利用核苷类逆转录酶抑制剂去除疫苗毒种中禽网状内皮组织增生症病毒污染

Eradication of REV contamination in attenuated vaccines seed by nucleoside reverse transcriptase inhibitors

使用污染有禽网状内皮组织增生症病毒(REV)的弱毒疫苗被认为是REV流行的原因之一,去除疫苗毒种中REV污染是生产合格疫苗的基础环节,本研究试图利用核苷类逆转录酶抑制剂类药物Azidothymidine(LAM)和Lamivudine(AZT)去除疫苗毒种中REV污染。首先通过在培养液中添加不同质量浓度AZT或LAM连续传代以及利用CCK-8测定细胞活性的方法,证实了在培养液中添加不超过5mg/L的抑制剂类药物对于DF-1细胞复制和活性没有影响。体外试验证实,当在培养液中分别添加5mg/L的AZT或LAM以及两者联合应用时均可以显著抑制REV在DF-1细胞上的复制,通过在新城疫弱毒疫苗中人为添加REV模拟AZT或LAM以及两者联合应用对疫苗毒种中REV污染的净化作用,结果显示经药物干预4代后即将REV去除,并且净化后毒种检验合格。本研究证实了AZT或LAM在体外对REV的抑制作用,并在人工模拟试验中成功净化了疫苗毒种中污染的REV。

REV is the most frequent exogenous virus that contaminates attenuated vaccines and the vaccine seed without REV contamination is the foundation for vaccine manufacture, the research attempted to eradicate the REV contamination in vaccine seed by the suppress of these drugs. Seri- al passages of immortalized chicken embryo fibroblast DF-1 cells were performed in the presence of different concentrations of azidothymidine（AZT） or lamivudine（LAM）, and cell activity was de- tected by a cell counting kit-8（CCK-8）. It was found that at quality concentrations less than 5 mg/L AZT or LAM had no effect on DF-1 cell replication. Compared with the control group, 5 mg/L AZT or LAM significantly inhibited REV replication in DF-1 cells. The REV contamination in NDV vaccine seed was eradicate after a four passage under 5 mg/L AZT or LAM or combination with 3 mg/L AZT and LAM. This method will facilitate the eradication of REV contamination of attenuated vaccine seed.