The Periodic Acid‐Schiff Reaction in Neutrophil Leukocytes in Chronic Myeloproliferative Diseases A microspectrophotometric study

Abstract

Microspectrophotometric quantification of the PAS reaction has been carried out in about 4500 individual mature neutrophil leukocytes from 20 normal persons, and from 15 patients with chronic myelocytic leukaemia (CML), 18 patients with myeloid metaplasia, and 17 patients with polycythaemia vera (PV). The reaction is considered to be a quantitative measure of the amount of PAS reactive material (PASMa) in neutrophils, most of it probably glycogen. Neutro‐phils from CML had a mean amount of PASMa that was 39 per cent below normal, the frequency distribution of PASMa per cell being approximately normal in individual cases. Neutrophils from myeloid metaplasia usually had elevated mean amounts of PASMa and considerably elevated variance between cells within subjects. Neutrophils from PV had a mean amount of PASMa that was 33 per cent above normal, with a highly augmented variance between cells within subjects. There was no correlation between the number of nuclear lobes and the amount of PASMa per cell in any diagnostic group. The quantified PAS reaction is considered to be of diagnostic importance for the differentiation of CML and myeloid metaplasia.

Stimulus‐specific defect in oxidative metabolism of polymorphonuclear granulocytes in polycythemia vera

“…Whether the PV granulocyte is activated or not is controversial. Some, but not all, previous studies have suggested that resting PV PMNs may have higher oxidative metabolism, phagocytosis and glycogen metabolism (1)(2)(3)(4)(5). We could not detect any deviation from normal PMNs with regard to spontaneous CL.…”

“…Thus, Wagner (1) found an increased glycogen content in PMNs from patients with PV, and a 1.5 times greater rate of glycogen turnover was reported by Luganova & Seitz (2). A mean increase of 33% in PAS-reactive material in PV granulocytes was noted by Gahrton (3). Enhanced phagocytosis of yeast (4) and of polystyrene particles ( 5 ) has also been shown.…”

Cytochemical Variants of Neutrophil Leukocyte Populations in Chronic Myelocytic Leukaemia

“…The control material consisted of 20 normal subjects previously reported (Gahrton 1966~). Both blood and bone marrow neutrophils were studied repeatedly in one of these cases.…”

“…P A S staining. The McManus PAS reaction (Mc Manus & Mowry 1960) was performed as previously modified (Gahrton 1964(Gahrton , 1966: 0.5 % periodic acid for 60 minutes; distilled water for 3 minutes (1 minute in each of three rinses); Schiff reagent (Fuchsin-Sulphurous Acid Test Solution, Harleco, Hartmann-Leddon Company, Philadelphia, Pa.) for 60 minutes; sulphuric acid rinse (10 % sodium metabisulphite 6 ml., normal hydrochloric acid 5 ml., distilled water 100 ml.) for 6 minutes (2 minutes in each of three rinses); rinsed in running tap water for 5 minutes; rinsed in distilled water for 30 seconds.…”

“…The extinction values were automatically integrated over the neutrophil area and registered for each neutrophil as the total extinction at 546 mp, E,,,NL, a measure of the amount of PASMa (Gahrton 1964(Gahrton , 1966d. All values were related to a normal, standardized control stained and measured at the same time the control was representative of the mean of twenty normal subjects as previously reported (Gahrton 1966). 30 to 45 neutrophils were measured in each case.…”