新冠病毒特异性TCR 及其表位鉴定

新冠肺炎全球爆发，严重危害人类健康。虽然多种药物和疫苗已经投
入使用，但新出现的新冠病毒突变株能够逃逸最初感染或疫苗接种所建立
的中和抗体。研究发现，大部分自然感染或疫苗接种诱导的特异性T 细胞
应答能够维持对突变株的反应性，防止突变株感染造成的重症化或者死亡，
提示了特异性T 细胞反应的免疫保护作用。因此，新冠病毒特异性T 细胞
受体（T cell receptor，TCR）及其抗原表位的鉴定，将有助于更加深入地
理解新冠病毒特异性的T 细胞反应，为未来新冠疫苗设计提供更多参考依
据。
本课题通过分析六例新冠病人的支气管肺泡灌洗液的单细胞TCR 组库
测序数据，选取了每个病例中克隆数最多的一个TCR，将其体外表达在已敲
除内源性TCR 的J76 细胞表面，最后使用多肽库鉴定了TCR 的抗原特异性
及其表位。前期，我们利用两个已知表位的特异性TCR 成功构建了一个TCR
体外表达平台。我们通过在J76 细胞中过表达CD8 分子以及NFAT 启动子
控制下的Luciferase 元件优化该平台。我们应用此平台鉴定出了一个新冠病
人支气管肺泡灌洗液来源的新冠特异性TCR C152-1，并筛选到其表位为新
冠病毒N 蛋白的N81 和N82。
特异性TCR 的鉴定将为新冠病毒的TCR-T 免疫细胞治疗提供重要依据，
而抗原表位的鉴定可以为疫苗的设计提供重要参考，提高疫苗对人群的广谱
保护效力。

The emerging severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2) threatens public health. Although a variety of drugs and vaccines
have been approved, the SARS-CoV-2 variants have been shown to escape from
neutralizing antibody responses elicited by current vaccines or infection. Recent
studies have shown that the SARS-CoV-2-specific T cell responses induced by
the initial infection or vaccination could maintain responsiveness to variants and
provide protection against severe SARS-CoV-2 infection, indicating an
immunoprotective effect of SARS-CoV-2-specific T cells. The identification of
SARS-CoV-2-specific TCRs and their epitopes will contribute to a deeper
understanding of the SARS-CoV-2-specific T cell response, which will guide the
vaccine design in the future.
Here we analyzed the single-cell TCR repertoire sequencing data of
bronchoalveolar lavage fluid (BALF) from six COVID-19 patients. We first
selected the TCRs with the largest clone. Then, the TCRs were expressed on the
surface of J76 cells with knockout of TCR. Finally, we explored TCR specificity,
followed by screening for the epitope using peptide pool. Previoudly, we
constructed an in vitro TCR expression platform through utilizing two
SARS-CoV-2-specific TCRs with known epitopes. We optimized this TCR
expression platform by over-expressing CD8 molecules and a NFAT
promoter-Luciferase element in J76 cell line. Eventually, we identified a
SARS-CoV-2-specific TCR C152-1 from BALF of a COVID-19 patient and
screened out N81 and N82 as the epitopes of this TCR.
The discovery of elite SARS-CoV-2-specific TCRs can facilitate the
development of TCR-T immunotherapy for COVID-19. The identification of
conserved epitopes present in variants can provide more information for
broad-spectrum vaccine design, improving vaccination effectsin the population.
 

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