摘要:

AbstractA novel method for labeling resident peritoneal macrophages (MO) by injection of a dye into the peritoneal cavity is described. The dye, which fluoresces green, is selectively taken up by the resident Mø. Dye labeled cells can be further characterized by labeling of cell surface antigens with monoclonal antibodies (Mabs) and phycoerythrin conjugated second antibody. After such labeling with the Mabs F4/80 or Mac 1 the resident Mø were labeled by both the green dye and the red Mab markers, while recruited Mø or neutrophils were labeled with just the red Mab; the two populations of cells were readily distinguished by two‐color flow cytometry. This technique enabled identification of resident and recruited Mø in each animal without the use of radioisotopes, irradiation, or bone marrow ablation. Sufficient numbers of cells can be analyzed from each animal so that Individual animals could be evaluated.We found no adverse effects of this labeling technique on expression of cell surface antigens or Mø mediated cytotoxicity. We did find evidence that the i.p. injection induced a mild inflammation in the peritoneal cavities of animals injected with either the dye or the balanced salt solution vehicle. Examination of the intracellular staining pattern indicated that the label rapidly sequestered in the cytoplasm of the Mø, possibly in the lysosomes. Dye solubility studies showed that the dye was partially soluble at the concentration used for in vivo labeling. We hypothesize that the Mø labeling occurred by a combination of phagocytosis of dye aggregates and endocytosis of labeled plasma membrane.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.387

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractIn previous studies on the respiratory burst of neutrophils, it was observed that the small, water‐soluble quinone, duroquinone, activated the respiratory burst in rat peritoneal neutrophils but not in human blood neutrophils. Here we report studies which indicate that a selective diminution of certain granule enzymes, gelatinase and azocaseinase but not elastase, occurs during elicitation in the rat. Neutrophils circulating in rat blood have about twice the amount of gelatinase and azocaseinase activity as compared to elicited peritoneal cells, and the respiratory burst of circulating cells is stimulated to only about 20% by duroquinone. The significance of these results is considered in terms of activation of the neutrophil respiratory burst.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.398

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractPorcine intravascular macrophages were isolated by perfusion of the pulmonary vasculature with 0.1% collagenase solution. The isolated cells formed intercellular adhesion plaques with endothelial cells when incubated with porcine pulmonary artery, aorta, and corneal cups. Intercellular adhesion plaques were focal junctionlike membrane specializations consisting of paired submembranous amorphous densities subjacent to 15‐20 nm gaps between parallel apposing cell membranes. The intermembranous space was filled with moderately electron dense, finely granular material. Adhesion plaques formed in 4‐8 hours and resembled the adhesion plaques formed between pulmonary intravascular macrophages and endothelium in vivo. Alveolar macrophages and peripheral blood monocytes did not form intercellular adhesion plaques with endothelial cells. Intravascular macrophages had histologic and ultrastructural features of macrophages, were alpha naphthyl butyrate esterase positive, adhered to plastic coverslips after 1 hour of incubation, and were smaller than alveolar macrophages and endothelial cells. The formation of intercellular adhesion plaques in vivo and in vitro by cells with morphologic and histochemical features of macrophages distinguishes intravascular macrophages from monocytes and alveolar macrophages.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.403

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractThe synthetic diacylglycerols (DG), sn‐1,2‐dihexanoylglycerol (diC6), sn‐1,2‐dioctanoyl‐glycerol (diC8), and 1‐oleoyl‐2‐acetylglycerol (OAG) stimulated the release of granule constituents from and superoxide anion (Of) generation by human polymorphonuclear neutrophils (PMN). The DGs did not induce a rise in the cytosolic‐free calcium concentration ([Ca2+]i), as monitored by the fluorescence of PMNs loaded with the fluorescent CA2+indicator, Fura‐2. DiC6, diC8, and OAG inhibited PMN degranulation elicited with the receptor‐specific ligands, N‐formyl‐methionyl‐leucyl‐phenylalanine (FMLP), acetylsn‐glyceryl‐3‐phosphorylcholine (AGEPC), and 5(S),12(R)‐dihydroxy‐6,14‐cis‐8,10‐trans‐eicosatetraenoic acid (LTB4) and the calcium ionophore, A23187. In contrast to their inhibitory effects on granule exocytosis, diC6, diC8and OAG enhanced FMLP‐, AGEPC‐, LTB4and A23187‐stimulated O2‐production. Activation of the respiratory burst with phorbol 12‐myristate 13‐acetate (PMA) was unaffected by the DGs. DiC8inhibited the rise in [Ca2+]ielicited with FMLP, LTB4, and AGEPC; this effect, as well as the DG‐mediated suppression of degranulation, could be reversed with the protein kinase C (PKC) inhibitor, 1‐(‐5‐isoquinolinesulfonyl)‐2‐methylpiperazine hydrochloride (H‐7). These data indicate that in addition to possessing the intrinsic capacity to activate PMNs, DG may function in a PKC‐mediated autoregulatory mode to influence PMN activation in a response‐specific manner by affecting certain components of receptor‐coupled and receptor‐independent signal transduction systems in a stimulus‐specific manner.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.411

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractBinding of a potent chemotactic formyl tetrapeptide, formylmethionyl‐leucyl‐phenyl‐alanyl‐phenylalanine (fMet‐Leu‐Phe‐Phe), to the formyl peptide receptors on the rabbit neutrophil was assessed by two approaches. A tritiated preparation of fMet‐Leu‐Phe‐Phe was used for direct binding studies, whereas indirect studies comprised an assessment of the ability of the formyl tetrapeptide to competitively inhibit the binding of35S‐labeled formylmethionyl‐leucyl‐phenylalanine. These two approaches yielded analogous results. The formyl tetrapeptide fMet‐Leu‐Phe‐Phe showed rapid and saturable binding to the same chemotactic receptors as the less potent formyl tripeptides with which it was compared. Its equilibrium‐binding pattern, however, was different: fMet‐Leu‐Phe‐Phe showed a homogeneous binding pattern, in contrast to the heterogeneity seen with the less potent compounds. The relative potencies for high‐affinity binding of the two standard formyl tripeptides and fMet‐Leu‐Phe‐Phe correlated well with their relative potencies for stimulating the biological response of degranulation; the relative potencies for low‐affinity binding correlated less well.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.420

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractTumor necrosis factor (TNF) was found in the lung lavage fluids ofLegionella pneumophila‐infected mice within 24 hr of intratracheal (i.t.) inoculation. Since this cytokine has been reported to activate polymorphonuclear leukocyte (PMN) function, the effect of TNF on the in vitro bactericidal capacity of PMN‐enriched cultures was determined. Murine thioglycollate‐elicited PMN which were treated with recombinant human TNF demonstrated augmented killing ofL. pneumophilabacteria in vitro. Furthermore, treatment of PMN suspensions with cytokine‐containing lung lavage fluid was found to enhance the bactericidal activity of PMN. The addition of anti‐cachectin/TNF antibodies partially abrogated the stimulatory effects of the lavage fluid, suggesting that in vivo activation of PMN during the course of infection was likely, and that TNF was partially responsible for the enhanced bactericidal activity. In vivo treatment of animals with TNF resulted in significant protection of the animals from mortality. Furthermore, the rate of clearance of bacteria from the lung tissues of infected mice was increased in those animals treated with TNF, and correlated with the ability of this cytokine to protect the animals. These data suggest that the induction of TNF byLegionellabacteria during infection are involved in the non‐specific host defense mechanisms, and that PMN activated by the TNF may be instrumental in clearing the organism from infected lung tissues, thereby protecting the animal.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.429

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe investigated the effect of lipopolysaccharide (LPS) incorporated into phospholipid vesicles (liposomes) on the induction of macrophage‐mediated tumor cytotoxicity and tumor necrosis factor (TNF) secretion. The incorporation ofSalmonella minnesotarough (Re)‐LPS into multilamellar or small unilamellar vesicles (liposomes) resulted in an 100‐to 1,000‐fold reduction in its potency to activate both the macrophage cell line RAW 264.7 and murine thioglycolate elicited peritoneal macrophages to become cytotoxic for L929 and P815 tumor cells. Liposomal LPS was also a 100‐ to 1,000‐fold less potent inducer of TNF secretion from RAW 264.7 cells. Cytokines secreted by the activated macrophages contributed to the cytotoxic effect on the L929 cells but not the P815 cell line. Human recombinant TNF was not cytotoxic for either cell line but was cytostatic for the L929 cell line. Morphological examination of the cells after uptake of fluorescent, free, and liposomal LPS revealed that both forms were internalized by the endocytic pathway. This, together with the considerably reduced potency of liposomal LPS to induce tumor cytotoxicity and TNF secretion, suggests that the interaction of the hydrophobic part of the lipid A moiety of LPS with the macrophage plasma membrane is needed to optimally activate these cells. Incorporation of LPS into liposomes effectively abrogates this interaction.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.436

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractTo assess the effects of advanced age on the nonspecific antimicrobial activity of resident alveolar macrophages (AM), superoxide anion (O2‐) release and the phagocytic and bactericidal capacity of cells from three genetically distinct murine strains were evaluated. In initial experiments, resident AM, obtained by bronchoalveolar lavage of pathogen‐free adult female CD‐1 mice and studied in suspension, were found to produce O2‐spontaneously and in response to phorbol myristate acetate (PMA) snd unopsonized zymosan. Maximum quantities of O2‐were released following stimulation with 1 μg/ml PMA and by a particle‐to‐cell ratio of 100:1 with zymosan; responses to the agonists peaked between 60 and 90 min. Resident AM obtained from pathogen and disease‐free senescent (18‐26‐month‐old) female C57BL/6, BALB/c, and DBA/2 mice released significantly more O2‐in response to both PMA and zymosan than did cells secured from adult (4‐8‐month‐old) animals. In vivo, the capacity of AM from adult and senescent animals to phagocytose Streptococcus pneumoniae (unencapsulated strain) andStaphylococcus aureuswas comparable, and although the cells from the senescent mice tended to be more efficient in their ability to kill internalized bacteria, statistically significant differences between the two groups were not observed. The results of these studies indicate that the enhanced susceptibility of the senescent host to infection of the lower respiratory tract cannot be attributed to age‐related changes in the nonspecific antimicrobial activity of resident AM.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.445

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractStaining rat tissues with a battery of 15 lectin‐horseradish peroxidase conjugates showed that macrophages contain glycoconjugates possessing terminal α, β‐galactose, N‐acetylgalactosamine, fucose, and N‐acetylneuraminic acid, plus two terminal disaccharides. Dissimilar binding of lectins by different phagocyte populations in the same or different organs evidenced variability in glycoconjugates according to the location of the macrophages. With a group of four lectins, macrophages stained most intensely in lung, next strongest in splenic red pulp and lymph node sinuses, and weakest in skin and liver. Two populations of macrophages were newly recognized in spleen on the basis of content of fucose‐rich glycoconjugate. These included a necklace‐like band of macrophages at the border between marginal zone and germinal center and distinctive macrophages dispersed throughout the marginal zone. In lymph nodes, phagocytes stained strongly in the germinal centers and weakly in sinuses for glycoconjugate with N‐linked oligosaccharides and conversely for glycoconjugate with terminal β‐galactose. Variable lectin binding indicated heterogeneity of thymic macrophages. Lectin cytochemistry offers increased sensitivity for detecting macrophages in tissue sections, provides selective staining that shows the prevalence and distribution of the phagocytes and differentiates macrophages into separate subtypes.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.455

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractIn 1978, McKeever et al. [10], studying lepromatous tissues fromMycobacterium leprae‐infected armadillos by cytochemical and electronmicroscopic technics, found a subpopulation of macrophages containing peroxisomes (probably monocytes), but not infected with bacilli, and a subpopulation of macrophages lacking peroxisomes and infected with bacilli. Both infected and noninfected macrophages displayed acid phosphatase activity that was more intense and less precisely localized in heavily infected and vacuolated macrophages than in moderately or noninfected cells. Then, in 1983, Gomez Estrada et al [5], looking for the presence of myeloperoxidase (MPO) in histiocytes, Kupffer cells, and PMN leukocytes of nodular lepromatous patients, found abundantM. lepraein the phagocytes of the lesions but they did not find MPO activity within these cells (except in one case of reactional leprosy). They also found that MPO activity was present in Kupffer's cells and in PMN of patients and controls.

ISSN:0741-5400    

DOI:10.1002/jlb.43.5.468

出版商:Wiley    

年代:1988

数据来源: WILEY