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Related Experiment Video

Updated: Jun 26, 2026

Determination of Regulatory T Cell Subsets in Murine Thymus, Pancreatic Draining Lymph Node and Spleen Using Flow Cytometry
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Rabbit Immune Cell Function: In Vitro Assays for Immunological Assessment Studies Using Flow Cytometry.

Tamiris C Sardinha1, Philipe P L Pereira1, Tamires A R Gomes1

  • 1Research Centre for Veterinary Toxicology (CEPTOX), Department of Veterinary Pathology, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Pirassununga 13635-900, SP, Brazil.

Biotech (Basel (Switzerland))
|June 25, 2026
PubMed
Summary

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This summary is machine-generated.

Standardized flow cytometry protocols were developed for rabbit immune function assessment. These methods enable robust evaluation of oxidative burst, phagocytosis, and lymphocyte proliferation in rabbits for safety studies.

Area of Science:

  • Immunology
  • Flow Cytometry
  • Animal Models

Background:

  • Rodent models present challenges for longitudinal studies due to repeated blood collection impacting animal welfare.
  • Rabbits offer a viable alternative for longitudinal studies owing to larger blood volumes and easier sampling.
  • Standardized assays for assessing rabbit immune function are currently limited.

Purpose of the Study:

  • To establish and optimize standardized flow cytometry protocols for evaluating key immune functions in rabbits.
  • To assess oxidative burst, phagocytosis, and lymphocyte proliferation in rabbits using flow cytometry.
  • To provide a framework for immunotoxicological and safety evaluation studies in rabbits.

Main Methods:

  • Oxidative burst and phagocytosis were analyzed in whole blood using DCFH and fluorescently labeled *Staphylococcus aureus*.
Keywords:
immunotoxicologylagomorph specieslymphocyte proliferationoxidative burstphagocytosis

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  • Lymphocyte proliferation was assessed in peripheral blood mononuclear cells (PBMCs) labeled with CFSE and stimulated with ConA.
  • Flow cytometry was utilized for simultaneous quantification of reactive oxygen species (ROS) generation, phagocytic uptake, and CFSE dilution.

    • Main Results:

    • Rabbit heterophils demonstrated significant ROS production (median 90.4%) and phagocytic activity (median 23.6%) upon *S. aureus* stimulation.
    • Phagocyte oxidative burst reached near-complete levels (median 99.1%) with PMA stimulation, aligning with mammalian neutrophils.
    • CFSE-based assays confirmed measurable lymphocyte proliferative responses to ConA, validating adaptive immune assessment.

    Conclusions:

    • Standardized flow cytometry protocols provide a reliable method for assessing innate and adaptive immune responses in rabbits.
    • These validated assays support the use of rabbits as a model for immunotoxicological and safety evaluations.
    • The developed protocols contribute to advancing the use of rabbits in preclinical research and comparative immunology.