Rowe W P, Baum S G, Pugh E, Hoggan M D
J Exp Med. 1965 Nov 1;122(5):943-54. doi: 10.1084/jem.122.5.943.
The titers of adenovirus and SV40 genetic carriers in hybrid preparations (E46(+)) can be quantitated by determining the percentage of cells showing neoantigens stainable with fluorescent antibody (FA) at 21 to 24 hours; both titers can be obtained with a single coverslip. The adenovirus and SV40 antigen-inducing titers so obtained are of the same order of magnitude in stock preparations of E46(+) grown in either African green monkey kidney (AGMK) or human embryonic kidney tissue culture (HEK), and are generally within one log of the infectivity titer. Quantitative studies of 50 degrees C heat inactivation, ultracentrifugation, and equilibrium density gradient centrifugation of E46(+) gave no indication that SV40 neoantigen induction could be dissociated from adenovirus, whereas adenovirus and SV40 virus grown as a mixed infection were readily dissociated by these procedures. Pretreatment of HEK cells with a medium containing 5-fluorouracil desoxyriboside (FUDR) did not affect induction of either adenovirus or SV40 neoantigen or development of cytopathic effects after infection with E46(+), but did prevent formation of Ad. 7 viral antigen.
杂种制剂(E46(+))中腺病毒和SV40基因载体的滴度可通过测定在21至24小时时显示可用荧光抗体(FA)染色的新抗原的细胞百分比来定量;两种滴度可通过单个盖玻片获得。在非洲绿猴肾(AGMK)或人胚肾组织培养(HEK)中生长的E46(+)储备制剂中,如此获得的腺病毒和SV40抗原诱导滴度处于相同数量级,并且通常在感染性滴度的一个对数范围内。对E46(+)进行50℃热灭活、超速离心和平衡密度梯度离心的定量研究表明,没有迹象表明SV40新抗原诱导可与腺病毒分离,而作为混合感染生长的腺病毒和SV40病毒很容易通过这些程序分离。用含有5-氟尿嘧啶脱氧核苷(FUDR)的培养基预处理HEK细胞,不影响E46(+)感染后腺病毒或SV40新抗原的诱导或细胞病变效应的发展,但确实可防止Ad. 7病毒抗原的形成。