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首页> 外文期刊>Journal of microbiology and biotechnology >Continuous Passaging of a Recombinant C-Strain Virus in PK-15 Cells Selects Culture-Adapted Variants that Showed Enhanced Replication but Failed to Induce Fever in Rabbits
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Continuous Passaging of a Recombinant C-Strain Virus in PK-15 Cells Selects Culture-Adapted Variants that Showed Enhanced Replication but Failed to Induce Fever in Rabbits

机译：在PK-15细胞中的重组C-菌株病毒的连续传递选择培养适应的变体，该变体显示出增强的复制，但未能诱导兔子发烧

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摘要

Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that RecCpp80(+10) failed to induce fever in rabbits, whereas RecCpp40(+10) caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.
机译：典型的猪瘟病毒（CSFV）是古典猪瘟的病因，这是一种高度传染性的疾病，导致猪业的重大经济损失。逐渐利用对CSFV的疫苗菌株，对CSFV的广泛使用疫苗菌株具有低的细胞培养效率，这限制了疫苗工业的生产率。在该研究中，构建了衍生自C-菌株的重组病毒，并在PK-15细胞中进行连续传代，其目的是获得高后代病毒产量。细胞适应的病毒变体RECCPP80比其亲本C-菌株更高1,000倍，但损失了诱导兔子发烧的能力。细胞适应的RECC变体的序列分析表明，在RECCPP80中固定至少六个核苷酸变化。在猪睾丸细胞中的RECCPP80变体的进一步适应导致较高的病毒产量而没有额外的突变。将这些残留物中的每一个引入野生型RECC主干显示中，位于E2的C末端区域中的一个突变M979R（T3310G）可能与细胞适应表型密切相关。兔接种显示Reccpp80（+10）未诱导兔子发烧，而RECCPP40（+10）导致与商业C-菌株疫苗相似的发热反应。总之，通过在其E2蛋白中引入特异性突变，C-菌株可以适应细胞培养。 RECCPP80中的突变导致兔子中发热反应丧失需要进一步调查。 PK-15细胞中的基于C菌株的重组病毒的连续传递可以增强其体外适应。 3310和3531的非同义突变可能在常规病毒繁殖的增强能力中发挥重要作用。这种发现可以帮助设计改进的C菌株，以提高商业疫苗的生产率降低。

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  • 作者

    Tong Chao; Chen Ning; Liao Xun; Yuan Xuemei; Sun Mengjiao; Li Xiaoliang; Fang Weihuan;

  • 作者单位

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

    Zhejiang Univ Inst Prevent Vet Med Zhejiang Prov Key Lab Prevent Vet Med Hangzhou 310058 Zhejiang Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;
  • 关键词

    Classical swine fever; C-strain; cell culture adaptation; fever in rabbits;

    机译：古典猪瘟;c-菌株;细胞培养适应;兔子发烧;

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