The accumulation of calcium by the polymorphonuclear leucocyte treated with staphylococcal leucocidin and its significance in the extrusion of protein

Woodin, A. M.; Wieneke, Antonnette A.

doi:10.1042/bj0870487

Cited by 63 publications

(42 citation statements)

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“…Moreover, cellular concentrations of both cyclic nucleotides, and thus cellular functions, can be influenced by autonomic neurohormones (5,8). Evidence for the existence of such a bidirectionally regulated biologic system in neutrophils was reported recently (4,15). For example, acetylcholine enhanced both the immunologic discharge of lysosomal enzymes and the accumulation of neutrophil cyclic GMP whereas epinephrine inhibited enzyme discharge and promoted the accumulation of cyclic AMP.…”

Section: Discussionmentioning

confidence: 94%

“…Several reports have illustrated the requirement of extracellular calcium for the selective discharge of lysosomal enzymes from granulocytes (3,15,16). The data in the present study reveal that extracellular calcium is required for the immunologic discharge of /3-glucuronidase from purified human neutrophils regardless of the type of immunologic stimulus employed.…”

Section: Discussionmentioning

confidence: 99%

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Mediation of Immunologic Discharge of Lysosomal Enzymes From Human Neutrophils by Guanosine 3',5'-Monophosphate

Ignarro

George

1974

The Journal of Experimental Medicine

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The purpose of this investigation was to elucidate the relationship of cyclic GMP and calcium to the immunologic discharge of lysosomal enzymes from purified human neutrophils. Contact of neutrophils with a variety of immunologic stimuli, including zymosan particles treated with either normal or rheumatoid arthritic (RA) serum, heat-aggregated (agg) IgG, particulate and immobilized agg IgG each treated with RA serum, and zymosan-treated serum, provoked the discharge of ß-glucuronidase, but not cytoplasmic lactate dehydrogenase, and stimulated the accumulation of cyclic GMP. Both enzyme release and elevation of cyclic GMP levels required the presence of extracellular calcium as neither cellular event proceeded in its absence. Cholinergic enhancement of the immunologic secretion of ß-glucuronidase from neutrophils by acetylcholine was associated with a concomitant accumulation of cyclic GMP. These actions of acetylcholine on neutrophils did not proceed in the absence of extracellular calcium. Whereas the concentrations of cyclic GMP in neutrophils were elevated by both immune reactants and a combination of the latter and acetylcholine, cyclic AMP levels remained unaltered. Thus, cyclic GMP, but not cyclic AMP, was associated with the immunologic and pharmacologic discharge of lysosomal enzymes from neutrophils. Contrariwise, cyclic AMP, but not cyclic GMP, was associated with inhibition of lysosomal enzyme release. For example, dibutyryl cyclic AMP and epinephrine inhibited the release of ß-glucuronidase from neutrophils that was elicited by each of the immune reactants tested. Moreover, cyclic AMP levels in the cells were elevated markedly in every instance that enzyme discharge was inhibited by epinephrine. Epinephrine did not alter the neutrophil concentrations of cyclic GMP at times when those of cyclic AMP were elevated. The data in this report constitute partial evidence that the immunologic discharge of lysosomal enzymes from human neutrophils is mediated or signaled by intracellular cyclic GMP and that calcium is linked to this stimulation of enzyme secretion.

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Mediation of Immunologic Discharge of Lysosomal Enzymes From Human Neutrophils by Guanosine 3',5'-Monophosphate

Ignarro

George

1974

The Journal of Experimental Medicine

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“…The presence of extracellular calcium ions was shown to be necessary for secretions in other tissues, such as secretion of catecholamines from adrenal chromaffin cells (26), acetylcholine from nerve endings (27), secretion of protein from submaxillary salivary gland (28), and protein release from leucocidine-treated leucocytes (29). Therefore, it is tempt ing to speculate as to whether a mechanism common with secretion processes in these tis sues is involved in the process of histamine release although available evidence is still lack ing for decisive conclution.…”

Section: Methodsmentioning

confidence: 99%

Calcium-Dependent Histamine Release With Degranulation From Isolated Rat Mast Cells by Adenosine 5'-Triphosphate

Sugiyama

1971

Jpn.J.Pharmacol

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A preliminary report on these findings has been published earlier (11). MATERIALS AND METHODSIsolation of mast cells. Mast cells were isolated from the peritoneal fluid of Wistar rats (male, 250-350 g) by the gum arabic density gradient centrifugation procedure which is a modification of Kimura's method (12). Gum arabic stock solution was prepared as follows: A solution of 32 g of gum arabicpowder dissolved in 90 ml of deionized water at 80°C was filtered through gauze. After centrifugation to remove insoluble sediment, the solution was diluted with deionized water to make 1.090 in specific gravity, and pH adjusted to 7.4 with saturated NaOH. NaC1 was added to give a final concentration 0.2%. This stock solution was kept at 4°C until use.

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“…The inorganic P content of the trichloroacetic acid extracts was determined. The concentration of inorganic P in the cell was calculated by assuming the extracellular volumes in the pellets of normal and leucocidin-treated cells to be 25 and 55% respectively (Woodin & Wieneke, 1963). Effect of antitoxin on the synergism of the two components of leucocidin.…”

mentioning

confidence: 99%

“…The determination and the unit of 1963 fi-glucuronidase is described by Woodin & Wieneke (1963). Inorganic, total and acid-labile phosphorus were determined by the Berenblum & Chain (1938) method after hydrolysis or digestion as described by Woodin (1961).…”

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confidence: 99%

The incorporation of radioactive phosphorus in the leucocyte during the extrusion of protein induced by staphylococcal leucocidin

Woodin¹,

Wieneke²

1963

Biochemical Journal

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It has been shown that treatment of the polymorphonuclear leucocyte with leucocidin stimulates the incorporation of radioactive phosphorus into the phospholipids of the cell (Woodin, 1962). The radioactivity of the precursors was not known and so it was uncertain if this represented an increased turnover. The present paper shows that the stimulated incorporation of radioactive phosphorus is found in the nucleotides and other acidsoluble compounds of the leucocidin-treated leucocyte. There is no evidence for a causal connexion between this stimulated incorporation and the extrusion of protein from the granules of the leucocyte. METHODSThe two components of leucocidin were crystallized as described by Woodin (1960); unless otherwise indicated they were added to cell suspensions to give 2 jig./108 cells.Polymorphonuclear leucocytes were obtained from peritoneal exudates of rabbits (Woodin, 1962) and will be referred to below as leucocytes. ATP and ADP were obtained from C. F. Boehringer und Soehne, Mannheim, Germany; GTP and GDP were obtained from the Sigma Chemical Co., St Louis, Mo., U.S.A. The solutions used for suspending the cells were Hanks (1948) Incorporation of 32p into the nucleotide& of the leucocyte.Tubes containing 5 x 109 cells in 10 ml. of Hanks solution were incubated for 10 min. at 370 and then leucocidin was added to one tube and 82p (0.05 ml. containing 107 counts/ min.) was added to both tubes. Incubation was continued for 10 min. and the tubes were cooled in ice. They were centrifuged at 0°for 5 min. at 2000 rev./min. and the supernatant was rejected. The cell pellets were extracted for 2 hr. at room temperature with 20 vol. of chloroformmethanol (2:1, v/v). The suspensions were centrifuged and the supernatants used for the preparation of the phospholipids (see below). The cell residues were washed with ether and dried in a stream of air. The powder was then extracted with 10 ml. of ice-cold HC104 (3 %, w/v), residual ether and occluded air being removed by evacuating the tubes until the cell residues no longer floated on the surface. Extraction was continued for 30 min. and then the suspensions were centrifuged at 20 000 rev./min. for 20 min. The slightly opalescent supernatant was passed down a column containing 100 mg. of Norit and 100 mg. of Celite (Threlfall, 1957), which was then washed three times with 3 ml. of water. The effluent from the column was collected; it contained the nucleotide-free, acid-soluble fraction. The nucleotides were eluted from the washed Norit-Celite column in 7 ml. of pyridine and freeze-dried. The solid was dissolved in a small volume of water and put on paper for chromatography. After separation as described below the specific activities of the nucleotides were determined.In a similar experiment, 2 x 109 leucocytes in 12 ml. of Hanks medium were incubated alone or with added leucocidin for 10 min. 32P (01 ml.; 2 x 108 counts/min.) was added and incubation at 37°continued for 20 sec. The suspensions of cells were then diluted with 75 ml. of icecold Hanks mediu...

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The accumulation of calcium by the polymorphonuclear leucocyte treated with staphylococcal leucocidin and its significance in the extrusion of protein

Cited by 63 publications

References 9 publications

Mediation of Immunologic Discharge of Lysosomal Enzymes From Human Neutrophils by Guanosine 3',5'-Monophosphate

Mediation of Immunologic Discharge of Lysosomal Enzymes From Human Neutrophils by Guanosine 3',5'-Monophosphate

Calcium-Dependent Histamine Release With Degranulation From Isolated Rat Mast Cells by Adenosine 5'-Triphosphate

The incorporation of radioactive phosphorus in the leucocyte during the extrusion of protein induced by staphylococcal leucocidin

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