PMID- 29698675
OWN - NLM
STAT- MEDLINE
DCOM- 20190111
LR  - 20190816
IS  - 1096-0007 (Electronic)
IS  - 0014-4835 (Linking)
VI  - 173
DP  - 2018 Aug
TI  - Human choroidal melanocytes express functional Toll-like receptors (TLRs).
PG  - 73-84
LID - S0014-4835(18)30063-0 [pii]
LID - 10.1016/j.exer.2018.04.014 [doi]
AB  - Toll-like receptors (TLRs) are a class of pattern recognition receptors that 
      sense highly conserved pathogen associated antigenic determinants, triggering an 
      innate immune response and subsequently instructing the adaptive immune system so 
      that together, the pathogen can be eliminated. TLRs are widely distributed in 
      human ocular tissues and cell types, and are active players in ocular 
      inflammation. To date, the presence and function of TLRs on human choroidal 
      melanocytes (HCMs), the most abundant choroidal cell type, have not been 
      characterized. The current study investigated the in vitro and in situ expression 
      and functional status of TLRs on HCMs. HCMs were isolated and cultured from 
      post-mortem human donor eyes, and displayed characteristic melanocyte morphology 
      and MART1 expression - a key melanocyte lineage marker up to passage 5 (P5). In 
      vitro experiments used P1 to P4 HCMs from different donor eyes. Initial 
      quantitative real-time PCR (qPCR) analysis revealed that HCMs (n = 3 donors) 
      expressed specific mRNA transcripts for TLR1-10 and MYD88 (a key adaptor protein 
      initiating the TLR signalling pathway). HCMs were stimulated with a set of 
      synthetic TLR specific agonists and the secretion of pro-inflammatory cytokines, 
      MCP-1 and IL-8, at 24 h measured by ELISA (n = 3 donors). The agonists Pam3CSK4 
      (TLR1/2), Poly I:C (TLR3), LPS (TLR4), Flagellin (TLR5), and FLS-1 (TLR2) induced 
      a significant increase in the production of MCP-1 and IL-8, compared to untreated 
      cells. Application of biotinylated Pam3CSK4 provided in vitro visualization of 
      receptor-agonist interactions for TLR1/2. We confirmed that cultured HCMs (n = 3 
      donors) expressed TLR1-6 protein using immunocytochemistry and confocal 
      microscopy. The expression and distribution of TLR 1-6 was also studied in human 
      choroid and retinal pigment epithelium (RPE) sections (n = 3 eyes) using 
      immunofluorescence and confocal microscopy. Strong TLR1-6 immunolabelling that 
      co-localized with melanocyte-dense areas (and RPE) was consistently observed; 
      intraluminal and blood vessel-related cells (including endothelial cells) also 
      expressed several TLRs. Taken together these observations show for the first time 
      that HCMs constitutively express a range of functional TLRs, and as such can 
      contribute to choroidal responses during infection and inflammation.
CI  - Copyright (c) 2018 Elsevier Ltd. All rights reserved.
FAU - Cioanca, Adrian V
AU  - Cioanca AV
AD  - School of Optometry and Vision Science, University of New South Wales, Sydney, 
      NSW 2052, Australia; Save Sight Institute, Discipline of Ophthalmology, 
      University of Sydney, Sydney NSW 2006, Australia. Electronic address: 
      valerin.cioanca@anu.edu.au.
FAU - McCluskey, Peter J
AU  - McCluskey PJ
AD  - Save Sight Institute, Discipline of Ophthalmology, University of Sydney, Sydney 
      NSW 2006, Australia. Electronic address: peter.mccluskey@sydney.edu.au.
FAU - Eamegdool, Steven S
AU  - Eamegdool SS
AD  - Save Sight Institute, Discipline of Ophthalmology, University of Sydney, Sydney 
      NSW 2006, Australia. Electronic address: seam6322@uni.sydney.edu.au.
FAU - Madigan, Michele C
AU  - Madigan MC
AD  - School of Optometry and Vision Science, University of New South Wales, Sydney, 
      NSW 2052, Australia; Save Sight Institute, Discipline of Ophthalmology, 
      University of Sydney, Sydney NSW 2006, Australia. Electronic address: 
      m.madigan@unsw.edu.au.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180423
PL  - England
TA  - Exp Eye Res
JT  - Experimental eye research
JID - 0370707
RN  - 0 (CCL2 protein, human)
RN  - 0 (CXCL8 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Interleukin-8)
RN  - 0 (MYD88 protein, human)
RN  - 0 (Myeloid Differentiation Factor 88)
RN  - 0 (RNA, Messenger)
RN  - 0 (Toll-Like Receptors)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Cells, Cultured
MH  - Chemokine CCL2/metabolism
MH  - Choroid/*metabolism
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Female
MH  - Gene Expression Regulation/*physiology
MH  - Humans
MH  - Interleukin-8/metabolism
MH  - Male
MH  - Melanocytes/*metabolism
MH  - Middle Aged
MH  - Myeloid Differentiation Factor 88/genetics
MH  - RNA, Messenger/genetics
MH  - Signal Transduction
MH  - Tissue Donors
MH  - Toll-Like Receptors/*genetics
OTO - NOTNLM
OT  - Human choroidal melanocytes
OT  - Infection
OT  - Inflammation
OT  - PAMPs
OT  - Pro-inflammatory cytokines
OT  - Toll-like receptors
EDAT- 2018/04/27 06:00
MHDA- 2019/01/12 06:00
CRDT- 2018/04/27 06:00
PHST- 2018/01/31 00:00 [received]
PHST- 2018/03/28 00:00 [revised]
PHST- 2018/04/21 00:00 [accepted]
PHST- 2018/04/27 06:00 [pubmed]
PHST- 2019/01/12 06:00 [medline]
PHST- 2018/04/27 06:00 [entrez]
AID - S0014-4835(18)30063-0 [pii]
AID - 10.1016/j.exer.2018.04.014 [doi]
PST - ppublish
SO  - Exp Eye Res. 2018 Aug;173:73-84. doi: 10.1016/j.exer.2018.04.014. Epub 2018 Apr 
      23.