摘要:

AbstractThe relationship between inter‐α inhibitor (IαI) and urinary proteinase inhibitor (UPI) was examined by comparing purified UPI with a proteolytic fragment of IαI (l'), and by demonstrating that inflammatory cells produce similar fragments under physiologic conditions. Purified I', derived by chymotrypsin digestion of IαI, was similar to UPI in apparent molecular weight (68,000‐69,000), amino acid composition, immunoreactivity, and inhibitory activity against trypsin, chymotrypsin, and neutrophil elastase. The production of similar inhibitory fragments by murine peritoneal macrophages, human neutrophils, and a murine mast cell line was quantified. Neutrophils were most efficient at proteolyzing IαI. Comparison of the pattern of IαI degradation by neutrophil preparations with that by pure enzymes, suggested that both elastase and cathepsin G mediate neutrophil proteolysis of IαI. These proteinases may thus be responsible for inflammation‐related increases in UPl‐like inhibitor levels in vivo.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.1

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractHuman eosinophilic cell line EoL‐1 was studied using luminol‐dependent chemiluminescence (CL) for the ability to produce a respiratory burst upon stimulation with 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) and N‐formyl‐L‐methionyl‐L‐leucyl‐L‐phenylalanine (fMLP). TPA, a potent activator of protein kinase C, induced a biphasic CL response in EoL‐1. Treatment of EoL‐1 with interferon‐γ (IFN‐γ) for 5 days dramatically enhanced TPA‐inducible CL, and IFN‐αA had a similar effect. Neither IFN‐γ nor IFN‐αA strongly inhibited EoL‐1 cell growth. Tumor necrosis factor (TNF) also enhanced TPA‐inducible CL response of EoL‐1 and, furthermore, was quite inhibitory to EoL‐1 cell growth. The effects of IFN‐γ and TNF were synergistic, whereas those of IFN‐αA and TNF were additive. Superoxide dismutase completely abrogated TPA‐induced CL, but sodium azide suppressed only the late phase of CL. EoL‐1 pretreated with IFN‐7, IFN‐αA, or TNF also became capable of producing CL response to a chemotactic peptide (fMLP). The effects of IFN‐γ and TNF were again synergistic. EoL‐1 cells treated with IFN‐γ, IFN‐αA, or TNF had abundant cytoplasm, but only TNF increased cells having distinct eosinophilic granules. IFN‐γ but not IFN‐αA enhanced the cytological effect of TNF. It was further demonstrated that treatment of EoL‐1 with IFN‐γ and TNF strongly increased the binding sites for phorbol diesters and also dramatically induced the surface expression of fMLP receptors. IFN‐γ had, however, little effect on the number or the ligand‐binding affinity of TNF receptors on EoL‐1. Thus, EoL‐1 may provide a useful experimental model for the study of differentiation and regulation of human eosinophils.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.10

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractInterleukin‐1 (IL‐1α and IL‐1β collectively) has been shown to be produced by a wide variety of cell types. The purpose of this study was to evaluate the ability of bovine polymorphonuclear leukocytes (PMNs) to synthesize and release IL‐1‐like cytokines and characterize the active molecule(s). Purified peripheral blood PMNs were cultured for various periods of time in the presence of opsonized zymosan particles. The resulting culture supernatants exhibited IL‐1 activity as determined by enhanced mitogen‐induced proliferation of the D10 G4.1 murine T‐helper cell line. Supernatants from nonstimulated PMNs or PMNs stimulated for less than 6 h did not enhance D10 G4.1 proliferation. The active molecule (PMNIL‐1) was isolated by using gel filtration high‐performance liquid chromatography (HPLC). Further characterization of the HPLC‐purtfied molecule by SDS‐PAGE and isoelectric focusing indicates bovine PMNIL‐1 has a molecular weight of 17.6 kd and a pl of 4.1.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.21

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractInterferon (IFN)‐induced tryptophan degradation, catalyzed by indoleamine 2,3‐dioxygenase (IDO), has been shown to mediate antimicrobial activity in epithelial cells. IDO activity has also been augmented in peripheral blood mononuclear cells (PBMC) treated with IFN or interleukin‐2 (IL‐2). The effector cells in this population have now been further characterized. PBMCs were isolated from normal donors, separated into monocyte and lymphocyte populations by plastic adherence, treated with IFN or IL‐2, and cultivated in medium supplemented with [3H]tryptophan. Culture supernatants were collected after a 48‐h incubation and fractionated by high‐performance liquid chromatography; radioactivity was determined in fractions corresponding to tryptophan and its metabolites. IFN‐γ and IFN‐β induced IDO activity only in monocytes (plastic‐adherent, nonspecific esterase‐positive PBMCs). The induction of IDO activity by IL‐2 required both monocytes and lymphocytes. Interaction was required between these populations for induction of IDO by IL‐2, due to production of IFN‐γ by T lymphocytes, with subsequent IFN‐γ‐mediated induction of IDO in monocytes. A number of myeloid cell lines as well as monocyte‐derived macrophages were also tested for their ability to be induced to degrade tryptophan in response to IFN treatment. Monocyte‐derived macrophages were found to retain their capacity to be induced by IFN‐γ and IFN‐β to degrade tryptophan after differentiation, and to possess seven times more IDO activity per cell than IFN‐induced monocytes. However, the presence of lipopolysaccharide (LPS) in the culture medium was required for the maximum induction of IDO activity by IFN‐β. Furthermore, higher concentrations of LPS were sufficient to induce IDO activity in macrophages in the absence of exogenous IFN.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.29

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractMacrophages are important positive and negative regulators of both primary and secondary antibody responses, and their activity may, in turn, be controlled by soluble mediators secreted by other cells. Fibronectin is a 440,000 dalton normal constituent of plasma and extracellular membranes that acts through macrophages to inhibit mitogen‐and alloantigen‐stimulated lymphoproliferation. We examined the effect of Fn on the antigen‐stimulated lymphoproliferative and antibody responses in cells from trinitrophenol‐derivitized keyhole limpet hemocyanin (TNP‐KHL) primed rats. Fn in concentrations equivalent to normal plasma levels inhibited TNP‐KLH‐stimulated lymphoproliferation by unseparated lymph node leukocytes. When the experiment was repeated using purified lymph node T cells and added thioglycollate‐induced peritoneal exudate macrophages or splenic adherent macrophages, Fn alone and TNP‐KLH alone stimulated lymphoproliferation, but in combination they were strongly inhibitory. The effect was not due to decreased lymphocyte viability in the presence of both TNP‐KLH and Fn. Nor was it due to complexes between TNP‐KLH and Fn or to a simple alteration in the kinetics of lymphoproliferation. Fn had to be present with the TNP‐KLH within the 1st hour of incubation. If macrophages were coincubated with TNP‐KLH and Fn for 24 h, washed, and added to enriched T cells, inhibition was equivalent to that seen with continuous coculture. Similarly, coculture of TNP‐KLH and Fn inhibited both total immunoglobulin and TNP‐KLH‐specific antibody synthesis at optimal concentrations of splenic adherent cells. However, at suboptimal levels of splenic macrophages, the combination was synergistic, stimulating more total immunoglobulin synthesis than either TNP‐KLH or Fn alone. These data suggest that the inhibitory effect was dependent upon the concentration and phenotype of macrophages present in culture.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.35

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractHuman promyelocytic cells (HL‐60) were labeled with35S‐sulfate and either3H‐glucosamine or3H‐serine as precursors. Accumulation of35S‐labeled macromolecules was approximately linear for up to 96 h, with a mean cell:medium ratio of 5.5:1, although activity/105viable cells reached a plateau level after 24 h. Virtually none of the cell‐associated proteoglycan was removed by trypsinization, consistent with a predominantly intracellular localization. Proteoglycan heterogeneity was investigated by DEAE‐Sephacel chromatography, isopyknic CsCI gradient centrifugation, and gel filtration chromatography. HL‐60 cells appeared to synthesize a single proteoglycan species, Kav = 0.46 on Sepharose CL‐4B and Kav = 0.32 on Sepharose CL‐6B, recovered primarily from the high‐density fractions of a dissociative CsCI gradient (ρ>1.40 g/l). Degradation products of lower charge density, lower buoyant density, and lower hydrodynamic size were also present, mainly in the cell pellets. The major proteoglycan was found to contain chondroitin sulfate chains of average Mr= 14.5 kD, yielding virtually 100% 4‐sulfated disaccharides on digestion with chondroitinase ABC. The proteoglycan was resistant to trypsin, chymotrypsin, plasmin, and papain, and the core protein Mr was approximately 20 kD by molecular sieve chromatography. Induction of HL‐60 cells with 0.15 dimethyl sulfoxide (DMSO) resulted in differentiation to a more mature granulocytic phenotype and was associated with a reduction in35S‐sulfate incorporation to 45% of control values or 32%, expressed as activity/105cells. Proteoglycans synthesized by DMSO‐treated cells were identical to those from untreated cells in terms of hydrodynamic size, glycosaminoglycan Mr, and sulfation.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.46

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractPrimary mouse hepatocytes were treated with the acute‐phase mediators interleukin‐1, Interleukin‐6, and glucocorticoids, singularly and in combination, in order to delineate the spectrum of proteins induced by the stimulatory factors. As found for rat and human liver cells, mouse hepatocytes responded to the cytokines by increasing production of acute‐phase proteins, which in mice include haptoglobin, α1‐acid glycoprotein, complement C‐3, serum amyloid A, and hemopexin. Serum amyloid A was unusual in that only the acidic peptide form responded to treatment with IL‐1 and IL‐6; the more basic form remained unchanged. In addition, an unidentified secretory protein was induced only by mixtures containing IL‐6. The present study shows that a combination of IL‐1, IL‐6, and glucocorticoids is required for regulation of acute‐phase plasma protein production in mouse liver cells.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.55

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractThe migration of polymorphonuclear leukocytes (PMN) in response to recombinant interleukin‐1 (IL‐1), tumor necrosis factor‐alpha (TNF), C5a, and f‐met‐leu‐phe‐lys (FMLPL) in vivo was studied using a mouse subcutaneous sponge implantation model. In this model sponges were implanted in C3H/OUJ mice, and 2 days later they were injected with the test sample. After varying times, sponges were removed and digested with collagenase, and total cell counts and differentials were enumerated. IL‐1 was found to stimulate a significant influx of PMN, which peaked at 6 hr and declined to near baseline levels by 24 hr. This response was dose‐dependent, with the greatest response observed when 5 units of IL‐1 were injected. When the IL‐1 concentration was increased to 10 U, the total number of PMN migrating into the sponge was decreased, compared with that observed with 5 U of IL‐1. The overall number of PMN migrating into the sponge 6 hr after injecting 5 U of IL‐1 averaged 269% of the number of PMN migrating randomly into the sponge. No difference in the total number of macrophages or lymphocytes in control or IL‐1‐injected sponges was observed in this time frame. Heat treatment of the IL‐1 at 90°C for 30 min ablated the response. Similar studies with TNF and C5a showed that both of these agents also stimulated an influx of PMN that peaked 6 hr postinjection. In contrast, FMLPL did not stimulate a PMN response. When IL‐1 and TNF were injected simultaneously, an additive response was observed. These data indicate that IL‐1, TNF, and C5a can all stimulate a PMN response in vivo and support the hypothesis that these substances are actively involved in the mobilization of PMN to inflammatory sites in vivo.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.62

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractThymic stromal cell lines, termed MRL104 and MRL28, have been isolated from long‐term liquid cultures of thymic stromal cells from MRL/1 mice. The capacities of these parental lines and derived clones to express la antigens and to produce cytokines involved in T‐cell proliferation and/or differentiation were investigated. Parental lines and their clones did not exhibit a typical fibroblastic, macrophage‐like, or epithelial appearance in electron as well as phase‐contrast micrographs. These thymic stromal cells seemed to differ from established fibroblast lines in that these thymic stromal cells expressed la antigens after exposure to interferon‐γ (IFN‐γ), whereas fibroblast lines did not They also appeared to differ from macrophage cell lines in that they lacked the expression of Mac‐1 antigens on their cell surface and produced no detectable level of interteukin‐1 (IL1) before or even after exposure to lipopolysaccharide. When these parental lines and its clones were tested for their ability to produce various types of cytokines, it was revealed that they were capable of producing colony‐stimulating factor (CSF), IL6, and thymic stroma‐derived T cell growth factor (TSTGF). which was recently described, but were unable to generate other lymphokines and IFNs. Thus these cell lines and clones represent unique features in that they have potentials to express la antigens and to produce CSF, IL6, and TSTGF. The biological significance for the expression of these features is discussed in the context of intrathymic T‐cell maturation and T‐cell repertoire selection.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.69

出版商:Wiley    

年代:1989

数据来源: WILEY

摘要:

AbstractInvestigation into the rnechanism(s) whereby monocytes‐macrophages are mobilized to the myocardium of coxsackievirus (CVB3)‐infected mice was approached by assessing the role of anti‐idiotypic antibodies against specific antiviral IgG molecules. Balb/c mice preinocuiated with polyclonal anti‐idiotypic, syngeneic antibodies resulted in significantly decreased myocarditis after virus challenge. On the other hand, oil‐induced peritoneal exudate cells obtained from virus‐exposed animals demonstrated increased chemotaxis in the presence of polyclonal anti‐Id preparations, whereas peritoneal exudate cells from normal animals did not Specificity of the anti‐Ids as confirmed by the binding of the syngeneic anti‐anti‐idiotypic antibodies (antibody 3) to solubilized viral antigen(s) and Fab fragments prepared from the anti‐idiotypic antibodies. Paradoxic biological effects of anti‐Id, demonstrated in vitro and in vivo, suggest possible and subtle immunoregulatton of the macrophage effector arm by anti‐ldiotypes during the course of virus infection.

ISSN:0741-5400    

DOI:10.1002/jlb.45.1.79

出版商:Wiley    

年代:1989

数据来源: WILEY