Le Doux J M, Morgan J R, Snow R G, Yarmush M L
Department of Chemical and Biochemical Engineering, Rutgers University, Piscataway, New Jersey 08854, USA.
J Virol. 1996 Sep;70(9):6468-73. doi: 10.1128/JVI.70.9.6468-6473.1996.
Using a model recombinant retrovirus encoding the Escherichia coli lacZ gene, we have found that medium conditioned with NIH 3T3 cells and packaging cell lines derived from NIH 3T3 cells inhibits infection. Most of the inhibitory activity was greater than 100 kDa and was sensitive to chondroitinase ABC digestion, which is consistent with the inhibitor being a chondroitin sulfate proteoglycan. Proteoglycans secreted by NIH 3T3 cells and purified by anion-exchange chromatography inhibited amphotropic retrovirus infection. Pretreatment of amphotropic retrovirus stocks with chondroitinase ABC boosted the level of transduction efficiency by more than twofold. The implications of these findings with respect to retrovirus-cell interactions and the production of high-titer retroviral stocks are discussed.
利用一种编码大肠杆菌β-半乳糖苷酶基因的重组逆转录病毒模型，我们发现用NIH 3T3细胞和源自NIH 3T3细胞的包装细胞系处理过的培养基会抑制感染。大部分抑制活性大于100 kDa，且对软骨素酶ABC消化敏感，这与该抑制剂为硫酸软骨素蛋白聚糖一致。NIH 3T3细胞分泌并经阴离子交换色谱法纯化的蛋白聚糖可抑制双嗜性逆转录病毒感染。用软骨素酶ABC预处理双嗜性逆转录病毒储备液可使转导效率提高两倍以上。本文讨论了这些发现对于逆转录病毒与细胞相互作用以及高滴度逆转录病毒储备液生产的意义。
