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Flow Cytometry of Urinary Leukocytes and Lymphocytes as a Biomarker of Renal Disease

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Biomarkers in Kidney Disease

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Abstract

Urine is normally nearly devoid of leukocytes. In various renal diseases however, immune cells can be observed in the urine. Flow cytometry is the state-of-the-art technique for analyzing the quantity and qualities of cells in suspension, making this method an ideal approach for investigating urinary immune cells. Increased amounts of urinary T cells and macrophages are routinely observed in inflammatory renal disease. In contrast, noninflammatory renal syndromes are not associated with an increase in urinary immune cell counts. The composition and the phenotype of the urinary immune cells are reminiscent of the infiltrating cells observed in the respective kidney biopsies, indicating that the urinary immune cells mirror local kidney inflammation. Therefore, they can be used as a “window into the kidney” to investigate the local cellular pathogenesis of renal diseases. Urinary T cells and macrophages have been probed as biomarkers in IgA nephritis, lupus nephritis, ANCA-associated glomerulonephritis, and renal transplant rejection, showing promising results in all entities.

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Abbreviations

ANCA:

Anti-neutrophil cytoplasmic antibody

Anti-HLA-DR:

Antibody staining HLA-DR MHCII molecules

Anti-URO-3:

Antibody specific for tubular epithelial cells

BKV:

BK-virus, member of the polyomavirus family, can cause infection of renal transplants

CCR7:

Chemokine receptor of the CC family among others expressed on naïve T cells

CD14:

Marker to identify classical monocytes and macrophages

CD16:

Surface molecule expressed by neutrophil granulocytes and a subset of monocytes/macrophages

CD3:

Identifies T cells which can be subdivided in CD3+CD4+ T helper T cells and CD3+CD8+ T killer T cells

CD4+ effector/memory T cells (EM T cells):

Once naïve T cells encounter their cognate antigen, they differentiate into effector/memory T cells

CD45RO:

Surface molecule expressed by memory T lymphocytes

CD54:

ICAM-1 adhesion molecule

CD56:

Marker expressed on NK T cells

CXCR3:

Chemokine receptor of the CXC family binds CXCL9, 10, and 11. Primarily expressed on Th1 T cells and mediates recruitment of these cells

DAPI:

Fluorescence stain that binds DNA visualizes cell nucleuses in immunofluorescence

Fas-Ligand (Fas-L):

Protein of the TNF family binding mediates apoptosis

GN:

Glomerulonephritis

SLEDAI:

Score reflecting the disease activity of SLE patients

LN:

Lupus nephritis

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Authors and Affiliations

  1. Klinik mit Schwerpunkt Nephrologie und internistische Intensivmedizin, Charité Berlin, Augustenburger Platz 1, 13353, Berlin, Germany

    Philipp Enghard

  2. Klinik mit Schwerpunkt Rheumatologie und klinische Immunologie, Charité Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Philipp Enghard & Jan Klocke

  3. Institut für Pathologie, Charité Berlin, Augustenburger Platz 1, 13353, Berlin, Germany

    Birgit Rudolph

  4. Rheumatologie, Universität Lübeck, Rateburger Allee 160, 23538, Lübeck, Germany

    Gabriela Riemekasten

Authors
  1. Philipp Enghard
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  2. Birgit Rudolph
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  3. Jan Klocke
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  4. Gabriela Riemekasten
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Corresponding author

Correspondence to Philipp Enghard .

Editor information

Editors and Affiliations

  1. Department of Biomedical Sciences, University of Westminster, London, United Kingdom

    Vinood B. Patel

Definitions

ANCA-associated glomerulonephritis

ANCA autoantibody-induced systemic vasculitis affecting the kidneys and causing renal inflammation.

Flow cytometry

Method for quantitative and qualitative analysis of large amounts of suspended cells. Usually, cells are stained with fluorochrome-coupled antibodies, which are detected using lasers and detectors for specific wavelengths.

Hematuria

The presence of blood in the urine.

IgA nephropathy

One of the most frequent forms of glomerulonephritis caused by deposition of IgA in the mesangium.

Lupus nephritis (LN)

Renal inflammation associated with the systemic autoimmune disease SLE.

Proliferative nephritis

Renal disease associated with the proliferation of cells in the glomeruli due to inflammation of the kidneys.

Proteinuria

The presence of elevated amounts of protein in the urine.

Renal transplant rejection

Immunologic reaction against the renal graft, a frequent cause of graft failure.

SLE

Systemic lupus erythematosus, a systemic autoimmune disease characterized by the presence of anti-nuclear autoantibodies and inflammation of several potential target organs.

Urine sediment

Microscopic analysis of the insoluble contents of urine after centrifugation.

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Enghard, P., Rudolph, B., Klocke, J., Riemekasten, G. (2015). Flow Cytometry of Urinary Leukocytes and Lymphocytes as a Biomarker of Renal Disease. In: Patel, V. (eds) Biomarkers in Kidney Disease. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-7743-9_35-1

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  • DOI: https://doi.org/10.1007/978-94-007-7743-9_35-1

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  • Online ISBN: 978-94-007-7743-9

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