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NLRP3 Inflammasome Activation in THP-1 Target Cells Triggered by Pathogenic Naegleria fowleri

Authors
Kim, JH | Sohn, HJ  | Yoo, JK | Kang, H | Seong, GS | Chwae, YJ  | Kim, K  | Park, S  | Shin, HJ
Citation
Infection and immunity, 84(9). : 2422-2428, 2016
Journal Title
Infection and immunity
ISSN
0019-95671098-5522
Abstract
Naegleria fowleri, known as the brain-eating amoeba, causes acute primary amoebic meningoencephalitis. During swimming and other recreational water activities, N. fowleri trophozoites penetrate the nasal mucosa and invade the olfactory bulbs, resulting in intense inflammatory reactions in the forebrain tissue. To investigate what kinds of inflammasome molecules are expressed in target cells due to N. fowleri infection, human macrophage cells (THP-1 cells) were cocultured with N. fowleri trophozoites in a noncontact system, and consequently, interleukin-1beta (IL-1beta) production was estimated. Caspase-1 activation and IL-1beta production from THP-1 cells by Western blotting and the culture supernatant by enzyme-linked immunosorbent assay analysis were observed at 3 h after cocultivation. In addition, the increased expression of ASC and NLRP3, which make up an inflammasome complex, was also observed at 3 h after cocultivation. To confirm the caspase-1 activation and IL-1beta production via the NLRP3 inflammasome in THP-1 cells triggered by N. fowleri trophozoites, THP-1 cells were pretreated with several inhibitors. The inhibition assay showed that CA-074 (a cathepsin B inhibitor), glybenclamide (an NLRP3 molecule inhibitor), and N-benzyloxycarbony-Val-Ala-Asp(O-methyl)-fluoromethylketone (Z-VAD-FMK: a caspase-1 inhibitor) reduced the levels of caspase-1 activation and IL-1beta production from THP-1 cells. This study suggests that N. fowleri infection induces the NLRP3 inflammasome, which activates caspase-1 and subsequently produces IL-1beta, thus resulting in inflammation.
MeSH

DOI
10.1128/IAI.00275-16
PMID
27297387
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
Ajou Authors
김, 경민  |  박, 선  |  손, 혜진  |  신, 호준  |  최, 용준
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