Effect of high glucose on human alveolar macrophage phenotype and phagocytosis of mycobacteria

Vance, Jesse; Santos, Andres; Sadofsky, Laura; Morice, Alyn; Cervantes, Jorge

Abstract

Purpose
Diabetes mellitus (DBM) reduces immunological activity and increases susceptibility to various infections, including tuberculosis (TB). Human alveolar macrophage (hAM) functions are altered in DBM.

Methods
To mimic hyperglycemic conditions in the lung alveolus, we co-cultured a hAM cell line (Daisy cell line) with human umbilical vein endothelial cells for 48 h in the presence of culture media alone, normal glucose (5 mM), and high glucose (22 mM). Using flow cytometry, immunophenotype characterization included cell surface markers CD 11c, CD14, CD16, CD86, CD163, CD169, CD206, CX3CR-1, CSF-1R, and matrix metalloproteinase-9 (MMP9). Phagocytic function was measured by immunofluorescence microscopy at 24 h after inoculation of cells with GFP-expressing Mycobacterium smegmatis.

Results
Direct exposure of AMs to high glucose and exposure in the co-culture system yield different results for the same phenotypic markers. MMP9 expression was increased under both conditions. CD169 and CX3CR1 expressions were decreased when AMs were exposed directly to high glucose but increased under co-culture. Immunofluorescence assay revealed that phagocytosis decreased in AMs when directly exposed to increased glucose levels from 2.5 mM to normal glucose (5 mM), yet AMs under co-culture did not show decreased phagocytosis until concentrations were raised to 25 mM.

Conclusion
Alteration in the expression of certain receptors may contribute to defective sentinel function of AMs, promoting susceptibility to TB in a diabetic host. Variability in cell surface marker expression under direct glucose exposure compared to exposure via co-culture reveals that cell signaling between endothelial cells and AMs may play a crucial role in the phenotypic expression of AMs.

Citation

Vance, J., Santos, A., Sadofsky, L., Morice, A., & Cervantes, J. (2019). Effect of high glucose on human alveolar macrophage phenotype and phagocytosis of mycobacteria. Lung, 197(1), 89–94. https://doi.org/10.1007/s00408-018-0181-z

• Journal Article Type: Article
• Acceptance Date: Nov 19, 2018
• Online Publication Date: Nov 24, 2018
• Publication Date: 2019-02
• Deposit Date: Dec 7, 2018
• Publicly Available Date: Nov 25, 2019
• Journal: Lung
• Print ISSN: 0341-2040
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 197
• Issue: 1
• Pages: 89–94
• DOI: https://doi.org/10.1007/s00408-018-0181-z
• Keywords: Diabetes; Alveolar macrophages; Tuberculosis; Mycobacteria
• Public URL: https://hull-repository.worktribe.com/output/1175473
• Publisher URL: https://link.springer.com/article/10.1007%2Fs00408-018-0181-z
• Contract Date: Dec 14, 2018