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Isolation, Transfection, and Culture of Primary Human Monocytes

Isolation, Transfection, and Culture of Primary Human Monocytes

Published on: December 16, 2019

MAIL regulates human monocyte IL-6 production.

Sudarshan Seshadri¹, Yashaswini Kannan, Srabani Mitra

¹Davis Heart and Lung Research Institute and Pulmonary, Allergy, Critical Care and Sleep Medicine Division, The Ohio State University, Columbus, OH 43210, USA.

Journal of Immunology (Baltimore, Md. : 1950)

|September 29, 2009

Summary

Molecule containing ankyrin repeats induced by LPS (MAIL) is crucial for IL-6 production in human monocytes. Suppressing MAIL significantly reduces IL-6, highlighting its role in inflammation.

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Area of Science:

Immunology
Molecular Biology
Cell Biology

Background:

Interleukin-6 (IL-6) is a cytokine involved in sepsis and cancer.
Human monocytes produce more IL-6 than monocyte-derived macrophages.
Molecule containing ankyrin repeats induced by LPS (MAIL) knockout animals show reduced IL-6 production.

Purpose of the Study:

To investigate the role of MAIL in human IL-6 production.
To explain the difference in IL-6 production between human monocytes and macrophages.
To determine if MAIL regulation is key to IL-6 production.

Main Methods:

Comparison of MAIL expression in human monocytes and macrophages stimulated with LPS.
Analysis of MAIL protein form, localization, and interaction with NF-kappaB.
Assessment of IL-6 production via luciferase assays and ELISA.
Silencing of MAIL using small interfering RNA (siRNA).

Main Results:

LPS-induced MAIL expression in monocytes is transient and suppressed during macrophage differentiation.
Human MAIL-L localizes to the nucleus and binds to the p50 subunit of NF-kappaB.
Monocytes produced significantly more IL-6 than macrophages.
MAIL suppression decreased IL-6 production in THP-1 cells and primary monocytes.
siMAIL suppressed enhanced IL-6 response to LPS and muramyl dipeptide.

Conclusions:

MAIL is a key regulator of IL-6 production in human monocytes.
MAIL plays a significant role in inflammation induced by Toll-like receptor and NOD-like receptor ligands.
MAIL regulation may explain differential IL-6 production between monocytes and macrophages.