PMID- 27258267
OWN - NLM
STAT- MEDLINE
DCOM- 20170323
LR  - 20181113
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 17
IP  - 6
DP  - 2016 Jun 1
TI  - Stable Toll-Like Receptor 10 Knockdown in THP-1 Cells Reduces TLR-Ligand-Induced 
      Proinflammatory Cytokine Expression.
LID - 10.3390/ijms17060859 [doi]
LID - 859
AB  - Toll-like receptor 10 (TLR10) is the only orphan receptor whose natural ligand 
      and function are unknown among the 10 human TLRs. In this study, to test whether 
      TLR10 recognizes some known TLR ligands, we established a stable TLR10 knockdown 
      human monocytic cell line THP-1 using TLR10 short hairpin RNA lentiviral particle 
      and puromycin selection. Among 60 TLR10 knockdown clones that were derived from 
      each single transduced cell, six clones were randomly selected, and then one of 
      those clones, named E7, was chosen for the functional study. E7 exhibited 
      approximately 50% inhibition of TLR10 mRNA and protein expression. Of all the 
      TLRs, only the expression of TLR10 changed significantly in this cell line. 
      Additionally, phorbol 12-myristate 13-acetate-induced macrophage differentiation 
      of TLR10 knockdown cells was not affected in the knockdown cells. When exposed to 
      TLR ligands, such as synthetic diacylated lipoprotein (FSL-1), lipopolysaccharide 
      (LPS), and flagellin, significant induction of proinflammatory cytokine gene 
      expression including Interleukin-8 (IL-8), Interleukin-1 beta (IL-1beta), Tumor 
      necrosis factor-alpha (TNF-alpha) and Chemokine (C-C Motif) Ligand 20 (CCL20) 
      expression, was found in the control THP-1 cells, whereas the TLR10 knockdown 
      cells exhibited a significant reduction in the expression of IL-8, IL-1beta, and 
      CCL20. TNF-alpha was the only cytokine for which the expression did not decrease in 
      the TLR10 knockdown cells from that measured in the control cells. Analysis of 
      putative binding sites for transcription factors using a binding-site-prediction 
      program revealed that the TNF-alpha promoter does not have putative binding sites for 
      AP-1 or c-Jun, comprising a major transcription factor along with NF-kappaB for TLR 
      signaling. Our results suggest that TLR10 is involved in the recognition of 
      FSL-1, LPS, and flagellin and TLR-ligand-induced expression of TNF-alpha does not 
      depend on TLR10.
FAU - Le, Hai Van
AU  - Le HV
AD  - Department of Life Science, Gachon University, Seongnam, Kyeonggi-Do 461-701, 
      Korea. levan.hus@gmail.com.
FAU - Kim, Jae Young
AU  - Kim JY
AD  - Department of Life Science, Gachon University, Seongnam, Kyeonggi-Do 461-701, 
      Korea. jykim85@gachon.ac.kr.
LA  - eng
PT  - Journal Article
DEP - 20160601
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Cytokines)
RN  - 0 (Inflammation Mediators)
RN  - 0 (Ligands)
RN  - 0 (NF-kappa B)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Toll-Like Receptor 10)
RN  - 0 (Toll-Like Receptors)
RN  - 0 (Transcription Factor AP-1)
SB  - IM
MH  - Cell Line
MH  - Cytokines/genetics/*metabolism
MH  - Gene Expression Regulation
MH  - Gene Knockdown Techniques
MH  - Gene Silencing
MH  - Humans
MH  - Inflammation Mediators/*metabolism
MH  - *Ligands
MH  - Monocytes/metabolism
MH  - NF-kappa B/metabolism
MH  - Promoter Regions, Genetic
MH  - Protein Binding
MH  - RNA, Small Interfering/genetics
MH  - Toll-Like Receptor 10/*genetics/metabolism
MH  - Toll-Like Receptors/genetics/metabolism
MH  - Transcription Factor AP-1/metabolism
PMC - PMC4926393
OTO - NOTNLM
OT  - THP-1
OT  - knockdown
OT  - pro-inflammatory cytokines
OT  - toll-like receptor 10
EDAT- 2016/06/04 06:00
MHDA- 2017/03/24 06:00
PMCR- 2016/06/01
CRDT- 2016/06/04 06:00
PHST- 2016/02/10 00:00 [received]
PHST- 2016/05/24 00:00 [revised]
PHST- 2016/05/26 00:00 [accepted]
PHST- 2016/06/04 06:00 [entrez]
PHST- 2016/06/04 06:00 [pubmed]
PHST- 2017/03/24 06:00 [medline]
PHST- 2016/06/01 00:00 [pmc-release]
AID - ijms17060859 [pii]
AID - ijms-17-00859 [pii]
AID - 10.3390/ijms17060859 [doi]
PST - epublish
SO  - Int J Mol Sci. 2016 Jun 1;17(6):859. doi: 10.3390/ijms17060859.