High-purity preparation of HSV-2 vaccine candidate ACAM529 is immunogenic and efficacious in vivo.
High-purity preparation of HSV-2 vaccine candidate ACAM529 is immunogenic and efficacious in vivo.
复制标题
DOI：
10.1371/journal.pone.0057224
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Anderson SF
中科院分区：
文献类型：
--
作者：
Mundle ST;Hernandez H;Hamberger J;Catalan J;Zhou C;Stegalkina S;Tiffany A;Kleanthous H;Delagrave S;Anderson SF
生殖器疱疹是由单纯疱疹病毒2型（HSV-2）和单纯疱疹病毒1型（HSV-1）引起的性传播感染（STI）。HSV-2感染是终身的，尽管该疾病具有高度流行的性质，但与医疗保健系统的重大成本和社会耻辱相关。例如，HSV-2血清阳性与血清阴性成人的比例在美国约为1/5，在撒哈拉以南非洲的一些地区大于4/5。使用适用于GMP生产的细胞重新衍生复制缺陷型疫苗株病毒dl 5 -29，并将其重命名为ACAM 5 - 29。先前报道用dl 5 -29免疫在小鼠和豚鼠中均具有保护性，然而这些研究是用使用不能用于临床材料生产的方法纯化的疫苗进行的。在这里，我们描述的方法，作为一个主要的步骤，准备ACAM 529可能是适合在人类中测试。ACAM 529可以从反式互补细胞系AV 529克隆19（AV 529 -19）的感染细胞培养物中收获，而无需机械细胞破碎。然后可以通过新的纯化方案相对于宿主细胞DNA和蛋白质纯化ACAM 529，所述新的纯化方案包括内切核酸酶处理、深层过滤、阴离子交换色谱和超滤/渗滤（UF/DF）的组合。所得病毒保留感染性，并且相对于宿主细胞DNA和蛋白质，其纯度比通过超离心纯化的ACAM 529高200倍。此外，我们描述了一个并排的比较层析纯化的ACAM 529与蔗糖琼脂糖纯化的ACAM 529，这表明，这两种制剂是同样的免疫原性和保护性，当在体内测试。
Genital herpes is a sexually transmitted infection (STI) caused by herpes simplex virus 2 (HSV-2) and to a lesser extent herpes simplex virus 1 (HSV-1). Infection by HSV-2 is life-long and is associated with significant cost to healthcare systems and social stigma despite the highly prevalent nature of the disease. For instance, the proportion of HSV-2 seropositive to seronegative adults is approximately 1 in 5 in the US and greater than 4 in 5 in some areas of sub-Saharan Africa. The replication-defective vaccine strain virus dl5-29 was re-derived using cells appropriate for GMP manufacturing and renamed ACAM529. Immunization with dl5-29 was previously reported to be protective both in mice and in guinea pigs, however these studies were performed with vaccine that was purified using methods that cannot be scaled for manufacturing of clinical material. Here we describe methods which serve as a major step towards preparation of ACAM529 which may be suitable for testing in humans. ACAM529 can be harvested from infected cell culture of the trans-complementing cell line AV529 clone 19 (AV529-19) without mechanical cell disruption. ACAM529 may then be purified with respect to host cell DNA and proteins by a novel purification scheme, which includes a combination of endonuclease treatment, depth filtration, anion-exchange chromatography and ultrafiltration/diafiltration (UF/DF). The resultant virus retains infectivity and is ∼ 200-fold more pure with respect to host cell DNA and proteins than is ACAM529 purified by ultracentrifugation. Additionally, we describe a side-by-side comparison of chromatography-purified ACAM529 with sucrose cushion-purified ACAM529, which shows that both preparations are equally immunogenic and protective when tested in vivo.
登录
查看更多内容
影响因子：
6.4
作者：
Hoshino, Yo;Pesnicak, Lesley;Cohen, Jeffrey I.
通讯作者：
Cohen, Jeffrey I.
影响因子：
3.8
作者：
Jiang, Canping;Ataai, Mohammad;Glorioso, Joseph C.
通讯作者：
Glorioso, Joseph C.
影响因子：
--
作者：
Cattamanchi, Ashok;Posavad, Christine M.;Koelle, David M.
通讯作者：
Koelle, David M.
DOI：
10.1016/0732-8893(88)90015-6
发表时间：
1988-11-01
影响因子：
2.9
作者：
ARENS, M;SWIERKOSZ, E;DILWORTH, V
通讯作者：
DILWORTH, V
影响因子：
3.1
作者：
Fisman, DN;Lipsitch, M;Goldie, SJ
通讯作者：
Goldie, SJ