Detection of interleukin -1β from isolated human lymphocyte in response to lipopolysaccharide and lipoteichoic acid

AIM: To detect the interleukin -1β levels from single and pooled isolated human lymphocytes in response to lipolysaccharide and lipoteichoic acid. MATERIALS AND METHODS: Blood collected from healthy individuals (O +ve, A +ve, B +ve, and AB +ve) were subjected to gradient centrifugation to isolate lymphocytes. Different lymphocyte concentrations were used for in vitro pyrogen assay. Lymphocytes isolated were challenged with 5 EU of Gram negative (LPS) and 1 μg/μl of Gram positive (LTA) pyrogens in vitro and the inflammatory cytokine, Interleukin 1β (IL-1β) release was measured by Sandwich ELISA method. RESULTS: The results indicated that the release of IL-1β increases immediately after the initiation of incubation and reaches a maximum at 4 to 6(th) hour and then stabilizes for both the pyrogens. Furthermore, IL-1β release by 5 EU of LPS and 1 μg/μl of LTA is dependent on lymphocytes concentration. It was also observed that the difference in blood group did not interfere with the IL-1β release. CONCLUSION: The isolated lymphocyte system can be used as an alternative to the in vivo rabbit pyrogen assay.