Abstract: PO0192
A Novel Flow Cytometry Approach Identifies Kidney Mononuclear Phagocyte Subsets Involved in Mouse Kidney Injury Models
Session Information
- AKI Mechanisms - 2
October 22, 2020 | Location: On-Demand
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Nordlohne, Johannes, Bayer AG, Wuppertal, Germany
- Hulsmann, Ilona, Bayer AG, Wuppertal, Germany
- Schwafertz, Svenja, Bayer AG, Wuppertal, Germany
- Eitner, Frank, Bayer AG, Wuppertal, Germany
- Becker, Michael S., Bayer AG, Wuppertal, Germany
Background
Mononuclear phagocytes (MNPs) are heterogenous in phenotype and function, which reflects their double-edged role as drivers of inflammation and repair after kidney injury. Dissection of this complex network of cells into functional subunits has been challenging and more granular approaches could help to identify relevant subsets in preclinical kidney injury models. Here we used a novel flow cytometric approach to phenotypically and functionally dissect renal MNPs and perform a thorough comparison of MNP dynamics between two different kidney injury models.
Methods
The dynamic regulation of MNP subsets was monitored over 10d in two frequently used murine kidney injury models: ischemia reperfusion injury (IRI) and unilateral ureter obstruction (UUO). Using flow cytometric markers F4/80, CD11b and CD11c, kidney MNPs were phenotypically divided into five distinct subsets, which were further subdivided into functional subsets of proinflammatory M1-like (CD86+MHCII+CD206-) and regulatory M2-like (CD206+) cells.
Results
Three of the five renal MNP subsets were heavily contributing to both M1- and M2-like cell pools in both IRI and UUO, highlighting their functional multimodality regarding for example in vitro phagocytosis. The F4/80high MNP subset contributed most M2-like cells as from day 3 with a comparable MNP profile in both models. However, M1-like cells from two CD11bhigh subsets spiked 24h after IRI, while this spike was shifted to day 3 in the UUO model, which had a temporary early influx of M1-like F4/80high cells after 3h in turn. After 10d, total MNP numbers were decreasing in the UUO model, while M2-like F4/80high cells persisted in IRI kidneys.
Conclusion
Our novel flow cytometric approach unravels functional multimodality among MNP subsets and identifies distinct subsets responsible for an earlier M1-response and a more persistent M2-response in IRI compared to UUO. These results might support preclinical model selection and disease understanding in kidney injury.
Funding
- Commercial Support – Bayer AG