摘要:

AbstractSeveral in vitro functions of neonatal neutrophils (N‐PMN) have been reported to be deficient and may be functionally related to the increased susceptibility of the newborn to infection. To evaluate an in vitro event corresponding to one of the early steps in the sequence of inflammation, we used zymosan‐activated plasma as a source of activated complement fragments (Cf) and measured adherence of normal and Cf‐stimulated bovine N‐PMN to columns of Sephadex G‐25. Adherence of control N‐PMN and adult PMN (A‐PMN) was comparable. When N‐PMN and A‐PMN were stimulated with a subaggregating dose of Cf, both responded with similar increases in adhesiveness. The stimulatory effect of Cf on N‐PMN adhesiveness could be inhibited by pre‐incubation of the N‐PMN with either steroidal (0.05 mM dexamethasone) or non‐steroidal (32 mM phenylbutazone) anti‐inflammatory drugs. Ultrastructural observations correlated well with the results of the adhesiveness assays, and morphometric evaluation revealed an increase in the sectional circumference of Cf‐stimulated N‐PMN. Control cells were round with few short cytoplasmic projections, whereas Cf‐stimulated cells exhibited marked shape irregularity, polarity, and prominent organelle‐free lamellipodia development. There was a highly significant (P<0.001) increase in the measured circumference of Cf‐stimulated cells. Thus, N‐PMN were highly responsive to Cf stimulation, developed morphologic and functional changes indistinguishable from Cf‐stimulated A‐PMN, and were sensitive to pharmacologic inhibition.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.465

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractTwo activators of calcium and phospholipid dependent protein kinase (protein kinase C), the tumor promoter, 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) and the synthetic diacylglycerol, 1‐oleoyl‐2‐acetylglycerol (OAG), were compared as chemotactic agents for mouse peritoneal macrophages. Both of these compounds were found to induce Chemotaxis in the macrophages to a similar extent in a time and dose dependent manner. Induction of Chemotaxis was observed in the concentration range of 10‐100 nM for TPA and 25‐250 μM for OAG. Two structurally related syntheticsn1,2‐diacylglycerols, 1,2‐dioctanoylglycerol (diC8) and 1,2‐didecanoylglycerol (diC10), were also found to be chemotactic for macrophages, while monoacylglycerol (2‐monoolein) was inactive. Of the diacylglycerols, OAG was found to be the most active followed by dlC8 and diC10. In contrast to TPA, the synthetic diacylglycerols had no effect on superoxide anion release by the cells, suggesting that the mechanism of superoxide anion release by TPA in macrophages is distinct from Chemotaxis. Phorbol‐12,13‐diacetate, a biologically inactive phorbol ester analog that inhibits the binding of TPA to its cellular receptors, inhibited macrophage Chemotaxis induced by TPA, the synthetic diacylglycerols, and the complement fragment, C5a. Taken together, our results suggest that Chemotaxis in macrophages may be mediated by activation of protein kinase C.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.474

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractA computer‐assisted approach has been designed to analyze and quantitate polymorphonuclear leukocyte (PMN) Chemotaxis. This approach involves a rapid, objective, and semiautomated (user‐directed) image‐analysis system that is video‐ and microscope‐based. The entire system consists of a microvideo set‐up that is put on line with a Digital DEC‐LSI‐11/73 microcomputer, interfaced with a Datacube analog‐digital/digital‐analog converter. Video signals of PMN movement are digitized by the system at a resolution of 240 pixels vertically by 320 pixels horizontally (at 256 gray levels) and stored in a 76,800‐byte frame buffer. The digitized data are stored for later use or utilized immediately for image segmentation, image display, movement, and morphometric computations for each PMN in a maximum phase field (at 645 × high dry) of 50 PMNs at 10‐second intervals. The digitized data are used for computation of cell perimeter, surface area, optical density, contour‐ratio, position, speed, and direction of locomotion with the utilization of micro‐image‐analysis programs written in FORTRAN and MACRO assembly language, with the computer operating under RT‐11/TSX +. The reliability, objectivity, and reproducibility of measurements made with this quantitative approach have been tested by comparing with manual‐tracing measurements of PMN movement. A correlation factor of 0.99 has been obtained. However, the quantitative‐microscopic approach is much faster, more objective, less tedious, and much easier to operate than the conventional manual‐tracing method.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.481

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractHL‐60 promyelocytic cells acquire the surface expression of the Mo3e antigenic determinant after exposure to PMA or compounds that raise intracellular concentrations of cyclic AMP (dibutyryl cyclic AMP or a combination of cholera toxin and IBMX). The expression of Mo3e by these stimulated HL‐60 cells coincides with the development of features of monocyte‐macrophage differentiation (characteristic morphology, nonspecific esterase activity, and respiratory burst activity). During in vitro monocyte‐macrophage differentiation, HL‐60 cells become responsive to migration inhibitory factor (MIF); the MIF responsiveness of differentiated HL‐60 cells is blocked by anti‐Mo3e monoclonal antibody. These findings further support the relationship between the expression of Mo3e and the cellular response to MIF.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.492

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe mouse macrophage (MΦ) cell line IC‐21 preferentially ingests a subpopulation of homologous red blood cells (MRBC) from normal mice. This subpopulation presumably bears the so‐called transfusion lesion, a consequence of damage acquired during the drawing and processing of blood. To determine if all damaged MRBC were recognized by a common receptor site on IC‐21 MΦ, we prepared suspensions of MRBC damaged in vitro by treatment with tannic acid and compared the phagocytic uptake of these cells with those bearing the transfusion lesion. Trypsin treatment of IC‐21 MΦ rendered them unable to recognize MRBC bearing the transfusion lesion; but it had no effect on the uptake of tannic acid‐damaged MRBC, showing that IC‐21 MΦ have separate recognition sites for these two populations of damaged MRBC.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.500

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractHuman large granular lymphocytes (LGL), which account for virtually all natural killer activity, and T cells were separated from low and high density fractions of Percoll gradients, respectively. We were unable to detect interleukin 2 receptors (IL 2R) on fresh LGL and T cells using flow cytometric analysis with anti‐Tac monoclonal antibody or radiolabeled probes with [125l]anti‐Tac. IL 2R messenger ribonucleic acid was not observed in fresh LGL or T cells. Although Phytohemagglutinin induced the expression of IL 2R on purified LGL and T cells, recombinant IL 2 (rIL 2) alone induced IL 2R messenger ribonucleic acid, IL 2R, and proliferation of LGL but not of T cells. The high level of cytotoxicity of cultured LGL against K562 cells was directly dependent on rIL 2. When T cells were costimulated by Phytohemagglutinin and rIL 2 for 3 days, only very low levels of cytotoxicity were generated. Proliferation and cytotoxicity against K562 cells inhibited the culture of LGL in the presence of anti‐Tac antibody for 3 days. LGL began to grow more rapidly after 5 days in culture with rIL 2 alone. When rIL 2 were removed from growing LGL for 1 day, proliferation completely stopped, and cytotoxicity was no longer detected. These data indicate that rIL 2 induces IL 2R expression in fresh LGL at the transcriptional level, promoting growth and enhancing cytotoxicity. More importantly, the presence of rIL 2 is necessary and sufficient to induce proliferation of resting LGL and to maintain the growth of LGL with potent lytic activity.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.505

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractWe have previously reported that human peripheral blood polymorphonuclear leukocytes can be separated into at least six volume dependent fractions using counterflow centrifugal elutriation (CCE). Larger volume was shown to correlate with increased superoxide release with either the chemotactic peptide F‐met‐leu‐phe (FMLP) or the tumor promotor phorbol myristate acetate (PMA). To help explain these findings we have examined volume dependent PMN fractions (VDPF) for PMN functions felt to be correlated to PMN age, ie, alkaline phosphatase activity, phagocytosis of opsonized particles, adherence to plastic, and directed movement to FMLP and zymosan activated serum. PMN alkaline phosphatase activity was found to be low in the smallest PMNs, increasing in the PMNs of intermediate size, and then decreasing again in the largest PMN functions. A similar relationship was noted for PMN phagocytic activity. No significant difference among VDPF in adherence to plastic or Chemotaxis was found. Furthermore, no difference among VDPF was seen for receptor number or binding affinity of the ligands FMLP or phorbol dibutyrate. Total cellular activity of the NADPH dependent oxidase was, however, 200% greater in the largest compared to the smallest VDPF. Because of the lack of consistent correlation between “age” related PMN function and PMN volume, it is unlikely that PMN volume represents a manifestation of PMN age. It is likely that the increased oxidase activity seen among larger VDPF accounts for the more rapid oxidative burst previously noted.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.518

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractProtein synthetic patterns of murine peritoneal macrophages were analyzed by two‐dimensional Polyacrylamide gel electrophoresis (2D PAGE) of35S methionine‐labeled proteins. While the protein synthetic patterns exhibited by resident, inflammatory, and activated macrophages had numerous common features that distinguished them from the other normal non‐macrophage cell types examined, unique proteins also characterized each macrophage population from the others. The accumulation by resident macrophages of proteins 23, 25, and 37 distinguished them from elicited cells, as did the former's more abundant synthesis of proteins 54 and 52. The protein synthetic patterns of inflammatory thioglycollate‐ and proteose peptone‐elicited macrophages were strikingly similar, save for the former's greater levels of accumulation of proteins 14 and 28, and the letter's more pronounced expression of p23.5. Peritoneal macrophages elicited by treatment with heat‐killedPropionibacteriumacnes, the live, attenuated Mycobacteriumbovisstrain BCG,Listeria monocytogenes, and the protozoan flagellateTrypanosoma rhodesiense, all exhibited tumoricidal activity in 16‐h or 72‐h functional assays. They shared a common protein synthetic profile that differentiated them from the synthetic patterns characteristic of the non‐tumoricidal resident and inflammatory macrophages. These tumoricidal macrophages were unique in synthesizing a protein(s) of approximate molecular weight 26,000 daltons. A time‐course study employingP. acnes‐activated peritoneal macrophages indicated that p26 accumulation decayed with tumoricidal capacity as a function of time in culture, although no direct correlation between lytic activity and p26 expression could be definitively established. Peritoneal macrophages elicited with proteose peptone were not directly tumoricidal but were rendered so upon in vitro treatment with nanogram amounts of bacterial lipopolysaccharide. The accumulation of low levels of p26 by the newly explanted proteose peptone‐elicited macrophages suggests the possibility that this protein characterizes macrophage populations primed as well as triggered for tumoricidal activity.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.527

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractReceptor‐mediated ingestion was examined in macrophages derived from a canine model of the adult respiratory distress syndrome (ARDS). The results showed that Fc‐mediated ingestion by alveolar macrophages (AM) and macrophages from lung parenchyma (PM) was significantly diminished when compared with their respective controls. Pulsing all the experimental groups with lipopolysaccharide (LPS) for 1 hr in vitro failed to either enhance the response or return the activity to levels achieved by control cells. In parallel studies, an analysis of C3b‐mediated ingestion showed that both the experimental AM and PM performed this function only at a magnitude equal to the control cells. Similar responses were observed when an LPS pulse was performed. Although there was a reduction in Fc‐mediated ingestion and an apparent restraint of the C3b‐mediated ingestion, both AM and PM expressed a significantly enhanced ability to spread. These results suggested that the canine model of ARDS alters at least one select macrophage function that may be important to subsequently protect the host. Such disturbances in the cellular immune response may contribute to the progression of infection and lung pathology associated with this disease process.

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.539

出版商:Wiley    

年代:1987

数据来源: WILEY

ISSN:0741-5400    

DOI:10.1002/jlb.41.6.i

出版商:Wiley    

年代:1987

数据来源: WILEY