Intermedin (Adrenomedullin 2) Plays a Protective Role in Sepsis by Regulating T- and B-Cell Proliferation and Activity

Abstract

Background: Sepsis is the major cause of death in intensive care units. We previously found that intermedin (IMD), a calcitonin family peptide, can protect against sepsis by dynamically repairing vascular endothelial junctions and can ameliorate the inflammatory response by inhibiting the infiltration of macrophages in peripheral tissues. The effects of IMD on inflammatory and immune responses indicate that IMD may play a role in immunity. However, whether IMD affects immune cell development, differentiation and response to infection remains unclear.

Methods: IMD-knockout (Adm2-/-) mice were generated in our previous work. Wild-type IMD-KO mice were subjected to sham or cecal ligation and puncture (CLP) surgery, and bone marrow cells were obtained for RNA sequencing (RNA-Seq) analysis. The RNA-Seq results were verified by real-time RT-PCR. The effect of IMD KO or IMD rescue on the septic mice was explored using mild and severe infection models induced by CLP surgery at different levels of severity, and the survival outcomes were analyzed using Kaplan-Meier curves and the log-rank test. The mechanism underlying the effects of IMD in T/B cell proliferation and differentiation were investigated by PCR, Western blot (WB), and cell proliferation assays and flow cytometry analysis.

Results: RNA-Seq showed that IMD-KO mice exhibited a primary immunosuppression phenotype characterized by a marked decrease in the expression of T- and B-cell function-related genes. This immunosuppression made the IMD-KO mice vulnerable to pathogenic invasion, and even mild infection killed nearly half of the IMD-KO mice. Supplementation with the IMD peptide restored the expression of T/B-cell-related genes and significantly reduced the mortality rate of the IMD-KO mice. IMD is likely to directly promote T- and B-cell proliferation through ERK1/2 phosphorylation, stimulate T-cell differentiation via Ilr7/Rag1/2-controled T cell receptor (TCR) recombination, and activate B cells via Pax5, a transcription factor that activates at least 170 genes needed for B-cell functions.

Conclusion: Together with previous findings, our results indicate that IMD may play a protective role in sepsis via three mechanisms: protecting the vascular endothelium, reducing the inflammatory response, and activating T/B-cell proliferation and differentiation. Our study may provide the first identification of IMD as a calcitonin peptide that plays an important role in the adaptive immune response by activating T/B cells and provides translational opportunities for the design of immunotherapies for sepsis and other diseases associated with primary immunodeficiency.

Note:
Funding declaration: This work was supported by the National Natural Science Foundation of China (81971811 and 82172142 to F.X.), Sichuan Science and Technology Program (2020YFS0203 to W.Z.), Key Research and Development Project of Science and Technology Department of Sichuan Province (2021YFS0003 to Yan Kang), and National Key R&D Program of China (2022YFC2504500 to Yan Kang).

Conflicts of Interest: None

Ethical Approval: All animal procedures were approved (approval no: 2021309A) by Animal Ethics Committee of West China Hospital, Sichuan University. All animal experiments were performed according to institutional and national guidelines.

Keywords: sepsis, Intermedin (IMD), Adrenomedullin 2 (ADM2), adaptive immune response, T cells, B cells