摘要:

Three macrophage cell lines from bone marrow cells of C3H/HeN mice were isolated by successive transfer of the cells in culture with L‐cell‐conditioned medium (LCM) or WEHI‐3 cell‐conditioned medium (WEHI‐3CM). These cell lines, which express Fc receptors, are involved in Fc‐mediated phagocytosis and possess nonspecific esterase activity. Two (BDM‐1 and BDM‐2) of three cell lines show dependency for growth on either macrophage colony‐stimulating factor (M‐CSF) (CSF‐1) or granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and do not respond to interleukin 3 (IL‐3). The third clone (BDM‐3) proliferates in response to IL‐3 as well as to GM‐CSF and weakly responds to M‐CSF and to interleukin 4 (IL‐4). GM‐CSF, in combination with the suboptimal concentration of M‐CSF, acted synergistically on the proliferation of BDM‐1 cells. The tumor‐promoting phorbol diester, 12‐o‐tetradecanoyl‐phorbol‐13‐acetate (TPA) also acted synergistically with the three CSFs (IL‐3, GM‐CSF, and M‐CSF) to stimulate the proliferation of BDM‐1 cells. The synergistic effect was observed when cells were pretreated with TPA and subsequently stimulated with IL‐3. The calcium ionophore A23187 enhanced the proliferation of BDM‐1 cells costimulated with TPA and IL‐3. These factor‐dependent macrophage cell lines should be useful for studying signal transduction mechanisms in the regulation of cell growth.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.465

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

We have previously reported that natural killer (NK) lineage cells are progressively inactivated during tumor development by prostaglandin E2(PGE2) secreted by host macrophages; that this facilitates spontaneous tumor metastases, which can be prevented by chronic indomethacin therapy (CIT); and that CIT combined with multiple rounds of interleukin 2 (IL‐2) can cure experimental metastases and activate all killer lineage cells in situ including NK cells, lymphokine‐activated killer (LAK) cells, and tumoricidal macrophages. The present study tested whether PGE2secreted by tumor‐bearing host macrophages exerts pansuppressor effects against the activation of T cells, NK cells, LAK cells, and tumoricidal macrophages from normal splenic cell populations. Macrophages isolated from CBA mice bearing 21‐day intraperitoneal Ehrlich ascites tumors (EAT) or C3H/HeJ mice bearing 21‐day subcutaneous T58 mammary adenocarcinomas were added (± 10−5M indomethacin, or a monoclonal anti‐PGE2ab) to syngeneic splenic lymphocytes to examine the effects on 1) polyclonal activation (3‐d3H‐thymidine [3H‐TdR] uptake) with concanavalin A (Con A); 2) one‐way (CBA α BALB/C or C3H α BALBC) MLR (5‐d3H‐TdR uptake) and subsequent CTL generation (tested against51Cr‐labeled Con A blasts of the stimulator phenotype); 3) NK activity (after 24‐h co‐culture) against YAC‐1 targets; 4) generation of LAK cell activity (in the presence of 200 or 2,000 units recombinant IL‐2 for 3 or 5 days), tested against NK‐sensitive and NK‐resistant targets. Similar effects were also noted on the generation of tumoricidal activity in normal splenic macrophages cultured for 3 days in the presence of LPS. Normal splenic macrophages added under the same conditions served as controls. Effects of pure PGE2or PGF2(10−6M) were also examined on these activation events. Results revealed that tumor‐host‐derived macrophages (but not normal macrophages) markedly suppressed all these activation events and this suppression was abrogated nearly totally by indomethacin and totally by anti‐PGE2ab, indicating its mediation by PGE2. This finding ran parallel with high levels of PGE2production by tumor‐host‐derived but not normal splenic macrophages. Pure PGE2but not PGF2αmimicked these suppressor effects. While tumoricidal activity was generated in normal macrophages in the presence of LPS, IL‐2, or IFN‐γ or their various combinations (which led to further augmentation), these agents required the presence of indomethacin to generate significant killer activity in tumor‐host‐derived macrophages. These results indicate that macrophages in tumor‐bearing hosts exert pansuppressor effects against the activation of all tumoricidal effector lineages inclusive of T, NK, LAK cells, and killer macrophages.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.474

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

This study examines the effect of epithelial permeability on 1) the passage of the chemoattractant tritiated formyl methionyl‐leucyl‐phenylalanine (3H‐fMLP; m.w. 437) and 2) the migration of leukocytes. In addition, it also compares the kinetics of neutrophil and monocyte transepithelial migration. As we had demonstrated with neutrophils (Milks et al.:Journal of Cell Biology96:1241, 1983), when the permeability of the epithelium decreased, the accumulation of3H‐fMLP and the emigration of monocytes also decreased. When neutrophils and monocytes traversed epithelia with similar permeability, neutrophil accumulation was at least ninefold greater than that of monocytes at 30, 60, and 90 min. During the same time intervals, the number of neutrophil invasion sites/mm epithelium exceeded the number of monocyte invasion sites by at least fivefold, with approximately twice as many neutrophils as monocytes traversing each invasion site. These studies demonstrate that epithelial permeability affects the passage of the chemoattractant and the emigration of leukocytes. In addition, the enhanced ability with which neutrophils traverse occluding junctions compared to monocytes helps to explain, at least in part, the more rapid accumulation of neutrophils at inflammatory lesions.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.485

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

The development of macrophages from blood monocytes from 22 patients with untreated Hodgkin's disease and 20 normal subjects has been studied at intervals over a 6‐day period of suspension culture in the presence of autologous serum and lymphocytes. Morphometric measurements were made on electron micrographs and the results subjected to multivariate and univariate analyses of variance. The cells from the Hodgkin's group showed highly significant differences from normal. The whole‐cell volume, surface area, and cell membrane excess, as well as the mitochondrial volume and surface area, showed smaller increases over the period of culture, and the normal increase in mitochondrial profile numbers was not seen. It would appear that although the patients' monocytes transformed into macrophages in culture, their development was seriously deranged.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.493

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

Stimulation of the respiratory burst of murine bronchoalveolar macrophages obtained by lung lavage was studied using four different stimuli and different assay conditions. One soluble stimulus, phorbol myristate acetate (PMA), two intracellular particles, zymosan andBlastomyces dermatitidisconida, and one extracellular particle,B. dermatitidisyeast, were incubated with either freshly obtained macrophages in suspension or 2‐ and 48‐hour macrophage monolayers. Suspension cultures were incubated with stimuli for 90 minutes and monolayers for 10 minutes before O2−was assayed. PMA did not elicit O2−production in macrophage suspensions or 2‐hour macrophage monolayers, but 48‐hour macrophage monolayers exhibited a 13‐fold increase above control values (P= .0001). On the other hand, zymosan elicited an increase in O2−production in both suspensions and monolayers, although monolayers incubated for 48 hours produced almost fourfold more O2−than the other systems (P= .025). Opsonization had no effect on the ability of zymosan to elicit respiratory burstB. dermatitidisconidia resulted in a two‐ to threefold increase in O2−production in macrophage suspensions and a five‐ to eightfold increase in 48‐hour monolayers, representing significantly less respiratory burst stimulation than with either zymosan or PMA. Similarly,B. dermatitidisyeasts demonstrated similar submaximal stimulation of O2−, 3–4 times that over control values, and again this was less than zymosan and PMA. We conclude that 1) freshly obtained murine bronchoalveolar macrophages do not respond to PMA with an increase in O2−production, but that responsiveness is evident after 48 hours of incubation in monolayers; 2)B. dermatitidisconidia and yeasts do not stimulate respiratory burst activity to the same degree as zymosan or PMA; and 3) opsonization of zymosan is not necessary for stimulation of the murine bronchoalveolar macrophage oxidative burst, confirming previous data that functional complement receptors are not present on these cells.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.500

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

The present study was undertaken to explore the possible causes of ultraviolet radiation (UVR)‐induced disappearance of ATPase‐positive, epidermal Langerhans cells (LC).Monodelphis domesticawas used because it has the capacity for photoreactivation of UVR‐induced pyrimidine dimers in epidermal DNA. Single, 330 J/m2(ears) or 500 J/m2(back) UVR exposures (FS‐40 sunlamps) reduced the numbers of ATPase‐positive epidermal LC inM. domesticaears to ~15% of those in unirradiated ears and ~37% of those in unirradiated dorsal skin. Immediate 90‐minute exposures to photoreactivating light (PRL, 320–400 nm) post‐UVR reversed the effects of the UVR, resulting in ATPase‐positive LC numbers not being significantly different from controls. Exposure to PRL immediately proceeding UVR did not prevent ATPase‐positive LC disappearance. The photoreactivation of UVR‐induced ATPase‐positive LC disappearance indicates that DNA damage (pyrimidine dimers) is involved in the loss of ATPase‐positive LC.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.508

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

The ability of MulFN‐β and MulFN‐γ to potentiate the development of tumoricidal activity in proteose peptone‐elicited murine peritoneal macrophages was investigated. Macrophages were stimulated with increasing concentrations of either MulFN‐β or MulFN‐γ, alone and in combination, in the presence of 1 ng/ml lipopolysaccharide (LPS). The priming activities attributable to the interferons (IFNs) were quantified using the dose‐response curves obtained for these samples. The priming activity observed for mixtures of MulFN‐β and MulFN‐γ was greater than that expected if MulFN‐β and MulFN‐γ had acted in an additive manner. Isobologram analysis of data obtained when macrophages were stimulated with combinations of IFNs demonstrated that MulFN‐β and MulFN‐γ acted synergistically to prime macrophages for tumor cell killing. The greatest degree of synergy was observed when macrophages were stimulated with suboptimal and nearly equivalent concentrations of each class of IFN. Further studies demonstrated that macrophages stimulated with combinations of MulFN‐β and MulFN‐γ were more sensitive to the trigger signal provided by LPS than were cells primed with either IFN alone. Thus, the synergistic effects observed were quantitative in nature in that macrophages perceived combinations of MulFN‐β and MulFN‐γ as having higher priming activities than expected.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.514

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

Gangliosides have been prepared from resting murine thymocytes and splenic T cells. Profoundly different two‐dimensional thin layer chromatography (2D TLC) patterns were observed between these two cell types. Thymocytes contained 28–30 discrete gangliosides of which eight represented major gangliosides. Splenic T lymphocytes from both strains had much simpler patterns, with six to seven major gangliosides and 12–13 minor gangliosides. Computerized analysis of the thymocyte ganglioside patterns between LPS‐responder C3H/HeN mice and lipopolysaccharide (LPS)‐hyporesponsive C3H/HeJ mice revealed no significant difference in the major gangliosides. However, with splenic T cell gangliosides, there is a striking difference in the relative proportion of three homologous gangliosides between the two strains. Consistent with previous observations on macrophage gangliosides, the ratio of N‐acetylneuraminic acid‐containing ganglioside to N‐glycolylneuraminic acid‐containing ganglioside was higher in both thymocytes and T‐cells from the LPS‐responder strain. These results show that sialic acid‐containing glycolipids from thymocytes and T lymphocytes between endotoxin responder and hyporesponder strains manifest small but significant changes. These differences are present in unstimulated cell populations and may represent a manifestation of the Lps gene.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.521

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

The amyloid P‐component (AP), a ubiquitous component of amyloid fibrils, is also a plasma protein and a connective tissue constitutent. Its proximity to elastin, in particular, suggested that AP might serve to protect elastic tissue from hydrolytic enzymes. The inhibition of pancreatic elastase by AP has been reported. In the present study, the effects of AP on human neutrophil elastase andPseudomonaselastase were investigated, and AP was shown to interfere with the cleavage of soluble elastin. As indicated by Michaelis‐Menten analysis, AP is acting as a noncompetetlve inhibitor. C‐reactive protein, which is structurally similar to AP, had no effect on either elastase. AP was also found to inhibit the degradation of secondary amyloid fibrils by neutrophil elastase when these structures were first partially purified and then reexposed to AP.AP's ability to inhibit elastase was compared with alpha‐1 antitrypsin in the presence and absence of oxidizing agents. These substances, which are released by inflammatory cells, are known to abrogate alpha‐1 antitrypsin's anti‐protease capacity. This contributes to elevated levels of free proteases in the circulation and extravascular spaces during severe inflammation. AP is not susceptible to oxidation and remains a functional inhibitor under these conditions. The potential role of AP as an elastase inhibitor is discussed.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.529

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

The adhesive glycoprotein Mac‐1 (CD11b/CD18) of the CD11/CD18 complex contributes to multiple neutrophil inflammatory functions. Activation of neutrophils by chemotactic stimuli results in a rapid, protein synthesis‐independent increase in surface Mac‐1 derived from incompletely defined intracellular compartments. Therefore, we developed a novel quantitative lectin immunoblot technique to define intracellular pools of Mac‐1 in subcellular neutrophil fractions resolved on discontinuous Percoll gradients. In cavitates of unstimulated neutrophils, 30% and 26% of total Mac‐1 was identified in β [1.10 gm/ml; vitamin B12binding protein (vit B12B.P.)‐rich] or pre‐γ (1.07 gm/ml; vit B12B.P.‐poor) granular fractions, respectively, whereas 24% was associated with the plasma membrane‐rich γ (1.06 gm/ml) fractions. N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP) stimulation (10−8M, 15 min, 37°C) significantly diminished Mac‐1 in pre‐γ (‐18% of total,P<0.05) but not β fractions (+6% of total). Under these conditions, the content of Mac‐1 in γ fractions increased 13% in association with four‐ to eightfold increase in surface Mac‐1 expression (OKM‐1 binding). These findings suggest that chemotactic stimuli increase plasma membrane and/or surface Mac‐1 on human neutrophils by mobilizing a novel intracellular granule pool.

ISSN:0741-5400    

DOI:10.1002/jlb.44.6.535

出版商:Wiley    

年代:1988

数据来源: WILEY