肿瘤抗原肽的HLA限制性确认和诱导反应性T细胞杀瘤能力评估*

doi:10.3969/j.issn.1671-2587.2024.04.001

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (4): 433-439.DOI: 10.3969/j.issn.1671-2587.2024.04.001

• 论著 • 下一篇

肿瘤抗原肽的HLA限制性确认和诱导反应性T细胞杀瘤能力评估^*

杨颖¹, 李子涵², 丁旭苹³, 张嘉敏¹, 李勤¹, 郭忠慧¹, 赵俸涌¹, 杨启修¹, 王晨¹, 路丽明³, 朱自严¹

¹上海市血液中心,上海 200051;
²上海交通大学医学院附属胸科医院,上海 200030;
³上海交通大学医学院,上海 200025

收稿日期:2024-06-25 出版日期:2024-08-20 发布日期:2024-09-23
通讯作者: 杨颖,主要从事输血医学、免疫和微生物学、免疫血液学等领域的研究,探讨输血、干细胞移植、细胞治疗等方面的新疗法、疗效评估和副反应预防及其机制,（E-mail）yangyinghla@hotmail.com。
作者简介:杨颖,主要从事输血医学、免疫和微生物学、免疫血液学等领域的研究,探讨输血、干细胞移植、细胞治疗等方面的新疗法、疗效评估和副反应预防及其机制,（E-mail）yangyinghla@hotmail.com。
基金资助:
*本课题受中国输血协会威高基金重点基金（No.CSBT-MWG-2020-01）资助

Identification of HLA Restriction for Neoantigens and Evaluation of Anti-tumor Ability of Neoantigen Induced Reactive T cells

YANG Ying¹, LI Zihan², DING Xuping³, ZHANG Jiamin¹, LI Qin¹, GUO Zhonghui¹, ZHAO Fengyong¹, YANG Qixiu¹, WANG Chen¹, LU Liming³, ZHU Ziyan¹

¹Shanghai Blood Center, Shanghai, 200051;
²Shanghai Chest Hospital, Shanghai Jiaotong University, 200030;
³Shanghai Jiaotong University, School of Medicine, Shanghai 200025

Received:2024-06-25 Online:2024-08-20 Published:2024-09-23

摘要/Abstract

摘要： 目的了解肿瘤抗原肽的HLA限制性以及其诱导的T细胞能否杀伤肿瘤细胞,探索无关供者来源细胞用于过继性抗肿瘤T细胞治疗。方法使用16个肿瘤抗原肽诱导18名无关供者外周血单个核细胞（PBMC）分化为反应性T细胞,并分析HLA型别;利用NetMHC数据库预测肽和HLA分子亲和力;选择HLA-A2限制性的肿瘤抗原肽诱导第二组17名无关供者的PBMC进行杀瘤实验,反应性T细胞作为效应细胞,T2细胞及肿瘤抗原肽同源的肿瘤细胞作为靶细胞,测量LDH(乳酸脱氢酶)释放量或者RTCA(实时无标记细胞分析仪)检测效应细胞杀瘤效率,比较HLA-A2+和A2-T细胞杀瘤效率。结果筛出和HLA-A2具有高亲和力的肿瘤抗原肽LM7,可诱导5/11 HLA-A2+为反应性T细胞,其中HLA-A2+纯合子则为3/3,而HLA-A2-者则为2/7。LM7诱导反应性T细胞杀伤肿瘤百分比A2+组明显强于A2-组（60.72±11.28 vs 47.2±4.46,P=0.03）。结论本研究显示NetMHC预测对于纯合子样品更有帮助,肿瘤抗原肽LM7具有HLA-A2限制性,可诱导部分HLA-A2+PBMC分化为反应性T细胞,可杀伤肿瘤,应对供者进行HLA筛选并分析其细胞功能,其诱导的反应性T细胞可作为过继性T细胞抗肿瘤治疗的细胞来源。

关键词: HLA, 限制性, 肿瘤新抗原, 反应性T细胞, 过继性T细胞免疫治疗

Abstract: Objectives To explore the possibility to utilize unrelated donors' cells as sources for adoptive T cell anti-tumor therapy, thus we need to identify HLA restriction for neoantigens and evaluate neoantigen induced reactive T cells' cytolysis ability. Methods 16 neoantigen peptides were used to induce 18 unrelated donors' (group 1) PBMC (peripheral blood mononuclear cells) into reactive T cells, and HLA typing was performed, the affinity between peptides' and HLA molecule was predicted by NetMHC database, then neoantigen with HLA-A2 restriction was selected to induce another group (group 2) of 17 unrelated donors' PBMC into reactive T cells, these cells would act as effectors, tumor cell line T2 cells or the tumor cells with the same source of this neoantigen would act as targets either, LDH release tests and RTCA (real time celluar analysis) were performed to evaluate the effectors' anti-tumor ability. Then these results were compared between HLA-A2+ and HLA-A2- samples. Results NetMHC database predicted neoantigen LM7 showed high affinity with HLA-A2+ antigens, 5/11 of HLA-A2+ samples can successfully be induced into reactive T cells, including 3/3 homozygous HLA-A2+ samples, while 2/7 of HLA-A2- samples with more reactive T cells. Induced T cells from A2+ samples showed higher percentages of tumor cells killed than those A2- samples(60.72±11.28 vs 47.2± 4.46, P=0.03). Conclusions Our study suggests prediction by NetMHC is more helpful in homozygous samples, neoantigens LM7 is confirmed as HLA-A2 restrictive, which can induce some HLA-A2+ PBMC into reactive T cells which can kill tumor cells more efficiently, unrelated donors should be screened not only by HLA-typing but also cell function tested, thus induced reactive T cells can take the roles as the source for adoptive anti-tumor T cells therapy.

Key words: HLA restriction, Neoantigen, Reactive T cell, Adoptive T cell therapy

中图分类号:

R446

杨颖, 李子涵, 丁旭苹, 张嘉敏, 李勤, 郭忠慧, 赵俸涌, 杨启修, 王晨, 路丽明, 朱自严. 肿瘤抗原肽的HLA限制性确认和诱导反应性T细胞杀瘤能力评估^*[J]. 临床输血与检验, 2024, 26(4): 433-439.

YANG Ying, LI Zihan, DING Xuping, ZHANG Jiamin, LI Qin, GUO Zhonghui, ZHAO Fengyong, YANG Qixiu, WANG Chen, LU Liming, ZHU Ziyan. Identification of HLA Restriction for Neoantigens and Evaluation of Anti-tumor Ability of Neoantigen Induced Reactive T cells[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(4): 433-439.

参考文献

[1] KENNEDY L B,SALAMA A K S. A review of cancer immunotherapy toxicity[J]. CA Cancer J Clin,2020,70(2):86-104.
[2] ROSENBERG S A,RESTIFO N P.Adoptive cell transfer as personalized immunotherapy for human cancer[J]. Science,2015,348(6230):62-68.
[3] ROSENBERG S A,RESTIFO N P,YANG J C,et al.Adoptive cell transfer:a clinical path to effective cancer immunotherapy[J]. Nat Rev Cancer,2008,8(4):299-308.
[4] GROSS G,WAKS T,ESHHAR Z.Expression of immunoglobulin-T-cell receptor chimeric molecules as functional receptors with antibody-type specificity[J]. Proc Natl Acad Sci U S A,1989,86(24):10024-10028.
[5] IMAI C,MIHARA K,ANDREANSKY M,et al.Chimeric receptors with 4-1BB signaling capacity provoke potent cytotoxicity against acute lymphoblastic leukemia[J]. Leukemia,2004,18(4):676-684.
[6] SHARPE A H,PAUKEN K E.The diverse functions of the PD1 inhibitory pathway[J]. Nat Rev Immunol,2018,18(3):153-167.
[7] OTT P A,HU-LIESKOVAN S,CHMIELOWSKI B,et al. A phase ib trial of personalized neoantigen therapy plus anti-PD-1 in patients with advanced melanoma,non-small cell lung cancer,or bladder cancer[J]. Cell,2020,183(2):347-362.e24.
[8] KAST F,KLEIN C,UMAÑA P,et al. Advances in identification and selection of personalized neoantigen/T-cell pairs for autologous adoptive T cell therapies[J]. Oncoimmunology,2021,10(1):1869389.
[9] HU Z T,OTT P A,WU C J.Towards personalized,tumour-specific,therapeutic vaccines for cancer[J]. Nat Rev Immunol,2018,18(3):168-182.
[10] LEKO V,ROSENBERG S A.Identifying and targeting human tumor antigens for T cell-based immunotherapy of solid tumors[J]. Cancer Cell,2020,38(4):454-472.
[11] DAVIS L,TARDUNO A,LU Y C.Neoantigen-reactive T cells:the driving force behind successful melanoma immunotherapy[J]. Cancers (Basel),2021,13(23):6061.
[12] ANAGNOSTOU V,SMITH K N,FORDE P M,et al.Evolution of neoantigen landscape during immune checkpoint blockade in non-small cell lung cancer[J]. Cancer Discov,2017,7(3):264-276.
[13] CHEN F J,ZOU Z Y,DU J,et al.Neoantigen identification strategies enable personalized immunotherapy in refractory solid tumors[J]. J Clin Invest,2019,129(5):2056-2070.
[14] HUANG R C,ZHAO B,HU S,et al.Adoptive neoantigen-reactive T cell therapy:improvement strategies and current clinical researches[J]. Biomark Res,2023,11(1):41.
[15] OTT P A,HU Z T,KESKIN D B,et al.An immunogenic personal neoantigen vaccine for patients with melanoma[J]. Nature,2017,547(7662):217-221.
[16] LIU C X,SHAO J,DONG Y B,et al.Advanced HCC patient benefit from neoantigen reactive T cells based immunotherapy:a case report[J]. Front Immunol,2021,12:685126.
[17] ZACHARAKIS N,CHINNASAMY H,BLACK M,et al.Immune recognition of somatic mutations leading to complete durable regression in metastatic breast cancer[J]. Nat Med,2018,24(6):724-730.
[18] PARKHURST M R,ROBBINS P F,TRAN E,et al.Unique neoantigens arise from somatic mutations in patients with gastrointestinal cancers[J]. Cancer Discov,2019,9(8):1022-1035.
[19] 杨颖,路丽明,李勤,等.肿瘤新抗原肽诱导献血者PBMC特异反应性T细胞[J].临床输血与检验,2022,24(5):565-573.
[20] CHEN Y,XUE S A,BEHBOUDI S,et al.Ex vivo PD-L1/PD-1 pathway blockade reverses dysfunction of circulating CEA-specific T cells in pancreatic cancer patients[J]. Clin Cancer Res,2017,23(20):6178-6189.
[21] JENSEN K K,ANDREATTA M,MARCATILI P,et al.Improved methods for predicting peptide binding affinity to MHC class II molecules[J]. Immunology,2018,154(3):394-406.
[22] HOOF I,PETERS B,SIDNEY J,et al.NetMHCpan,a method for MHC class I binding prediction beyond humans[J]. Immunogenetics,2009,61(1):1-13.
[23] JURTZ V,PAUL S,ANDREATTA M,et al.NetMHCpan-4.0:improved peptide-MHC class I interaction predictions integrating eluted ligand and peptide binding affinity data[J]. J Immunol,2017,199(9):3360-3368.
[24] ATZIN-MÉNDEZ J A,LÓPEZ-GONZÁLEZ J S,BÁEZ R,et al. Expansion of quiescent lung adenocarcinoma CD8+ T cells by MUC1-8-mer peptide-T2 cell-β2 microglobulin complexes[J]. Oncol Rep,2016,35(1):33-42.
[25] BOSSI G,GERRY A B,PASTON S J,et al.Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells[J]. Oncoimmunology,2013,2(11):e26840.
[26] RESTIFO N P,DUDLEY M E,ROSENBERG S A.Adoptive immunotherapy for cancer:harnessing the T cell response[J]. Nat Rev Immunol,2012,12(4):269-281.
[27] DE MATTOS-ARRUDA L,VAZQUEZ M,FINOTELLO F,et al. Neoantigen prediction and computational perspectives towards clinical benefit:recommendations from the ESMO Precision Medicine Working Group[J]. Ann Oncol,2020,31(8):978-990.

肿瘤抗原肽的HLA限制性确认和诱导反应性T细胞杀瘤能力评估^*

Identification of HLA Restriction for Neoantigens and Evaluation of Anti-tumor Ability of Neoantigen Induced Reactive T cells

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