PMID- 30777078
OWN - NLM
STAT- MEDLINE
DCOM- 20200401
LR  - 20200401
IS  - 1479-5876 (Electronic)
IS  - 1479-5876 (Linking)
VI  - 17
IP  - 1
DP  - 2019 Feb 18
TI  - High throughput sequencing of T-cell receptor repertoire using dry blood spots.
PG  - 47
LID - 10.1186/s12967-019-1796-4 [doi]
LID - 47
AB  - BACKGROUND: Immunology research, particularly next generation sequencing (NGS) of 
      the immune T-cell receptor beta (TCRbeta) repertoire, has advanced progression in 
      several fields, including treatment of various cancers and autoimmune diseases. 
      This study aimed to identify the TCR repertoires from dry blood spots (DBS), a 
      method that will help collecting real-world data for biomarker applications. 
      METHODS: Finger-prick blood was collected onto a Whatman filter card. RNA was 
      extracted from DBS of the filter card, and fully automated multiplex PCR was 
      performed to generate a TCRbeta chain library for next generation sequencing (NGS) 
      analysis of unique CDR3s (uCDR3). RESULTS: We demonstrated that the dominant 
      clonotypes from the DBS results recapitulated those found in whole blood. 
      According to the statistical analysis and laboratory confirmation, 40 of 2-mm 
      punch disks from the filter cards were enough to detect the shared top clones and 
      have strong correlation in the uCDR3 discovery with whole blood. uCDR3 discovery 
      was neither affected by storage temperatures (room temperature versus - 20  degrees C) 
      nor storage durations (1, 14, and 28 days) when compared to whole blood. About 
      74-90% of top 50 uCDR3 clones of whole blood could also be detected from DBS. A 
      low rate of clonotype sharing, 0.03-1.5%, was found among different individuals. 
      CONCLUSIONS: The DBS-based TCR repertoire profiling method is minimally invasive, 
      provides convenient sampling, and incorporates fully automated library 
      preparation. The system is sensitive to low RNA input, and the results are highly 
      correlated with whole blood uCDR3 discovery allowing study scale-up to better 
      understand the relationship and mutual influences between the immune and 
      diseases.
FAU - Wu, Shang-Gin
AU  - Wu SG
AD  - Department of Internal Medicine, National Taiwan University Hospital, National 
      Taiwan University, Taipei, 10002, Taiwan.
AD  - Department of Internal Medicine, National Taiwan University Cancer Center, 
      National Taiwan University, Taipei, 10672, Taiwan.
FAU - Pan, Wenjing
AU  - Pan W
AD  - HudsonAlpha Institute for Biotechnology, 601 Genome Way, Huntsville, AL, 35806, 
      USA.
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA.
FAU - Liu, Hongna
AU  - Liu H
AD  - iCubate Inc., Huntsville, AL, 35806, USA.
FAU - Byrne-Steele, Miranda L
AU  - Byrne-Steele ML
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA.
FAU - Brown, Brittany
AU  - Brown B
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA.
FAU - Depinet, Mollye
AU  - Depinet M
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA.
FAU - Hou, Xiaohong
AU  - Hou X
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA.
FAU - Han, Jian
AU  - Han J
AD  - HudsonAlpha Institute for Biotechnology, 601 Genome Way, Huntsville, AL, 35806, 
      USA. jhan@irepertoire.com.
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA. 
      jhan@irepertoire.com.
FAU - Li, Song
AU  - Li S
AD  - iRepertoire Inc., 800 Hudson Way Suite 2319, Huntsville, AL, 35806, USA. 
      sli@irepertoire.com.
LA  - eng
PT  - Journal Article
DEP - 20190218
PL  - England
TA  - J Transl Med
JT  - Journal of translational medicine
JID - 101190741
RN  - 0 (Complementarity Determining Regions)
RN  - 0 (Receptors, Antigen, T-Cell, alpha-beta)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Complementarity Determining Regions/genetics
MH  - Dried Blood Spot Testing/*methods
MH  - High-Throughput Nucleotide Sequencing/*methods
MH  - Humans
MH  - Preservation, Biological
MH  - RNA/isolation & purification
MH  - Receptors, Antigen, T-Cell, alpha-beta/*genetics
MH  - Temperature
PMC - PMC6379990
OTO - NOTNLM
OT  - CDR3
OT  - Diversity
OT  - Dry blood spots
OT  - Next-generation sequence
OT  - T cell receptor
OT  - TCR repertoire
EDAT- 2019/02/20 06:00
MHDA- 2020/04/02 06:00
PMCR- 2019/02/18
CRDT- 2019/02/20 06:00
PHST- 2019/01/08 00:00 [received]
PHST- 2019/02/09 00:00 [accepted]
PHST- 2019/02/20 06:00 [entrez]
PHST- 2019/02/20 06:00 [pubmed]
PHST- 2020/04/02 06:00 [medline]
PHST- 2019/02/18 00:00 [pmc-release]
AID - 10.1186/s12967-019-1796-4 [pii]
AID - 1796 [pii]
AID - 10.1186/s12967-019-1796-4 [doi]
PST - epublish
SO  - J Transl Med. 2019 Feb 18;17(1):47. doi: 10.1186/s12967-019-1796-4.