Adenylate pool and energy charge in human lymphocytes and granulocytes irradiated at 632 nm (HeNe Laser)

Aim of this report was to investigate the adenylate pool and the energy charge in human white blood cells exposed to increasing time (15, 30 and 60 mm) of HeNe laser treatment. EDTA treated human blood diluted 1:1 whith 0.88% KCI was added (1:5) with NaCI-dextran solution to allow sedimentation of red blood cells. 6 ml of white cells floating in the supernatant were layered on 3 ml of Lymphoprep in plastic tubes and each tube was centrifuged (from 50 to 5000 x g for 5 mm). Granulocytes were concentrated in the lower phase, whilst lymphocytes were in the intermediated phase. After further purification cytological homogeneity was tested by a cell counter. Granulocytes and lymphocytes were irradiated at +22°C with HeNe (Space, Valfivre equipment). On these population ATP was tested by luminometric procedure, the adenylate pool was separated by HPLC (Jasco) on neutralyzed perchloric extracts. ATP concentration increases in lymphocytes (+63.9%, p < 0.01) and in granulocytes (+25.0%, p < 0.05) after 60 mm irradiation. The adenylate pool (tested by HPLC) does not change significatively in lymphocytes or granulocytes after 30 mm irradiation, whilst in 60 mm irradiated lymphocytes and granulocytes a significative increment was observed in nucleotide concentration. No changes were observed in energy charge according to Atkinson.