Figure 1From: Reduced levels of reactive oxygen species correlate with inhibition of apoptosis, rise in thioredoxin expression and increased bovine leukemia virus proviral loadsSpontaneous apoptosis and ROS production in short term cultures. A) Peripheral blood mononuclear cells (PBMCs) from BLV-infected (n = 13) and control (n = 6) sheep were isolated and cultivated for 24 h. B cells were labeled using anti-IgM monoclonal antibody (clone 1H4) and FITC-conjugated rabbit anti-mouse and fixed in ethanol. After staining with PI, hypodiploid cells (sub-G1 population) considered to be apoptotic were quantified by flow cytometry. Data are presented as the means of apoptotic rates ± standard deviation. ** denotes the statistical significance according to the non-paired Student's t test p < 0.01. B) PBMCs from BLV-infected (n = 13) and non-infected sheep (n = 6) were seeded in 24-well plates at a density of 106 cells/ml and incubated for 30 min at 37°C with 10 μM of CM-H2DCFDA. After 24 h of culture, B cells were stained using anti-IgM monoclonal (clone Pig45) and Alexa Fluor 647-conjugated donkey anti-mouse antibodies. The intracellular ROS levels were determined by flow cytometry and are presented as the mean fluorescence intensities (± standard deviation) of cellular chloromethyldichlorofluorescein (CM-DCF) within B and non-B lymphocyte populations. *** denotes the statistical significance according to the non paired Student's t test p < 0.001. C) Correlation between apoptotic rates and ROS levels measured in ex vivo cultures. Apoptotic rates and ROS levels were determined by flow cytometry as described in panels A and B, respectively. A correlation coefficient R2 = 0.4097 was calculated from the linear regression analysis on percentages of apoptotic B cells and means of ROS fluorescence intensities. p < 0.05 denotes the statistical significance according to the non parametric Spearman test.Back to article page