NADI REACTION OF MYELOID LEUCOCYTES AND VITAMIN K

The stable Nadi reaction of myeloid leucocyte granules has been reported to be a non-enzymatic reaction because of the stability against fixation and heating for 10min at 100°C. Dimethyl-p-phenylenediamine and α-naphthol solutions are oxidized spontaneously, and a combination of these solutions gives a fine Nadi reaction. Freshly prepared regents are not so effective. Therefore, the chemical mechanism of Nadi reaction seems to be based on a dehydrative condensation of the two reagents.The present study demonstrated that some naphthoquinone may be a Nadi reaction factor in the granules of leucocytes obtained from the peritoneal cavity of rat by peptone stumulation. It was proved that the reaction in these granules was affected by acetone. Although about 10% of leucocytes showed a positive reaction to treatment with pure acetone for 2hr (probably eosinophiles), all of the reactivity was lost when the treatment was continued 24hr longer. But the reaction was restored by vitamin K. However, vitamin K had no effect when leucocytes were treated with 96% acetone. This experiment suggests the presence of a naphthoquinone-like substance, which may be bound to phospholipids.A clear liquid substance of high viscosity was extracted from leucocytes with 100% acetone. This was not soluble in alkali or water but was soluble in ethanol, chloroform and acid, and slightly soluble in methanol. This liquid became faint yellowish brown in color when exposed to light. In ultraviolet spectrophotometry, this acetone-extracted substance showed fine absorption peaks at 256 and 325mμ. This absorption spectrum corresponds with that of naphthoquinones, i. e. vitamin K.