Identification of Tumoricidal TCRs from Tumor-Infiltrating Lymphocytes by Single-Cell Analysis

被引:31
作者
Shitaoka, Kiyomi [1 ]
Hamana, Hiroshi [1 ]
Kishi, Hiroyuki [1 ]
Hayakawa, Yoshihiro [2 ]
Kobayashi, Eiji [1 ]
Sukegawa, Kenta [1 ,3 ]
Piao, Xiuhong [1 ]
Lyu, Fulian [1 ]
Nagata, Takuya [3 ]
Sugiyama, Daisuke [4 ]
Nishikawa, Hiroyoshi [4 ,5 ]
Tanemura, Atsushi [6 ]
Katayama, Ichiro [6 ]
Murahashi, Mutsunori [7 ]
Takamatsu, Yasushi [8 ]
Tani, Kenzaburo [9 ]
Ozawa, Tatsuhiko [1 ]
Muraguchi, Atsushi [1 ]
机构
[1] Grad Sch Med & Pharmaceut Sci Med, Dept Immunol, Toyama, Japan
[2] Univ Toyama, Inst Nat Med, Div Pathogen Biochem, Toyama, Japan
[3] Grad Sch Med & Pharmaceut Sci Med, Dept Surg & Sci, Toyama, Japan
[4] Nagoya Univ, Grad Sch Med, Dept Immunol, Nagoya, Aichi, Japan
[5] Natl Canc Ctr, Div Canc Immunol, Exploratory Oncol Res & Clin Trial Ctr, Res Inst, Kashiwa, Chiba, Japan
[6] Osaka Univ, Grad Sch Med, Dept Dermatol, Suita, Osaka, Japan
[7] Kyushu Univ Hosp, Dept Adv Cell & Mol Therapy, Fukuoka, Japan
[8] Fukuoka Univ, Dept Internal Med, Div Med Oncol Hematol & Infect Dis, Fukuoka, Japan
[9] Univ Tokyo, Inst Med Sci, Project Div ALA Adv Med Res, Tokyo, Japan
关键词
RECEPTOR ALPHA-CHAIN; T-CELLS; BETA-CHAIN; MOUSE T; ANTIGEN; EXPRESSION; CANCER; PD-1; REPERTOIRE; REVEALS;
D O I
10.1158/2326-6066.CIR-17-0489
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
T-cell receptor (TCR) gene therapy is a promising next-generation antitumor treatment. We previously developed a single-T-cell analysis protocol that allows the rapid capture of paired TCR alpha and beta cDNAs. Here, we applied the protocol to analyze the TCR repertoire of tumor-infiltrating lymphocytes (TIL) of various cancer patients. We found clonally expanded populations of T cells that expressed the same clonotypic TCR in 50% to 70% of CD137(+)CD8(+) TILs, indicating that they responded to certain antigens in the tumor environment. To assess the tumor reactivity of the TCRs derived from those clonally expanded TILs in detail, we then analyzed the CD137(+)CD8(+) TILs from the tumor of B16F10 melanoma cells in six C57BL/6 mice and analyzed their TCR repertoire. We also found clonally expanded T cells in 60% to 90% of CD137(+)CD8(+) TILs. When the tumor reactivity of dominant clonotypic TCRs in each mouse was analyzed, 9 of 13 TCRs induced the secretion of IFN gamma in response to, and showed killing of, B16F10 cells in vitro, and 2 of them showed strong antitumor activity in vivo. Concerning their antigen specificity, 7 of them reacted to p15E peptide of endogenous murine leukemia virus-derived envelope glycoprotein 70, and the rest reacted to tumor-associated antigens expressed on EL4 lymphoma as well as B16 melanoma cells. These results show that our strategy enables us to simply and rapidly obtain the tumor-specific TCR repertoire with high fidelity in an antigen- and MHC haplotype-independent manner from primary TILs. (C) 2018 AACR.
引用
收藏
页码:378 / 388
页数:11
相关论文
共 39 条
[21]   Analysis of the Paired TCR α- and β-chains of Single Human T Cells [J].
Kim, Song-Min ;
Bhonsle, Latika ;
Besgen, Petra ;
Nickel, Jens ;
Backes, Anna ;
Held, Kathrin ;
Vollmer, Sigrid ;
Dornmair, Klaus ;
Prinz, Joerg C. .
PLOS ONE, 2012, 7 (05)
[22]   A new cloning and expression system yields and validates TCRs from blood lymphocytes of patients with cancer within 10 days [J].
Kobayashi, Eiji ;
Mizukoshi, Eishiro ;
Kishi, Hiroyuki ;
Ozawa, Tatsuhiko ;
Hamana, Hiroshi ;
Nagai, Terumi ;
Nakagawa, Hidetoshi ;
Jin, Aishun ;
Kaneko, Shuichi ;
Muraguchi, Atsushi .
NATURE MEDICINE, 2013, 19 (11) :1542-+
[23]   Autoimmunity against a defective ribosomal insulin gene product in type 1 diabetes [J].
Kracht, Maria J. L. ;
van Lummel, Menno ;
Nikolic, Tatjana ;
Joosten, Antoinette M. ;
Laban, Sandra ;
van der Slik, Arno R. ;
van Veelen, Peter A. ;
Carlotti, Francoise ;
de Koning, Eelco J. P. ;
Hoeben, Rob C. ;
Zaldumbide, Arnaud ;
Roep, Bart O. .
NATURE MEDICINE, 2017, 23 (04) :501-+
[24]   Immune Checkpoint Inhibitors: An Introduction to the Next-Generation Cancer Immunotherapy [J].
Lee, Lucy ;
Gupta, Manish ;
Sahasranaman, Srikumar .
JOURNAL OF CLINICAL PHARMACOLOGY, 2016, 56 (02) :157-169
[25]   DERIVATION OF A T-CELL HYBRIDOMA VARIANT DEPRIVED OF FUNCTIONAL T-CELL RECEPTOR ALPHA-CHAIN AND BETA-CHAIN TRANSCRIPTS REVEALS A NONFUNCTIONAL ALPHA-MESSENGER-RNA OF BW5147 ORIGIN [J].
LETOURNEUR, F ;
MALISSEN, B .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1989, 19 (12) :2269-2274
[26]   High-throughput identification of antigen-specific TCRs by TCR gene capture [J].
Linnemann, Carsten ;
Heemskerk, Bianca ;
Kvistborg, Pia ;
Kluin, Roelof J. C. ;
Bolotin, Dmitriy A. ;
Chen, Xiaojing ;
Bresser, Kaspar ;
Nieuwland, Marja ;
Schotte, Remko ;
Michels, Samira ;
Gomez-Eerland, Raquel ;
Jahn, Lorenz ;
Hombrink, Pleun ;
Legrand, Nicolas ;
Shu, Chengyi Jenny ;
Mamedov, Ilgar Z. ;
Velds, Arno ;
Blank, Christian U. ;
Haanen, John B. A. G. ;
Turchaninova, Maria A. ;
Kerkhoven, Ron M. ;
Spits, Hergen ;
Hadrup, Sine Reker ;
Heemskerk, Mirjam H. M. ;
Blankenstein, Thomas ;
Chudakov, Dmitriy M. ;
Bendle, Gavin M. ;
Schumacher, Ton N. M. .
NATURE MEDICINE, 2013, 19 (11) :1534-+
[27]   Cancer regression in patients after transfer of genetically engineered lymphocytes [J].
Morgan, Richard A. ;
Dudley, Mark E. ;
Wunderlich, John R. ;
Hughes, Marybeth S. ;
Yang, James C. ;
Sherry, Richard M. ;
Royal, Richard E. ;
Topalian, Suzanne L. ;
Kammula, Udai S. ;
Restifo, Nicholas P. ;
Zheng, Zhili ;
Nahvi, Azam ;
de Vries, Christiaan R. ;
Rogers-Freezer, Linda J. ;
Mavroukakis, Sharon A. ;
Rosenberg, Steven A. .
SCIENCE, 2006, 314 (5796) :126-129
[28]   Identification of broadly conserved cross-species protective Leishmania antigen and its responding CD4+ T cells [J].
Mou, Zhirong ;
Li, Jintao ;
Boussoffara, Thouraya ;
Kishi, Hiroyuki ;
Hamana, Hiroshi ;
Ezzati, Peyman ;
Hu, Chuanmin ;
Yi, Weijing ;
Liu, Dong ;
Khadem, Forough ;
Okwor, Ifeoma ;
Jia, Ping ;
Shitaoka, Kiyomi ;
Wang, Shufeng ;
Ndao, Momar ;
Petersen, Christine ;
Chen, Jianping ;
Rafati, Sima ;
Louzir, Hechmi ;
Muraguchi, Atsushi ;
Wilkins, John A. ;
Uzonna, Jude E. .
SCIENCE TRANSLATIONAL MEDICINE, 2015, 7 (310)
[29]   Isolation of T-Cell Receptors Specifically Reactive with Mutated Tumor-Associated Antigens from Tumor-Infiltrating Lymphocytes Based on CD137 Expression [J].
Parkhurst, Maria ;
Gros, Alena ;
Pasetto, Anna ;
Prickett, Todd ;
Crystal, Jessica S. ;
Robbins, Paul ;
Rosenberg, Steven A. .
CLINICAL CANCER RESEARCH, 2017, 23 (10) :2491-2505
[30]   Comprehensive assessment of T-cell receptor β-chain diversity in αβ T cells [J].
Robins, Harlan S. ;
Campregher, Paulo V. ;
Srivastava, Santosh K. ;
Wacher, Abigail ;
Turtle, Cameron J. ;
Kahsai, Orsalem ;
Riddell, Stanley R. ;
Warren, Edus H. ;
Carlson, Christopher S. .
BLOOD, 2009, 114 (19) :4099-4107