Identification and Quantitation of Leukocyte Populations in Human Kidney Tissue by Multi-parameter Flow Cytometry

Katrina Kildey; Becker M P Law; Kimberly A Muczynski; Ray Wilkinson; Healy Helen; Andrew J Kassianos

doi:10.21769/BioProtoc.2980

Identification and Quantitation of Leukocyte Populations in Human Kidney Tissue by Multi-parameter Flow Cytometry

Bio Protoc. 2018 Aug 20;8(16):e2980. doi: 10.21769/BioProtoc.2980.

Authors

Katrina Kildey^{1

2}, Becker M P Law^{1

2

3}, Kimberly A Muczynski⁴, Ray Wilkinson^{1

2

3

5}, Healy Helen^{1

2

5}, Andrew J Kassianos^{1

2

3

5}

Affiliations

¹ Conjoint Kidney Research Laboratory, Pathology Queensland, Brisbane, Queensland, Australia.
² Kidney Health Service, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia.
³ Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia.
⁴ Division of Nephrology, University of Washington, Seattle, Washington, USA.
⁵ Medical School, University of Queensland, Brisbane, Queensland, Australia.

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.

Keywords: Flow cytometry; Immunology; Kidney; Kidney biopsy; Leukocytes.