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Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging.

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Co-detection by indexing (CODEX) reveals how immune cell neighborhoods impact protein expression in healthy and lupus mouse spleens. This spatial cytometry approach uncovers novel cell interactions in disease.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Immune cell interactions are crucial for tissue function and disease.
  • Understanding spatial relationships in lymphoid tissues is complex.
  • Multiplexed imaging offers potential for detailed cellular analysis.

Purpose of the Study:

  • To apply co-detection by indexing (CODEX) for high-plex spatial profiling of murine spleens.
  • To investigate the impact of cellular neighborhoods on immune cell protein expression.
  • To compare splenic architecture and cell interactions in healthy versus autoimmune disease models.

Main Methods:

  • Utilized co-detection by indexing (CODEX), an iterative antibody visualization technique.
  • Employed DNA barcodes, fluorescent nucleotides, and in situ polymerization for indexing.
  • Developed an algorithmic pipeline for single-cell antigen quantification in dense tissues.

Main Results:

  • Observed a significant influence of cellular neighborhoods on immune cell receptor expression.
  • Characterized previously unappreciated splenic cell-interaction dynamics in MRL/lpr lupus mice.
  • Demonstrated de novo lymphoid tissue architecture analysis at single-cell and neighborhood levels.

Conclusions:

  • CODEX provides high-fidelity multiplexed spatial cytometry for quantitative tissue analysis.
  • Cellular neighborhood context profoundly affects immune cell phenotype.
  • This approach reveals distinct splenic cellular dynamics in autoimmune disease.