PMID- 35183540
OWN - NLM
STAT- MEDLINE
DCOM- 20220427
LR  - 20220506
IS  - 1096-0007 (Electronic)
IS  - 0014-4835 (Linking)
VI  - 218
DP  - 2022 May
TI  - Complement activation by RPE cells preexposed to TNFalpha and IFNgamma.
PG  - 108982
LID - S0014-4835(22)00063-X [pii]
LID - 10.1016/j.exer.2022.108982 [doi]
AB  - Age-related macular degeneration (AMD) has been associated with both complement 
      activation and increased levels of circulating cytokines. Here, we sougth to 
      investigate if cytokine-preexposure of retinal pigment epithelial (RPE) leads to 
      increased complement activation and deposition of membrane attack complex (MAC). 
      Primary human RPE and the ARPE19 cell line cultured in serum-free conditions were 
      preexposed to 100 ng/ml interferon-gamma (IFNgamma) and 20 ng/ml tumor necrosis 
      factor-alpha (TNFalpha) for 48 h followed by exposure to diluted serum from healthy 
      donors or complement factor B deficient (CFBd) serum for 70 min. Deposition of 
      membrane attack complexes (MAC) was examined by use of a MAC-ELISA kit and by 
      immunofluorescence. Eculizumab (anti-C5) was examined for its ability to prevent 
      deposition of MAC on RPE cells exposed to serum. Lactatdehydrogenase (LDH) and 
      thiazolyl blue tetrazolium bromide (MTT) assays were used to assess cellular 
      metabolism and survival. MAC was deposited only on RPE preexposed to both IFNgamma 
      and TNFalpha. Lack of complement factor B or inhibition of C5 abrogated the 
      MAC-deposition on RPE cells, while reconstitution of CFBd serum with CFB resulted 
      in MAC-deposition. MAC-deposition resulted in RPE-release of LDH, but unaltered 
      mitochondrial activity estimated by MTT. We conclude that preexposure of primary 
      RPE and ARPE19 with inflammatory cytokines promoted alternative pathway 
      activation of complement and deposition of MAC. This implies that circulating 
      inflammatory mediators may increase susceptibility to local complement activation 
      and MAC-deposition, which may represent an early event in the pathogenesis 
      leading to AMD development.
CI  - Copyright (c) 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.
FAU - Udsen, Maja
AU  - Udsen M
AD  - Department of Immunology and Microbiology, University of Copenhagen, Faculty of 
      Health and Medical Sciences, Copenhagen, Denmark.
FAU - Tagmose, Christian
AU  - Tagmose C
AD  - Department of Immunology and Microbiology, University of Copenhagen, Faculty of 
      Health and Medical Sciences, Copenhagen, Denmark.
FAU - Garred, Peter
AU  - Garred P
AD  - Laboratory of Molecular Medicine, Department of Clinical Immunology, Section 
      7631, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
FAU - Nissen, Mogens Holst
AU  - Nissen MH
AD  - Department of Immunology and Microbiology, University of Copenhagen, Faculty of 
      Health and Medical Sciences, Copenhagen, Denmark.
FAU - Faber, Carsten
AU  - Faber C
AD  - Department of Immunology and Microbiology, University of Copenhagen, Faculty of 
      Health and Medical Sciences, Copenhagen, Denmark; Department of Ophthalmology, 
      Copenhagen University Hospital, Rigshospitalet, Denmark. Electronic address: 
      carstenfaber@gmail.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220217
PL  - England
TA  - Exp Eye Res
JT  - Experimental eye research
JID - 0370707
RN  - 0 (Complement Membrane Attack Complex)
RN  - 0 (Retinal Pigments)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 82115-62-6 (Interferon-gamma)
RN  - EC 3.4.21.47 (Complement Factor B)
SB  - IM
MH  - Complement Activation/physiology
MH  - Complement Factor B/metabolism
MH  - Complement Membrane Attack Complex/metabolism
MH  - Humans
MH  - Interferon-gamma/metabolism/pharmacology
MH  - *Macular Degeneration/metabolism
MH  - Retinal Pigment Epithelium/metabolism
MH  - Retinal Pigments/metabolism
MH  - *Tumor Necrosis Factor-alpha/metabolism/pharmacology
OTO - NOTNLM
OT  - Complement
OT  - Interferons
OT  - Membrane attack complex
OT  - Retinal pigment epithelium
OT  - Tumor necrosis factor
EDAT- 2022/02/21 06:00
MHDA- 2022/04/28 06:00
CRDT- 2022/02/20 20:30
PHST- 2021/10/06 00:00 [received]
PHST- 2022/02/08 00:00 [revised]
PHST- 2022/02/14 00:00 [accepted]
PHST- 2022/02/21 06:00 [pubmed]
PHST- 2022/04/28 06:00 [medline]
PHST- 2022/02/20 20:30 [entrez]
AID - S0014-4835(22)00063-X [pii]
AID - 10.1016/j.exer.2022.108982 [doi]
PST - ppublish
SO  - Exp Eye Res. 2022 May;218:108982. doi: 10.1016/j.exer.2022.108982. Epub 2022 Feb 
      17.