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Published on: June 27, 2020
Identification and Quantitation of Leukocyte Populations in Human Kidney Tissue by Multi-parameter Flow Cytometry
Katrina Kildey1,2, Becker M P Law1,2,3, Kimberly A Muczynski4
1Conjoint Kidney Research Laboratory, Pathology Queensland, Brisbane, Queensland, Australia.
Insights
This study introduces a new flow cytometry method to precisely identify and count immune cells in kidney biopsies, improving diagnosis for kidney diseases like acute kidney injury (AKI) and chronic kidney disease (CKD). This advanced technique overcomes limitations of older methods for better characterization of immune cell populations.
Area of Science:
- Nephrology
- Immunology
- Biotechnology
Background:
- Inflammatory immune cells are implicated in the pathology of acute kidney injury (AKI) and chronic kidney disease (CKD).
- Current immunohistochemical (IHC) techniques limit the precise identification and characterization of distinct leukocyte populations in human kidney biopsies.
- Existing methods struggle to combine multiple markers needed to define specific immune cell subsets.
Purpose of the Study:
- To develop and validate a multi-parameter, flow cytometry-based approach for identifying and quantifying immune cell populations in human kidney biopsies.
- To overcome the limitations of IHC for detailed immune cell profiling in kidney disease.
Main Methods:
- Fresh human kidney tissue is processed into a single-cell suspension.
- Cells are labeled with antibody panels targeting specific leukocyte markers.
- Multi-parameter flow cytometry is used for acquisition and analysis of immune cell subpopulations.
Main Results:
- The developed flow cytometry approach enables accurate detection and quantitation of diverse leukocyte subpopulations from a single kidney biopsy specimen.
- This method allows for the use of comprehensive panels of cell-specific markers, surpassing IHC limitations.
- The technique provides a robust platform for immune cell enumeration in kidney disease.
Conclusions:
- The novel flow cytometry technique represents a significant advancement in identifying and enumerating immune cell subpopulations in human kidney diseases.
- This method offers a powerful complement to traditional histopathological examinations of kidney biopsies.
- Accurate immune cell characterization can enhance understanding and management of kidney diseases.
Abstract:
Inflammatory immune cells play direct pathological roles in cases of acute kidney injury (AKI) and chronic kidney disease (CKD). However, the identification and characterization of distinct populations of leukocytes in human kidney biopsies have been confounded by the limitations of immunohistochemical (IHC)-based techniques used to detect them. This methodology is not amenable to the combinations of multiple markers necessary to unequivocally define discrete immune cell populations. We have developed a multi-parameter, flow cytometric-based approach that addresses the need for panels of cell-specific markers in the identification of immune cell populations, allowing both the accurate detection and quantitation of leukocyte subpopulations from a single, clinical kidney biopsy specimen. In this approach, fresh human kidney tissue is dissociated into a single cell suspension followed by antibody-labeling and flow cytometric-based acquisition and analysis. This novel technique provides a major step forward in identifying and enumerating immune cell subpopulations in human kidney disease and is a powerful platform to complement traditional histopathological examinations of clinical kidney biopsies.
