Studies on the Collection of Leprosy Bacilli from Infected Tissues for Metabolic Experiments

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A simple and rapid method for the collection of leprosy bacilli from infected tissues was studied to obtain the purified bacilli available to the metabolic investigations.The results are summarized as follows:Murine leprosy bacilli were collected from leprous spleen of mice by repeated centrifugations, followed by the treatment of crude bacillary fraction with water adjusted to pH 10.5 and shaking at 37°C for 60 min. The bacilli obtained by this method were found to be almost free from host tissue components by the test for the acid phosphatase and β-glucuronidase activities which are involved in lysosomes of host tissues, in addition to the microscopic and electron microscopic analyses.The bacillary preparations thus obtained showed high activities of mycobacterial enzymes such as FAD dependent NADPH-cyt. c reductase and NADH-cyt. c reductase.The human leprosy bacilli, collected from leproma of armadillo by this method, were also pure microscopically and enzymatically as above and showed to maintain the activity to incorporate label from L-methionine-14CH3S into bacterial lipids.From these results, the influence of the alkaline treatments employed for the collection of bacilli from infected tissues on bacterial enzyme activities was discussed.
日本ハンセン病学会の論文