符合药品生产质量管理规范的严重急性呼吸综合征冠状病毒2细胞毒性T淋巴细胞的制造与表征

Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes.

作者信息

Chu Yaya, Milner Jordan, Lamb Margaret, Maryamchik Elena, Rigot Olivia, Ayello Janet, Harrison Lauren, Shaw Rosemarie, Behbehani Gregory K, Mardis Elaine R, Miller Katherine, Prakruthi Rao Venkata Lakshmi, Chang Hsiaochi, Lee Dean, Rosenthal Elana, Kadauke Stephan, Bunin Nancy, Talano Julie-An, Johnson Bryon, Wang Yongping, Cairo Mitchell S

机构信息

Department of Pediatrics, New York Medical College, Valhalla, New York, USA.

Department of Hematology/Oncology/Bone Marrow Transplant, Center for Childhood Cancer and Blood Diseases, Nationwide Children's Hospital, Columbus, Ohio, USA.

出版信息

J Infect Dis. 2023 Mar 28;227(6):788-799. doi: 10.1093/infdis/jiac500.

DOI:10.1093/infdis/jiac500

PMID:36583990

Abstract

BACKGROUND

Coronavirus 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).

METHODS

Donor 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.

RESULTS

Of 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 TEMRA cell subsets. SARS-CoV-2-vCTLs were polyfunctional with increased gene expression in T-cell function, interleukin, pathogen defense, and tumor necrosis factor superfamily pathways.

CONCLUSIONS

Highly functional SARS-CoV-2-vCTLs can be rapidly generated by direct cytokine enrichment (12 hours) from convalescent donors.

CLINICAL TRIALS REGISTRATION

NCT04896606.

摘要

背景

2019冠状病毒病（COVID-19）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淋巴细胞（vCTLs）可能为COVID-19治疗提供一种有前景的方法。我们旨在使用CliniMACS细胞因子捕获系统（CCS）筛选、制备和鉴定从COVID-19康复者供体中产生的SARS-CoV-2-vCTLs。

方法

通过用病毒肽刺激COVID-19康复者供体的外周血单个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。使用CliniMACS CCS制备临床级SARS-CoV-2-vCTLs。通过T细胞受体测序、质谱流式细胞术和转录组分析对富集的SARS-CoV-2-vCTLs进行表征。

结果

在康复者供体血样中，93%通过了临床制备的筛选标准。三次验证运行产生了富集的T细胞，其中79%（平均标准误差21%）为IFN-γ+ T细胞。SARS-CoV-2-vCTLs显示出高度多样化的T细胞受体库，记忆性CD8和CD4 T细胞均有增加，尤其是在CD8 TEM、CD4 TCM和CD4 TEMRA细胞亚群中。SARS-CoV-2-vCTLs具有多功能性，在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。