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Related Experiment Video

Updated: Jun 29, 2025

Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments
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Protocol for identifying stressed granulocytes from septic mice.

Yu Hao1, Can Zhang1, Fangyuan Li1

  • 1Biomedical Innovation Center, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China; Beijing Key Laboratory for Therapeutic Cancer Vaccines, Beijing 100038, China.

STAR Protocols
|April 3, 2024
PubMed
Summary
This summary is machine-generated.

Sepsis primes immune cells called granulocytes, enhancing their response to subsequent infections. This study details a mouse model to investigate this phenomenon and its impact on inflammation and survival.

Keywords:
Flow CytometryImmunologyModel Organisms

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Area of Science:

  • Immunology
  • Microbiology
  • Infectious Diseases

Background:

  • Sepsis can alter immune cell function, potentially affecting responses to subsequent infections.
  • Granulocytes play a critical role in the innate immune response to bacterial pathogens.

Purpose of the Study:

  • To present a reproducible murine model for studying sepsis followed by a secondary infection.
  • To investigate the functional changes in granulocytes following sepsis and their impact on host defense.

Main Methods:

  • Establishment of a murine model using cecal ligation and puncture (CLP) to induce sepsis.
  • Rechallenging mice with lipopolysaccharide (LPS) or Pseudomonas aeruginosa during the recovery phase.
  • Characterization of granulocyte phenotypes and assessment of their functional capacity.

Main Results:

  • The developed model allows for the study of sepsis-induced immune training in granulocytes.
  • Sepsis-conditioned granulocytes exhibit altered functional phenotypes.
  • The functional state of granulocytes influences the mortality of mice facing secondary infections.

Conclusions:

  • This protocol provides a valuable tool for investigating the complex interplay between sepsis, granulocyte function, and secondary infections.
  • Understanding sepsis-induced immune modulation in granulocytes is crucial for developing targeted therapies to improve patient outcomes.