Applications of Flow Cytometry in Veterinary Science


What is flow cytometry?
Applications of flow cytometry in veterinary medicine
Limitations and future prospects
references
Continue reading


The last few decades have completely transformed veterinary science; For example, the life expectancy of cats and dogs has increased to higher levels than ever before. Veterinary science not only improves the health and welfare of animals, but can also be vital to human health by monitoring and controlling zoonotic diseases. One technique that has gained popularity is flow cytometry, particularly in assessing the immunological status and development of animals.

Credit: Sergey Nivens/Shutterstock.com

Credit: Sergey Nivens/Shutterstock.com

What is flow cytometry?

Flow cytometry is a widely used technique to assess quantitative and qualitative aspects of cells, especially in hematology and immunology. Flow cytometry uses lasers to generate signals detected and converted into data that provide insight into the cells being analyzed. Flow cytometry uses expensive equipment and requires highly skilled people.

In the past 30 years there have been significant advances in flow cytometry that have enabled wider application and easier navigation. It has also become a more reliable and effective tool, making it a more attractive technique in biochemical analysis. This has led to its increasing use in veterinary medicine.

Applications of flow cytometry in veterinary medicine

Flow cytometry is used in many areas of veterinary medicine; Immunology, oncology, diagnosis of autoimmune diseases, quantification of reticulocytes, drug surveillance and rabies.

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Flow cytometry has played a crucial role in assessing immune function, particularly in dogs. A subset of suppressive CD4+ T cells was identified using flow cytometry. Some animals that expressed higher levels of suppressive T cells were less likely to develop autoimmune diseases. This discovery led to the conclusion that these cells are relevant to the mechanism of self-tolerance as they would regulate and limit the immune system and prevent it from overactivating and attacking its own cells. This better understanding of the role of immune cells is crucial for the treatment of various diseases.

A new application for flow cytometry is the pharmacodynamic monitoring of drugs. For some life-threatening conditions, earlier proper treatment greatly increases the chances of survival, hence a reliable drug monitoring technique is required.

Photo Credit: Babul Pants/Shutterstock.com

Photo Credit: Babul Pants/Shutterstock.com

Cyclosporine, a powerful drug that inhibits T-cell production, is used in veterinary medicine to treat inflammatory and autoimmune diseases. Cyclosporine prevents the formation of immune cytokines such as IFN-Y and IL2, which weakens the immune response. Pharmacodynamic monitoring is the process of observing the effects of drugs and making sure they are within therapeutic values.

A research group developed a cyclosporine assay to measure its effects on canine tissues and observed two items from these assays; At low doses, the drug suppressed IFN-Y, while at higher doses it suppressed both IFN-Y and IL2. In this assay, flow cytometry was able to ensure drug efficacy and identify new markers.

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The use of flow cytometry is not limited to treatment, but can also be used to diagnose conditions such as parasitemia (a parasitic blood infection) in dogs. Two main methods are currently used to diagnose this; View of a blood smear and PCR (polymerase chain reaction).

Using blood smears can be difficult because many parasites would be required for the test to give a positive result; leading to many false-negative results. PCR as a technique is very sensitive; even a few parasitized red blood cells would give a false positive result. Flow cytometry is a middle ground between these techniques and allows for a more reliable result.

A new approach to the detection of intracellular rabies virus antigens involves the use of flow cytometry. One of the benefits of using flow cytometry in this field would be to reduce the time it takes to confirm a viral infection. Therefore, this technique could be crucial for industries currently trying to produce rabies vaccines or antiviral drugs.

Credit: hedgehog94/Shutterstock.com

Credit: hedgehog94/Shutterstock.com

Flow cytometry has a promising future in oncology. Acute leukemia in dogs is rarely cured and has a poor prognosis; this is also due to the poor lineage differentiation of the cells. A classification of acute leukemia into the different lines would improve the prognosis. The traditional methods currently used to characterize cell lines are subjective in interpretation and difficult to quantify; therefore, flow cytometry would enable further developments in this field. Flow cytometry can assess multiple characteristics of a large number of cells using comparatively objective criteria.

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Limitations and future prospects

Although flow cytometry has a promising future in many areas of veterinary medicine, it has its own set of challenges that need to be addressed. Limitations include the high cost of the instruments and the need for highly qualified personnel. Flow cytometry also requires single cell suspensions as aggregates would give false results. However, when looking at the limitations, one cannot overlook the uses, and the importance of flow cytometry still remains. Some information gleaned from flow cytometry was used to improve and refine a quantitatively more robust reverse transcriptase PCR assay; this shows the wide range of applications of flow cytometry and its importance in future scientific research.

references

  • J., Thomason & Archer, Todd & Mackin, Andrew & Stokes, John & Pinchuk, Lesya. (2014). Applications of flow cytometry in veterinary research and small animal clinical practice. Journal of Veterinary Medicine and Research. 1. 1-9.
  • Reggeti, F., & Bienzle, D. (2011). Flow cytometry in veterinary oncology. veterinary pathology, 48(1), 223-235. https://doi.org/10.1177/0300985810379435
  • Molla, D., Z. Tekle, A. Ayisheshim, D. Kebede, A. Wondimu and WM Ahmed, 2019. Flow cytometry and its diagnostic application in animal health: A review. Global Vet., 21:1-7. DOI: 10.5829/idosi.gv.2019.01.07

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