Flow cytometric determination of oxidative burst activity of equine peripheral blood and bronchoalveolar lavage-derived leucocytes.

This research article presents the development of flow cytometric techniques to evaluate oxidative burst activity in various types of horse blood cells and lung cells. The techniques were found to be suitable and efficient, and the study identified differences in cell reactions to stimulants.

Development of Flow Cytometric Techniques

• The researchers developed flow cytometric techniques for the purpose of evaluating oxidative burst activity. Oxidative burst is a process where certain cells produce reactive oxygen species to kill pathogens.
• A key aspect of this research was focusing on equine peripheral blood neutrophils and lymphocytes, as well as bronchoalveolar lavage derived pulmonary alveolar macrophages and lymphocytes. In simpler terms, they evaluated cells from the bloodstream and cells from the lung tissue of horses.

Process of Evaluation and Findings

• The method involved measuring the oxidation of dichlorofluorescin, observed through the increased fluorescence of cells. The cells were stimulated with phorbol myristate acetate or other various stimulants.
• The authors found that flow cytometry was a suitable method for the evaluation of intracellular oxidation in all cell populations they tested. Also, they found this method to be rapid and that it does not require cell separation before analysis.
• The research found heterogeneous cell populations with differing responsiveness to phorbol myristate acetate-stimulated oxidative burst in both peripheral blood neutrophil and alveolar macrophage populations. This means that not all cells reacted in the same way to the stimulants, some showing different levels of oxidative burst activity.

Significance of the Study

• The characterisation of flow cytometric techniques for the evaluation of oxidative burst activity in equine peripheral blood and bronchoalveolar lavage-derived leucocytes is significant as it presents a direct, efficient way to study this cellular response. Such studies can help understand various pathologies in horses and potentially other species.