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A Consistent Method to Identify and Isolate Mononuclear Phagocytes from Human Lung and Lymph Nodes.

Sophie L Gibbings1, Claudia V Jakubzick2,3

  • 1Department of Pediatrics, National Jewish Health, Denver, CO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 30, 2018
PubMed
Summary

Researchers identified distinct mononuclear phagocyte subtypes in the human lung and lymph nodes, clarifying their roles and aiding future functional studies. This work advances understanding of pulmonary immune cells.

Keywords:
Alveolar macrophagesDendritic cellsHumanInterstitial macrophagesLungMonocytesMononuclear phagocytesPulmonary

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Area of Science:

  • Immunology
  • Cell Biology
  • Pulmonology

Background:

  • Mononuclear phagocytes (MPs), including macrophages, dendritic cells (DCs), and monocytes, play crucial roles in lung immunity.
  • While MP subtypes in mouse lungs and human blood are well-studied, human pulmonary and lymph node MPs remain poorly characterized.
  • Understanding these cells is vital for discerning their functions in both health and disease.

Purpose of the Study:

  • To comprehensively identify and characterize human pulmonary mononuclear phagocyte subtypes using cell surface markers.
  • To differentiate these cells in the lung and draining lymph nodes (LNs).
  • To provide a foundation for future functional studies of human pulmonary MPs.

Main Methods:

  • Systematic examination of cell surface markers on human pulmonary and lymph node cells from over 100 lungs.
  • Flow cytometry and multi-parameter analysis to identify distinct MP populations.
  • Comparison with known markers from mouse lung and human blood studies.

Main Results:

  • Consistently identified five extravascular pulmonary MP subtypes and three LN MP subtypes across diverse donors.
  • Found that human blood CD141+ DCs were absent in healthy lungs and LNs; CD141 expression was observed on monocytes.
  • Differentiated alveolar and interstitial macrophage subtypes and identified at least two DC subtypes (DEC205+, CD1c+) in the human lung and LNs.

Conclusions:

  • Established a robust method for identifying and distinguishing human pulmonary and lymph node MP subtypes.
  • The findings challenge previous assumptions about CD141+ DCs in the human lung and highlight monocytes' plasticity.
  • This work provides a critical resource for future research into the functional roles of human pulmonary immune cells.