摘要:

AbstractLittle is known regarding the interaction ofBordetella pertussiswith polymorphonuclear leukocytes (PMNL) or the role PMNL play as an initial line of defense against B.pertussisinfection. An in vitro system was developed to establish conditions for the study of phagocytosis and killing of virulentB. pertussisby human PMNL. Phagocytosis ofB. pertussisstrains BP504, BP165, and BP338 occurred by opsonization with anti‐B. pertussisantibody, while autologous normal human sera did not induce significant phagocytosis. In PMNL bacterial killing assays virulent B.pertussisstrains survived PMNL bactericidal activities whileEscherichia colicontrols were readily killed. Electron microscopy studies using acid phosphatase as a lysosomal marker strongly suggested that B. pertussis inhibits phagosome‐lysosome fusion in PMNL. These results indicate that virulent B.pertussisstrains are capable of surviving intracellularly within PMNL and that such survival may be due to inhibition of phagosome‐lysosome fusion.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.321

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractFragmentation of subendothelial matrix‐bound fibronectin by proteases released from stimulated leukocytes has been implicated in lung vascular injury. We studied the degradation of fibronectin bound to denatured collagen by inflammatory polymorphonuclear leukocytes (PMNL). Tissue culture wells coated with denatured collagen (gelatin) were pretreated with125I rat plasma fibronectin to allow for fibronectin binding prior to the addition of rat inflammatory PMNL. The release of both intact and fragmented fibronectin from the125l‐labelled artificial matrix was quantified following the addition of PMNL stimulated by the phagocytosis of opsonized zymosan as well as leukocyte elastase. Stimulated PMNL released three times more radiolabelled fibronectin from the denatured collagen surface during a 4 h incubation as compared with unstimulated PMNL. This pattern of125I‐fibronectin release could also be elicited by the addition of purified leukocyte elastase alone, in the absence of PMNL. The release of radiolabelled fibronectin by stimulated PMNL was blocked in a dose‐dependent manner by the addition of both methoxysuccinyl‐alanine‐alanine‐valine chloromethyl ketone (AAPVCK), a leukocyte elastase specific inhibitor as well as phenylmethylsulfonylfluoride (PMSF), a non‐specific serine protease inhibitor. Western blot analysis coupled with autoradiography confirmed the presence of fibronectin fragments in the medium after addition of PMNL or leukocyte elastase. Thelargemolecular weight fragments (60–200 kD) were not labelled, but thesmallermolecular weight fragments (<45 kD), derived from the artifical matrix, were labelled. Thus, fibronectin complexed with denatured collagen is susceptible to proteolytic degradation by stimulated inflammatory PMNL. Such a process may have a role in the pathogenesis of acute vascular injury following microvascular margination of activated blood leukocytes.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.331

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractHuman T‐cell cultures infected with human T‐lymphotropic virus type I (HTLV‐I) and interleukin‐2 (IL‐2)‐dependent for their continuous growth were treated with N‐methyl‐N'‐nitro‐N‐nitrosoguanidine (MNNG) and then maintained in the medium containing phorbol 12‐myristate 13‐acetate (TPA). Cells achieved independence from IL‐2 but became TPA‐dependent for continuous growth. Multiple ultraviolet (UV) irradiations of TPA‐dependent cells resulted in their autonomous growth. G‐band karyotype analysis revealed multiple chromosomal abnormalities that were seen in cells before and after MNNG treatment and UV irradiations, and those that were only seen in autonomously growing cells. Viral expression was found to be transiently enhanced in association with emergence of certain chromosomal changes. Exposure of HTLV‐I infected cells to certain mutagens may promote the occurrence of the specific rearrangement of cellular genes responsible for regulation of cellular and viral replication and may lead these cells to neoplastic transformation.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.341

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractThe actions of retroviral infections, aging, and cocaine and morphine injection on cytokine production were investigated in C57BL/6 female mice. Retroviral infection with LP‐BM5 murine leukemia virus was further developed as a model of murine acquired immunodeficiency syndrome (AIDS). The effects of cocaine and morphine on gammainterferon (IFN) and tumor necrosis factor (TNF) production in vivo and with isolated spleen cells were measured by a sandwich enzyme‐linked immunosorbent assay (ELISA) method. Serum IFN was generally not detected in any group except mice injected with saline and young mice infected with LP‐BM5 virus. Splenocytes from mice with murine AIDS produced less IFN when stimulated in vitro by ConA. In aged mice, IFN production by spleen cells was severely suppressed by retroviral infection. Cocaine had a tendency to suppress IFN production by stimulated cells in vitro. Morphine tended to reduce IFN production by spleen cells from retrovirally infected animals.The serum TNF level in mice with murine AIDS was elevated creating higher levels in morphine and morphine plus cocaine treated uninfected mice while cocaine injection eliminated serum TNF. When stimulated in vitro by lipopolysaccharides (LPS), splenocytes from mice with murine AIDS also produced more TNF than uninfected controls. TNF production in vitro and in vivo was significantly increased by retroviral infection. Therefore, results indicate that cocaine and retroviral infection modulate TNF and IFN production.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.349

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractSubstantial amounts of macromolecular activators of phagocytosis from platelets (MAPPs) were released in response to exposure of platelets to the specific agonists thrombin and collagen and to calcium ionophore A23187. Centrifugation of the platelets in culture medium also resulted in a release of MAPPs, but not when the platelets were frozen and thawed prior to centrifugation.In an experiment using outdated platelet concentrates, larger and smaller MAPPs (1‐MAPP and s‐MAPP, respectively) were purified from the thrombin stimulated supernatant (PRPr‐plasma) by sequential steps of ammonium sulfate precipitation, delipidation with ethylacetate, ConA‐Sepharose affinity chromatography, MONO Q anion exchange chromatography, and Superose 12 gel filtration. This procedure yielded 59,500‐fold and 63,600‐fold purified 1‐MAPP (0.95 mg) and s‐MAPP (0.41 mg), respectively, from 1,320 ml PRPr‐plasma containing 84,500 mg protein. By gradient polyacrylamide gel electrophoresis the respective molecular weights (MW) of 1‐MAPP and s‐MAPP were 290–320 kd and 140–160 kd; isoelectric points were 5.6 and 4.6. Both MAPPs activated neutrophil phagocytosis of IgG‐SRBC at concentrations in the range of 1 pM–100 nM. Indirect enzyme‐linked immunosorbent assay (ELISA) and comparisons of the concentrations required for phagocytosis activation suggested that the MAPPs were not derived from fibronectin.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.356

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractAlthough a substantial amount of information is available on pulmonary alveolar macrophages (PAMs), little is known about pulmonary intravascular macrophages (PIMs), a separate population of lung macrophages found apposed to the endothelium of pulmonary capillaries. We compared these two populations of lung immunocytes to determine their relative immunological activity. Our results suggest that PAMs are more phagocytic than PIMs; however, PIMs may be more efficient at lysing ingested bacteria than PAMs. Although similar in antibody‐dependent cellular cytotoxicity, PIMs are more spontaneously cytolytic than PAMs. Depending upon the effector:target cell ratio studied, the tumoricidal activity of PIMs was less than or equal to that of PAMs. Both cell populations produced the cytokines interleukin‐1 and tumor necrosis factor‐α at similar concentrations. These data suggest that PIMs are immunologically active, although the degree of activity may differ between PIMs and PAMs.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.364

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractWe previously demonstrated that the α1(l) polypeptide chain of collagen can bind and activate polymorphonuclear neutrophils (PMN). In the present experiments, performed in culture grade 96‐well plastic plates coated with collagen, fibronectin, or other proteins, adhesion was assessed by staining the adhering cells after 30 min with crystal violet and measuring absorbance at 560 nm, and activation of PMNs was assessed by measuring the amount of O2‐formed. Adhesion occurred at 17 and 37°C but activation at 37°C only. Monoclonal antibody anti‐CD 18 inhibited adhesion, showing that the receptor of collagen I on PMNs is a β2integrin. On the other hand, adhesion of PMNs to fibronectin was inhibited by monoclonal antibodies to CD18 and to CD11b.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.373

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractThe metrial gland and decidua basalis are uterine structures which, during pregnancy in mice and rats, contain bone marrow derived cells. The current study demonstrates that large numbers of bone marrow derived cells, identified by common leukocyte antigen (CLA) positivity, accumulate in the mesometrial uterus between days 6 and 10 and contribute significantly to the cellularity of both the metrial gland and the decidua basalis. The distribution of F4/80+cells (macrophages) is similar, suggesting that CLA+cells in the metrial gland and decidua basalis are macrophages. Disappearance of luminal epithelium in the mesometrial uterus between days 11 and 12 leads to regression of metrial gland and decidua basalis and coincident disappearance of CLA+and F4/80+cells. A second population of CLA+and F4/80+cells appears in association with the development of new uterine luminal epithelium which surrounds the fetus and placenta during the final week of pregnancy. These very large accumulations of macrophages invariably are related to presence of epithelium.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.381

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractGangliosides have been shown to act as immunoregulatory agents by altering proliferative responses of lymphocytes to both antigens and mitogens. Most early studies have utilized brain gangliosides and have required high concentrations. The role of endogenous gangliosides from macrophages has remained unexplored. In this study, thioglycollate‐elicited murine peritoneal macrophage gangliosides were purified and added to cultures of murine lymphocytes. Nanogram amounts caused a profound inhibition of LPS‐induced DNA synthesis of splenocytes and of purified B lymphocytes, without demonstrable cellular toxicity. No effect was seen using asialo‐GM1. This effect was present across a wide range of lipopolyssacharide (LPS) doses. Nanogram amounts of macrophage gangliosides also inhibited concanavalin A (ConA)‐mediated lymphocyte proliferation. Inhibition of LPS‐induced mitogenesis was present even if gangliosides were removed from the extracellular environment after 15–60 min of incubation prior to the addition of LPS. This inhibition was reversible with incubation of ganglioside pre‐treated lymphocytes in medium containing serum. These inhibitory properties of macrophage gangliosides are distinct from those found in studies using brain gangliosides, and support a potential role for macrophage gangliosides as negative modulators of lymphocyte proliferation.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.393

出版商:Wiley    

年代:1991

数据来源: WILEY

摘要:

AbstractNylon‐passed spleen cells were found to proliferate when cultured with syngeneic nonparenchymal adherent liver cells and their culture supernatants. The supernatants contained IL‐1, IL‐6, GM‐CSF, and IFN (α + β) activities but not IL‐2 and IL‐3 activities. The IFN level was higher in early culture sup (2–24 hr) than in later culture sup (48–72 hr). Proliferation was greatly increased by anti‐IFN (α + β) serum in the spleen cells cultured in the earlier sup. This antiserum increased the spleen cell proliferation only slightly in the later culture sup. This suggests that nonparenchymal liver cells produce two factors, one having a suppressor, and the other an enhancer action, with IFN being one of the suppressor factors. With culture time, DNA synthesis of spleen cells increased and IL‐2 and IL‐3 activities were generated in the culture sup. Cells proliferated during culture were found to be morphologically lymphocytes, granulocytes, and macrophages. The mechanisms by which nonparenchymal liver cells regulate the hematolymphoid system are discussed based on our observations.

ISSN:0741-5400    

DOI:10.1002/jlb.50.4.402

出版商:Wiley    

年代:1991

数据来源: WILEY