Increasing antimicrobial activity of macrophage to methicillin resistant staphylo-coccus aureus via TLR2 agonist-Pam3Csk4
10.3969/j.issn.1000-484X.2016.01.003
- VernacularTitle:Pam3 Csk4预处理增强巨噬细胞对耐甲氧西林金黄色葡萄球菌杀菌作用
- Author:
Yiguo CHEN
;
Linqiang DENG
;
Yong ZHANG
;
Hui CHEN
;
Xiaomei GUI
;
Nanjin ZHOU
;
Keng YUAN
;
Yanrong YU
;
Lizhi YU
;
Zhanghua XIONG
;
Xiaomu WU
;
Weiping MIN
- Publication Type:Journal Article
- Keywords:
Pam3Csk4;
MRSA;
TLR2;
Immunoregulation
- From:
Chinese Journal of Immunology
2016;(1):13-18
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate immune response of murine peritoneal macrophage challenging by methicillin-resistant S.aureus(MRSA)after pretreatment with Pam3Csk4(TLR2 agonist).Methods: Murine peritoneal macrophage was pretreated with Pam3Csk4(1 μg/ml).Following pretreatment 12 h later,heat-killed MRSA( HK-MRSA) was added and incubated for another 2 or 6 hours.The protein and mRNA level of TNF-α, IL-6 and IL-1 were determined by ELISA and Q-PCR, respectively.To estimate phagocytosis of macrophage,HK-MRSA/MSSA labeled with FITC( FITC-HK-MRSA/MSSA) were added to well and incubated for 30 min.After washing 5 times with PBS,intracellular FITC-HK-MRSA was detected by flow cytometry.To estimate antimicrobal activity of macrophage,live MRSA and MSSA were added to well and incubated at indication time,the CFU of s.aureus was estimated via a 10-fold serial dilution on agar media.cDNA was further quantitative assessed using primers for mouse FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 by Q-PCR .Results: Compared with saline-pretreated cell, the protein and mRNA level of TNF-α, IL-6 and IL-1 were markely reduced, respectively.However, both the phagocytosis and antimicrobal activity to S.aureus were significantly increased in macrophages pretreated with Pam3Csk4.Further study found that the macrophages had higher FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 expression at 6 h and 12 h post-stimulation Pam3Csk4.Conclusion: The results suggest that Pam3Csk4 could activate murine antimicrobal activity of peritoneal macrophage challenging by methicillin-resistant Saureus via increasing opsonophagocytosis in depended antibodies, complements manners.The results suggest Pam3Csk4 probably be a novel immunotherapy candidate against MRSA.
