Splenic T lymphocytes induce the formation of immunosuppressive neutrophils through IFN-γ in sepsis

Background: Despite many advances in treatment, the prognosis of patients with sepsis still remains poor. Polymorphonuclear leukocytes (PMNs) are the first line of defense against Infection. This study aimed to reveal the reason and mechanism of the production of PD-L1⁺ PMNs in sepsis.

Methods: Cecal ligation and perforation mouse model was established to simulate sepsis. And PMNs were treated for 4 h, 12 h with or without 100 ng/mL (IFN-γ) for further gene sequencing. PD-L1, PD-1, Ly6G, and CD3 were detected by multiplexed immunofluorescence. In addition, expression of PD-L1 and function of PMNs were assessed by flow cytometry. Serum and Cell Culture supernatant were measured with ELISA assays. Western blot was used to verify the JAK2/STAT1 pathway.

Results: Our study demonstrates that PMNs are the main immune cells with high expression of PD-L1 during sepsis, and these cells, therefore, play a critical role in immunosuppression. In vivo studies demonstrated a specific interaction between PD-L1⁺ PMNs and PD-1⁺ T cells. In vitro studies further demonstrated that IFN-γ induced the production of PD-L1⁺ PMNs through the JAK2/STAT1 pathway. In addition, Fedratinib, an inhibitor of JAK2, was shown to significantly reduce the expression of PD-L1 in neutrophils.

Conclusions: These data demonstrate that secretion of IFN-γ by splenic T lymphocytes induces the production of PD-L1 + PMNs through the JAK2/STAT1 pathway in sepsis.