PMID- 36466875
OWN - NLM
STAT- MEDLINE
DCOM- 20221206
LR  - 20221206
IS  - 1664-3224 (Electronic)
IS  - 1664-3224 (Linking)
VI  - 13
DP  - 2022
TI  - Rapid identification of CMV-specific TCRs via reverse TCR cloning system based on 
      bulk TCR repertoire data.
PG  - 1021067
LID - 10.3389/fimmu.2022.1021067 [doi]
LID - 1021067
AB  - Advances in next-generation sequencing (NGS) have improved the resolution of 
      T-cell receptor (TCR) repertoire analysis, and recent single-cell sequencing has 
      made it possible to obtain information about TCR pairs. In our previous study, 
      cytomegalovirus (CMV) pp65-specific T-cell response restricted by a single human 
      leukocyte antigen (HLA) class I allotype was observed in an individual. 
      Therefore, to effectively clone an antigen-specific TCR from these T cells, we 
      developed a TCR cloning system that does not require a single cell level. First, 
      we established the improved Jurkat reporter cell line, which was TCRalphabeta double 
      knock-out and expressed CD8alphabeta molecules. Furthermore, functional TCRs were 
      directly obtained by reverse TCR cloning using unique CDR3-specific PCR primers 
      after bulk TCR sequencing of activation marker-positive CD8 T cells by NGS. A 
      total of 15 TCRalpha and 14 TCRbeta strands were successfully amplified by PCR from cDNA 
      of 4-1BB-positive CD8 T cells restricted by HLA-A*02:01, HLA-A*02:06, 
      HLA-B*07:02, and HLA-B*40:06. The panels with combinations of TCRalpha and TCRbeta genes 
      were investigated using Jurkat reporter cell line and artificial 
      antigen-presenting cells (APCs). In two TCR pairs restricted by HLA-A*02:01, one 
      TCR pair by HLA-A*02:06, four TCR pairs by HLA-B*07:02, and one TCR pair by 
      HLA-B*40:06, their specificity and affinity were confirmed. The TCR pair of 
      A*02:01/1-1 showed alloreactivity to HLA-A*02:06. The one TCR pair showed a 
      higher response to the naturally processed antigen than that of the peptide pool. 
      This reverse TCR cloning system will not only provide functional information to 
      TCR repertoire analysis by NGS but also help in the development of TCR-T therapy.
CI  - Copyright (c) 2022 Hong, Pyo, Baek and Kim.
FAU - Hong, Cheol-Hwa
AU  - Hong CH
AD  - Department of Biomedicine and Health Sciences, College of Medicine, The Catholic 
      University of Korea, Seoul, South Korea.
AD  - Department of Microbiology, College of Medicine, The Catholic University of 
      Korea, Seoul, South Korea.
AD  - Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic 
      University of Korea, Seoul, South Korea.
FAU - Pyo, Hong-Seon
AU  - Pyo HS
AD  - Department of Biomedicine and Health Sciences, College of Medicine, The Catholic 
      University of Korea, Seoul, South Korea.
AD  - Department of Microbiology, College of Medicine, The Catholic University of 
      Korea, Seoul, South Korea.
FAU - Baek, In-Cheol
AU  - Baek IC
AD  - Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic 
      University of Korea, Seoul, South Korea.
FAU - Kim, Tai-Gyu
AU  - Kim TG
AD  - Department of Microbiology, College of Medicine, The Catholic University of 
      Korea, Seoul, South Korea.
AD  - Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic 
      University of Korea, Seoul, South Korea.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221118
PL  - Switzerland
TA  - Front Immunol
JT  - Frontiers in immunology
JID - 101560960
RN  - 0 (Receptors, Antigen, T-Cell)
SB  - IM
MH  - Humans
MH  - *Receptors, Antigen, T-Cell/genetics
MH  - Cell Membrane
MH  - DNA Repair
MH  - Cloning, Molecular
MH  - *Cytomegalovirus Infections
PMC - PMC9716090
OTO - NOTNLM
OT  - CMV
OT  - NGS - next generation sequencing
OT  - TCR - T cell receptor
OT  - artificial APC
OT  - reverse TCR cloning
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2022/12/06 06:00
MHDA- 2022/12/07 06:00
PMCR- 2022/01/01
CRDT- 2022/12/05 04:13
PHST- 2022/08/17 00:00 [received]
PHST- 2022/10/31 00:00 [accepted]
PHST- 2022/12/05 04:13 [entrez]
PHST- 2022/12/06 06:00 [pubmed]
PHST- 2022/12/07 06:00 [medline]
PHST- 2022/01/01 00:00 [pmc-release]
AID - 10.3389/fimmu.2022.1021067 [doi]
PST - epublish
SO  - Front Immunol. 2022 Nov 18;13:1021067. doi: 10.3389/fimmu.2022.1021067. 
      eCollection 2022.