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Integrate Imaging Flow Cytometry and Transcriptomic Profiling to Evaluate Altered Endocytic CD1d Trafficking
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Integrate Imaging Flow Cytometry and Transcriptomic Profiling to Evaluate Altered Endocytic CD1d Trafficking.

Manju Sharma1, Xiang Zhang2, Shouxiong Huang3

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This study introduces a novel imaging flow cytometry method to analyze endocytic protein changes in human dendritic cells (DCs) exposed to air pollutants. The findings reveal altered protein localization, indicating impaired cellular function.

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Area of Science:

  • Immunology
  • Cell Biology
  • Environmental Health

Background:

  • Analyzing endocytic protein alterations at the populational level is challenging.
  • Existing methods require single-cell imaging and complex statistical analysis.
  • Understanding pollutant effects on immune cells is crucial for public health.

Purpose of the Study:

  • To develop and validate an imaging flow cytometry approach for assessing endocytic protein changes in human dendritic cells (DCs).
  • To investigate the subcellular localization of cluster of differentiation 1d protein (CD1d) in DCs exposed to benzo[a]pyrene (BaP).
  • To correlate protein localization changes with transcriptomic alterations in endocytic gene expression.

Main Methods:

  • Utilized imaging flow cytometry to capture thousands of cell images.
  • Employed transcriptomic profiling (RNA-seq) to analyze gene expression.
  • Applied IDEAS and ImageJ-Fiji software for quantitative image analysis of protein colocalization (CD1d and Lamp1).

Main Results:

  • Developed a robust method to quantify protein colocalization at single and populational levels.
  • Observed enhanced colocalization of CD1d and Lamp1 in DCs exposed to benzo[a]pyrene.
  • Demonstrated a link between altered protein localization and impaired endocytic gene expression.

Conclusions:

  • The developed imaging flow cytometry and bioinformatic approach effectively measures protein colocalization in pollutant-exposed DCs.
  • Benzo[a]pyrene exposure induces significant alterations in CD1d subcellular localization and endocytic function in human DCs.
  • This study provides a valuable tool for investigating environmental pollutant impacts on cellular immunity.