BioNTech US, Inc., 40 Erie Street, Cambridge, MA 02139, USA.
Cell Rep Methods. 2023 Jan 13;3(1):100388. doi: 10.1016/j.crmeth.2022.100388. eCollection 2023 Jan 23.
CD4 T cells are critical to the immune system and perform multiple functions; therefore, their identification and characterization are crucial to better understanding the immune system in both health and disease states. However, current methods rarely preserve their phenotype, thus limiting our understanding of their functions. Here we introduce a flexible, rapid, and robust platform for CD4 T cell identification. By combining MHCII allele purification, allele-independent peptide loading, and multiplexed flow cytometry technologies, we can enable high-throughput personalized CD4 T cell identification, immunophenotyping, and sorting. Using this platform in combination with single-cell sorting and multimodal analyses, we identified and characterized antigen-specific CD4 T cells relevant to COVID-19 and cancer neoantigen immunotherapy. Overall, our platform can be used to detect and characterize CD4 T cells across multiple diseases, with potential to guide CD4 T cell epitope design for any disease-specific immunization strategy.
CD4 T 细胞对于免疫系统至关重要，具有多种功能；因此，对其进行鉴定和特征分析对于更好地了解健康和疾病状态下的免疫系统至关重要。然而，目前的方法很少能保留其表型，从而限制了我们对其功能的理解。在这里，我们介绍了一种灵活、快速和稳健的 CD4 T 细胞鉴定平台。通过结合 MHCII 等位基因纯化、与等位基因无关的肽加载和多色流式细胞术技术，我们可以实现高通量的个性化 CD4 T 细胞鉴定、免疫表型分析和分选。我们使用该平台与单细胞分选和多模态分析相结合，鉴定并表征了与 COVID-19 和癌症新抗原免疫治疗相关的抗原特异性 CD4 T 细胞。总体而言，我们的平台可用于检测和分析多种疾病中的 CD4 T 细胞，有可能为任何针对特定疾病的免疫策略指导 CD4 T 细胞表位设计。
