Immune Netw.  2011 Dec;11(6):424-427. 10.4110/in.2011.11.6.424.

The Role of Intracellular Receptor NODs for Cytokine Production by Macrophages Infected with Mycobacterium leprae

• Kang TJ
• Chae GT

• KMID: 2150731
• DOI: http://doi.org/10.4110/in.2011.11.6.424

Abstract

The nucleotide-oligomerization domain (NOD) proteins are members of the NOD-like receptor (NLR) family, which are intracellular and cytoplasmic receptors. We analyzed the role of NODs for cytokine production by macrophages infected with intracellular pathogen M. leprae, the causative agent of leprosy. Production of pro-inflammatory cytokines such as IL-1beta and TNF-alpha was inhibited in the presence of cytochalasin D, an agent blocking phagocytosis, suggesting that intracellular signaling was, partially, required for macrophage activation to M. leprae infection. Next, we investigated the role of NOD1 and NOD2 proteins on NF-kappaB activation and cytokine expression. Treatment with M. leprae significantly increased NF-kappaB activation and expression of TNF-alpha and IL-1beta in NOD1- and NOD2-transfected cells. Interestingly, their activation and expression were inhibited by cytochalasin D, suggesting that stimulation of NOD proteins may be associated with the enhancement of cytokine production in host to M. leprae.

Keyword

Mycobacterium leprae; Cytokine; Cytochalasin D; NOD

MeSH Terms

Cytochalasin D
Cytokines
Humans
Leprosy
Macrophage Activation
Macrophages
Mycobacterium
Mycobacterium leprae
NF-kappa B
Phagocytosis
Proteins
Receptors, Cytoplasmic and Nuclear
Tumor Necrosis Factor-alpha
Cytochalasin D
Cytokines
NF-kappa B
Proteins
Receptors, Cytoplasmic and Nuclear
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 The role of phagocytosis in cytokine productions in RAW264.7 cells infected with M. leprae. The cells (106) were incubated with cytochalasin D (5 uM) for 30 min and then treated M. leprae (MOI of 1 and 10) for 8 h, and supernatants were assayed for cytokines productions. Data are representative of at least three independent experiments, each performed in triplicates. *p<0.05, **p<0.01 vs. non-treatment (Non-Tx).

• 

  Figure 2 The role of phagocytosis and intracellular sensor, NOD proteins on the expression of pro-inflammatory cytokines in HEK cells infected with M. leprae. HEK cells were transfected with NOD1 or NOD2 for 8 hours with viable M. leprae at 10.0 MOI in absence or presence of cytochalasin D (cyto D). The cell extracts were used for measuring NF-κB activation (A). The total RNA was isolated and the relative mRNA levels for IL-1β (B) and TnF-α (C) were measured by real-time PCR. Data are representative of at least three independent experiments, each performed in triplicate. *p<0.05, **p<0.01 vs. non-treatment with cytochalasin D.

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