Pieniazek D, Pieniazek N J, Macejak D, Luftig R B
Department of Microbiology, Immunology and Parasitology, Louisiana State University Medical Center, New Orleans 70112-1393.
Virology. 1990 Sep;178(1):72-80. doi: 10.1016/0042-6822(90)90380-a.
It had been postulated that due to lack of growth of enteric adenovirus 41 (Ad41) on human primary cells and its growth on Graham-293 cells there was a defect in the Ad41 E1A region. However, we found as a result of careful evaluation of Ad41 growth on several primary cell lines (HEK, WI-38, or Detroit 551) that efficient viral multiplication is possible if the serum concentration in the medium used postinfection (p.i.) is kept between 0.2 and 1%. In contrast, only slight growth of Ad41 occurs in infected cells maintained in 5% serum and virtually no viral replication is found in infected cells cultivated in medium with 10% serum. The serum inhibitory effect appears limited to primary cells because no difference in Ad41 replication, as assayed by accumulation of Ad41 DNA, was found in infected continuous cell lines (HEp-2, 293) cultivated p.i. in either 1 or 10% FBS. Also, this effect appears specific for enteric adenoviruses, such as Ad41, since conventional adenoviruses, such as Ad5, grow well in both 1 and 10% FBS. The above results show that Ad41 can grow in a variety of primary cell lines, under specific culture conditions. In addition, we found that Ad41-infected primary cells grown in medium containing 0.2% serum had an increase in synthesis of the 70-kDa heat shock protein (HSP70) at about 6 hr p.i. and also Ad41 was able to complement the Ad5 E1A deletion mutant dl312. These results show that the E1A function of Ad41 is not impaired in infected cells.
据推测,由于肠道腺病毒41型(Ad41)在人原代细胞上无法生长,而能在Graham-293细胞上生长,所以Ad41的E1A区域存在缺陷。然而,我们通过仔细评估Ad41在几种原代细胞系(HEK、WI-38或Detroit 551)上的生长情况发现,如果感染后(p.i.)所用培养基中的血清浓度保持在0.2%至1%之间,病毒就能高效增殖。相比之下,在5%血清中培养的感染细胞中,Ad41只有轻微生长,而在10%血清培养基中培养的感染细胞中几乎没有病毒复制。血清抑制作用似乎仅限于原代细胞,因为在感染后于1%或10%胎牛血清(FBS)中培养的连续细胞系(HEp-2、293)中,通过检测Ad41 DNA积累量发现,Ad41复制没有差异。此外,这种作用似乎对肠道腺病毒如Ad41具有特异性,因为传统腺病毒如Ad5在1%和10% FBS中都能良好生长。上述结果表明,在特定培养条件下,Ad41能在多种原代细胞系中生长。此外,我们发现,在含有0.2%血清的培养基中生长的Ad41感染的原代细胞,在感染后约6小时,70 kDa热休克蛋白(HSP70)的合成增加,而且Ad41能够互补Ad5 E1A缺失突变体dl312。这些结果表明,Ad41在感染细胞中的E1A功能没有受损。