摘要:

AbstractFunctional and biochemical techniques were used to further characterize heterogeneity between rat Kupffer cells and peritoneal macrophages. Both macrophage cell types were found to phagocytize antibody coated sheep red blood cells in a time‐dependent manner. However, Kupffer cells were two to three times more phagocytic than were peritoneal macrophages. In contrast, the peritoneal cells released significantly more superoxide anion in response to the complement cleavage product, C5a and the phorbol ester tumor promoter, 12‐0‐tetradecanoyl‐phorbol‐13‐acetate, and produced more hydrogen peroxide than did the liver macrophages. Both cell types responded chemotactically to C5a. These results suggest that macrophages may develop specialized functions depending on the needs of their local environment. Using one and two dimensional SDS‐polyacrylamide gel electrophoresis, we also compared the production of newly synthesized proteins by Kupffer cells and peritoneal macrophages. In general, the macrophages were found to produce similar types and numbers of proteins with some exceptions. These included proteins that were unique to peritoneal macrophages and other proteins observed only in Kupffer cells. The production of these proteins in liver macrophages did not appear to correlate with levels of functional activation, but may be more related to the tissue origin of the cells.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.71

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractA simple and improved procedure is described for the isolation of human eosinophils from normal individuals with about 2% eosinophils in their peripheral blood. This method comprises a preincubation of a mixed granulocyte preparation with 10 nM fMLP for 10 min at 37° followed by a one‐step density centrifugation on isotonic Percoll. The recovery of eosinophils is 49 ± 4% at 89 ± 4% purity. Because of the relatively high rate of recovery, it is now possible to isolate eosinophils from blood samples as small as 20 ml.Because treatment with fMLP may alter the functional activity of the eosinophils, the following metabolic functions were tested: changes in cytosolic free Ca2+, oxygen consumption, chemiluminescence, chemotaxis, and leukotriene C4 formation. We found that 10 nM fMLP does not activate eosinophils in these assays, whereas 1 μM fMLP does (with the exception of chemotaxis). Furthermore, pretreatment of eosinophils with 10 nM fMLP did not influence the response to other stimuli in these assays. The usefulness of this method was evaluated by comparing it with three other previously described procedures. In our hands, only the method presented here enabled us to isolate eosinophils from normal individuals with about 2% eosinophils in their peripheral blood.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.79

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractTo test whether or not sialodacryoadenitis virus (SDAV) infection in rats affects pulmonary macrophage function, we intranasally inoculated pathogen‐free F344 rats with SDAV and collected alveolar and interstitial macrophages 5 d later. We assessed Fc receptor‐mediated attachment and phagocytosis by phase‐contrast microscopic examination of monolayers of alveolar and interstitial macrophages incubated with zymosan, nonopsonized sheep erythrocytes, or erythrocytes opsonized with rabbit antisheeperythrocyte IgG. Alveolar macrophages from virus‐infected rats had significantly (P≤ .05) lower indices of attachment and phagocytosis of opsonized erythrocytes than control macrophages, but there was no difference in attachment of zymosan particles. Interstitial macrophages were not affected. Alveolar macrophages from SDAV‐infected rats produced significantly less interleukin‐1 than those from control rats, as assessed by testing supematants from lipopolysaccharide‐stimulated macrophage cultures for induction of mouse thymocytes to take up tritiated thymidine. Effects of SDAV infection on lung macrophages could increase host susceptibility to other pathogens or complicate studies of respiratory tract immunity.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.87

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractThe effect of platelets and their products on in vitro responses of human neutrophils to f‐met‐leu‐phe (fMLP) and phorbol myristate acetate (PMA) was studied. Platelets produced a concentration‐dependent inhibition of neutrophil superoxide anion (O2‐) generation with maximum inhibition of the response to fMLP approaching 42.2 ± 5.4% and that to PMA approaching 85.9 ± 3.4%. Supernates of thrombin‐activated platelets produced maximal inhibition of neutrophil O2‐generation in response to fMLP (66.3 ± 4.2%) at a ratio of 10 platelet equivalents (PEQ) to 1 neutrophil but failed to affect the cells' response to PMA. In contrast, lysate of sonicated platelets Inhibited neutrophil O2‐generation in response to both fMLP (59.6 ± 2.0%) and PMA (90.1 ± 1.3 %). Biochemical characterization of platelet lysate identified at least two stimulus‐specific inhibitory activities which differ in size, thermal stability and enzyme sensitivity. Platelet lysate also maximally inhibited neutrophil degranulation and chemotaxis at a ratio of 20 PEQ to 1 neutrophil. Our findings indicate that physiologically achievable levels of platelets or their products modulate the response of neutrophils in a stimulus‐specific manner and are thereby capable of potentially limiting tissue damage which accompanies neutrophil activation during inflammation.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.93

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe have analyzed the characteristics and cellular sources of the T cell‐derived lymphokines that affect the proliferation and the oxidative metabolic capabilities of the U937 monocytic cell line. Although gamma‐interferon (gIFN) and tumor necrosis factor‐alpha (TNFa) can, in high doses, inhibit the proliferation of U937 cells, the predominant antiproliferative factor produced by activated CD4+ and CD8+ T cells has a MW=45‐55 Kd, is resistant to heat treatment, and is distinct and independent from gIFN, TNFa, and GM‐CSF. The inhibitory effect of this lymphokine on U937 cell growth requires an 18‐24‐hr induction period; thereafter, this growth arrest persists for up to 5 d, even in the absence of the factor. The lymphokine responsible for inducing oxidative metabolic capabilities in U937 cells is also a non‐gIFN, heat‐resistant, 45‐55‐Kd factor secreted by CD4+ and CD8+ cells, and we postulate that this differentiation factor is identical to the factor responsible for inhibiting U937 growth. These data demonstrate the prominent role of T cell‐derived factors distinct from gIFN, TNFa, or GM‐CSF in regulating the growth and functional capabilities of monocyte‐lineage cells. Furthermore, the data suggest it may be appropriate to distinguish monocyte activation, in which cells at a given maturational stage develop a heightened ability to perform a particular function, from changes in the functional repertoire of cells acquired as a consequence of lymphokine‐induced differentiation.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.101

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractReported herein are the results of studies demonstrating the utility of a chemically defined, serum‐free medium designated as AIM‐V (GIBCO) for the long‐term (>2 weeks) cultivation of functionally‐defined human macrophages. The AIM‐V medium is a mixture of HEPES‐buffered Dulbecco's Modified Eagle Medium and Ham's Nutrient Mixture F12 that had been supplemented with purified human albumin, transferrin, insulin, and a proprietary mixture of purified factors. Nonadherent macrophages for serial studies were generated in petri dishes with a Teflon liner that had been seeded with a heterogeneous population of peripheral blood mononuclear cells of healthy human adults. For comparison, cultures were initiated with serum‐free medium AIM‐V and medium supplemented with AB/Rh+serum or freshly collected autologous serum. Viability was defined by trypan blue dye, glass adherence, and phagocytosis of yeast. Macrophages were characterized by light microscopy, cytochemistry, and phenotypic analysis. Ultrastructural morphology was defined by scanning electron microscopy. These studies demonstrate that functionally defined human macrophages can be sustained in long‐term culture with the use of serum‐free medium that has not been augmented with mitogenic stimulants, growth‐promoting lymphokines/monokines, or differentiation‐inducing agents. Serum‐free medium AIM‐V, which has been approved for generating lymphokine, (i.e., interteukin‐2; IL‐2)‐activated killer cells (LAK) for IL‐2/LAK adoptive immunotherapy modalities, may also prove useful for studies defining and isolating regulatory proteins produced by activated monocytes and macrophages and for generating cytolytic macrophages for different antitumor regimens.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.111

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractFemale B6C3F1mice were given intraperitoneal injections of ammonium metavanadate (2.5 or 10 mg V/Kg), ammonium chloride, or sodium phosphate buffer every 3 days for 6 weeks. Resident peritoneal macrophages were harvested, lysed by freeze‐thawing, and the resulting cytolysate was assayed for total protein content and enzyme activities of glutathione reductase, glutathione peroxidase, and glucose‐6‐phosphate dehydrogenase. In addition, peritoneal macrophages were assayed for superoxide production using nitroblue tetrazolium reduction, as well as for intracellular levels of oxidized and reduced glutathione. Exposure of mice to vanadium resulted in a dose‐trend depression in the three macrophage enzyme activities as compared with the controls. Vanadium treatment resulted in a similar decrease in the production of superoxide anion, and an increase in levels of oxidized glutathione; however, the total glutathione pool (reduced plus oxidized forms) was not affected.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.122

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractCalcium is mobilized from intracellular stores during phagocyte activation and appears to be involved in stimulus‐response coupling in these and other cell types. Because the lysosome is a calcium‐sequestering organelle in the neutrophil and abnormal lysosome morphology is associated with defective neutrophil function in Chediak‐Higashi syndrome (CHS), we examined ATP‐dependent calcium uptake in neutrophil lysosomes from the beige mouse model of CHS. We present findings indicating that CHS lysosomes have an enhanced capacity for ATP‐dependent calcium uptake relative to control lysosomes. Kinetic analysis showed differences in Vmaxand in the Kmfor both ATP and calcium, suggesting that both the number of lysosomal calcium uptake pumps and their substrate affinity may be altered in CHS. We conclude that a genetically determined abnormality of a subcellular calcium transport system may contribute to the structural and functional defects of CHS cells.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.130

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractMacrophage differentiation is mediated by the action of a variety of environmental signals, such as cytokines and endotoxin. For in vitro analysis of macrophage differentiation, most studies utilize a basal tissue culture medium supplemented with fetal calf serum (FCS). As the composition of specific components within FCS varies enormously from lot to lot, one can never be certain that the differentiative effects observed in vitro are attributable solely to the exogenous signals provided to the cultures. In this study, primary macrophages were cultured in a basal medium supplemented either with FCS or a compositionally defined supplement, HL‐1™, and a spectrum of differentiative functions (i.e., induction of antiviral activity, Fc receptor‐mediated phagocytosis, Ia antigen expression, and tumoricidal activity) were measured following stimulation with exogenous signals. The results indicate that the use of serum‐free, defined media may provide an important approach to dissect and characterize the differentiative signals which are operative in macrophage activation.

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.136

出版商:Wiley    

年代:1988

数据来源: WILEY

ISSN:0741-5400    

DOI:10.1002/jlb.44.2.143

出版商:Wiley    

年代:1988

数据来源: WILEY