Mukhopadhyay A, Mukhopadhyay S N, Talwar G P
National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India.
J Biotechnol. 1994 Aug 15;36(2):177-82. doi: 10.1016/0168-1656(94)90053-1.
Replication of vaccinia virus carrying beta-hCG gene at the 'tk' locus was studied in microcarrier culture of Vero cell. The yields of recombinant vaccinia virus and its expressed product (beta-hCG) were influenced by the physiological state of the host cell, at which it was infected. Cells infected with the virus between the 3rd and 6th day gave maximum amount of virus and beta-hCG in a parallel manner, the yields of which were 2.3 x 10(9) pfu per 10(6) cells and 450 ng per 10(6) cells, respectively. The yields were reduced by 33-36% in the virus-infected stationary culture of the 8th day. The physiological state of the host cell has been quantified by the cellular activity parameter 'P' (mu(t)/mu max) in the scale of 1 to 0. The value of 'P' was highest in the exponentially growing culture, which reduced with decline in growth rate. The cellular energy generation has been related with the parameter 'P'. ATP generation was reduced to 55% in the stationary culture on the 8th day (P = 0). Also, the adsorption of virus was reduced by nearly 55% in extremely slow growing or stationary culture. Host cells at moderate growth rate (P > or = 0.15) were found to be optimum for maximizing the yield of viral products.
在Vero细胞的微载体培养中研究了携带β-人绒毛膜促性腺激素（β-hCG）基因的痘苗病毒在“tk”位点的复制情况。重组痘苗病毒及其表达产物（β-hCG）的产量受宿主细胞被感染时的生理状态影响。在第3天至第6天之间被病毒感染的细胞以平行方式产生的病毒和β-hCG量最多，其产量分别为每10⁶个细胞2.3×10⁹个空斑形成单位（pfu）和每10⁶个细胞450纳克。在第8天病毒感染的静止培养物中，产量降低了33%至36%。宿主细胞的生理状态已通过细胞活性参数“P”（μ(t)/μ max）在1至0的范围内进行量化。“P”值在指数生长培养物中最高，随生长速率下降而降低。细胞能量产生与参数“P”相关。在第8天的静止培养物中，三磷酸腺苷（ATP）产生降低至55%（P = 0）。此外，在生长极慢或静止培养物中，病毒吸附减少了近55%。发现中等生长速率（P≥0.15）的宿主细胞最适合使病毒产物产量最大化。
