Related Experiment Video
Updated: Mar 25, 2026
Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages
Published on: April 20, 2018
Immortalized MH-S cells lack defining features of primary alveolar macrophages and do not support mouse pneumovirus
Todd A Brenner1, Tyler A Rice1, Erik D Anderson1
1Inflammation Immunobiology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, United States.
Abstract:
The SV-40-transformed MH-S cell line maintains some, but not all, features of primary alveolar macrophages (AMs) from BALB/c mice. We show here that MH-S cells produce inflammatory cytokines IL-6 and CXCL10 in response to challenge with Gram-positive Lactobacillus reuteri, and to TLR2 and NOD2 ligands Pam3CSK4 and MDP, respectively. In contrast, although wild-type AMs are infected in vivo by pneumonia virus of mice (PVM), no virus replication was detected in MH-S cells. Interestingly, the surface immunophenotype of MH-S cells (CD11c(+)Siglec F(-)) differs from that of wild-type AMs (CD11c(+) Siglec F(+)) and is similar to that of immature AMs isolated from granulocyte macrophage-colony stimulating factor (GM-CSF) gene-deleted mice; AMs from GM-CSF(-/-) mice also support PVM replication. However, MH-S cells do not express the GM-CSF receptor alpha chain (CD116) and do not respond to GM-CSF. Due to these unusual features, MH-S cells should be used with caution as experimental models of AMs.
Insights
The MH-S cell line, a model for alveolar macrophages (AMs), exhibits altered inflammatory responses and lacks pneumonia virus of mice (PVM) replication. Researchers caution its use due to significant differences from primary AMs.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Alveolar macrophages (AMs) are crucial for lung immunity.
- The SV-40-transformed MH-S cell line is used as a model for AMs.
- Understanding AMs' response to pathogens and their cellular characteristics is vital.
Purpose of the Study:
- To evaluate the MH-S cell line as a model for primary mouse alveolar macrophages (AMs).
- To compare the inflammatory cytokine production and viral replication capabilities of MH-S cells versus primary AMs.
- To investigate the surface immunophenotype and response to granulocyte-macrophage colony-stimulating factor (GM-CSF) in MH-S cells.
Main Methods:
- Stimulation of MH-S cells and primary AMs with Lactobacillus reuteri, Pam3CSK4, and MDP.
- Infection of MH-S cells and primary AMs with pneumonia virus of mice (PVM).
- Flow cytometry analysis of cell surface markers (CD11c, Siglec F, CD116).
- Assessment of cytokine production (IL-6, CXCL10).
Main Results:
- MH-S cells produced inflammatory cytokines IL-6 and CXCL10 in response to bacterial ligands but did not support PVM replication.
- MH-S cells exhibited a distinct surface immunophenotype (CD11c(+)Siglec F(-)) compared to wild-type AMs (CD11c(+)Siglec F(+)).
- MH-S cells do not express the GM-CSF receptor alpha chain (CD116) and do not respond to GM-CSF, despite similarities to immature AMs.
Conclusions:
- The MH-S cell line partially mimics primary AMs' inflammatory responses but differs significantly in viral susceptibility and surface markers.
- The lack of PVM replication and distinct immunophenotype suggest limitations of MH-S cells as a comprehensive AM model.
- Caution is advised when using MH-S cells to model alveolar macrophage functions due to their unique characteristics.
