CULTIVATION OF VIRULENT TREPONEMA-PALLIDUM IN TISSUE-CULTURE
CULTIVATION OF VIRULENT TREPONEMA-PALLIDUM IN TISSUE-CULTURE
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DOI：
10.1128/iai.32.2.908-915.1981
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发表时间：
1981-01-01
影响因子：
3.1
通讯作者：
MOECKLI, RA
中科院分区：
文献类型：
--
作者：
FIELDSTEEL, AH;COX, DL;MOECKLI, RA
在一系列的7个实验中，T.在1.5%O2的气氛下，在常规单层培养中生长的棉尾兔上皮组织培养细胞（Sf 1 Ep）的表面上显示出苍白球附着和复制。接种106 T.梅毒螺旋体中，密螺旋体的数量增加到8 × 10 - 6之间。106和2.59 ×107.收获的微生物的活力范围为86- 97%。T.苍白球持续增加，通常在孵育的第9-12天之间达到稳定水平，增加范围高达100倍，平均49倍。似乎存在约2倍的倍增上限。108，不考虑接种物，其范围为106-108 T.苍白球同时对含T. pallidum以获得复制的进一步证据。当接种量为106-107时，观察到密螺旋体DNA显著增加，如预期的那样，在前一组中增加最大。每个密螺旋体的DNA量也有极好的相关性; 106个密螺旋体的平均值为2.5 × 106。106和107组的平均值为3.46 × 106。10-14，3.28 ×10-14和2.79 ×10-14 g/密螺旋体（3.14 ±. 0.72. times. 10-14 g/密螺旋体）。在每个实验中，从用106 T.在孵育7天后，对苍白球进行培养以测试毒力。在所有情况下，微生物都是有毒的;平均6-7个微生物产生肿胀、硬化、含密螺旋体的病变。扫描电子显微镜显示，在复制过程中，在细胞表面形成许多密螺旋体的小菌落。
In a series of 7 experiments, the virulent Nichols strain of T. pallidum was shown to attach and replicate on the surface of tissue culture cells of cottontail rabbit epithelium (Sf1Ep) growing in conventional monolayer cultures under an atmosphere of 1.5% O2. Five days after inoculation of 106 T. pallidum, the number of treponemes had increased to between 8 .times. 106 and 2.59 .times. 107. The viability of harvested organisms ranged from 86-97%. The number of T. pallidum continued to increase, generally reaching a plateau between days 9-12 of incubation, with increases ranging up to 100-fold and averaging 49-fold. There appeared to be a ceiling of multiplication of about 2 .times. 108 irrespective of the inoculum, which ranged from 106-108 T. pallidum. Concurrent DNA assays were performed on the cultures containing T. pallidum to obtain further evidence of replication. Significant increases in treponemal DNA were observed when the inocula ranged from 106-107, with the greatest increases, as might be expected, in the former group. There was also excellent correlation in the amount of DNA per treponeme; the averages for the 106, 2.5 .times. 106, and 107 groups were 3.46 .times. 10-14, 3.28 .times. 10-14 and 2.79 .times. 10-14 g/treponeme, respectively (3.14 .+-. 0.72 .times. 10-14 g/treponeme). In each experiment, organisms were harvested from the group inoculated with 106 T. pallidum after 7 days of incubation to test for virulence. In all instances, the organisms were virulent; erythematous, indurated, treponeme-containing lesions were produced from an average of 6-7 organisms. Scanning electron microscopy revealed that during the course of replication many microcolonies of treponemes formed on the surface of the cells.