Immune Netw.  2015 Dec;15(6):319-324. 10.4110/in.2015.15.6.319.

Expression of Nucleotide-oligomerization Domain (NOD) and Related Genes in Mouse Tissues Infected with Mycobacterium leprae

• Kim TH
• Chae GT
• Kang TJ

• KMID: 2132706
• DOI: http://doi.org/10.4110/in.2015.15.6.319

Abstract

The nucleotide-oligomerization domain (NOD) is an important molecule involved in host defense against bacterial infection. To study the role of NODs in the host response to Mycobacterium leprae, we measured the mRNA levels of NODs and related genes in infected mouse tissues. The mRNA expression of NOD1, NOD2, caspase-1 and ASC was increased in mouse footpads. Whereas NOD2 expression in macrophages was increased at 2 and 24 h post-infection with M. leprae, there was no expression of NOD1 at these time points. An increase in caspase-1 expression was observed at 2 h and continued at 24 h. However, the expression of ASC was increased only at the early time point. The expression of caspase-1 is regulated by NOD2-dependent pathway in established HEK 293. Our results suggest NOD2, rather than NOD1, may be associated with the host response to M. leprae and that caspase-1 activation is essential for the host response.

Keyword

Mycobacterium leprae; NOD2; Caspase-1; ASC

MeSH Terms

Animals
Bacterial Infections
Macrophages
Mice*
Mycobacterium leprae*
Mycobacterium*
RNA, Messenger
RNA, Messenger

Figure

• 

  Figure 1 Caspase-1 activation and IL-1β production in RAW264.7 cells infected with M. leprae. Macrophages (106) were incubated with caspase-1 inhibitor (Ac-YVAD-CMK) for 1 h and then treated with LPS (100 ng/ml) and M. leprae (MOI of 10) for 18 h, and the supernatants and cell lysates were assayed for caspase-1 activity (A) and IL-1β (B), respectively. Data are representative of at least three independent experiments, each performed in triplicate. **p<0.01.

• 

  Figure 2 The expression of NOD-related genes in mouse footpads infected with M. leprae by determined by real-time PCR. Data are representative of at least three independent experiments, each performed in triplicate. *p<0.05; **p<0.01 vs. no stimulation.

• 

  Figure 3 The expression of NOD-related genes (A; NOD1, B; NOD2, C; caspase-1, D; ASC) in RAW 264.7 cells infected with M. leprae determined by real-time PCR. Data are representative of at least three independent experiments, each performed in triplicate. *p<0.05; **p<0.01 vs. no stimulation.

• 

  Figure 4 Caspase-1 expression in HEK cells transfected with NOD1 and NOD2. HEK cells were transfected with NOD1 or NOD2, or co-transfected with both NODs and stimulated for 24 hours with viable M. leprae at 10.0 MOI. Data are representative of at least three independent experiments, each performed in triplicate. *p<0.05; **p<0.01 vs. empty vector.

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