Itami Nanase, Kondo Yoko, Tademoto Sayuri, Ito Daisuke, Fukumoto Soji, Otsuki Hitoshi
Division of Medical Zoology, Department of Microbiology and Immunology, School of Medicine, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.
Technical Department, Tottori University, Yonago 683-8503, Japan.
Yonago Acta Med. 2020 Jan 24;63(1):34-41. doi: 10.33160/yam.2020.02.005. eCollection 2020 Feb.
Trichinellosis is a serious zoonosis with a worldwide distribution. Fecund adult worms in the intestine release newborn larvae (NBL) that enter the general circulation from 4 days post infection (dpi). Alternatively activated macrophages in the peritoneal cavities and the diaphragms in infected mice have been reported. However, a role of newborn larvae is poorly understood.
The total numbers of peritoneal macrophages in mice infected with 500 muscle-stage larvae were counted during early infection and then total RNA was extracted. Peritoneal macrophages from uninfected C57BL/6 mice were incubated with IL-4 or LPS as a control, or co-cultured with live NBL, and peritoneal macrophages were obtained from mice injected with live or frozen dead NBL into peritoneal cavity. Total RNA was extracted from these macrophages. Two types of gene expression, classical and alternative activation, were examined in the macrophages and diaphragms of the infected mice using semi-quantitative reverse transcription-PCR.
The number of peritoneal macrophages in infected mice increased significantly. mRNA peak expression of alternative activation markers, Ym1 and arginase-1 (Arg1), was confirmed in the peritoneal macrophages and in diaphragm of mice around 15 dpi, while mRNA expression of classical activation markers, TNFα, IP-10, and iNOS was not detected. Injection of live NBL into the peritoneal cavities induced mRNA expression of Ym1 and Arg1 in the peritoneal macrophages of mice 9 dpi. However, dead NBL did not induce such gene expression. Alternative activation was not detected in the peritoneal macrophages co-cultured with NBL in vitro.
Gene expression of alternative activation makers, Ym1 and Arg1, was confirmed in the peritoneal macrophages and diaphragms of mice infected with . However, gene expression of classical activation markers was not detected. Live NBL induced an alternative activation of peritoneal macrophages in vivo, but not in vitro.
旋毛虫病是一种严重的人畜共患病,分布于世界各地。肠道内成熟的成虫释放新生幼虫(NBL),感染后4天起这些幼虫进入体循环。已有报道称感染小鼠的腹腔和膈肌中存在替代活化的巨噬细胞。然而,新生幼虫的作用尚不清楚。
对感染500条肌幼虫的小鼠在感染早期进行腹腔巨噬细胞总数计数,然后提取总RNA。将未感染的C57BL/6小鼠的腹腔巨噬细胞用IL-4或LPS作为对照进行孵育,或与活的NBL共培养,并从向腹腔内注射活的或冻融死亡的NBL的小鼠获取腹腔巨噬细胞。从这些巨噬细胞中提取总RNA。使用半定量逆转录PCR检测感染小鼠的巨噬细胞和膈肌中两种类型的基因表达,即经典活化和替代活化。
感染小鼠的腹腔巨噬细胞数量显著增加。在感染后约15天,在小鼠的腹腔巨噬细胞和膈肌中证实了替代活化标志物Ym1和精氨酸酶-1(Arg1)的mRNA峰值表达,而未检测到经典活化标志物TNFα、IP-10和诱导型一氧化氮合酶的mRNA表达。在感染后9天,向腹腔内注射活的NBL可诱导小鼠腹腔巨噬细胞中Ym1和Arg1的mRNA表达。然而,死亡的NBL未诱导这种基因表达。在体外与NBL共培养的腹腔巨噬细胞中未检测到替代活化。
在感染旋毛虫的小鼠的腹腔巨噬细胞和膈肌中证实了替代活化标志物Ym1和Arg1的基因表达。然而,未检测到经典活化标志物的基因表达。活的NBL在体内可诱导腹腔巨噬细胞的替代活化,但在体外则不能。