scholarly journals Isolation of Human Polymorphonuclear Leukocytes (Granulocytes) from a Leukocyte-Rich Fraction

2002 ◽  
Vol 2 ◽  
pp. 1393-1396 ◽  
Author(s):  
John Graham
Keyword(s):  
Peripheral Blood ◽  
Polymorphonuclear Leukocytes ◽  
Human Peripheral Blood ◽  
Human Polymorphonuclear Leukocytes

Human peripheral blood polymorphonuclear leukocytes (PMNs) or granulocytes from a leukocyte-rich plasma (LRP) are banded at an interface between two layers of iodixanol. If the denser layer of iodixanol is omitted the PMNs may alternatively be pelleted. The procedure can be adapted to blood from other species by small changes to the density of the two iodixanol layers. The method works optimally with EDTA- or citrate-anticoagulated blood.

scholarly journals Thromboxane generation by human peripheral blood polymorphonuclear leukocytes.

1978 ◽  
Vol 148 (3) ◽  
pp. 787-792 ◽  
Author(s):  
I M Goldstein ◽  
C L Malmsten ◽  
H Kindahl ◽  
H B Kaplan ◽  
O Rådmark ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Arachidonic Acid ◽  
Thin Layer ◽  
Peripheral Blood ◽  
Polymorphonuclear Leukocytes ◽  
Cytochalasin B ◽  
Human Peripheral Blood ◽  
Thromboxane B2 ◽  
Cyclooxygenase Inhibitor

Human peripheral blood polymorphonuclear leukocytes were stimulated to generate thromboxane B2 in a time- and concentration-dependent fashion upon exposure to serum-treated zymosan particles. Conversion by stimulated PMN of [14C] arachidonic acid to [14C]thromboxane B2 was confirmed by thin-layer radiochromatography, radio-gas chromatography, and mass spectrometry. Generation of thromboxane B2 was independent of platelet contamination and could be inhibited by the cyclooxygenase inhibitor, indomethacin. Cells rendered incapable of ingesting particles by treatment with cytochalasin B generated comparable amounts of thromboxane B2. These results suggest that human peripheral blood polymorphonuclear leukocytes synthesize thromboxanes in response to surface stimulation independently of phagocytosis.

scholarly journals Inhibitory effect of LY 255283 on the synthesis of leukotriene B4and thromboxane A2in human peripheral blood polymorphonuclear leukocytes and monocytes

1993 ◽  
Vol 2 (6) ◽  
pp. 407-409 ◽  
Author(s):  
M. Ugur ◽  
M. Melli
Keyword(s):  
Receptor Antagonist ◽  
Peripheral Blood ◽  
Polymorphonuclear Leukocytes ◽  
Human Peripheral Blood ◽  
Calcium Ionophore ◽  
Inhibitory Effect ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Human Monocytes

LY 255283 [(1-(5-ethyl-2-hydroxy-4-(6-methyl-6-)1H-tetrazol-5-yl)-heptyloxy) phenyl)ethanone], a specific leukotriene B4(LTB4) receptor antagonist, inhibited the production of LTB4in human peripheral blood polymorphonuclear leukocytes (PMNL) and in monocytes activated by calcium ionophore A23187. In human monocytes activated by ionophore it inhibited also the production of thromboxane B2(TXB2). The effect of LY 255283 on 5-lipoxygenase (5-LO) and LTA4hydrolase activities which catalyse the production of LTB4and LTA4has not been studied yet. It is thought that LY 255283 may inhibit the production of LTB4and TXA2by antagonising the effect of ionophore-induced LTB4on 5-lipoxygenase and cyclooxygenase in human peripheral blood PMNL and monocytes.

scholarly journals Microtubule dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes.

1978 ◽  
Vol 76 (2) ◽  
pp. 439-447 ◽  
Author(s):  
B R Burchill ◽  
J M Oliver ◽  
C B Pearson ◽  
E D Leinbach ◽  
R D Berlin
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Reduced Glutathione ◽  
Human Peripheral Blood ◽  
Microtubule Assembly ◽  
Glutathione Metabolism ◽  
Resting Cells ◽  
Tertiary Butyl ◽  
Thin Sections ◽  
Butyl Hydroperoxide ◽  
Human Polymorphonuclear Leukocytes

Glutathione oxidants such as tertiary butyl hydroperoxide were shown previously to prevent microtubule assembly and cause breakdown of preassembled cytoplasmic microtubules in human polymorphonuclear leukocytes. The objectives of the present study were to determine the temporal relationship between the attachment and ingestion of phagocytic particles and the assembly of microtubules, and simultaneously to quantify the levels of reduced glutathione and products of its oxidation as potential physiological regulators of assembly. Polymorphonuclear leukocytes from human peripheral blood were induced to phagocytize opsonized zymosan at 30 degrees C. Microtubule assembly was assessed in the electron microscope by direct counts of microtubules in thin sections through centrioles. Acid extracts were assayed for reduced glutathione (GSH) and oxidized glutathione (GSSG), by the sensitive enzymatic procedure of Tietze. Washed protein pellets were assayed for free sulfhydryl groups and for mixed protein disulfides with glutathione (protein-SSG) after borohydride splitting of the disulfide bond. Resting cells have few assembled microtubules. Phagocytosis induces a cycle of rapid assembly followed by disassembly. Assembly is initiated by particle contact and is maximal by 3 min of phagocytosis. Disassembly after 5-9 min of phagocytosis is preceded by a slow rise in GSSG and coincides with a rapid rise in protein-SSG. Protein-SSG also increases under conditions in which butyl hydroperoxide inhibits the assembly of microtubules that normally follows binding of concanavalin A to leukocyte cell surface receptors. No evidence for direct involvement of GSH in the induction of assembly was obtained. The formation of protein-SSG, however, emerges as a possible regulatory mechanism for the inhibition of microtubule assembly and induction of their disassembly.

scholarly journals Effects of glutathione-oxidizing agents on microtubule assembly and microtubule-dependent surface properties of human neutrophils.

1976 ◽  
Vol 71 (3) ◽  
pp. 921-932 ◽  
Author(s):  
J M Oliver ◽  
D F Albertini ◽  
R D Berlin
Keyword(s):  
Concanavalin A ◽  
Peripheral Blood ◽  
Polymorphonuclear Leukocytes ◽  
Human Peripheral Blood ◽  
Microtubule Assembly ◽  
Human Neutrophils ◽  
Hexose Monophosphate ◽  
Con A ◽  
Oxidizing Agents ◽  
The Face

In human peripheral blood polymorphonuclear leukocytes and lymphocytes, GSH-oxidizing agents promote the movement of surface-bound concanavalin A (Con A) into caps and inhibit the assembly of microtubules (MT) that is normally induced by Con A binding. Con A capping and inhibition of MT assembly occur when GSH levels in cell suspensions are decreased by 30-70%, and return to GSH to control levels is accompanied by the appearance of cytoplasmic MT and by inhibition of the capping response with Con A. Oxidation of GSH markedly stimulates the hexose monophosphate shunt, and regeneration of GSH occurs rapidly. The data indicate that MT cannot be assembled or maintained in the face of decreased GSH levels. Thus, GSH homeostasis becomes critical during physiological events such as phagocytosis which simultaneously induce the assembly of MT and the production of agents like H2O2 that can oxidize GSH.

scholarly journals THE INTERACTION IN VITRO BETWEEN POLYMORPHONUCLEAR LEUKOCYTES AND MYCOPLASMA

1971 ◽  
Vol 134 (6) ◽  
pp. 1417-1430 ◽  
Author(s):  
Michael S. Simberkoff ◽  
Peter Elsbach
Keyword(s):  
Peripheral Blood ◽  
Polymorphonuclear Leukocytes ◽  
Human Peripheral Blood ◽  
Peritoneal Exudate ◽  
Specific Antiserum ◽  
E Coli ◽  
14Co2 Production ◽  
Stimulation Of

The interaction, between mycoplasma (PPLO) and human or rabbit leukocytes was examined in vitro. Upon incubation of M. hominis or M. arthritidis for 2 hr with rabbit peritoneal exudate granulocytes or leukocytes from human peripheral blood, no killing of mycoplasma was observed either in the presence or absence of type-specific antiserum. However, 14CO2 production from glucose-1-14C was stimulated up to 10-fold in the presence of live or heat-killed PPLO. The extent of stimulation depended upon the number of organisms and the presence of type-specific antiserum. The stimulation of 14CO2 production seems not because of tight adherence of PPLO to the leukocytes, since PPLO were quantitatively recovered in the medium after sedimenting the granulocytes. The enhanced conversion of medium lysolecithin to cellular lecithin that accompanies phagocytosis of polystyrene particles was significantly reduced when PPLO were also present. Mycoplasma alone elicited no stimulation of lecithin formation. Killing of E. coli, a microorganism readily engulfed and killed by leukocytes in vitro, was diminished when the leukocytes were preincubated with mycoplasma. These findings indicate that M. hominis and M. arthritidis are not ingested by granulocytes to any detectable extent, but that these organisms affect the leukocytes' metabolism and also impair phagocytosis of E. coli.

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