Endotoxin exposure alters viability and apoptotic potential of buffalo neutrophils

Buffalo which contributes towards the social well being of millions of South East Asian farmers commonly suffer from various infectious diseases. There is paucity of literature on the innate immune response of buffalo specially the cells like neutrophils which are the first line of defense. Buffalo neutrophils were isolated by using Hi Sep LSM-1077 along with Dextran and viability was determined by trypan blue exclusion test. Presence of myeloperoxidase suggested presence of neutrophils. The freshly isolated neutrophils showed a purity of 98.4% with viability up to 98.44 ± 0.35%. There was a significant decrease (p<0.05) in the number of viable neutrophils after 24 hr, 48 hr and 72 hr post-incubation in the control group. LPS challenge showed significantly (p<0.05) more number of viable neutrophils at 48 and 72 hr compared to non LPS group. Buffalo neutrophils undergoing apoptosis showed condensation of nucleus and presence of apoptotic bodies with intact cell membrane. LPS challenge resulted a significant decrease in the number of apoptotic neutrophils compared to control group at different time intervals. It is the first data on the viability and apoptotic potential of neutrophils of water buffalo suggesting that LPS treatment activated the neutrophils which led to significant reduction in apoptosis and increased life span of neutrophils as compared to untreated cells.