Development of a next-generation sequencing protocol for the canine T cell receptor beta chain repertoire

IF 1.4 3区农林科学 Q4 IMMUNOLOGY

Veterinary immunology and immunopathology Pub Date : 2023-12-12 DOI:10.1016/j.vetimm.2023.110702

Cindy L. Zuleger , Rene Welch Schwartz , Irene M. Ong , Michael A. Newton , David M. Vail , Mark R. Albertini

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Abstract

Profiling the T cell receptor (TCR) repertoire using next-generation sequencing has become common in both human and translational research. Companion dogs with spontaneous tumors, including canine melanoma, share several features, e.g., natural occurrence, shared environmental exposures, natural outbred population, and immunocompetence. T cells play an important role in the adaptive immune system by recognizing specific antigens via a surface TCR. As such, understanding the canine T cell response to vaccines, cancer, immunotherapies, and infectious diseases is critically important for both dog and human health. Off-the-shelf commercial reagents, kits and services are readily available for human, non-human primate, and mouse in this context. However, these resources are limited for the canine. In this study, we present a cost-effective protocol for analysis of canine TCR beta chain genes. Workflow can be accomplished in 1–2 days starting with total RNA and resulting in libraries ready for sequencing on Illumina platforms.

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为犬类 T 细胞受体 beta 链序列制定下一代测序方案

在人类研究和转化研究中，使用新一代测序技术分析 T 细胞受体 (TCR) 基因库已变得十分普遍。患有自发性肿瘤（包括犬黑色素瘤）的伴侣犬有几个共同的特征，如自然发生、共同的环境暴露、天然外交种群和免疫能力。T 细胞通过表面 TCR 识别特定抗原，在适应性免疫系统中发挥着重要作用。因此，了解犬 T 细胞对疫苗、癌症、免疫疗法和传染病的反应对犬和人类的健康都至关重要。在这方面，现成的商业试剂、试剂盒和服务可随时用于人类、非人灵长类动物和小鼠。然而，这些资源对于犬类来说却十分有限。在本研究中，我们提出了一种经济有效的犬 TCR beta 链基因分析方案。从总 RNA 开始，工作流程可在 1-2 天内完成，并生成可在 Illumina 平台上测序的文库。

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来源期刊

Veterinary immunology and immunopathology 农林科学-免疫学

CiteScore

3.40

自引率

5.60%

发文量

审稿时长

70 days

期刊介绍： The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.