Steiniger Sebastian C J, Glanville Jacob, Harris Douglas W, Wilson Thomas L, Ippolito Gregory C, Dunham Steven A
Zoetis VMRD Global Therapeutics Research, 333 Portage Street, Kalamazoo, MI 49001, USA.
Distributed Bio Inc, San Francisco, CA, USA.
Biologicals. 2017 Mar;46:81-87. doi: 10.1016/j.biologicals.2017.01.004. Epub 2017 Jan 26.
Next-Generation Sequencing combined with bioinformatics is a powerful tool for analyzing the large number of DNA sequences present in the expressed antibody repertoire and these data sets can be used to advance a number of research areas including antibody discovery and engineering. The accurate measurement of the immune repertoire sequence composition, diversity and abundance is important for understanding the repertoire response in infections, vaccinations and cancer immunology and could also be useful for elucidating novel molecular targets. In this study 4 individual domestic cats (Felis catus) were subjected to antibody repertoire sequencing with total number of sequences generated 1079863 for VH for IgG, 1050824 VH for IgM, 569518 for VK and 450195 for VL. Our analysis suggests that a similar VDJ expression patterns exists across all cats. Similar to the canine repertoire, the feline repertoire is dominated by a single subgroup, namely VH3. The antibody paratope of felines showed similar amino acid variation when compared to human, mouse and canine counterparts. All animals show a similarly skewed VH CDR-H3 profile and, when compared to canine, human and mouse, distinct differences are observed. Our study represents the first attempt to characterize sequence diversity in the expressed feline antibody repertoire and this demonstrates the utility of using NGS to elucidate entire antibody repertoires from individual animals. These data provide significant insight into understanding the feline immune system function.
下一代测序技术与生物信息学相结合是分析表达的抗体库中大量DNA序列的有力工具,这些数据集可用于推进包括抗体发现和工程在内的多个研究领域。准确测量免疫库序列组成、多样性和丰度对于理解感染、疫苗接种和癌症免疫学中的库反应很重要,也可能有助于阐明新的分子靶点。在本研究中,对4只家猫(Felis catus)进行了抗体库测序,生成的IgG的VH序列总数为1079863条,IgM的VH序列为1050824条,VK序列为569518条,VL序列为450195条。我们的分析表明,所有猫都存在相似的VDJ表达模式。与犬类库相似,猫类库由单个亚组主导,即VH3。与人类、小鼠和犬类对应物相比,猫的抗体互补决定区显示出相似的氨基酸变异。所有动物都表现出相似的VH CDR-H3谱倾斜,与犬类、人类和小鼠相比,观察到明显差异。我们的研究首次尝试表征表达的猫抗体库中的序列多样性,这证明了使用二代测序技术来阐明单个动物的整个抗体库的实用性。这些数据为理解猫免疫系统功能提供了重要见解。