CHAPTER 65 COMPOSITION AND METABOLISM OF NEUTROPHILS
CHAPTER 65 COMPOSITION AND METABOLISM OF NEUTROPHILS
Composition of Neutrophils
Water and Electrolytes
Amino Acids, Peptides, and Proteins
Nucleotides and Nucleic Acids
Vitamins and Cofactors
Metabolism of Neutrophils
Dna and Rna Metabolism
Neutrophils are highly specialized differentiated cells, and details of their specialized metabolic pathways are given in Chap. 67 and Chap. 72. This chapter deals with the composition of granulocytes, their content of water, electrolytes, carbohydrates, amino acids, peptides, proteins, lipids, nucleic acids, vitamins, and cofactors. The housekeeping metabolic pathways of neutrophils for aerobic and anaerobic energy metabolism, and DNA, nucleotide, and lipid metabolism are also reviewed.
Acronyms and abbreviations that appear in this chapter include: ATP, adenosine triphosphate; cAMP, cyclic adenosine monophosphate; cGMP, cyclic guanosine monophosphate; GM-CSF, granulocyte-macrophage colony-stimulating factor; 5-HETE, 5-hydroxyeicosatetraenoic acid; 5-HPETE, 5-hydroperoxy-6,8,11,14-eicosatetraenoic acid; 15-HPETE, 15-hydroperoxy-5,8,11,13-eicosatetraenoic acid; LAP, leukocyte alkaline phosphatase; LTA4, leukotriene A4; PAF, platelet-activating factor; SE, standard error; SRS, slow-reactivity substance.
COMPOSITION OF NEUTROPHILS
Many of the measurements of the composition of leukocytes were performed at a time when those carrying out the analyses did not appreciate that the white cells of the blood were heterogeneous in origin and function. Thus, many of the data pertain to leukocytes as a whole, not to isolated neutrophils. Often, granulocytes were studied rather than neutrophils. In many cases, however, the content of analytes is similar in neutrophils and other white blood cells, and the best data available are presented here and expressed as values in neutrophils, recognizing that in some cases the values may be distorted by the presence of other leukocytes in the mixtures analyzed.
WATER AND ELECTROLYTES
Approximately 82 percent of the leukocyte weight is water.1 There is a remarkable paucity of data regarding the electrolyte content of neutrophils. The often quoted 1929 study of Endres and Herget1 was carried out on mixed leukocytes from the blood of horses obtained at a slaughterhouse. They found an average of 2610 mg (113 mmol) sodium, 889 mg (22.7 mmol) potassium, 72 mg (1.8 mmol) calcium, 10.3 mg (0.18 mmol) iron, 2487 mg (70.2 mmol) chloride, and 299 mg (9.65 mmol) inorganic phosphate per liter of leukocytes. The copper content of neutrophils has been reported to average 4.69 nmol/109 cells,2 zinc 109.2 nmol/109 cells2 and 50.16 nmol/109 cells,3 and magnesium 3.11 fmol/cell.4 There is little selenium in neutrophils, the median concentration having been reported as less than 0.0075 µmol/109 cells.5 Otherwise, electrolyte determinations on human leukocytes appear to have been limited to leukemic cells and to pus.6
The rate of metabolism of glucose by neutrophils is affected by insulin in diabetics but not in normal subjects.7,8 The neutrophil is particularly rich in glycogen. The concentration of this complex polysaccharide has been reported to average 7.36 mg/109 cells.9,10 and 11
AMINO ACIDS, PEPTIDES, AND PROTEINS
The concentrations of most amino acids are higher in neutrophils than is the surrounding plasma.12 The amino acid concentration in neutrophils is summarized in Table 65-1. The reduced glutathione content of neutrophils is 9.8 nmol per 107 cells.13
TABLE 65-1 UNBOUND AMINO ACID CONCENTRATIONS IN LEUKOCYTES (LYMPHOCYTES INCLUDED)
The protein content of the neutrophil is 74.2 ± 3.1 (mean ± 1 SE) mg/109 cells.14 These proteins include those of the structural matrix of the neutrophil; proteins required for its locomotion, chemotactic properties, and adhesiveness; and the many granule proteins with bactericidal, hydrolytic, and inflammatory functions. These proteins are described in detail in Chap. 64, Chap. 67, and Chap. 72.
As in other cells, the plasma membrane and the membranes of the intracellular organelles are rich in lipids. Five percent of the wet weight of neutrophils is lipid, which is distributed among various classes, as shown in Table 65-2.15,16,17,18 and 19 The rare polyphosphoinositides are of special interest as sources of inositol 1,4,5-trisphosphate (a calcium-releasing mediator) and diacylglycerol (which activates protein kinase C).20,21 The main glycolipid of neutrophils is lactosylceramide.22
TABLE 65-2 LIPID COMPOSITION OF NEUTROPHILS
NUCLEOTIDES AND NUCLEIC ACIDS
The levels of nucleotides in the neutrophils are summarized in Table 65-3.23,24
TABLE 65-3 NUCLEOTIDES IN LEUKOCYTES (LYMPHOCYTES INCLUDED)
Neutrophils contain all the forms of RNA needed for protein synthesis: transfer RNA, ribosomal RNA, and messenger RNA.27,28 The DNA content of neutrophils is identical to that of all other haploid cells, at 0.7 pg DNA phosphorus per cell.29
VITAMINS AND COFACTORS
The average folic acid content of packed leukocytes of normal subjects was 0.1 µg/ml of packed leukocytes, and about 20 percent of this was free and the remainder conjugated.30 The cocarboxylate content is 340 µg/1011 cells,31 pyridoxal phosphate 0.24–0.38 ng/106 cells,32 thiamine 67.5 ± 4.1 µg/100 ml,33 ascorbic acid 16.5 ± 5.1 mg/100 ml,34 and folate 92 ng/ml.35
METABOLISM OF NEUTROPHILS
The Main Glycolytic (Embden-Meyerhoff) Pathway The main energy-producing pathway in the neutrophil is glycolysis, resulting in the conversion of glucose to lactate.36,37 and 38 When intact or homogenized leukocytes are incubated with glucose uniformly labeled with 14C, about 80 percent of the radioactivity is recovered in lactic acid. Glycolysis is inhibited by cortisol.7 The activities of the glycolytic enzymes of neutrophils are summarized in Table 65-439,40 and 41; in some cases the conditions under which the neutrophils are disrupted have a significant effect on the activities measured.40 Hexokinase is the rate-limiting enzyme of glycolysis in normal neutrophils.37 The rate of glycolysis is not altered during phagocytosis,38 but ATP levels, normally 1.9 nmol/106 cells, fall to 0.8 nmol/106 cells. Both the glycogen stores of neutrophils and the glucose of the plasma can serve as the source of glucose. Galactose, mannose, and fructose can also be metabolized by leukocytes.43
TABLE 65-4 GLYCOLYTIC AND RELATED ENZYME ACTIVITIES IN NEUTROPHILS
The Hexose Monophosphate Shunt Pathway Neutrophils also metabolize glucose by way of the hexose monophosphate shunt,44,45 and 46 and this accounts for some of the oxygen consumption of the cells. In resting cells, the amount of glucose metabolized via this route amounts to only 2 to 3 percent of the total glucose consumed by the cell.45,46 and 47 The operation of the hexose monophosphate shunt, however, is of special importance to the neutrophil, because it is this pathway that provides the NADPH needed for the generation of microbicidal oxidants (see Chap. 67).
Glycogen Metabolism Neutrophils contain a large quantity of glycogen (see above), arising mostly from glucose; there is little net synthesis from substrates at the triose phosphate level. Glycogen turnover increases when these cells are deprived of glucose, especially if they are engaged in phagocytosis, but resynthesis occurs when adequate glucose is added.38,48,49 During phagocytosis by glucose-starved cells, glycogen phosphorylase activity rises, but phosphorylase kinase and glycogen synthase levels remain unchanged.48 Glycogen first appears in myelocytes and increases with cell maturation.50
Neutrophils consume 0.15 µmol oxygen per 107 cells in the absence of glucose and 0.015 µmol oxygen per 107 cells in the presence of glucose.51 Oxygen consumption by neutrophils is influenced by a wide variety of physiologic and pathologic stimuli.52 In addition to phagocytosis (see Chap. 67), these include thyroid hormone, CO2 tension,53 glucose concentration,54 serum,55 pyrogens,56 complement components, chemotactic peptides, and immune complexes.57 A number of chemicals depress neutrophil respiration, including saponin, thiouracil, chloramphenicol, cyanide, fluoroacetate, malonate, and p-hydroxymercuribenzoate. Other compounds, such as ascorbic acid and dinitrophenol, increase O2 consumption.52
Few mitochondria are found in mature neutrophils,58 and mitochondrial respiration accounts for only 5 percent of the glucose consumed by the neutrophil.59,60 Because of the efficiency of mitochondrial ATP synthesis, however, it furnishes nearly half the ATP generated by the cell. The following Krebs cycle enzymes have been detected in leukocytes: isocitric dehydrogenase, aconitase, fumarase, and malic dehydrogenase.61,62 In addition, the metabolically related enzymes glutamate-oxaloacetate aminotransferase and glutamate-pyruvate aminotransferase are also found in neutrophils.63 The four enzymes necessary for gluconeogenesis were not detected in leukocytes.
DNA AND RNA METABOLISM
DNA polymerase is most active in early neutrophil precursors.64 Activity diminishes with cell maturation and is barely detectable in mature cells. Consistent with this finding, the myelocyte is the most mature neutrophil precursor that can still incorporate thymidine into DNA and undergo mitosis.65,66 Like other cells, neutrophils synthesize RNA using DNA as a template.67,68 Earlier studies using the incorporation of [14C]uridine into RNA as a measure of RNA synthesis were difficult to interpret because they were carried out with mixed populations of cells.67 However, Northern blotting has indicated unequivocally that neutrophils have the capacity to synthesize specific messenger RNA.69,70
Mature neutrophils and neutrophil precursors incorporate labeled amino acids into proteins68,71,72,73 and 74 and have been shown to synthesize fibronectin.70 Protein synthesis also seems to play a role in receptor recycling by neutrophils.75 Once proteins are synthesized, they undergo extensive posttranslational modification and are sorted into the appropriate organelle.76
Many of the studies on nucleotide biosynthesis have been conducted in mixed cell populations. Conclusions from such studies regarding nucleotide biosynthesis in neutrophils must be regarded as provisional. Leukocytes are capable of de novo biosynthesis of pyrimidines. The enzymes of pyrimidine biosynthesis (aspartate carbamyltransferase, dihydroorotase, dihydroorotic dehydrogenase, and orotidylic decarboxylase) are found in normal leukocytes (predominantly neutrophils).77 The failure to demonstrate 14C-glycine incorporation into the acid-insoluble nucleotide pool in normal leukocytes suggested that, in contradistinction to their ability to carry on pyrimidine biosynthesis de novo, these cells are incapable of the earlier steps of purine synthesis.78 In addition to de novo pyrimidine synthesis, ribo- and deoxyribonucleotides can also be formed via the “salvage” pathway through the kinase-catalyzed interaction of ATP with nucleosides and deoxynucleosides (cytidine, uridine, deoxycytidine, deoxyuridine, and thymidine).79,80,81,82,83 and 84 The enzyme that catalyzes the conversion of ribonucleotides to deoxyribonucleotides, however, has not been detected in normal neutrophils.
The presence of ribonuclease and deoxyribonuclease in lysosomal granules of leukocytes85,86 suggests that these organelles are involved in the breakdown of exogenous and/or endogenous nucleic acids. Ribonuclease activity is 10 times higher in mature neutrophils than in blast forms.87 In addition, nucleotidases,88 several isoenzymes of acid phosphatase,89 and a nucleoside deaminase have been described.83 Mature neutrophils, however, contain only very low levels of 5′-nucleotidase and adenosine deaminase.
One of the most extensively investigated neutrophil enzymes is LAP. Leukocyte alkaline phosphatase is a zinc-containing phosphomonoesterase with a pH optimum near 10 that catalyzes the hydrolysis of a wide variety of phosphoester substrates.90,91 The activity of LAP, which is limited to the neutrophilic series, first appears in myelocytes and rapidly increases with maturation of the cell to the segmented polymorphonuclear neutrophil.92 Glucocorticoids markedly increase the activity in normal leukocytes, probably by induction of the enzyme, which may explain the high LAP activity observed during infections.93 Although the in vivo function of LAP remains uncertain, the assay of this activity has found many clinical applications. Marked changes in LAP activity are observed in chronic myelocytic leukemia and other myeloproliferative disorders, as well as in certain other conditions, including idiopathic thrombocytopenic purpura, infectious mononucleosis, aplastic anemia, and sarcoidosis.94,95,96 and 97
Cyclic 3′,5′-cAMP is present in the human neutrophil.98 This “second messenger” is involved in the activation of leukocyte glycogen phosphorylase. The synthesis of cAMP is catalyzed by adenyl cyclase and its degradation by cAMP phosphodiesterase, both of which are found in normal neutrophils.89 The accumulation of cAMP in the leukocyte is stimulated by epinephrine, prostaglandin E, and adenyl cyclase.99,100 A transient rise in cAMP levels (duration 2–5 min) is also seen after exposure of neutrophils to inflammatory agonists such as formylated oligopeptides or immune complexes, but the cyclic nucleotide appears to have only a minor effect on neutrophil function.101 The cytosol of neutrophils contains a protein kinase that is stimulated by cAMP.102 These cells also contain histone phosphatases, which dephosphorylate the product of the protein kinase reaction.103 A reduced responsiveness of b-receptor function for isoproterenol (Isuprel) in leukocytes of patients suffering from acute bronchial asthma has been reported.104,105 In asthmatic patients in remission, this response was within normal limits.106
There has been little study of cGMP in neutrophils; exposure of neutrophils to inflammatory mediators caused no change in their levels of cGMP.101
Early studies revealed that lipid biosynthesis, as measured by the incorporation of [14C]acetate, takes place in neutrophils. Two-thirds of the radioactivity was incorporated into neutral lipids and the remainder into phospholipids.107 Neutrophils also incorporated [2-14C]acetate and [2-14C]mevalonate into squalene but not into sterols.108 Younger neutrophils, as found in infection, had lower rates of incorporation of labeled acetate into lipids.109
The phosphatidic acid pathway incorporating fatty acids into neutral lipids is operative in these cells.110 The incorporation of fatty acids into lysophospholipids also occurs in neutrophils, leading to the formation of diacylglyceryl phosphocholine and diacylglyceryl phosphoethanolamine.111 Phagocytosis is accompanied by a threefold increase in the acylation of exogenous lysolecithin, leading to a net increase in phospholipid. PAF is synthesized by replacing the 2-acyl group (usually arachidonate) in 1-alkyl-2-acylglycerol phosphocholine with acetate.112
Acetyl CoA carboxylase, the first enzyme required for the synthesis of long-chain fatty acids, has been found in myeloblasts but not in mature neutrophils. The latter cells, however, retain the capability of elongating the chains of preformed fatty acids.113
A number of lipolytic activities are present in human neutrophils. One of these, a triacylglycerol acylhydrolase acting on lipoprotein and chylomicron substrates, has been purified.114 A cholesterylesterase activity is also associated with this enzyme. Fatty acid ester hydrolases have been described.115 Several phospholipases are found in neutrophils.116,117 Their activation occurs upon stimulation of the neutrophil, leading to the production of signal-transducing chemicals and lipid mediators.
Arachidonic acid is the precursor of a group of lipid mediators that play important roles in the regulation of a wide range of biological responses.118 It is released from phospholipids by phospholipase A2,119 activated by the exposure of neutrophils to such stimuli as opsonized zymosan, calcium ionophore, or chemotactic factors.119,120 and 121 Lipid mediators are then produced from the liberated arachidonic acid by either a cyclooxygenase- or a lipoxygenase-catalyzed oxidation (Fig. 65-1). In the neutrophil, oxidation by lipoxygenase exceeds that by cyclooxygenase,112,119,122,123 and it may be that the arachidonic acid activates the lipoxygenase.124
FIGURE 65-1 Production of lipid mediators from arachidonic acid. Above, production of thromboxane A2 and prostacyclin. Below, production of leukotrienes A4, B4, and C4.
Cyclooxygenase (prostaglandin synthetase) catalyzes the conversion of arachidonic acid into the cyclic endoperoxides PGG2 and PGH2, which in turn are isomerized into the prostaglandins PGE2, PGD2, and PGF2.118 PGE2 is a major mediator of the inflammatory process, dilating and permeabilizing small blood vessels to give rise to edema and erythema.104 PGH2 may also be converted to the unstable vasoconstrictor thromboxane A2, which rapidly hydrolyzes to thromboxane B2,125 which is vasoinactive but chemotactic.
The most important lipoxygenase in neutrophils is 5-lipoxygenase.126,127 This enzyme catalyzes the oxidation of arachidonic acid to 5-HPETE and its subsequent conversion to leukotriene A4 (LTA4), the unstable parent compound of the leukotrienes, a group of lipid tbmediators with major effects on the inflammatory process (Fig. 65-2). nbLTA4 may add glutathione to form LTC4,128 whose peptide bonds may subsequently be successively hydrolyzed to yield LTD4, containing a cysteine-glycine dipeptide, and LTE4, containing cysteine only. Together, leukotrienes C4, D4, and E4 constitute the activity known formerly as the SRS of anaphylaxis.129,130,131 and 132
FIGURE 65-2 Structure and formation of the cysteinyl leukotrienes. Leukotriene C4, produced by the reaction of glutathione with leukotriene A4, as shown in Fig. 65-1, is converted to leukotrienes D4 and E4 by the successive removal of the terminal amino acids from the peptide chain. GGTP, g-glutamyl tripeptidase.
Alternatively, LTA4 may be hydrolyzed to generate LTB4,133,134 a potent chemotactic factor and neutrophil activator.135,136 LTB4 production by neutrophils is induced by a number of stimuli137,138,139 and 140 whose effects on its production are further regulated by the growth factor GM-CSF.138,139,140 and 141 LTB4 is inactivated by neutrophil P450 cytochrome(s), which catalyze successive oxidations at the x omega position to yield 20-OH-LTB4, LTB4-20-carboxaldehyde, and finally LTB4-20-carboxylic acid.142,143 and 144
The enzymes responsible for leukotriene production can also oxidize C20-D3 (c-linolenic) and C20-D5 fatty acids, leading to the LTA3 and A5 series of leukotrienes. Because they are less potent than the A4 series of leukotrienes, the A3 and A5 leukotrienes can act as anti-inflammatory agents, partially antagonizing the effects of the A4 series.145,146
The hydroperoxyl group of 5-HPETE is sometimes reduced to a hydroxyl group before conversion to LTA4 can take place. This reduction yields a major product, 5-HETE,147 and a minor product, 12-HETE (an isomer of 12,L-hydroxy-5,8,10,14-eicosatetraenoic acid). Both 5- and 12-HETE have chemotactic properties and stimulate the release of lysozyme from neutrophils.148
Neutrophils also contain a 15-lipoxygenase that converts arachidonic acid to 15-HPETE.126 Subsequent oxidation of 15-HPETE by 5-lipoxygenase followed by hydrolysis of the resulting epoxide gives rise to the lipoxins, a family of C-20 fatty acids containing four conjugated double bonds and three hydroxyl groups.128,148 These, too, are inflammatory mediators, with effects that are similar in general but different in particular from those of the leukotrienes.
Endres G, Herget L: Mineralzusammensetzung der Bluplättchen und weissen Blutkörperchen. Z Biol 88:451, 1929.
Williams NR, Rajput-Williams J, West JA, Nigdikar SV, Foote JW, Howard AN: Plasma, granulocyte and mononuclear cell copper and zinc in patients with diabetes mellitus. Analyst 120:887, 1995.
Prasad AS, Mantzoros CS, Beck FW, Hess JW, Brewer GJ: Zinc status and serum testosterone levels of healthy adults. Nutrition 12:344, 1996.
Loun B, Astles R, Copeland KR, Sedor FA: Intracellular magnesium content of mononuclear blood cells and granulocytes isolated from leukemic, infected, and granulocyte colony-stimulating factor-treated patients. Clin Chem 41:1768, 1995.
Rukgauer M, Zeyfang A, Uhland K, Kruse-Jarres JD: Isolation of corpuscular components of whole blood for the determination of selenium in blood cells. J Trace Elem Med Biol 9:130, 1995.
Rigas DA: Electrolyte nitrogen and water content of human leukemic leukocytes: Relation to cell maturity. J Lab Clin Med 58:234, 1961.
Rauch HC, Loomis ME, Johnson ME, Favour CB: In vitro suppression of polymorphonuclear leukocyte and lymphocyte glycolysis by cortisol. Endocrinology 68:375, 1961.
Martin SP, McKinney GR, Green R, Becker C: The influence of glucose, fructose, and insulin on the metabolism of leukocytes of healthy and diabetic subjects. J Clin Invest 32:1171, 1953.
Scott RB, Cooper LW: Glycogen in human peripheral blood leukocytes: I. Characteristics of the synthesis and turnover of glycogen in vitro. J Clin Invest 47:344, 1968.
Scott RB, Still WJS, Cooper LW: Glycogen in human peripheral blood leukocytes: II. The macromolecular state of leukocyte glycogen. J Clin Invest 47:353, 1968.
Esman V: The glycogen content of WBC from diabetic and nondiabetic subjects. Scand J Lab Invest 13:134, 1961.
McMenamy RH, Lund CC, Neville GJ, Wallach DFH: Studies of unbound amino acid distributions in plasma, erythrocytes, leukocytes and urine of normal human subjects. J Clin Invest 39:1675, 1960.
Thornalley PJ, Bellavite P: Modification of the glyoxalase system during the functional activation of human neutrophils. Biochim Biophys Acta 931:120, 1987.
Beutler E, Kuhl W: Unpublished 1991.
Gottfried EL: Lipids of human WBC: Relation to cell type. J Lipid Res 8:321, 1967.
Gottfried EL: Lipid patterns of leukocytes in health and disease. Semin Hematol 9:241, 1970.
Boyd EM: The lipid content of the white blood cells in normal young women. J Biol Chem 101:623, 1933.
Boyd EM, Stephens DJ: A comparison of lipid composition with differential count of the white blood cells. Proc Soc Exp Biol Med 33:558, 1936.
Kidson C: Relation of leucocyte lipid metabolism to cell age: studies in infective leucocytosis. Br J Exp Pathol 42:597, 1961.
Nishizuka Y: Studies and perspectives of protein kinase C. Science 233:305, 1986.
Berridge MJ, Irvine RF: Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature 312:315, 1984.
Symington FW, Murray WA, Bearman SI, Hakomori S-I: Intracellular localization of lactosylceramide, the major human neutrophil glycophingolipid. J Biol Chem 262:11356, 1987.
Willoughby HW, Waisman HA: Nucleic acid precursors and nucleotides in normal and leukemic blood: I. Comparison of formic acid chromatograms. Cancer Res 17:942, 1957.
Silber R, Gabrio BW, Huennekens FM, Albrecht M: Studies on normal and leukemic leukocytes: III. Pyridine nucleotides. J Clin Invest 41:230, 1962.
Noyes BE, Mevarech M, Stein R, Agarwal KL: Detection and partial sequence analysis of gastrin mRNA by using an oligodeoxynucleotide probe. Proc Natl Acad Sci USA 76:1770, 1979.
Löhr GW, Waller HD: Zellstoffwechsel und Zellalterung. Klin Wochenschr 37:833, 1959.
Silber R, Unger KW, Ellman L: RNA metabolism in normal and leukaemic WBC: Further studies on RNA synthesis. Br J Haematol 14:261, 1968.
Tryfiates GP, Laszlo J: Human leukemic polyribosomes. Proc Soc Exp Biol Med 124:1125, 1967.
Garcia AM, Iorio R: Studies on DNA in WBC and related cells of mammals: V. The fast green histone and the fuelgen-DNA content of rat WBC. Acta Cytol 12:46, 1968.
Swendseid ME, Bethell FH, Bird OD: The concentration of folic acid in leukocytes. Observations on normal subjects and persons with leukemia. Cancer Res 11:864, 1951.
Smits G, Florijn E: The aneurinpyrophosphate content of red and white blood corpuscles in the rat and in man, in various states of aneurin provision and in disease. Biochim Biophys Acta 3:44, 1949.
Boxer GE, Pruss MP, Goodhart RS: Pyridoxal-5-phosphoric acid in whole blood and isolated leukocytes of man and animals. J Nutr 63:623, 1957.
Burch HB, Bessey OA, Love RH, Lowry OH: The determination of thiamine and thiamine phosphates in small quantities of blood and blood cells. J Biol Chem 198:477, 1952.
Barkhan P, Howard AN: Distribution of ascorbic acid in normal and leukaemic human blood. Biochem J 70:163, 1958.
Hoffbrand AV, Newcombe BFA: Leucocyte folate in vitamin B12 and folate deficiency and in leukaemia. Br J Haematol 13:954, 1967.
Beck WS, Valentine WN: The aerobic metabolism of leukocytes in health and leukemia: I. Glycolysis and respiration. Cancer Res 12:818, 1952.
Beck WS: A kinetic analysis of the glycolytic rate and certain glycolytic enzymes in normal and leukemic leukocytes. J Biol Chem 216:333, 1955.
Borregaard N, Herlin T: Energy metabolism of human neutrophils during phagocytosis. J Clin Invest 70:550, 1982.
Lane TA, Beutler E, West C, Lamkin GE: Glycolytic metabolism of stored granulocytes. Transfusion 21:717, 1981.
Fauth U, Schlechtriemen T, Heinrichs W, Puente-Gonzalez I, Halmagyi M: The measurement of enzyme activities in the resting human polymorphonuclear leukocyte—Critical estimate of a method. Eur J Clin Chem Clin Biochem 31:5, 1993.
Löhr GW, Waller HD: Zellstoffwechsel und Zellalterung. Klin Wochenschr 37:833, 1959.
Beutler E, West C: Unpublished, 1993.
Stjernholm RL, Burns CP, Hohnadel JH: Carbohydrate metabolism by leukocytes. Enzyme 13:7, 1972.
Sbarra AJ, Karnovsky ML: The biochemical basis of phagocytosis: I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem 234:1355, 1959.
Beck WS: Occurrence and control of the phosphogluconate oxidation pathway in normal and leukemic leukocytes. J Biol Chem 232:271, 1958.
Stjerholm R, Manek RC: Carbohydrate metabolism in leukocytes: XIV. Regulation of pentose cycle activity and glycogen metabolism during phagocytosis. J Reticuloendothel Soc 8:550, 1970.
Wood HG, Katz J, Landau BR: Estimation of pathways of carbohydrate metabolism. Biochem J 338:809, 1963.
Borregaard N, Juhl H: Activation of the glycogenolytic cascade in human polymorphonuclear leucocytes by different phagocytic stimuli. Eur J Clin Invest 11:257, 1981.
Scott RB: Glycogen in human peripheral blood leukocytes: I. Characteristics of the synthesis and turnover of glycogen in vitro. J Clin Invest 47:344, 1968.
Wachstein M: The distribution of histochemically demonstrable glycogen in human blood and bone marrow cells. Blood 4:54, 1949.
Martin SP, McKinney GR, Green R: The metabolism of human polymorphonuclear leukocytes. Ann NY Acad Sci 59:996, 1955.
Cline MJ: The White Cell, Harvard, Cambridge, 1975.
Bicz W: The influence of carbon dioxide tension on the respiration of normal and leukemic leukocytes: I. Influence on endogenous respiration. Cancer Res 20:184, 1960.
McKinney GR, Martin SP, Rundles RW, Green R: Respiration and glycolytic activities of human leukocytes in vitro. J Appl Physiol 5:355, 1953.
McLeod J, Rhoads C: Metabolism of leukocytes in Ringer-phosphate and in serum. Proc Soc Exp Biol Med 41:268, 1939.
Cline MJ, Melmon KL, Davis WC, Williams HE: Mechanism of endotoxin interaction with human leukocytes. Br J Haematol 15:539, 1968.
Strauss BS, Stetson CA Jr: Studies on the effect of certain macromolecular substances on the respiratory activity of the leucocytes of peripheral blood. J Exp Med 112:652, 1960.
Bessis M: Cytology of the Blood and Blood-Forming Organs, Grune & Stratton, New York, 1956.
Foster JM, Terry ML: Studies on the energy metabolism of human leukocytes: I. Oxidative phosphorylation by human leukocyte mitochondria. Blood 30:168, 1967.
Cheson DB, Curnutte JT, Babior BM: The oxidative killing mechanisms of the neutrophil, in Progress in Clinical Immunology, vol 3, p 1. Grune & Stratton, New York, 1977.
Tanaka KR, Valentine WN: Aconitase activity of human leukocytes. Acta Haematol 26:12, 1961.
Tanaka KR, Valentine WN: Fumarase activity of human leukocytes and erythrocytes. Blood 17:328, 1961.
Belfiore F, Borzi V, LoVecchio L, Napoli E, Rabuazzo AM: Enzyme activities of NADPH forming metabolic pathways in normal and leukemic leukocytes. Clin Chem 21:880, 1925.
Rabinowitz Y: DNA polymerase and carbohydrate metabolizing enzyme content of normal leukemic glass column separated leukocytes. Blood 27:470, 1966.
Bond VP, Fliedner TM, Cronkite EP, Rubine JR, Brecher G, Schork PK: Proliferative potentials of bone marrow and blood cells studied by in vitro uptake of H3-thymidine. Acta Haematol 21:1, 1959.
Rubine JR, Cronkite EP, Bond VP, Fliedner TM: The metabolism and fate of tritiated thymidine in man. J Clin Invest 39:909, 1960.
Cline MJ: Isolation and characterization of RNA from human leukocytes. J Lab Clin Med 68:33, 1966.
Torelli V, Torelli G, Cadossi R: Double stranded ribonucleic acid in human blast cells. Eur J Cancer 11:117, 1975.
Ezekowitz RAB, Orkin SH, Newburger PE: Recombinant interferon gamma augments phagocyte superoxide production and X-chronic granulomatous disease gene expression in X-linked variant chronic granulomatous disease. J Clin Invest 80:1009, 1987.
La Fleur M, Beaulieu AD, Kreis C, Poubelle P: Fibronectin gene expression in polymorphonuclear leukocytes. Accumulation of mRNA in inflammatory cells. J Biol Chem 262:2111, 1987.
Weisberger AS, Suhrland LS, Griggs RC: Incorporation of radioactive L-cystine and L-methionine by leukemic leukocytes in vitro. Blood 9:1095, 1954.
Weisberger AS, Levine B: Incorporation of radioactive L-cystine by normal and leukemic leukocytes in vivo. Blood 9:1082, 1954.
Baker WH, Zamecnik PC, Stephenson ML: In vitro incorporation of C14-DL-leucine into normal and leukemic white cells. Blood 12:822, 1957.
Granelli-Piperno A, Vassalli JD, Reich E: RNA and protein synthesis in human peripheral blood polymorphonuclear leukocytes. J Exp Med 149:284, 1979.
Woodman RC, Curnutte JT, Babior BM: Evidence that de novo protein synthesis participates in a time-dependent augmentation of the chemotactic peptide-induced respiratory burst in neutrophils: Effects of recombinant human colony stimulating factors and dihydrocytochalasin B. Free Radical Biol Med 5:355, 1988.
Gullberg U, Andersson E, Garwicz D, Lindmark A, Olsson I: Biosynthesis, processing and sorting of neutrophil proteins: insight into neutrophil granule development. Eur J Haematol 58:137, 1997.
Smith LH Jr, Baker FA: Pyrimidine metabolism in man: I. The biosynthesis of orotic acid. J Clin Invest 38:798, 1959.
Scott JL: Human leukocyte metabolism in vitro: I. Incorporation of adenine-8-C14 and formate-C14 into the nucleic acids of leukemic leukocytes. J Clin Invest 41:67, 1962.
Wilmanns W: Thymidine kinase in normal and leukemic myeloid cells [translated]. Klin Wochenschr 45:505, 1967.
Bianchi PA: Thymidine phosphorylation and deoxyribonucleic acid synthesis in human leukaemia cells. Biochim Biophys Acta 55:547, 1962.
Marsh JC, Perry S: Thymidine catabolism by normal and leukemic human leukocytes. J Clin Invest 43:267, 1964.
Silber R, Gabrio BW, Huennekens FM: Studies on normal and leukemic leukocytes: VI. Thymidylate synthetase and deoxycytidylate deaminase. J Clin Invest 42:1913, 1963.
Silber R: Regulatory mechanism in human leukocyte: I. Feedback control of deoxycytidylate deaminase. Blood 29:896, 1967.
Coleman CN, Stoller RG, Chabner BA: Properties of cytidine kinase enzyme from human leukemic granulocytes. Blood 46:791, 1975.
Barnes JM: The enzymes of lymphocytes and polymorphonuclear leucocytes. Br J Exp Pathol 21:261, 1940.
Cohn ZA, Hirsch JG: The isolation and properties of specific cytoplasmic granules of rabbit polymorphonuclear leukocytes. J Exp Med 112:983, 1960.
Silber R, Unger KW, Keller J, Bertino JR: RNA metabolism of normal and leukemic leukocytes: II. Ribonuclease. Blood 29:57, 1967.
Swenseid ME, Wright PD, Bethell FH: Variations in nucleotidase activity of leukocytes in normal and pathologic conditions. J Lab Clin Med 40:515, 1952.
Li CY, Yam LT, Lam KW: Acid phosphatase isoenzyme in human leukocytes in normal and pathologic conditions. J Histochem Cytochem 18:473, 1970.
Follette JH, Valentine WN, Hardin EB, Lawrence JS: A comparison of human phosphate activity toward sodium beta-glycerophosphate, adenosine 5′-phosphate, and glucose-1-phosphate. Blood 14:415, 1959.
Trubowitz S, Feldman D, Morgenstern SW, Hunt VM: The isolation, purification and properties of the alkaline phosphatase of human leukocytes. Biochem J 80:369, 1961.
Valentine WN, Beck WS: Biochemical studies on leukocytes: I. Phosphatase activity in health, leukocytosis, and myelocytic leukemia. J Lab Clin Med 38:39, 1951.
Valentine WN, Follette JH, Solomon DH, Reynolds J: The relationship of leukocyte alkaline phosphatase to “stress,” to ACTH, and to adrenal 17-OH-corticosteroids. J Lab Clin Med 49:723, 1957.
Wachstein M: Alkaline phosphatase activity in normal and abnormal human blood and bone marrow. J Lab Clin Med 31:1, 1946.
Cline MJ: Metabolism of the circulating leukocyte. Physiol Rev 45:674, 1965.
Hayhoe FGJ, Quaglino D, Doll R: The Cytology and Cytochemistry of Acute Leukemias. HM Stationery Office, London, 1964.
Garg S, Silber R: Decreased leukocyte alkaline phosphatase in monocytic leukemia. Am J Clin Pathol 58:668, 1972.
Mittal CK: Measurements of cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in polymorphonuclear leukocytes. Methods Enzymol 132:428, 1986.
Scott RE: Effects of prostaglandins, epinephrine and NaF on human leukocyte, platelet and liver adenyl cyclase. Blood 35:514, 1970.
Ishitoya J, Takenawa T: Potentiation of PGE1-induced increase in cyclic AMP by calmodulin-dependent processes. J Immunol 138:1201, 1987.
Smolen JE, Korchak HM, Weissmann G: Increased levels of cyclic adenosine-3′,5′-monophosphate in human polymorphonuclear leukocytes after surface stimulation. J Clin Invest 65:1077, 1980.
Huang CK, Mackin WM, Bormann BJ, Becker EL: Cyclic AMP receptor protein and cyclic AMP-dependent protein kinase activity in rabbit peritoneal neutrophils. J Reticuloendothel Soc 34:413, 1983.
Tsung PK, Sakamoto T, Weissmann G: Protein kinase and phosphatases from human polymorphonuclear leukocytes. Biochem J 145:437, 1975.
Parker CW, Baumann ML, Huber MG: Alterations in cyclic AMP metabolism in human bronchial asthma: II. Leukocyte and lymphocyte responses to prostaglandins. J Clin Invest 52:1336, 1973.
Parker CW, Huber MG, Baumann ML: Alterations in cyclic AMP metabolism in human bronchial asthma: III. Leukocyte and lymphocyte responses to steroids. J Clin Invest 52:1342, 1973.
Alston WC, Patel KR, Kerr JW: Response of leukocyte adenyl cyclase to isoprenaline and effects of alpha blocking drugs in extrinsic bronchial asthma. Br Med J 1:90, 1974.
Marks PA, Gellhorn A, Kidson C: Lipid synthesis in human leukocytes, platelets, and erythrocytes. J Biol Chem 235:2579, 1960.
Fogelman AM, Seager J, Edwards PA, Hokom M, Popjak G: Cholesterol biosynthesis in human lymphocytes, monocytes, and granulocytes. Biochem Biophys Res Commun 76:167, 1977.
Chanock SJ, Faust LR, Barrett D, et al: O2– production by B lymphocytes lacking the respiratory burst oxidase subunit of p47-phox after transfection with an expression vector containing a p47-phox cDNA. Proc Natl Acad Sci USA 89:10174, 1992.
Elsbach P: Lipid metabolism by phagocytes. Semin Hematol 9:227, 1972.
Wang P, Waite M, Dechatelet LR: Membrane lipid metabolism of bacillus Calmette-Guérin-induced rabbit alveolar macrophages. Biochim Biophys Acta 487:163, 1977.
Nieto ML, Velasco S, Sanchez Crespo M: Modulation of acetyl-Coa: 1-alkyl-2-lyso-sn-glycero-3-phosphocholine (lyso-PAF) acetyltransferase in human polymorphonuclears: The role of cyclic AMP-dependent and phospholipid-sensitive, calcium-dependent protein kinases. J Biol Chem 263:4607, 1988.
Majerus PW, Lastra R: Fatty acid biosynthesis in human leukocytes. J Clin Invest 46:1596, 1967.
Elsbach P, Kayden HJ: Chylomicron lipid-splitting activity in homogenates of rabbit polymorphonuclear leukocytes. Am J Physiol 209:765, 1965.
Dienstle F, Sailer S, Sandhager F, Braunsteiner H: Lipid activity in leucocytes and macrophages. Blood 24:607, 1964.
Elsbach P, Weiss J: Lipid Metabolism by Phagocytic Cells in the Reticulendothelial System, edited by AJ Sbarra, RR Strauss. Plenum, New York, 1980, p. 91.
Pai J-K, Siegel MI, Egan RW, Billah MM: Phospholipase D catalyzes phospholipid metabolism in chemotactic peptide-stimulated HL-60 granulocytes. J Biol Chem 263:12472, 1988.
Samuelsson B, Goldyne M, Granstrom E, Hamberg M, Hammarstrom S, Malmsten C: Prostaglandins and thromboxanes. Annu Rev Biochem 47:997, 1978.
Walsh CE, Waite BM, Thomas MJ, Dechatelet LR: Release and metabolism of arachidonic acid in human neutrophils. J Biol Chem 256:7228, 1981.
Sellmayer A, Strasser T, Weber PC: Differences in arachidonic acid release, metabolism and leukotriene B4 synthesis in human polymorphonuclear leukocytes activated by different stimuli. Biochim Biophys Acta 927:417, 1987.
Godfrey RW, Manzi RM, Clark MA, Hoffstein ST: Stimulus-specific induction of phospholipid and arachidonic acid metabolism in human neutrophils. J Cell Biol 104:925, 1987.
Borgeat P, Samuelsson B: Transformation of arachidonic acid by rabbit polymorphonuclear leukocytes. Formation of a novel dihydroxyeicosatetraenoic acid. J Biol Chem 254:2643, 1979.
Bokoch GM, Reed PW: Stimulation of arachidonic acid metabolism in the polymorphonuclear leukocytes by an N-formylated peptide. Comparison with ionophore A23187. J Biol Chem 255:10223, 1980.
Dusi S, Poli G, Berton G, Catalano P, Fornasa CV, Peserico A: Chronic granulomatous disease in an adult female with granulomatous cheilitis. Evidence for an X-linked pattern of inheritance with extreme lyonization. Acta Haematol 84:49, 1990.
Moncada S, Ferreira SH, Vane JR: Prostaglandins, aspirin-like drugs and the oedema of inflammation. Nature 246:217, 1978.
Samuelsson B, Dahlen S-E, Lingren J-A, Rouzer CA, Serhan CN: Leukotrienes and lipoxins: Structures, biosynthesis, and biological effects. Science 237:1171, 1987.
Rouzer CA, Samuelsson B: On the nature of the 5-lipoxygenase reduction in human leukocytes: Enzyme purification and requirement for multiple stimulatory factors. Proc Natl Acad Sci USA 82:6040, 1985.
Samuelsson B, Dahlen S-E, Lindgren J-A, Rouzer CA, Serhan CN: Leukotrienes and lipoxins: Structures, biosynthesis and biological effects. Science 237:1171, 1987.
Orning L, Hammarstrom S, Samuelsson B: Leukotriene D: A slow reacting substance from rat basophilic leukemia cells. Proc Natl Acad Sci USA 77:2014, 1980.
Bach MK, Brashler JR, Hammarstrom S, Samuelsson B: Identification of leukotriene C-1 as a major component of slow reacting substance from rat mononuclear cells. J Immunol 125:115, 1980.
Bach MK, Brashler JR, Hammarstrom S, Samuelsson B: Identification of a component of rat mononuclear cell SRS as leukotriene D. Biochem Biophys Res Commun 93:1121, 1980.
Samuelsson B, Hammarstrom S, Murphy RC, Borgeat P: Leukotrienes and slow-reacting substance of anaphylaxis (SRS-A). Allergy 35:375, 1980.
Radmark O, Malmsten C, Samuelsson B, Goto G, Marfat A, Corey EJ: Leukotriene A. Isolation from human polymorphonuclear leukocytes. J Biol Chem 255:11828, 1980.
Evans JF, Dupuis P, Ford-Hutchinson AW: Purification and characterization of leukotriene A4 hydrolase from rat neutrophils. Biochim Biophys Acta 840:43, 1985.
Ford-Hutchinson AW, Bray MA, Cunningham FM, Davidson EM, Smith MJH: Isomers of leukotriene B4 possess different biological potencies. Prostaglandins 21:143, 1981.
Goldman DW, Gifford LA, Olson DM, Goetzl EJ: Transduction by leukotriene B4 receptors of increases in cytosolic calcium in human polymorphonuclear leukocytes. J Immunol 135:525, 1985.
Dahinden CA, Zingg J, Maly FE, de Weck AL: Leukotriene production in human neutrophils primed by recombinant human granulocyte/macrophage colony-stimulating factor and stimulated with the complement component C5A and FMLP as second signals. J Exp Med 167:1281, 1988.
Fitzharris P, Cromwell O, Moqbel R, et al: Leukotriene B4 generation by human neutrophils following IgG-dependent stimulation. Immunology 61:449, 1987.
Roubin R, Elsas PP, Fiers W, Dessein AJ: Recombinant human tumour necrosis factor (rTNF)2 enhances leukotriene biosynthesis in neutrophils and eosinophils stimulated with the Ca2+ ionophore A23187. Clin Exp Immunol 70:484, 1987.
Weisbart RH, Kwan L, Golde DW, Gasson JC: Human GM-CSF primes neutrophils for enhanced oxidative metabolism in response to the major physiological chemoattractants. Blood 69:18, 1987.
Kelleher D, Bloomfield FJ, Lenehan T, Griffin M, Geighery C, McCann SR: Chronic granulomatous disease presenting as an oculomucocutaneous syndrome mimicking Beheçt’s syndrome. Postgrad Med J 62:489, 1986.
Marcus AJ: The eicosanoid in biology and medicine. J Lipid Res 25:1511, 1984.
Soberman RJ, Harper TW, Murphy RC, Austen KF: Identification and functional characterization of leukotriene B4 20-hydroxylase of human polymorphonuclear leukocytes. Proc Natl Acad Sci USA 82:2292, 1985.
Sumimoto J, Takeshige K, Minakami S: Characterization of human neutrophil leukotriene B4 omega-hydroxylase as a system involving a unique cytochrome P-450 and NADPH-cytochrome P-450 reductase. Eur J Biochem 172:315, 1988.
Lee TH, Hoover RL, Williams JD, et al: Effect of dietary enrichment with eicosapentaenoic and docosahexaenoic acids on in vitro neutrophil and monocyte leukotriene generation and neutrophil function. N Engl J Med 312:1217, 1985.
Payan DG, Wong MY, Chernov-Rogan T, et al: Alterations in human leukocyte function induced by ingestion of eicosapentaenoic acid. J Clin Immunol 6:402, 1986.
Borgeat P, Hamberg M, Samuelsson B: Transformation of arachidonic acid and homo-gamma-linoleic acid by rabbit polymorphonuclear leukocytes: Monohydroxy acids from novel lipoxygenases. J Biol Chem 251:7816, 1976.
Stenson WF, Parker SW: Monohydroxyeicosatetraenoic acids (HETEs) induce degranulation of human neutrophils. J Immunol 124:2100, 1980.
Serhan CN, Fiore S, Levy BD: Cell-cell interactions in lipoxin gener- ation and characterization of lipoxin A4 receptors. Ann N Y Acad Sci 744:166, 1994.
Copyright © 2001 McGraw-Hill
Ernest Beutler, Marshall A. Lichtman, Barry S. Coller, Thomas J. Kipps, and Uri Seligsohn