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           On the basis that ozone is a very rective and potenially toxic gas, it has been a common
           wisdom to use a fairly narrow range of ozone concentration anong 5 and 40µg/ml of blood.
           This was based on empirical data, that low ozone concentrations are immunostimulatory
           while high concentration are suppressive. After having clarfied that an important
           mechanism of action of ozone is to induce cytokine productions by mononuclear cells,
           we could define a reliable end-point and correlate ozone concentration and cytokine levels
           after a suitable incubation of blood. The main aim of this research was to achivev an effective
           immunostimulation with the least toxic effects by measuring the level of ozone-induced hemolsis
           (below 3.5%), possible formation of metahemoglobin (always absent), morphologic damage
           (absent below 80µg/ml of ozone) as evaluated by electron microscopy, plasma level of lipid
           hydroperoxides (increasing 3 fold after ozonization with 90µg/ml ozone/ml of blood but rapidly
           returning to base line values) and intraerythrocytic reduced glutathion levels never below
           10 % and repidly restored. Unexpectedly and contrary to ozone dosage usually used
           in autohemotherapy (from 5 to 40µg/ml O₃/ml of blood), we found that we could
           raise the ozone level and the most effective concentrations without toxicity are ranging
           between 50 and 80µg/ml depending upon individual plasma levels of anti-oxidant compounds.
           The concept of correlating the production of cytokines of blood mononuclear cells versus ozone
           concentration has thus represented a crucial advantage and it has become an indispensable
           end-point

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