Kidney Week

Abstract: TH-PO422

Neonatal Kidney Stem/Progenitor Cells (nKSPCs) Downregulate Activation of Human Neutrophils In Vitro

Session Information

• Development, Organoids, Injury, and Regeneration
  October 24, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 600 Development, Stem Cells, and Regenerative Medicine

Authors

• Chen, Junyu, Amsterdam UMC Locatie AMC, Amsterdam, Noord-Holland, Netherlands

Junyu Chen, MSc

• Gouwy, Mieke, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

Mieke Gouwy

• Proost, Paul, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

Paul Proost, MA, PhD

• Akalay, Sara, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

Sara Akalay, MD

• Lantermans, Hildo C., Amsterdam UMC Locatie AMC, Amsterdam, Noord-Holland, Netherlands

Hildo C. Lantermans, MSc

• Van den Heuvel, Lambertus P.W.J., Radboud Universiteit, Nijmegen, Gelderland, Netherlands

Lambertus P.W.J. Van den Heuvel, PhD

• Arcolino, Fanny Oliveira, Amsterdam UMC Locatie AMC, Amsterdam, Noord-Holland, Netherlands

Fanny Oliveira Arcolino, PhD

• Levtchenko, Elena, Amsterdam UMC Locatie AMC, Amsterdam, Noord-Holland, Netherlands

Elena Levtchenko, MD, PhD

Group or Team Name

• Translational Paediatric Nephrology.

Background

Neonatal Kidney Stem/Progenitor Cells (nKSPC), a novel type of kidney progenitor cell, isolated from urine of preterm neonates, have demonstrated immunoregulatory capacity and can inhibit proliferation of T-lymphocytes in mixed lymphocyte reaction. Our group is testing nKSPC as a potential source of cell therapy in the context of kidney transplantation. Neutrophils play important roles in ischemia-reperfusion injury (IRI), a common occurrence in kidney transplantation. In this study, we examined the capacity of nKSPC to inhibit human neutrophils in vitro.

Methods

Freshly isolated neutrophils activated by TNF-α were co-cultured with nKSPC at varying ratios (10:1 to 500:1). CD16 and CD66b as markers related to neutrophil activation were measured by Fluorescence-Activated Cell Sorting (FACS). Apoptosis was analyzed by Annexin V assay in FACS. Assessing neutrophil-produced reactive oxygen species (ROS), we employed a Luminol assay under multiple stimuli (TNF-α, IL-1β, fMLF, PGN). Neutrophil chemotaxis was investigated by Boyden chamber assay.

Results

Neutrophils, stimulated by TNF-α, exhibit increased CD66b and decreased CD16 expression, indicating activation. TNF-α also induced apoptosis of neutrophils. Co-culturing with nKSPC significantly reduced TNF-α-induced neutrophil activation and apoptosis. TNF-α, IL-1β, fMLF, and PGN stimulation promoted ROS production in neutrophils, and a co-culture with nKSPC inhibited this induction. Neutrophil chemotaxis induced by TNF-α was not inhibited by nKSPC.

Conclusion

Our in vitro data show the immunomodulatory capacity of nKSPC on neutrophils with decreased activation, reduction of apoptosis and abolishing ROS production.

Funding

• Government Support – Non-U.S.