Expression of inflammasome-related molecules from U87MG cells cocultured with Naegleria fowleri in the non-contact system

Naegleria fowleri, a free living amoeba widespread in moist soil, water and sediment, exist as a virulent pathogen causing fetal primary amoebic meningoencephalitis (PAM) in experimental animals and in humam. Inflammasomes are large intracellular multiprotein complexes that play a central role in innate immunity. They detect and respond to a large range of pathogen-associated molecular patterns (PAMPs), including bacterial flagellin, and damage-associated molecular patterns (DAMPs), such as uric acid crystals. inflammasomes comprise a member of the NOD-like receptor (NLR) family, such as NLRP3 and IPAF, and are defined by the NLR protein that they contain. The NLR protein recruits the inflammasome-adaptor protein, ASC, which in turn interacts with caspase-1 leading to its activation. Consequently caspase-1 promotes the maturation of the proinflammatory cytokines such as, interleukin IL-1β and IL-18. Inflammasome activation is crucial for host defense to pathogens, but recent studies have also found a role for the inflammasomes in the pathogenesis of several inflammatory diseases such as inflammatory bowel disease, rheumatoid arthritis and microorganism infection. To observe the induction of inflammasome-related molecules from target cells cocultured with N. fowleri trophozoites in a non-contact system, U87MG cells were cultured with TRAIL, LPS and amoebic trohozoites for 30, 60 and 180 mins, respectively. By Western blotting and ELISA analysis, amounts of IL-1β and NLRP3 protein from U87MG cells cocultured with N. fowleri trophozoites were increased a time-dependent manner. Amount of caspase-1 was increased at 60 min. On the other hand, when the glybenclamide which is a specific-NLRP3 inhibitor was treated on the coculture system, the expression of IL-1β, NLRP3 and caspase-1 were down-regulated. Thus, this study showed that the target cells (U87MG cells) cocultured with N. fowleri trophozoites in a non-contact system secret some inflammasomes, NRLP3, caspase-1 and IL-1β, which may play an important role on the inflammation response.

Author dc:creator

• 심, 문희

Contributors dc:contributor

• 신, 호준
• 대학원 의생명과학과
• 201124279

• 파울러자유아메바
• inflammasome
• U87MG세포
• NLRP3
• IL-1β
• Caspase-1

Language dc:language

ko

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000014698
000000014698

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/8593