Kidney Week

Abstract: TH-PO0988

Dynamic Single-Cell Redox Profiling Enables Functional Immune Monitoring in Kidney Transplantation

Session Information

• Transplantation: Basic - Immune Biology, Tissue Injury, and Emerging Technologies
  October 22, 2026 | Location: Exhibit Hall A, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Transplantation

• 2001 Transplantation: Basic

Authors

• Goerlich, Nina, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Nina Goerlich, MD, DrMed

• Espinar Barranco, Laura, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Laura Espinar Barranco

• Ningoo, Mehek, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Mehek Ningoo, MS, PhD

• Metzke, Diana, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Diana Metzke

• Gisbert Vilanova, Cayetana, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Cayetana Gisbert Vilanova

• Arzig, Joram, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Joram Arzig

• Mirkheshti, Pouneh, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Pouneh Mirkheshti

• Klocke, Jan, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Jan Klocke, MD

• Cravedi, Paolo, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Paolo Cravedi, MD, PhD

• Fueyo-González, Francisco, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Francisco Fueyo-González, PhD

• Enghard, Philipp, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany

Philipp Enghard, DrMed

• Fribourg, Miguel, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Miguel Fribourg, PhD

Background

Reliable tools to assess functional immune status in kidney transplant recipients are lacking. Intracellular glutathione (GSH) reflects the balance between oxidative stress, metabolism, and antioxidant capacity. We developed GLedBlue, a fast reversible fluorescent biosensor enabling real-time single-cell quantification of intracellular GSH, capturing basal redox state and inducible redox capacity.

Methods

We performed single-cell redox profiling by flow cytometry in peripheral blood immune cells from kidney transplant recipients and healthy controls (n=61). Redox state (baseline GSH) and redox capacity (response to oxidative challenge) were quantified and integrated into a two-dimensional redox map. Analyses included cross-sectional comparison, longitudinal sampling before and after transplantation, and assessment before and after erythropoietin (EPO) therapy, complemented by transcriptomic analyses.

Results

GLedBlue resolved distinct immune cell–specific redox signatures. In transplant recipients, T cells exhibited preserved redox state but reduced redox capacity, most pronounced in CD4+ T cells (p=0.043), whereas monocytes and B cells showed increased basal redox state (p<0.01). Redox mapping enabled intuitive visualization of these shifts across immune populations. Longitudinal profiling demonstrated declining redox capacity in CD4+ and CD8+ T cells following transplantation and initiation of immunosuppression (p=0.012 and p=0.014), while redox state remained stable. EPO therapy induced selective redox changes in monocytes, with reduced redox state (p=0.017), without affecting T or B cells. This response was supported by transcriptomic signatures and recapitulated in vitro in a dose-dependent manner.

Conclusion

Single-cell GSH profiling using GLedBlue is feasible in clinical samples and enables functional immune monitoring by capturing basal and inducible redox dynamics. The redox map framework reveals cell-specific immune signatures, tracks longitudinal changes, and detects selective therapeutic responses, identifying monocytes as responders to EPO therapy.

Funding

• Other NIH Support