Sequential changes in human polymorphonuclear leukocytes after urate crystal phagocytosis. An electron microscopic study

Schumacher, H. R.; Phelps, Paulding

doi:10.1002/art.1780140411

Cited by 77 publications

(27 citation statements)

References 22 publications

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“…But sequential study of phagocytes exposed to RISU crystals for varying times before fixation showed that a phagosome was formed by inversion of the plasma membrane as expected, but that breaks in the sac soon occurred (19). T h e crystal appeared free in the cytoplasm at this point.…”

Section: Crystal-induced Inflammationmentioning

confidence: 60%

An overview of cellular and molecular mechanisms in crystal-induced inflammation

McCarty

Kozin

1975

Arthritis & Rheumatism

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Section: Crystal-induced Inflammationmentioning

confidence: 60%

An overview of cellular and molecular mechanisms in crystal-induced inflammation

McCarty

Kozin

1975

Arthritis & Rheumatism

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“…6. Direct rupture of dye-filled, MSU-containing lysosomes into the cytosol can be observed in living cells (see above), and ultrastructural studies (14,38) indicate that lysosomal membrane breaks and diffuse reaction products of acid phosphatase and myeloperoxidase are observed in cells that have ingested MSU. These discontinuities or enzyme redistributions are not observed following uptake of other particles (14).…”

Section: Discussionmentioning

confidence: 98%

“…Uptake of crystalline MSU, but not of other particles (zymosan, immune complexes, latex, calcium pyrophosphate dihydrate), leads to cell death of leukocytes (2, 5,13,14). Cell death occurs after a latent period and is associated with release of lysosomal hydrolases together with cytoplasmic LDH.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore Schumacher and Phelps (14) have shown by means of electron micrographs that, whereas MSU crystals newly ingested by human neutrophils are soon located within membrane-bounded organelles, the crystals subsequently lie free in the damaged cytosol and their limiting membranes disappear progressively. Based on Allison's original model of silica-induced inflammation (15), on our in vitro studies, and on the suggestions of Wallingford and McCarty (12), we came to the conclusion that MSU crystals cause acute gouty inflammation by disrupting the internal membranes of the secondary lysosomes of leukocytes by a kind of "perforation from within" (1).…”

Section: Mechanisms Of Lysosomal Enzyme Release From Leukocytes IV Imentioning

confidence: 99%

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Mechanisms of lysosomal enzyme release from leukocytes

Hoffstein

Weissmann

1975

Arthritis & Rheumatism

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In order to determine the possible mechanisms whereby interactions between phagocytic cells and crystals of monosodium urate (MSU) lead to cell death with simultaneous release of both cytoplasmic and lysosomal enzymes, phagocytic leukocytes of the smooth dogfish shark Mustelus canis were studied by means of light and electron microscopy, and biochemistry. Lysosomes of these cells can be stained supravitally with toluidine blue and are large enough (0.7-0.8 I*) to be clearly resolved with the light microscope. Light microscopic observations showed that of cells exposed to MSU 87% of those containing visible ingested crystals died within 1 hour, whereas 92% of adjacent cells in the same wet mount without such crystals survived. Cell death occured after a latent period of 10-15 minutes following fusion of lysosomes with crystal-containing phagosomes. Electron microscopic examination of both dogfish and human leukocytes exposed to MSU for more than 1 hour and then fixed in situ revealed occasional discontinuities or ruptures in secondary lysosome membranes. Endogenous peroxidase activity could be cytochemically localized in primary and secondary lysosomes and in the cytoplasm adjacent to such ruptured secondary lysosomes. It was not seen adjacent to primary lysosomes, a result indicating that the cytoplasmic reaction product was not a diffusion artifact. T o exclude the possibility that crystals were exercising their affect primarily upon the plasma membrane, suspensions of dogfish buffy coat cells were incubated with cytochalasin B (5 pg/ml, 10 minutes), which inhibits phagocytosis but not exocytosis of lysosomal enzymes b y stimulated phagocytes. Whereas cells exposed to MSU crystals released 30% of their content of lysosomal pglucuronidase activity and 28% of their cytoplasmic lactate dehydrogenase (LDH) activity within 3 hours, preincubation with cytochalasin B reduced the release of LDH activity within that period to 6% but reduced the release of p-glucuronidase activity only to 20%.Preincubation with 10-3 M cyclic adenosine monophosphate (CAMP) and theophylline M), which inhibit lysosomal fusion, reduced the release of both LDH and p-glucuronidase activities to 7% and 6% respectively. Cells that were preincubated with both cytochalasin B and CAMP + theophylline released only 1% LDH activity and 4% pglucuroni-

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“…The mechanism of cell destruction is indicated in studies of sequential ultrastructural changes in polymorphonuclear leukocytes after phagocytosis of urate crystals (7,14), and of the interaction of urate crystals with model membrane systems-erythrocytes (15), isolated lysosomes, and artificial anionic liposomes (16). It appears that crystals of monosodium urate, like those of silica (1 7), can disrupt cholesterol-rich membranes, perhaps when hydrogen bonding is allowed to occur between "phenolic" hydrogens of the crystal and lone-pair electrons on the proton-acceptor oxygen of membrane phosphatidylcholine phosphate esters.…”

mentioning

confidence: 99%

The phlogistic potential of urate in solution

Malawista

Blaricom

Cretella

et al. 1979

Arthritis & Rheumatism

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Physiologic concentrations of urate in solution had a clear and dosagedependent suppressive effect on the concomitant uptake of radioiodinated human serum albumin (13'EHSA) during the ingestion of particles (bacteria) by human polymorphonuclear leukocytes (PMN). The effect was rapid, reversible on washing the cells, and demonstrable over a wide range of incubation times and ratios of particles to PMN. In this method, the incubated cells are washed, and most surface radioactivity is thereby removed, the amount of label trapped during particle ingestion then usually provides a sensitive measure of phagocytosis. However, no evidence for an effect of urate on phagocytosis was found by use of several other methods for the assessment of particle u p take, including 1) direct microscopic counts of cell-associated bacteria, 2) studies in which the bacteria were radiolabeled (instead of the media), and 3) the quantitative recovery of surviving bacteria from the media and from disrupted cells. An alternative explanation for the results with '"I-HSA is that urate interferes reversibly with the binding of albumin to the cell membrane, including those portions that become internalized during ~~ From the Section of Rheumatology, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut.Supported in part by grants from the USPHS (AM-10493, AM-19742, AM-5639, AM-07107), the Arthritis Foundation, and the Kroc Foundation of Santa Ynez, California. Part of this work has appeared in abstract (I).Stephen E. Malawista phagocytosis, thereby creating the false impression of less phagocytosis, when there is really only less trapped label (i.e., albumin). In fact, "on-phagocytizing leukocytes-which do not transport albumin across the cell membraneincubated in serum-bder with added urate and with '"I-HSA and then drained (not washed) had much less cell-associated radioactivity than controls without added urate. Washing removed the difference (and most of the label from both groups). Crystals of urate and of silica are known to kill the cells that ingest them by disrupting the membranes of phagolysosomes, but only after protective protein has been digested. In the presence of dissolved urate, this inflammatory mechanism might be augmented by the relative lack of protective, membrane-associated protein. Indeed, as measured by the release of lactic dehydrogenase, injury to cells given silica crystals was augmented considerably in the presence of dissolved urate.The persistent defect that characterizes gout is the supersaturation of body fluids with sodium urate and the propensity of that urate to form crystals, especially in joints and skin. Superimposed upon the persistent defect are periodic inflammatory paroxysms: acute gouty arthritis. A great deal of work has dealt with how urate crystals, through their interaction with phagocytic cells, contribute to the propagation and amplification of gouty inflammation (for review, see reference 1); one aspect of that interaction-the effect of crystals on membrane i...

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Sequential changes in human polymorphonuclear leukocytes after urate crystal phagocytosis. An electron microscopic study

Cited by 77 publications

References 22 publications

An overview of cellular and molecular mechanisms in crystal-induced inflammation

An overview of cellular and molecular mechanisms in crystal-induced inflammation

Mechanisms of lysosomal enzyme release from leukocytes

The phlogistic potential of urate in solution

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