Advantages of Beagle PBMCs
In the 1950s and 1960s, extensive use of beagles in toxicology studies, particularly in assessing the effects of pharmaceuticals and environmental exposures, started. Since then, the breed became widely recognized as a standard test subject for new drugs due to its genetic and physiological characteristics that allowed data to translate effectively to human outcomes.
Beagle PBMCs are instrumental in clinical research because they provide a consistent and ethical source of immune cells that closely resemble human immune function which is helpful when studying immune response, developing therapeutic measures and evaluating drug safety. These are the advantages of using beagle PBMCs.
Genetic Similarity to Humans
Their genetic and physiological similarities with humans make beagle PBMCs ideal for investigating human-like immune behavior especially in preclinical studies.
Consistent Immune Profile with Standardized Protocols
The immune profile of beagles have been well characterized so results are predictable and reproducible. Experimental protocols have also been standardized making results more reliable.
Reliable Cryopreservation and Viability
Beagle PBMCs respond to cryopreservation well so cells can be stored for future studies or experiment replication. Post-thawing viability of beagle PBMCs is also high, increasing the reliability of in vitro assays.
Comparable to Non-Human Primate (NHP) Models
PBMCs from beagles have been used as a complementary model when non-human primates cannot be used due to ethical or financial restrictions.
Autoimmune and Inflammatory Disorders
Beagles are known to be susceptible to specific autoimmune diseases so their PBMCs offer a good opportunity to study the mechanisms behind these conditions.
Ethical and Consistent Supply
Beagles are widely used in research, thus beagle PBMCs offer an ethical alternative to intrusive whole-animal experiments. Since beagles are easy to collect samples from due to their calm temperament, a steady supply of PBMCs for long-term studies is an advantage.
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Beagle PBMC Preparation
Animal Blood Collection
Dilution of Blood
Beagle PBMC-Based Assays
PBMCs are a versatile sample type in preclinical research due to their role in the immune system. Assays using PBMCs help assess immune function, response to therapies, and disease pathophysiology.
Animal PBMCs are also commonly used in preclinical Safety, Toxicology and Translational research to help select the right in vivo model for late preclinial studies.
The following are common PBMC-based assays in preclinical research.
Flow Cytometry
This assay quantifies and analyzes various immune cell subsets within PBMCs, such as T cells, B cells, NK cells, and monocytes. Flow cytometry is widely used to determine immune status in diseases like HIV or cancer, evaluate immune responses to therapies, and track cell phenotypes in clinical trials.
ELISPOT (Enzyme-Linked Immunospot) Assay
ELISPOT measures the frequency of cytokine-secreting cells, indicating immune activation. It is often used in vaccine trials or infectious disease research to assess cellular immune responses by quantifying cytokines like IFN-γ, which indicates T-cell activation.
Proliferation Assays
These assays measure the ability of PBMCs to proliferate in response to specific antigens or mitogens. They are used in immunological research to assess immune responsiveness in autoimmune diseases, vaccine trials, or immunodeficiencies. Proliferation assays help determine immune system activation and potential deficiencies in cell-mediated immunity.
Intracellular Cytokine Staining (ICS)
ICS is used to detect cytokine production within individual cells using flow cytometry. This assay helps identify specific functional responses, such as Th1, Th2, or Th17 responses, by measuring cytokines like IL-2, IFN-γ, and TNF. It’s particularly valuable in vaccine and immunotherapy studies.
Published Research Using Beagle PBMCs
The study by Razmara et al. (2023) investigates the use of peripheral blood mononuclear cells (PBMCs) from beagle donors in a clinical trial for allogeneic adoptive natural killer (NK) cell therapy targeting dogs with naturally occurring melanoma.
Key Highlights of Beagle PBMCs
Cell Expansion and Function
PBMCs were used as a source for NK cell expansion without prior CD5 depletion, yielding robust NK cells with enhanced activation and cytotoxicity-related gene expression (e.g., GZMB, NCR3). This approach showed improved NK cell functionality compared to traditional methods.
Clinical Outcomes
The treatment was safe, with no significant adverse effects observed. A notable case reported a dog surviving 445 days post-treatment, indicating potential efficacy of the therapy.
Genomic Insights
Single-cell sequencing revealed sustained NK cell activity and uncovered previously unrecognized similarities between NK and CD8 T cell gene expression. This suggests potential new biomarkers and mechanisms underlying immune responses in canine models.
Implications for Immunotherapy:
The study highlights PBMC-expanded NK cells as a promising avenue for cancer immunotherapy in veterinary and potentially translational contexts. It also underscores the value of genomic profiling in optimizing cell-based therapies.