Immune cells carry out some of their functions by means of cell-to-cell contact and via membrane receptors or antigens that are expressed at the cellular surface. Some molecules appear at various stages of cell differentiation or activation. In the present study, we recorded the following molecules: CD4, CD8, CD19, CD56, CD71, CD97, CD134, and CD173.
Phenotypic analysis of the various cell populations was carried out by immunofluorescence by means of flow cytometry with double or triple labeling. This technique constitutes a useful tool for cell identification and characterization. Following intravenous peripheral blood collection, 50 μl of blood were incubated for 10 minutes with 3 μl of the following combinations of monoclonal antibodies (Pharmigen):
CD4-PE + CD8-FITC + CD19-Cy5
CD8-FITC + CD56-PE
CD71-FITC + CD97-PE
CD134-FITC + CDw137-PE
Afterwards, 1 mL of a hematolysis solution (
Quicklysis, Cytognos) was added and the blood solution was kept in the dark for 5 minutes, after which time 10,000 cells were subjected to cytometric analysis.
Immunofluorescence analysis was carried out by means of a Vantage (Becton Dickinson)
FACS flow cytometer, equipped with an argon laser (488 nm) that excites the FITC, PE, and Cy5 fluorchromes, emitting at 520, 575, and 667 nm, respectively.
Reig-Ferrer A., Ferrer-Cascales R., Santos-Ruiz A., Campos-Ferrer A., Prieto-Seva A., Velasco-Ruiz I., Fernandez-Pascual M.D, & Albaladejo-Blazquez N. (2014). A relaxation technique enhances psychological well-being and immune parameters in elderly people from a nursing home: A randomized controlled study. BMC Complementary and Alternative Medicine, 14, 311.