RRC ID 52566
Author Kosmidou C, Efstathiou NE, Hoang MV, Notomi S, Konstantinou EK, Hirano M, Takahashi K, Maidana DE, Tsoka P, Young L, Gragoudas ES, Olsen TW, Morizane Y, Miller JW, Vavvas DG.
Title Issues with the Specificity of Immunological Reagents for NLRP3: Implications for Age-related Macular Degeneration.
Journal Sci Rep
Abstract Contradictory data have been presented regarding the implication of the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome in age-related macular degeneration (AMD), the leading cause of vision loss in the Western world. Recognizing that antibody specificity may explain this discrepancy and in line with recent National Institutes of Health (NIH) guidelines requiring authentication of key biological resources, the specificity of anti-NLRP3 antibodies was assessed to elucidate whether non-immune RPE cells express NLRP3. Using validated resources, NLRP3 was not detected in human primary or human established RPE cell lines under multiple inflammasome-priming conditions, including purported NLRP3 stimuli in RPE such as DICER1 deletion and Alu RNA transfection. Furthermore, NLRP3 was below detection limits in ex vivo macular RPE from AMD patients, as well as in human induced pluripotent stem cell (hiPSC)-derived RPE from patients with overactive NLRP3 syndrome (Chronic infantile neurologic cutaneous and articulate, CINCA syndrome). Evidence presented in this study provides new data regarding the interpretation of published results reporting NLRP3 expression and upregulation in RPE and addresses the role that this inflammasome plays in AMD pathogenesis.
Volume 8(1)
Pages 461
Published 2018-1-11
DOI 10.1038/s41598-017-17634-1
PII 10.1038/s41598-017-17634-1
PMID 29323137
PMC PMC5764999
MeSH Alu Elements Animals Antibodies / analysis* Antibody Specificity Cell Line DEAD-box RNA Helicases / genetics Disease Models, Animal Gene Deletion Humans Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / metabolism Macular Degeneration / genetics Macular Degeneration / immunology* Mice NLR Family, Pyrin Domain-Containing 3 Protein / immunology* Retinal Pigment Epithelium / cytology Retinal Pigment Epithelium / metabolism* Ribonuclease III / genetics THP-1 Cells
IF 4.011
Times Cited 18
Human and Animal Cells 454E2(HPS0077)