METABOLIC MAINTENANCE & GROWTH OF MYCOBACTERIUM LEPRAE

研究将用麻风分枝杆菌疫苗进行， 新鲜收获，从受感染的nu/nu小鼠的脚垫和 富含活性生物体，以探索最佳的生物物理和 维持新陈代谢的营养参数 麻风杆菌使用以下敏感剂组合 代谢分析：a)细胞内腺苷的定量 三磷酸(ATP)，b)将14C-棕榈酸酯掺入PGL-1， C)测量从14C-14C释放的14CO2的速率 棕榈酸。这些净能量指标的综合使用 状态、合成代谢和分解代谢活动将克服 使用单一代谢分析的固有局限性。 初步研究表明，酸性pH最适(5.5-6.5)， 持续维护气态环境的要求 氧浓度低于环境(2.5-10%)代谢 维护和体外药敏试验 条件明显优于采用封闭系统的 PH为中性。这种方法应该逐步地导致 发展一种支持长期发展的媒介 麻风杆菌的维持和随后的真正生长。较少 那么完全的成功应该仍然可以显著改善 抗麻风杆菌药物活性的体外培养基 通过代谢活动的变化进行评估。

Studies will be carried out with inocula of Mycobacterium leprae, freshly harvested, from the infected foot pads of nu/nu mice and enriched for viable organisms, to explore optimal biophysical and nutritional parameters for the metabolic maintenance of the leprosy bacillus using the following combination of sensitive metabolic assays: a) quantitation of intracellular adenosine triphosphate (ATP), b) incorporation of 14C-palmitate into PGL-1, c) measurement of the rate of 14CO2 released from 14C- palmitate. The combined use of these indicators of net energy status, anabolic and catabolic activity will overcome the limitations inherent in using a single metabolic assay. Preliminary studies indicate an acidic pH optimum (5.5-6.5) and a requirement for continuous maintenance of a gaseous environment with an oxygen concentration below ambient (2.5-10%) Metabolic maintenance and in vitro drug susceptibility testing under these conditions are markedly superior to closed systems employing neutral pH. This approach should lead to the step-wise development of a medium which will support long-term maintenance and subsequently genuine growth of M. leprae. Less than total success should still allow significant improvement in an in vitro medium in which drug activity against M. leprae could be assessed via changes in metabolic activity.