摘要:

AbstractThis study shows that normal human large granular lymphocytes (LGL) secrete tumor necrosis factor (TNF) in response toMycobacterium avium‐intracellularecomplex (MAI). Percoll density gradient fractionation of peripheral mononuclear cells showed TNF activity in the fractions corresponding to LGL and not T cells, even when 5% monocytes were added to the T lymphocytes for accessory function. TNF release was not abrogated by treatment of the crude LGL preparations with anti‐Leu M3. ‐CD4. and ‐CD8 antibodies (Ab) plus complement (C), but was abrogated by anti‐CD16 and ‐CD2 Ab, as expected. Interestingly, anti‐HLA‐DR monoclonal antibody (mAb) treatment significantly diminished TNF activity from LGL, but maintained natural killer (NK) cell function unmodified as opposed to CD2 ‐ and CD16‐ cell depletion. Panning studies demonstrated that TNF secretion upon MAI stimulation resided only in the HLA‐DR+ LGL and not the DR LGL population. These results indicate that normal fresh HLA‐DR+ LGL, as well as monocytes, are also responsible for rapid TNF secretion during early MAI infection. These DR ‐ cells appear to be distinct from those expressing NK function.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.529

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractPolymorphonuclear leukocyte (PMN) respiratory burst was stimulated by heterologous antibodies against PMN granule proteins but not by control antibodies. Fluorescence‐activated cell sorter (FACS) analysis of activated PMN demonstrated the presence of two primary granule proteins, proteinase 3 (PR‐3) and cationic protein 57 (CAP‐57) at the membrane surface. The presence of myeloperoxidase (MPO) at the cell surface of primed and unprimed PMN was confirmed by immunoelectron microscopy. Priming doses of recombinant tumor necrosis alpha (rTNFα) enhanced the rate of superoxide (O2‐) production by these antibodies and increased the amount of surface protein accessible to these antibodies. Anti‐neutrophil cytoplasmic autoantibodies (ANCA) with specificities for PMN granule proteins are present in patients with Wegener's granulomatosis, polyarteritis nodosa, and idiopathic and crescentic glomerulonephritis. The demonstration that antibodies against granule proteins activate PMN supports the hypothesis that the vasculitis seen in these diseases is due in part to PMN mediated oxidative injury following PMN stimulation by ANCA.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.539

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractSkin test anergy, the failure to produce a delayed type hypersensitivity (DTH) response, is associated with an increase in infection‐related complications and death usually due to multiple organ failure (MOF). Refractory intravascular activation of polymorphonuclear neutrophils (PMNs) has been implicated in the development of MOF. We studied 20 critically ill surgical patients with life threatening infections to determine if PMN intravascular activation was present and how this affected essential PMN functions such as exudation. The 11 anergic patients had a more intense inflammatory response to their infection. Plasma lactoferrin was 6.1 ± 0.3 μg/ml in anergic patients compared to 3.9 ± 1.5 in reactivesP<0.05, accompanied by reduced total primary (3.3 ± 1.9 vs 4.7 ± 2.1 μg/106PMNP<0.01) and secondary (2.8 ± 0.4 vs 5.0 ± 0.9 μg/106PMNP<0.01) granule content, respectively. In vitro superoxide production following 100 ng/ml PMA stimulation was 0.44 ± 0.1 in anergics vs 0.36 ± 0.1 nmol/μg PMN protein in reactivities,P<0.05. PMN chemotaxis was 8.2 ± 0.6 PMNs/HPF in anergics compared to 10.2 ± 1.6 PMNs/HPF in reactivesP<0.05, accompanied by decreased PMN delivery to skin blister windows (3.2 ± 1.4 vs 4.5 ± 1.9 × 107PMN/ml, respectively,P<0.05). We conclude that critically ill anergic surgical patients have increased intravascular PMN activation, which may contribute to oxygen‐derived tissue damage in the vascular space, as well as a deficient delivery of effector cells in areas of bacterial invasion. This may lead to inability to clear the inflammatory signals which set up the vicious circle of MOF leading to death.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.547

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractThe oxidative burst of rainbow trout(Oncorhynchus mykiss)phagocytes was previously found to be differentially modulated by adrenocorticotropic hormone (ACTH) and the catecholamine receptor agonists phenylephrine and isoproterenol. From data obtained using both luminol‐enhanced chemiluminescence (LECL) and ferricytochrome C (cyt C) reduction to measure oxidative burst kinetics, we postulated that the observed modulation was mediated by affects on enzymes responsible for the production and metabolism of superoxide anion. Using exogenous superoxide dismutase (SOD) and catalase as scavengers, nitroprusside to poison endogenous SOD, and an assay for hydrogen peroxide, we have tested our postulates by exploiting the differences with which various reactive oxygen intermediates influence LECL and cyt C reduction. The ability of ACTH to potentiate both assays of the oxidative burst appears due to its enhancing influence on the production of superoxide. Phenylephrine, an α‐adrenergic receptor agonist, appears to enhance the activity of endogenous SOD, whereas isoproterenol, a β‐adrenergic receptor agonist, may suppress SOD activity. This work reveals how components of the natural immune system may be regulated by products of the neuroendocrine system. Also, lymphocyte‐derived ACTH may provide a novel pathway for lymphoid regulation of inflammation.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.554

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractAn epithelial cell line, IT‐45R1, has been established from a thymus of normal Wistar rat and was found to produce growth factor which stimulated the proliferation of bone marrow cells. This growth factor induced the formation of colonies composed of macrophages, granulocytes, or both, in semi‐solid medium and stimulated the proliferation of an interleukin 3 (IL3)/granulocyte‐macrophage colony‐stimulating factor (GM‐CSF)‐dependent clone. Neither IL3‐dependent clone nor IL6‐dependent clone responded to IT‐45R1 factor. Additionally, IT‐45R1 factor acted on both rat and mouse bone marrow cells but not on human bone marrow cells. Molecular weight of IT‐45R1 factor was 30 kD and its isoelectric point was 4.5. To determine whether IT‐45R1 factor is GM‐CSF or not, Northern blot analysis and neutralization with anti‐mouse GM‐CSF antibody were carried out. IT‐45R1 expressed GM‐CSF mRNA, but neither M‐CSF nor IL6 transcripts. However, antiserum specific for mouse GM‐CSF did not neutralize IT‐45R1 factor. Taken together, a rat thymic epithelial cell line, IT‐45R1, constitutively produces GM‐CSF or GM‐CSF‐like growth factor.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.561

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractThe neutrophil serine proteinases elastase and cathepsin G produce connective tissue injury, the extent of which depends on the balance between these enzymes and their inhibitors. The most important of these inhibitors is α1‐proteinase inhibitor, a member of a superfamily of homologous proteins known as serpins. Neutrophil cytosol inhibited the activities of human neutrophil elastase and cathepsin G in a dose‐dependent fashion. To demonstrate formation of an enzyme‐inhibitor complex, we combined125l‐elastase or125l‐cathepsin G with neutrophil cytosol or α1‐proteinase inhibitor and analyzed the products by polyacrylamide gel electrophoresis. Unbound elastase and cathepsin G each migrated to an apparent molecular weight of 25 kDa. In the presence of cytosol from neutrophils both radiolabeled enzymes migrated with a relative size of 68 kDa, whereas in the presence of α1‐proteinase inhibitor the relative size was 85 kDa. Enzyme‐inhibitor complexes were stable in sodium dodecyl sulfate at 100° but were dissociated by hydrolysis in ammonium hydroxide (1.5 mol/L) at 37°. Formation of each complex was prevented by pretreatment of elastase or cathepsin G with diisopropylfluorophosphate, indicating that the inhibitor binds to the active site of the enzyme. Exposure of either α1‐proteinase inhibitor or neutrophil cytosol to the myeloperoxidase‐H2O2‐halide system prevented complex formation, suggesting the presence of an oxidizable amino acid at the binding site of the inhibitor. By electrophoretic analysis, the molecular weight of the cytosolic inhibitor was 43 kDa and neutrophils contained approximately 1 attomol of inhibitor per cell. The isoelectric points of the elastase and cathepsin G inhibitor were 5.5–5.9 and inhibitors of the two proteinases coeluted using size exclusion chromatography. These data demonstrate that human neutrophil cytosol contains a single serpin‐like protein that inhibits elastase and cathepsin G. The inhibitor may be important in protecting the intracellular environment from proteolytic injury during degranulation.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.568

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractThe drug 3′‐azido‐3′‐deoxythymidine (AZT), a synthetic thymidine analogue, has been used clinically in the management of acquired immune deficiency syndrome (AIDS). The drug is an effective antiviral agent due to its ability to block reverse transcriptase activity. This action of AZT was demonstrated in the Rauscher leukemia virus (RLV)‐induced murine erythroleukemia model system. Unfortunately, associated with AZT has been the development of hematopoietic toxicity manifested by anemia, neutropenia, and overall bone marrow suppression. Hematopoietic growth factors (GM‐CSF, erythropoietin), cytokines (interleukin‐1), and agents known to potentiate hematopoiesis (lithium) have been demonstrated to modulate drug and/or radiation‐induced hematopoietic toxicity. We report the results of further studies designed to investigate the ability of GM‐CSF, erythropoietin, interleukin‐1, and lithium to modulate AZT toxicity on murine hematopoietic granulocyte‐macrophage (CFU‐GM), megakaryocytic (CFU‐Meg), and erythroid (BFU‐E) progenitors cultured from bone marrow and spleen cells from mice infected with RLV. Hematopoietic progenitors from either normal or RLV‐infected animals when exposed to AZT demonstrated concentration‐dependent toxicity and differed for each progenitor with BFU‐E being the most sensitive (ID50concentration, 5 × 10‐9M) and CFU‐GM the least sensitive (ID50concentration, 5 × 10‐5M). As has been previously demonstrated using normal murine hematopoietic progenitors, when cultured with RLV‐infected marrow or spleen cells, addition of GM‐CSF, Meg‐CSF or erythropoietin failed to inhibit AZT toxicity in vitro on CFU‐GM, CFU‐Meg, and BFU‐E, respectively. However, in the presence of interleukin‐1 (recombinant human IL‐1α, 30 ngm) or lithium chloride (ultra‐pure, 1.0 mM), AZT toxicity CFU‐GM, CFU‐Meg, and BFU‐E cultured from RLV‐infected marrow or spleen cells was reduced. These results further demonstrate interleukin‐1 and lithium are effective in modulating the toxic action of ACT on hematopoietic progenitors and that RLV‐infected animals serve as a useful viral model system to study the effect of agents capable of modulating hematopoiesis in the presence of the anti‐viral drug AZT.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.580

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractTo further define the urtrastructural events associated with the killing ofCandida albicansby human neutrophils, four methods were used: (1) the periodate‐thiocarbohydrazidesilver proteinate (PA‐TCH‐SP) staining of vicinal‐glycol‐containing complex carbohydrates; (2) the localization of thermostable immunodeterminants of the yeast cell wall, mannans or mannoproteins, using monospecific antibodies and a protein A‐gold complex (monAb‐gold); (3) the localization of mannose residues with concanavalin A labeled with gold particles (Con A‐gold); (4) the localization of chitin oligomers using wheat germ agglutinin and ovomucoid labeled with gold particles (WGA‐gold). The mannan‐rich cell wall layers were progressively lost as shown by altered PA‐TCH‐SP reactivity and a diffuse pattern of staining with Con A‐gold and monAb‐gold. The de novo appearance of conspicuous amounts of glycogen‐like particles near the plasmalemma and in the cell wall was interpreted as evidence of a reparative process of the yeast cell wall. Chitin was seemingly unaltered and readily demonstrated by the WGA‐gold in the wall remnants of ghost cells.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.587

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractAlthough rat peritoneal neutrophils in the presence of cytochalasin B demonstrate superoxide (O2‐) responses to the chemotactic peptide N'‐formyl‐met‐leu‐phe (FMLP), neither elicited rat peritoneal macrophages nor rat alveolar macrophages show an O2‐response to FMLP (in the presence or absence of cytochalasin B), although a good O2‐response to opsonized zymosan is demonstrated by both types of macrophages. Using Fura‐2 loaded cells, peritoneal macrophages failed to show an increase in intracellular calcium after exposure to FMLP, f‐nor‐leu‐phe, F‐met‐met‐met, or F‐norleu‐leu‐phe‐norleu‐lys. FMLP also failed to induce elevations in intracellular calcium in alveolar macrophages. In3H‐FMLP binding studies, the lack of responsiveness of peritoneal and alveolar macrophages was associated with the lack of FMLP receptors on these cell types, in striking contrast to the presence of functional receptors on rat neutrophils.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.600

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractBurn trauma is associated with alterations of various components of host defenses, including impaired neutrophil functions. In an animal model of experimental thermal injury, we studied if the modifications of cellular reactivity result from alterations in signalling systems by comparing polyphosphoinositide breakdown, particularly the production of inositol phosphates (IP, IP2IP3), in healthy and burned rat polymorphonuclear neutrophil leukocytes (PMNs).Neutrophil activators such as N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP) and serum‐opsonized zymosan increased in vitro production of inositol phosphates in PMNs from healthy rats. The immunomodulator RU 41740 had no effect by itself, but decreased the stimulating effect of fMLP and zymosan. In PMNs from burned rats, the stimulating effects of fMLP and zymosan were decreased, while RU 41740 stimulated inositol phosphate generation. In vivo treatment with RU 41740 inhibited the activation of phosphoinositide metabolism by fMLP or zymosan in healthy rat PMNs. Similar treatment of burned rats after injury restored the stimulating effect of fMLP and zymosan on inositol phosphate accumulation in PMNs.Thus, RU 41740 can modulate fMLP and zymosan receptor‐mediated signal transduction, inducing an attenuation of the phosphatidylinositol hydrolysis response. After burn injury, when the activating effects of fMLP and zymosan are inhibited, RU 41740 can, on the contrary, stimulate phospholipase C‐mediated polyphosphoinositide turnover and the formation of intracellular messengers such as IP3.These data show that RU 41740 has different effects on polyphosphoinositide metabolism in rat PMNs, according to the physiological and pathological state of the animals. Interestingly, it has a beneficial action on the post‐burn decrease in PMN reactivity.

ISSN:0741-5400    

DOI:10.1002/jlb.50.6.607

出版商:Wiley    

年代:1991

数据来源: WILEY