Lack of RNase L attenuates macrophage functions.

Abstract

BACKGROUND:

Macrophages are one of the major cell types in innate immunity against microbial infection. It is believed that the expression of proinflammatory genes such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and cyclooxygenase-2 (Cox-2) by macrophages is also crucial for activation of both innate and adaptive immunities. RNase L is an interferon (IFN) inducible enzyme which is highly expressed in macrophages. It has been demonstrated that RNase L regulates the expression of certain inflammatory genes. However, its role in macrophage function is largely unknown.

METHODOLOGY:

Bone marrow-derived macrophages (BMMs) were generated from RNase L(+/+)and (-/-) mice. The migration of BMMs was analyzed by using Transwell migration assays. Endocytosis and phagocytosis of macrophages were assessed by using fluorescein isothiocyanate (FITC)-Dextran 40,000 and FITC-E. coli bacteria, respectively. The expression of inflammatory genes was determined by Western Blot and ELISA. The promoter activity of Cox-2 was measured by luciferase reporter assays.

CONCLUSIONS/FINDINGS:

Lack of RNase L significantly decreased the migration of BMMs induced by M-CSF, but at a less extent by GM-CSF and chemokine C-C motif ligand-2 (CCL2). Interestingly, RNase L deficient BMMs showed a significant reduction of endocytic activity to FITC-Dextran 40,000, but no any obvious effect on their phagocytic activity to FITC-bacteria under the same condition. RNase L impacts the expression of certain genes related to cell migration and inflammation such as transforming growth factor (TGF)-β, IL-1β, IL-10, CCL2 and Cox-2. Furthermore, the functional analysis of the Cox-2 promoter revealed that RNase L regulated the expression of Cox-2 in macrophages at its transcriptional level. Taken together, our findings provide direct evidence showing that RNase L contributes to innate immunity through regulating macrophage functions.

• Authors: Xin Yi, Chun Zeng, Hongli Liu, Xiaoli Chen, Ping Zhang, Boo Seok Yun, Ge Jin, Aimin Zhou
• Journal: PloS one (PLoS One) Vol. 8 Issue 12 Pg. e81269 ( 2013) ISSN: 1932-6203 [Electronic] United States
• PMID: 24324683 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Cytokines
• Ptgs2 protein, mouse
• Cyclooxygenase 2
• Endoribonucleases
• 2-5A-dependent ribonuclease

Topics

• Animals
• Cell Movement
• Cyclooxygenase 2 (genetics, metabolism)
• Cytokines (biosynthesis)
• Endocytosis
• Endoribonucleases (deficiency, metabolism)
• Macrophages (cytology, enzymology, metabolism)
• Mice
• Mice, Inbred C57BL
• Phagocytosis
• Promoter Regions, Genetic (genetics)
• Transcription, Genetic