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Lymphatic Cannulation for Lymph Sampling and Molecular Delivery

David C. Zawieja, Sangeetha Thangaswamy, Wei Wang, Raquel Furtado, Cristina C. Clement, Zachary Papadopoulos, Marco Vigano, Eric A. Bridenbaugh, Lello Zolla, Anatoliy A. Gashev, Jonathan Kipnis, Gregoire Lauvau and Laura Santambrogio
J Immunol October 15, 2019, 203 (8) 2339-2350; DOI: https://doi.org/10.4049/jimmunol.1900375
David C. Zawieja
*Department of Medical Physiology, Texas A&M Health Science Center, Temple, TX 76504;
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  • ORCID record for David C. Zawieja
Sangeetha Thangaswamy
†Department of Pathology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
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Wei Wang
*Department of Medical Physiology, Texas A&M Health Science Center, Temple, TX 76504;
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Raquel Furtado
‡Department of Microbiology and Immunology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
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Cristina C. Clement
†Department of Pathology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
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Zachary Papadopoulos
§Center for Brain Immunology and Glia, School of Medicine, University of Virginia, Charlottesville, VA 22908;
¶Department of Neuroscience, School of Medicine, University of Virginia, Charlottesville, VA 22908;
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Marco Vigano
†Department of Pathology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
‖Orthopaedic Biotechnology Lab, Galeazzi Orthopaedic Institute for Care and Scientific Research, 20161 Milan, Italy; and
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Eric A. Bridenbaugh
*Department of Medical Physiology, Texas A&M Health Science Center, Temple, TX 76504;
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Lello Zolla
‖Orthopaedic Biotechnology Lab, Galeazzi Orthopaedic Institute for Care and Scientific Research, 20161 Milan, Italy; and
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Anatoliy A. Gashev
*Department of Medical Physiology, Texas A&M Health Science Center, Temple, TX 76504;
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Jonathan Kipnis
§Center for Brain Immunology and Glia, School of Medicine, University of Virginia, Charlottesville, VA 22908;
¶Department of Neuroscience, School of Medicine, University of Virginia, Charlottesville, VA 22908;
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Gregoire Lauvau
‡Department of Microbiology and Immunology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
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Laura Santambrogio
†Department of Pathology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
‡Department of Microbiology and Immunology, Albert Einstein College of Medicine, Montefiore Medical Center, New York, NY 10461;
#Department of Agricultural and Forest Sciences, University La Tuscia, 01100 Viterbo, Italy
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Key Points

  • Lymph carries the “omic,” vesicular, and immune cell signature of the draining organs.

  • Lymph analysis provides precise and “undiluted” biochemical and cellular information.

  • A protocol for lymph collection from mouse and rat lymphatics is reported.

Abstract

Unlike the blood, the interstitial fluid and the deriving lymph are directly bathing the cellular layer of each organ. As such, composition analysis of the lymphatic fluid can provide more precise biochemical and cellular information on an organ's health and be a valuable resource for biomarker discovery. In this study, we describe a protocol for cannulation of mouse and rat lymphatic collectors that is suitable for the following: the “omic” sampling of pre- and postnodal lymph, collected from different anatomical districts; the phenotyping of immune cells circulating between parenchymal organs and draining lymph nodes; injection of known amounts of molecules for quantitative immunological studies of nodal trafficking and/or clearance; and monitoring an organ’s biochemical omic changes in pathological conditions. Our data indicate that probing the lymphatic fluid can provide an accurate snapshot of an organ’s physiology/pathology, making it an ideal target for liquid biopsy.

Footnotes

  • This work was supported by National Institutes of Health Grants AG045223 and AI137198 (to L.S.) and 1U01HL123420 (to D.C.Z. and A.A.G.).

  • The online version of this article contains supplemental material.

  • Abbreviations used in this article:

    DSS
    dextran sulfate sodium
    FDR
    false discovery rate
    IPA
    ingenuity pathway analysis
    LC
    liquid chromatography
    MS
    mass spectrometry
    ppm
    part per million
    UHPLC
    ultra-high performance LC.

  • Received April 1, 2019.
  • Accepted August 8, 2019.
  • Copyright © 2019 by The American Association of Immunologists, Inc.
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The Journal of Immunology: 203 (8)
The Journal of Immunology
Vol. 203, Issue 8
15 Oct 2019
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Lymphatic Cannulation for Lymph Sampling and Molecular Delivery
David C. Zawieja, Sangeetha Thangaswamy, Wei Wang, Raquel Furtado, Cristina C. Clement, Zachary Papadopoulos, Marco Vigano, Eric A. Bridenbaugh, Lello Zolla, Anatoliy A. Gashev, Jonathan Kipnis, Gregoire Lauvau, Laura Santambrogio
The Journal of Immunology October 15, 2019, 203 (8) 2339-2350; DOI: 10.4049/jimmunol.1900375

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Lymphatic Cannulation for Lymph Sampling and Molecular Delivery
David C. Zawieja, Sangeetha Thangaswamy, Wei Wang, Raquel Furtado, Cristina C. Clement, Zachary Papadopoulos, Marco Vigano, Eric A. Bridenbaugh, Lello Zolla, Anatoliy A. Gashev, Jonathan Kipnis, Gregoire Lauvau, Laura Santambrogio
The Journal of Immunology October 15, 2019, 203 (8) 2339-2350; DOI: 10.4049/jimmunol.1900375
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