[Cytochemical demonstration and characterization of acid mucopolysaccharides in normal granulocytic series]

Metachromatic granulations are seen in myeloblasts, promyelocytes, myelocytes, metamyelocytes and young mature cells, after staining by all metachromatic dyes, except methylene blue. Furthermore by screening at various pHs granular metachromasia is detectable starting at pH 3.2 in 6% of case, at pH 4 in 78% of cases, at pH 4.5 in 16% of cases. The use of toluidine blue 10(-4) M in solutions at varying pH, after methylation and after alkali demethylation demonstrate the acid sulfated mucopolysaccharide react within the pH range (3.2), 4, 4.5, 5; persistence of non-alcohol resistant granular metachromasia above pH 5.5 suggests the presence of granules containing acid mucopolysaccharides staining as the result of possessing --COOH radicals. This data was verified by other histochemical techniques (Alcian blue and PAS staining) which have shown positive granules. All the data on the metachromatic, alcian blue and PAS positive granulations, have been carefully confirmed by chemical and enzymatic reactions. The results of this study led us to the conclusion that azurophilic granulations are made of a sulfated polysaccharide of chondroitinsulfate group. In the more immature cells alcian blue positive granules (mucopolysaccharide of acid hyaluronic type) and PAS positive granules (polysaccharides with a low sulfation level) can be demonstrated. From this data, it is suggested that azurophilic granulations are the final event of an evolutive process of polysaccharide material, fundamentally based on a progressive sulfation.