Surface‐active substances and the liberation of enzymes from rabbit polymorphonuclear leucocytes

Abstract

Much can be learned of the function of the white cell by studying its constituent enzymes. Our present knowledge of white-cell enzymes has been reviewed in papers by Barnes (1940) and Rebuck (1947 Rossiter, 1948, esterases (Rossiter & Wong, 1949) and, more recently, the /3-glucuronidases of rabbit polymorphonuclear leucocytes. A preliminary report of these experiments has already appeared . METHODSCell preparation8. Polymorphonuclear leucocytes were obtained from the peritoneal cavity of the rabbit by the method of de Haan (1918). A warm sterile solution of 0.9% NaCl (300-500 ml.) was run into the peritoneal cavity and drained off 3-4 hr. later into a beaker containing a few grains of heparin. Alternatively, the solution was left in the animal overnight and a further quantity was run in the following morning and withdrawn immediately. In each instance the resulting opalescent fluid contained 2000-16,000 leucocytes/cu.mm., at least 95 % of which were polymorphonuclear.Alkaline pho8phatase e8timation. Alkaline phosphatase was determined by the method of King & Armstrong (1934) in which the phenol, liberated by the hydrolysis of disodium phenyl phosphate, is estimated by the method of Folin & Ciocalteu (1927). Sufficient magnesium chloride was added to each tube to make the final magnesium concentration 0 001 M. Details of the test have been published by Cram & Rossiter (1949). The results were recorded as the amount of phenol (in mg.) liberated, under the standard conditions of the test, by 100 ml. enzyme in 1 hr.

The Isolation and Properties of the Specific Cytoplasmic Granules of Rabbit Polymorphonuclear Leucocytes

“…Initial experiments with a variety of these enzymes gave erratic results in terms of both recovery and specific activity of the enzymes. Previous reports by Rossiter (26) had emphasized that the enzymatic activity of rabbit leucocytes could be increased considerably by the use of surface-active agents. This treatment was associated with the "solubilization" of cytoplasmic components and was thought to be related to the release of enzymes from structural elements.…”

Leukocyte Alkaline Phosphatase Cytochemistry: Applications and Methods*

“…4 Kay's conclusions were soon confirmed by a number of investigators using quantitative biochemical techniques. [5][6][7][8][9][10][11] It is ironic that Fiessinger and Boyer12 came to an opposite opinion regarding the presence of alkaline phosphatase activity in leukocytes, because they were unable to detect an increase in serum alkaline phosphatase after adding to the serum a large number of leukocytes obtained from the buffy coat of a patient with chronic myelogenous leukemia. For a more complete review of the early biochemical studies on LAPA, reference should be made to the publication by Valentine and Beck13 in 1951.…”