Innate immune TLR7 signaling mediates platelet activation and platelet-leukocyte aggregate formation in murine bacterial sepsis

Abstract

Thrombocytopenia is a common complication in sepsis and is associated with higher mortality. Activated platelets express CD62P, which facilitates platelet-leukocyte aggregate (PLA) formation and contributes to thrombocytopenia in sepsis. We have reported that thrombocytopenia in murine sepsis is partly attributable to TLR7 signaling, but the underlying mechanism is unclear. In the current study, we tested the hypothesis that TLR7 mediates platelet activation and PLA formation during sepsis. In vitro, whole blood from WT mice treated with loxoribine, a TLR7 agonist, exhibited a dose-dependent increase in activated platelets compared to the control (PBS with 0.05% DMSO) or loxoribine-treated TLR7^−/− whole blood. In a murine model of sepsis, there was a significant increase in platelet activation and PLA formation 24 hours after cecal ligation and puncture (CLP) as evidenced by double positive expression of CD41⁺/CD62P⁺ and CD45⁺/CD62P⁺, respectively. The sepsis-induced PLA formation was significantly attenuated in TLR7^−/− mice. Finally, in ex-vivo experiments, plasma isolated from septic mice induced WT platelet activation, but such effect was significantly attenuated in platelets deficient of TLR7. These findings demonstrate a pivotal role of TLR7 signaling in platelet activation and PLA formation during bacterial sepsis.

Keywords:

• Innate immunity
• platelet leukocyte aggregates
• platelets
• sepsis
• TLR7
• Toll-like receptor 7

Acknowledgements

We would like to acknowledge Dr. Magali Fontaine of the University of Maryland School of Medicine for project discussion. We would also like to acknowledge the University of Maryland School of Medicine Center for Innovative Biomedical Resources, Flow Cytometry Core– Baltimore, Maryland.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Conceptualization, B.W. and W.C.; Methodology, B.W., L.Z., and W.C.; Investigations, B.W., and J.Z.; Manuscript writing, B.W., L.Z., and W.C.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/09537104.2022.2107627

Additional information

Funding

This work was supported in part by the National Institutes of Health (NIH), United States, grants K08HL153784 (B.W.), R35GM124775 (L.Z.), and R35GM140822 (W.C.), by Mentored Research Training Grant (2019) from the Foundation in Anesthesia Education and Research (B.W.), and by Frontiers in Anesthesia Research Award from International Anesthesia Research Society (W.C.)