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Lactate Exposure Promotes Immunosuppressive Phenotypes in Innate Immune Cells

Abstract

Introduction

Lactate secreted by tumors is not just a byproduct, but rather an active modulator of immune cells. There are few studies aimed at investigating the true effect of lactate, which is normally confounded by pH. Such a knowledge gap needs to be addressed. Herein, we studied the immunomodulatory effects of lactate on dendritic cells (DCs) and macrophages (MΦs).

Methods

Bone marrow-derived innate immune cells were treated with 50 mM sodium lactate (sLA) and incubated for 2 days or 5 days at 37 °C. Controls included media, lipopolysaccharide (LPS), MCT inhibitors (α-cyano-4-hydroxycinnamic acid and AR-C15585). Flow cytometric analysis of immune phenotypes were performed by incubating cells with specific marker antibodies and viability dye. Differential expression analyses were conducted on R using limma-voom and adjusted p-values were generated using the Bejamini-Hochberg Procedure.

Results

Lactate exposure attenuated DC maturation through the downregulation of CD80 and MHCII expression under LPS stimulation. For MΦs, lactate exposure resulted in M2 polarization as evidenced by the reduction of M1 markers (CD38 and iNOS), and the increase in expression of CD163 and Arg1. We also revealed the role of monocarboxylate transporters (MCTs) in mediating lactate effect in MΦs. MCT4 inhibition significantly boosted lactate M2 polarization, while blocking of MCT1/2 failed to reverse the immunosuppressive effect of lactate, correlating with the result of gene expression that lactate increased MCT4 expression, but downregulated the expression of MCT1/2.

Conclusions

This research provides valuable insight on the influence of metabolic products on tumor immunity and will help to identify novel metabolic targets for augmenting cancer immunotherapies.

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ACKNOWLEDGMENTS

This work was supported by the National Institutes of Health (Grants R35125012 and R01AI139399).

Author Contributions

RS contributed to the design and execution of experiments, analysis of data, and compilation of the manuscript. BT and HH contributed to the execution of experiments and analysis of data. NP contributed to the analysis of RNA sequencing data and discussion on gene expression. RA provided input on experimental protocols. JSL contributed to the design and execution of experiments, analysis of data, manuscript compilation and has primary responsibility for the content of the manuscript.

Conflict of interest

Rapeepat Sangsuwan, Bhasirie Thuamsang, Noah J. Pacifici, Hyunsoo Han, Svetlana Miakicheva, Riley Allen, and Jamal S. Lewis have no conflicts of interest to disclose.

Research Involving Human and Animal Rights

C57BL/6 and BALB/cByJ mice were purchased from Jackson Laboratories and were housed in specific pathogen-free environment conditions at the University of California, Davis TRACS facility and used according to the UC Davis Institutional Animal Care and Use Committee (IACUC).

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Sangsuwan, R., Thuamsang, B., Pacifici, N. et al. Lactate Exposure Promotes Immunosuppressive Phenotypes in Innate Immune Cells. Cel. Mol. Bioeng. 13, 541–557 (2020). https://doi.org/10.1007/s12195-020-00652-x

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Keywords

  • Lactate
  • Dendritic cells
  • Macrophages
  • Immune escape
  • Cancer
  • Immunosuppressive