Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes.
Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes.
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符合良好生产规范的 SARS-COV-2 细胞毒性 T 淋巴细胞的制造和表征。
DOI：
10.1093/infdis/jiac500
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Rosenthal
中科院分区：
文献类型：
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作者：
Chu,Yaya;Milner,Jordan;Lamb,Margaret;Maryamchik,Elena;Rigot,Olivia;Ayello,Janet;Harrison,Lauren;Shaw,Rosemarie;Behbehani,GregoryK;Mardis,ElaineR;Miller,Katherine;PrakruthiRaoVenkata,Lakshmi;Chang,Hsiaochi;Lee,Dean;Rosenthal
背景2019 年冠状病毒病 (COVID-19) 是由严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 引起。 SARS-CoV-2 病毒特异性细胞毒性 T 细胞淋巴细胞 (vCTL) 可以为 COVID-19 治疗提供一种有前途的方式。我们的目的是使用 CliniMACS 细胞因子捕获系统 (CCS) 筛选、制造和表征恢复期 COVID-19 供体产生的 SARS-CoV-2–vCTL。方法通过用病毒肽刺激恢复期 COVID-19 供体外周血单核细胞并使用流式细胞术鉴定干扰素 γ (IFN-γ)+CD4 和 CD8 T 细胞来进行供体筛选。临床级 SARS-CoV-2–vCTL 使用 CliniMACS CCS 制造。通过T细胞受体测序、质谱流式分析和转录组分析对富集的SARS-CoV-2-vCTL进行表征。结果恢复期供血样本中，93%通过了临床生产的筛选标准。三个验证运行产生了富集的 T 细胞，其中 79%（平均值 21% 的标准误差）为 IFN-γ+T 细胞。 SARS-CoV-2-vCTL 显示出高度多样化的 T 细胞受体库，记忆 CD8 和 CD4 T 细胞均增强，特别是在 CD8 TEM、CD4 TCM 和 CD4 TEMRA 细胞亚群中。 SARS-CoV-2–vCTL 具有多功能性，T 细胞功能、白细胞介素、病原体防御和肿瘤坏死因子超家族途径中的基因表达增加。结论通过恢复期供体的直接细胞因子富集（12 小时）可以快速生成高功能的 SARS-CoV-2–vCTL。临床试验注册 NCT04896606。
BackgroundCoronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 virus-specific cytotoxic T-cell lymphocytes (vCTLs) could provide a promising modality in COVID-19 treatment. We aimed to screen, manufacture, and characterize SARS-CoV-2–vCTLs generated from convalescent COVID-19 donors using the CliniMACS Cytokine Capture System (CCS).MethodsDonor screening was done by stimulation of convalescent COVID-19 donor peripheral blood mononuclear cells with viral peptides and identification of interferonγ (IFN-γ)+CD4 and CD8 T cells using flow cytometry. Clinical-grade SARS-CoV-2–vCTLs were manufactured using the CliniMACS CCS. The enriched SARS-CoV-2–vCTLs were characterized by T-cell receptor sequencing, mass cytometry, and transcriptome analysis.ResultsOf the convalescent donor blood samples, 93% passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T cells that were 79% (standard error of the mean 21%) IFN-γ+T cells. SARS-CoV-2–vCTLs displayed a highly diverse T-cell receptor repertoire with enhancement of both memory CD8 and CD4 T cells, especially in CD8 TEM,CD4 TCM, and CD4 TEMRAcell subsets. SARS-CoV-2–vCTLs were polyfunctional with increased gene expression in T-cell function, interleukin, pathogen defense, and tumor necrosis factor superfamily pathways.ConclusionsHighly functional SARS-CoV-2–vCTLs can be rapidly generated by direct cytokine enrichment (12 hours) from convalescent donors.Clinical Trials RegistrationNCT04896606.