Abstract
Background
IL-10 knockout (KO) mice can be protected against experimental autoimmune encephalomyelitis (EAE) with low-dose estrogen (E2) treatment similar to wild type (WT) mice, indicating that IL-10 is not required for E2-induced EAE protection. Our previous study demonstrated that E2 treatment induced an increase in programmed death ligands 1 (PD-L1) and 2 (PD-L2) on monocytes and macrophages in the periphery and within the CNS. In this study, we selectively inhibited the function of PD-L1 and PD-L2 to evaluate their critical role in maintaining E2-induced protection against EAE in IL-10-KO mice.
Methods
This study used female IL-10 KO mice pre-treated with either E2 or sham pellets seven days prior to induction of EAE and subsequently treated with Vehicle or antibodies to PD-L1, PD-L2 or respective isotype controls. Mice were scored daily for EAE severity over 21 days post-EAE induction. Cells from the spleen and brain were evaluated by flow cytometry.
Results
Differences in EAE severity were assessed in E2 and sham pre-treated IL-10-KO mice treated with α-PD-L1 or α-PD-L2 antibodies over the course of disease compared to treatment with Vehicle or isotype control antibodies. The results revealed real-time development of severe EAE in E2-pre-treated IL-10-KO mice treated with α-PD-L1 but not α-PD-L2 antibodies, mediated in part by increased percentages of activated CD74+CD11b+ myeloid cells in spleen and brain as well as splenic B-cells, T-cells and CD73+ cells.
Conclusion
These results demonstrate unequivocally that PD-L1 but not PD-L2 was required to retain the inhibitory effects of E2 on clinical EAE scores in female IL-10-KO mice and further implicate the emergence of the MIF/CD74 axis as a contributing pathogenic mechanism.
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Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
The authors would like to thank the VAPORHCS Veterinary staff for housing and maintaining mouse environments.
Funding
This work was supported by the National Institute of Allergy and Infectious Diseases R21AI148490 (HO) and R42AI122574 (AV), the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development Merit Review Award 2I01 BX000226 (AV), BLR&D Merit Review for Pre-IND studies of Drugs and Biologics Award 5I01 BX005112 (AV), Senior Research Career Scientist Award 1IK6BX004209 (AV). The contents do not represent the views of the Department of Veterans Affairs or the United States Government.
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HO contributed to experimental design and wrote and reviewed the manuscript. DL performed experiments, collected and analyzed data, and helped with manuscript preparation. RMR and AAV reviewed and helped with manuscript preparation. All authors read and approved the final manuscript.
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Offner, H., Lockwood, D., Meza-Romero, R. et al. PD-L1 is required for estrogen-induced protection against severe EAE in IL-10 deficient mice1. Metab Brain Dis 38, 589–599 (2023). https://doi.org/10.1007/s11011-022-01129-8
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DOI: https://doi.org/10.1007/s11011-022-01129-8