Nowotny A, Keler T, Pham P H, Kovats E, Aiello A, Shonekan O, Friedman H, Johnson A G
T.W. Evans Museum and Dental Institute, University of Pennsylvania, Philadelphia 19104.
J Biol Response Mod. 1988 Jun;7(3):296-308.
White-type polysaccharide preparation (WPS) obtained from Serratia marcescens bacteria by hot 0.2 N acetic acid extraction was shown to have antitumor effects. These were manifested by enhanced resistance to the take of TA3 transplantable murine adenocarcinoma and by the induction of regression of Meth A sarcoma in mice. Optimal conditions for the liberation and isolation of these substances were sought to achieve the highest antitumor activity and the lowest endotoxin (ET) content. Simultaneously, the activities of the WPS preparations were tested in various tests which are frequently used as in vitro correlates of in vivo antitumor effects, such as the activation of macrophage cytotoxicity, activation of natural killer (NK) cells, and tumor necrosis factor (TNF) generation. We found that the enhanced resistance to the take of TA3 tumor correlated with ET content of the WPS preparations. Preparations with reduced or no ET content showed diminishing activity in this assay or were without any measurable effect. The induction of TNF production and NK activation did not show such close relationship with the ET content. This was particularly evident if testing WPS samples obtained after 60 or 120 min hydrolysis at 90 degrees C. The greatest discrepancy was found between ET content and the Meth A regression induction. Samples with no detectable ET content and no activity in the macrophage, NK, or TNF tests were potent inducers of Meth A regression. Partial purification of such WPS samples could be achieved and a preparation was obtained with high Meth A regression capacity. Preliminary chemical analysis of this preparation showed 25.5% amino acid, 53.7% neutral carbohydrate, less than 0.4% KDO, 0.8% hexosamine, less than 0.1% phosphorous, and less 1.0% long-chain carboxylic acid content. The above chemical analytical data are not consistent with designating such preparations as ET or ET derivatives, such as Lipid A or its split products. This conclusion was confirmed by the lack of endotoxic properties as determined by biological assays on this preparation.
通过用0.2N热乙酸从粘质沙雷氏菌中提取获得的白色型多糖制剂（WPS）显示出具有抗肿瘤作用。这些作用表现为对TA3可移植性小鼠腺癌接种的抵抗力增强，以及诱导小鼠Meth A肉瘤消退。为了实现最高的抗肿瘤活性和最低的内毒素（ET）含量，人们寻求这些物质释放和分离的最佳条件。同时，在各种经常用作体内抗肿瘤作用体外相关指标的试验中测试了WPS制剂的活性，例如巨噬细胞细胞毒性的激活、自然杀伤（NK）细胞的激活以及肿瘤坏死因子（TNF）的产生。我们发现对TA3肿瘤接种抵抗力的增强与WPS制剂的ET含量相关。ET含量降低或没有ET含量的制剂在该试验中活性降低或没有任何可测量的效果。TNF产生的诱导和NK激活与ET含量没有如此密切的关系。如果测试在90℃水解60或120分钟后获得的WPS样品，这一点尤为明显。在ET含量和Meth A消退诱导之间发现了最大差异。在巨噬细胞、NK或TNF试验中没有可检测到的ET含量且没有活性的样品是Meth A消退的有效诱导剂。可以实现此类WPS样品的部分纯化，并获得具有高Meth A消退能力的制剂。对该制剂的初步化学分析显示其氨基酸含量为25.5%，中性碳水化合物含量为53.7%，KDO含量小于0.4%，己糖胺含量为0.8%，磷含量小于0.1%，长链羧酸含量小于1.0%。上述化学分析数据与将此类制剂指定为ET或ET衍生物（如脂多糖A或其裂解产物）不一致。通过对此制剂进行生物测定确定其缺乏内毒素特性，这证实了这一结论。
