted to the right with higher E : T ratios; the EC(50) values (OD values showing HD halfway between maximal and minimal HD) obtained for 50 : 1 or 100 : 1 were higher than for 5 : 1 (P < 0.01). Using the same E : T ratio, interspecies differences were observed; for 5 : 1, lower EC(50) values were obtained for A. flavus and the zygomycete species. In conclusion, PMN-induced HD decreases with increasing biomass. This effect is both species-dependent and E : T ratio-dependent.
以往的研究尚未系统评估真菌生物量对多形核白细胞(PMN)诱导的丝状真菌菌丝损伤(HD)的影响。我们推测真菌生物量是影响PMN诱导的HD的一个重要因素。将烟曲霉、黄曲霉、土曲霉、米根霉、少根根霉、伯氏小克银汉霉、多育赛多孢和茄病镰刀菌各一个菌株,每毫升含2×10⁴个分生孢子,分别培养六个不同时间段,得到生物量值在0.01至0.1光密度(OD,405nm)之间。以5:1、10:1、20:1、50:1和100:1的效应物-靶标(E:T)比例加入多形核白细胞,并通过XTT [2,3-双-(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-羧基苯胺]代谢测定法评估HD。菌丝损伤随生物量增加而降低,符合S形(E(max))模型(中位数R²: 0.87)。在80次比较中的64次中,0.01 OD时的菌丝损伤比0.1 OD时的HD高出两倍以上(P < 0.01)。S形曲线随E:T比例升高向右移动;50:1或100:1时获得的EC₅₀值(显示HD在最大和最小HD之间的OD值)高于5:1时(P < 0.01)。使用相同的E:T比例时,观察到种间差异;对于5:1,黄曲霉和接合菌的EC₅₀值较低。总之,PMN诱导的HD随生物量增加而降低。这种效应既依赖于物种,也依赖于E:T比例。