Department of Microbiology and Immunology, University of Melbourne, at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
World Health Organization (WHO) Collaborating Centre for Reference and Research on Influenza, The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
PLoS One. 2020 May 29;15(5):e0233794. doi: 10.1371/journal.pone.0233794. eCollection 2020.
The domestic ferret (Mustela putorius furo) provides a critical animal model to study human respiratory diseases. However immunological insights are restricted due to a lack of ferret-specific reagents and limited genetic information about ferret B and T cell receptors. Here, variable, diversity and joining genes within the ferret kappa, lambda and heavy chain immunoglobulin loci were annotated using available genomic information. A multiplex PCR approach was derived that facilitated the recovery of paired heavy and light chain immunoglobulin sequences from single sorted ferret B cells, allowing validation of predicted germline gene sequences and the identification of putative novel germlines. Eukaryotic expression vectors were developed that enabled the generation of recombinant ferret monoclonal antibodies. This work advances the ferret as an informative immunological model for viral diseases by allowing the in-depth interrogation of antibody-based immunity.
家兔（Mustela putorius furo）为研究人类呼吸道疾病提供了重要的动物模型。然而，由于缺乏针对雪貂的试剂以及有关雪貂 B 和 T 细胞受体的遗传信息有限，因此免疫方面的研究受到限制。在这里，利用现有基因组信息对雪貂κ、λ和重链免疫球蛋白基因座内的可变、多样性和连接基因进行了注释。开发了一种多重 PCR 方法，该方法可从单个分选的雪貂 B 细胞中回收配对的重链和轻链免疫球蛋白序列，从而验证预测的胚系基因序列并鉴定可能的新胚系。还开发了真核表达载体，能够生成重组雪貂单克隆抗体。这项工作通过允许深入研究基于抗体的免疫反应，推动了家兔作为一种有意义的病毒性疾病免疫模型。
