摘要:

AbstractHuman alveolar macrophages and peripheral blood monocytes were obtained from smoking and nonsmoking normal volunteers. The macrophages and monocytes were incubated in vitro with bacterial lipopolysaccaride (LPS). The oxidative metabolic response of these cells was measured by superoxide anion production. Macrophages from smokers were suppressed in their superoxide anion response to LPS activation as compared to macrophages from nonsmokers. Monocytes from smokers and nonsmokers were not different. The cytotoxic properties of these macrophages and monocytes were assessed by an in vitro3H‐thymidine release assay against various allogeneic target cells. Macrophages and monocytes exposed to LPS were rendered tumoricidal. Macrophages from nonsmokers appeared to generate greater cytotoxic activity than macrophages from smokers. Macrophages from both smokers and nonsmokers were cytotoxic for three different tumorigenic cell lines but not for a nontumorigenic cell line. Monocytes from smokers and nonsmokers were not different in cytotoxic activity. We conclude that macrophages from both smokers and nonsmokers can be activated after exposure to LPS; however, macrophages from smokers may be slightly suppressed in their responses.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.313

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractFeline peritoneal cells were collected by lavage with Isotonic saline without the use of irritants or need for euthanasia of the cats. Macrophages were purified by centrifugation on Percoll followed by selective adherence. Although few macrophages could be obtained from an initial lavage, a second lavage performed on the same cat 9‐11 days later yielded six times as many macrophages as the first lavage, providing sufficient numbers of cells for characterization and infection experiments. Macrophages from these subsequent lavages were not more functionally activated in phagocytosis assays than the resident macrophages from the initial lavage, and they were equally susceptible to infection with feline infectious peritonitis virus (FIPV). Infected cultures produced peak titers of 105.0TCID50per ml, and FIPV antigen was detected in a small subset (0.1‐1.0%) of cells by indirect immunofluorescence. The FlPV‐infected cells were identified as macrophages by their characteristic morphology and ability to phagocytize rhodamine‐labeled latex beads. The successful isolation of large numbers of unactivated feline macrophages will permit in vitro studies of feline coronavirus‐macrophage interactions that otherwise would not have been possible. Such studies will undoubtedly provide valuable insights Into the pathogenesis of feline infectious peritonitis, an invariably fatal disease of domestic and exotic cats.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.319

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractPulmonary alveolar macrophages (PAM) and peripheral blood monocytes (PBMO) of the miniature swine can be converted to cytolytically active effector cells by treating with phorbol myristate acetate (PMA) as determined by enhancement of cytotoxicity to various target cells. Kinetics of the PMA‐activated PAM and PBMO in cytotoxicity show that the effective PMA concentration ranges from 10 to 1,000 ng/ml. Induction of cytotoxic macrophages and monocytes occurred as early as 30 min and to their maximum cytotoxicity after 1 hr exposure to PMA and the enhanced cytotoxic activity persisted up to 24 to 40 hr when PMA was removed by washing after 1 hr exposure, but prolonged exposure to PMA for more than 6 hr resulted in a drastic decrease of cytolytic activities suggesting the prolonged exposure to PMA causes macrophages and monocytes to become refractory to PMA stimulation. Target cells displayed varying degrees of cytotoxic sensitivity to the PMA‐activated PAM and PBMO; PRBC, SRBC, and K562 were sensitive, WEHI‐164 and U937 were relatively sensitive, and SB was very resistant to these activated effector cells. The mechanisms of PMA‐induced cytotoxicity could largely be divided into two categories. One was the H2O2mediated killing as shown by complete reduction of cytotoxicity after adding catatase in the assay. The other was the proteases mediated cytolysis, which could be blocked by protease inhibitors, Phenyl methyl sulfonyl fluoride (PMSF), and N‐ α‐p‐tosyl‐L‐lysine chloromethyl ketone (TLCK). H2O2was the only mediator produced in large enough quantities from PBMO to kill target cells, whereas PAM could produce both mediators (H2O2and proteases). PRBC, SRBC, and K562 appeared to be killed by H2O2produced by PAM and PBMO. In contrast, U937 and WEHI‐164 appeared to be killed by proteases in PAM mediated cytolysis but by H2O2in PBMO‐mediated cytolysis. These results suggest that the observed cytolytic mechanisms can be differed by type of target cells as well as the source of mononuclear phagocytes within the individual animal.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.329

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractMerocyanine 540 (MC540) is a fluorescent probe that binds preferentially to membranes with loosely packed lipids. When combined with flow cytometry, it provides a novel methodology for rapidly and quantitatively assessing lipid organization in individual leukocytes. Analysis of cells stained simultaneously with MC540 and 1‐[4‐trimethylam‐moniumphenyl]‐6‐phenyl‐1,3,5‐hexatriene, to normalize for surface area, revealed that all leukocytes in peripheral blood bind equivalent amounts of dye per unit surface area. This result indicates that the lipids of the plasma membranes of all types of circulating cells are organized similarly. Upon activation with appropriate stimuli, lymphocytes, monocytes, and neutrophils all bound increased amounts of dye per unit surface area, indicating a change in lipid organization to a less‐ordered state. Cells stained with MC540 were sorted on the basis of their fluorescence intensity yielding populations homogeneous with respect to lipid organization. Thus not only can MC540 and flow cytometry be combined for analyzing the organization of lipids in individual leukocytes, but sorted populations can be obtained for further biochemical, structural, and functional analyses.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.337

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractPreincubation of human neutrophils with recombinant tumor necrosis factor alpha has previously been shown by us to enhance superoxide production of neutrophils in response to the chemotactic peptide formyl‐methionyl‐leucyl‐phenylalanine, and the phorbol ester, phorbol myristate acetate. In this study, we further investigate the biochemical basis for this enhancement. We found that in neutrophils, TNF by itself does not induce: (1) an influx of sodium, (2) an alteration in activity or translocation of the calcium and phospholipid dependent protein kinase (C‐kinase), or (3) a release of arachidonic acid from preloaded cells. TNF did, however, induce a time‐ and concentration‐dependent increase in the phosphorylation of several neutrophil proteins, with the most dramatic concentration dependent increase in a 64,000 Da protein. Finally, the enhancement of O2production by pretreatment of neutrophils with TNF was found to be independent of a pertussis toxin‐sensitive guanine nucleotide regulatory protein.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.345

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractThe effects of three retinoids, all‐trans‐retinoic acid (RA), 13‐cis‐retinoic acid (cRA), and N‐(4‐hydroxyphenyl) ratinamide (4‐HPR), on macrophage function were evaluated. In vitro, RA, cRA, and 4‐HPR caused a greater than twofold increase in phagocytosis of IgG‐sensitized bovine erthrocytes (IgG‐ORBC) by a mouse macrophage cell line (RAW). Significant increases in phagocytosis were produced by retinoid concentrations as low as 2 x 10‐10M. RA also significantly increased phagocytosis of IgG‐sensitized ORBC by BALB/c peritoneal macrophages In vitro. The ability of RAW macrophages to bind IgG‐ORBC was significantly increased by 10‐6to 10‐14M RA. The potentiation of myogenic responses of spleen cells to Con A and PWM by RA was relatively independent of macrophage function, i.e., splenocytes that were macrophage‐depleted were responsive to the potentiating effects of RA. The effects of retinoids on T‐cell‐dependent B‐cell mitogenesis induced by PWM appeared not to be dependent on their previously reported capacity to alter prostaglandin synthesis. Treatment of spleen cells with 10‐6M indomethacin did not abolish the potentiating effects of RA. However, RA in a dose‐dependent fashion increased IL‐1 activity at the level of the target T‐cell. The greatest potentiation of IL‐1 activity was at 10‐8M RA. These results show that retinoids can modulate macrophage function at two different levels: potentiation of phagocytosis and potentiation of IL‐1 activity at the level of the T‐cell.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.353

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe evaluated the ability of normal human peripheral blood monocytes and polymorphonuclear leucocytes (PMNL) isolated from patients with acquired immunodeficiency syndrome (AIDS) or AIDS‐related conditions (ARC) to migrate toward a chemoattractant. Migration in blind‐well chambers was compared to that under agarose. Chemotaxis results obtained from both assays for PMNL were similar, however there was a difference in the results for monocyte Chemotaxis. PMNL isolated from patients with AIDS, but not ARC, exhibited decreased spontaneous and directed chemotaxis when assessed in blind‐well chambers and under agarose. Spontaneous and directed Chemotaxis in blind‐well chambers of AIDS patients' monocytes was normal. Directed migration of monocytes from ARC patients was greater than that of control, but spontaneous migration was comparable. Under agarose, spontaneous migration was depressed in monocytes of AIDS patients, while migration toward the attractant was depressed in those of ARC patients.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.361

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractResident peritoneal macrophages (Mø) were labeled in situ by intraperitoneal (i.p.) injection of the green fluorescent cell tracking dye PKH‐1. After immunofluorescence staining with Mø specific monoclonal antibodies (Mabs) and phycoerythrin (PE) second antibody, the resident Mø were labeled with both the green dye and red Mab label, while recruited Mø were labeled only with the red Mab tag. These populations were distinguished by two‐color flow cytometry.PKH‐1 labeled resident peritoneal Mø were followed for 1‐49 days in mice that received no further treatment (steady state). Dye labeled Mø were still detectable after 49 days in vivo, although their green fluorescence intensity had decreased steadily over time. The decrease in dye intensity was limited to Mø, as the fluorescence intensity of PKH‐1 labeled peritoneal lymphocytes did not change.Resident Mø populations were clearly separated from recruited Mø by the intensity of their staining with PKH‐1 for up to 28 days. No decrease in the number of resident (dye labeled) peritoneal Mø was observed over 1‐28 days. These data indicate that resident peritoneal Mø were not replaced by recruited blood monocytes in the steady state.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.367

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe investigated the capacity of counterflow‐isolated human monocytes to independently synthesize thromboxane B2(TxB2) and prostaglandin E2(PGE2) when stimulated with bacterial lipopolysaccharide (LPS). Independent metabolism was confirmed by establishing different specific activities (dpm/ng) of TxB2and PGE2released from LPS‐treated cells. For metabolites released during the initial 2‐hr treatment period, the specific activity of PGE2was approximately threefold higher than that of TxB2regardless of labeling with [3H] arachidonic acid (AA) or [14C]AA. Cells that were pulse‐labeled for 2 hr with [3H]AA demonstrated a decreasing PGE2specific activity over 24 hr, whereas the TxB2specific activity remained unchanged. In contrast, cells continuously exposed to [14C]AA demonstrated an increasing TxB2specific activity that approached the level of PGE2by 24 hr. These results suggest the presence of at least 2 cyclooxygenase metabolic compartments in counterflow‐isolated monocytes. Although freshly isolated monocytes have been reported to contain variable numbers of adherent platelets, additional experiments demonstrated that counterflow‐isolated platelets are not capable of releasing elevated levels of TxB2or PGE2when treated with LPS. It is proposed from these findings that at least two subsets of monocytes exist in peripheral blood that can be distinguished on the basis of independent conversion of AA to TxB2and PGE2.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.376

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractSerum‐free cultured macrophages could be stimulated for lucigenin‐dependent chemiluminescence by platelet activating factor (PAF) and phorbol myristate acetate (PMA). Stimulation with PMA resulted in a desensitization against PAF, whereas prestimulation with PAF had no influence on a following response caused by PMA. The PAF analogue, 1‐O‐octadecyl‐2‐O‐methyl‐rac‐glycero‐3‐phosphocholine (Et‐18‐OCH3), did not induce chemiluminescence by itself and desensitized the cells against PAF, like substimulating concentrations of PAF. PAF and PMA responsiveness was rapidly modulated in a similar manner during adherence of the cells to polystyrene tubes. At higher concentrations, Et‐18‐OCH3as well as lysophosphatidylcholines potentiated PMA‐induced chemiluminescence. The PAF analogue was most effective. Although PMA‐induced chemiluminescence was stimulated at least 5‐fold by Et‐18‐OCH3, this compound Increased the PMA‐induced activation of protein kinase C only 1.39‐fold. The priming effect of Et‐18‐OCH3was not reduced in the absence of extracellular Ca2+and after cell membrane depolarisation.

ISSN:0741-5400    

DOI:10.1002/jlb.44.5.385

出版商:Wiley    

年代:1988

数据来源: WILEY