Granulocyte colony-stimulating factor induced T-cell hyporesponsiveness via modulation of CD177⁺S100A^hi neutrophils in unexplained recurrent pregnancy loss

Abstract

Background: Numerous studies have demonstrated the promising efficacy of granulocyte colony-stimulating factor (G-CSF) in the treatment of couples with unexplained recurrent pregnancy loss (URPL) during early pregnancy. While neutrophils are recognised as the main effectors mediating immunoregulation, their G-CSF-mobilised phenotype and mechanisms regulating maternal–fetal immunity remain unclear.

Methods: Single-cell RNA sequencing (scRNA-seq) and single-cell T-cell receptor sequencing (scTCR-seq) were conducted to uncover the immune reconstitution dynamics of peripheral blood under G-CSF stimulation. Integrative analysis of transcriptomic-proteomic profiles with functional validation revealed a unique immunomodulatory neutrophil population. Further, we used spatial transcriptomics, flow cytometry and immunohistochemistry to explore the spatial distribution characteristics of this population at the maternal–fetal interface, and validated its therapeutic efficacy in animal models.

Results: G-CSF-mobilised peripheral blood (G-PB) displayed immune hyporesponsiveness. Unique neutrophils expressing high levels of CD177 and the S100A gene family expanded substantially in response to G-CSF. These neutrophils exhibited a comparatively immature morphology and impaired T-cell responses via contact-dependent arginase 1 release, as well as upregulation of T-cell immune checkpoints. A reduction of CD177⁺S100A^hi neutrophils was observed in both peripheral blood and decidua of URPL patients relative to healthy pregnant women. Functional validation in abortion-prone murine models confirmed that exogenous supplementation of G-CSF or adoptive transfer of CD177⁺S100A^hi neutrophils could successfully improve the pregnancy outcomes.

Conclusion: G-CSF played a crucial regulatory role in improving pregnancy outcomes by selectively expanding CD177⁺S100A^hi neutrophils with polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) properties, providing a solid theoretical foundation for the treatment of patients with URPL using G-CSF.

Keywords

G-CSF / PMN-MDSCs / scRNA-seq / scTCR-seq / URPL

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