摘要:

AbstractAn anti‐macrophage monoclonal antibody designated TRPM‐3 was produced using thioglycollate‐elicited rat peritoneal macrophages as immunogen. By the immunoper‐oxidase method, TRPM‐3 was found to be specific for certain macrophage populations, such as marginal zone macrophages and marginal metallophils of the spleen, sinus macrophages of the lymph nodes, and omentum macrophages. No epithelial cells, peripheral blood monocytes, granulocytes, or lymphocytes had reacted with TRPM‐3. Immunoelectron microscopically, reaction to TRPM‐3 was clearly demonstrated on the plasma membrane of splenic marginal zone macrophages, lymphatic sinus macrophages, omentum macrophages, and peritoneal macrophages. Accordingly, TRPM‐3 was considered to have recognized the particular membrane antigen expressed by restricted macrophage populations.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.187

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe interaction of highly purified recombinant human tumor necrosis factor‐alpha (rTNF‐α) with human polymorphonuclear neutrophils (PMNs) was investigated. Binding of125l‐rTNF‐α to PMN reached maximum levels in 30 min at 37°C and in 2 h at 4°C. Scatchard analysis of competitive binding data indicated approximately 6000 receptor sites per cell and a Kdof 1.37 nM. Binding data at 37°C indicated a rapid internalization of rTNF‐α. Following this receptor‐mediated interaction, recombinant TNF‐α was found to inhibit the migration of PMNs under agarose and to enhance PMN production of superoxide anion (O‐2) in a dose‐dependent manner. Furthermore, rTNF‐α‐activated PMNs caused a marked disruption of human umbilical‐vein‐derived endothelial cell monolayers and caused inhibition of their proliferative activities. These data substantiate the role of TNF‐α as an activator of PMN functions and indicate that PMN/TNF‐α/endothelial cell interactions may play a major role in inflammatory reactions.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.196

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe proliferation activity of adherent, tumor‐associated macrophages (TAM) from three related murine mammary carcinoma lines was measured by fluorescence‐activated cell sorter (FACS) analysis of the incorporation of 5′‐Bromo‐2′deoxyuridine (BrdU) in vivo during a 1 hr period prior to tumor removal. The tumor lines examined were line 67 which is nonmetastatic following either subcutaneous (SC) or intravenous (IV) injection, line 168 which colonizes the lung after IV injection but not SC, and line 66 which can colonize the lung from either site.The percentage of total cells identified morphologically as macrophages were similar for tumors from lines 66 and 67 (41% and 38%, respectively), as compared to line 168 tumors, in which, after digestion, 27% of cells were identified as macrophages (P<0.05).Adherent TAM from line 66 tumors had the greatest percentage of BrdU incorporating cells, with an average of 22%. This was statistically significant (P<0.01) from TAM from tumors of line 67, which were 13% positive. The percentage of adherent TAM from line 168 tumors (18% positive) was also significantly different from 67 TAM (P<0.01).These results demonstrate a possible correlation between the percentage of TAM undergoing replication and the ability of the host tumor to colonize the lung. There is no apparent relationship between the percentage of TAM in replication and the number of macrophages associated with a tumor.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.205

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractCompounds with estrogenic activity cause initial toxic responses in the bone marrow characterized by hypocellularity and stem cell myelotoxicity. To elucidate the biochemical nature of these toxic responses, bone marrow cells were collected from mice treated with pharmacological doses of estrogenic chemicals, separated into enriched cell populations, and the enzymatic responses of the individual cell types characterized. Female B6C3F1mice were injected s.c. with five daily doses of 0.07‐5.6 μmoles diethylstilbestrol (DES) or 17‐β estradiol. At 4 days posttreatment body and organ weights were recorded and bone marrow was collected for enumeration and assay of stem cell proliferative responses and enzyme analyses. Treatment with higher dose levels of either estrogenic chemical caused equivalent thymic atrophy, but DES resulted in greater liver and spleen hypertrophy than estradiol. Hexose monophosphate shunt dehydrogenase enzymes in unfractionated bone marrow cells were more sensitive to inhibition by lower estrogen doses than representative enzymes from glycolysis of the Kreb's Cycle, and on an equimolar basis were inhibited to a greater extent by DES than by estradiol. Enzyme analyses after density gradient cell separation indicated that 70‐80% of the hexose monophosphate shunt enzyme activity in bone marrow from untreated mice occurred in the enriched band of cells containing predominantly granulocyte‐macrophages. The majority of the enzyme inhibition induced by DES treatment could also be ascribed to this cellular population. Furthermore, it was shown that DES had a greater inhibitory effect on the proliferative capacity of the committed stem cells than on the multipotential stem cell population, and the main response was again expressed in the enriched band of cells containing predominantly granulocyte‐macrophage precursors. Preliminary endocrine ablation experiments indicated estrogen inhibition of hexose monophosphate shunt enzyme activity was independent of the adrenal and the ovary, but was mediated through the thymus at lower estrogen concentrations.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.212

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractPrevious studies on the mechanism of leukocyte traversal of basement membranes showed that rabbit peritoneal exudate polymorphonuclear cells (PMN) preferentially used laminin, a major constituent of basement membrane, to attach to another component, type IV collagen. PMN also responded chemotactically to nanomolar levels of laminin. We have now determined that PMN possess a receptor for laminin. Scatchard analysis using125I laminin indicates a single class of saturable high affinity binding sites (kd = 6.15 nM/L) on PMN and 3.6 x 104sites per cell. A chymotryptically derived fragment of laminin, C1, which lacks both the long arm and the globular end regions of the short arm (i.e., matrix binding sites) but retains the laminin receptor binding region, gave similar results. Immunoperoxidase studies using monoclonal antibodies to the laminin receptor (mAbLR) indicated the presence of the receptor on the surface of PMN. These cells responded chemotactically to nanomolar levels of C1and laminin, a result consonant with binding data. PMN chemotaxis to a formyl peptide was markedly inhibited by mAbLR, suggesting that the laminin receptor may be required for PMN chemotaxis in general. Our results suggest that PMN extravasation across basement membranes is aided both by reversible attachment of the cells to laminin in the matrix and by chemotaxis to a gradient of soluble intact and possibly degraded laminin. These characteristics have much in common with those of highly metastatic tumor cells.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.220

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractMice carrying the beige mutation (bg/bg) on a C57BI/6 background were challenged withHistoplasma capsulatum. bg/bgmice had higher mortality and higher lung tissue fungal counts in their lungs than eitherbg/+ or C57BI/6 mice challenged with equal inocula. Immunologic studies showed thatbg/bgmice developed normal delayed‐type hypersensitivity (DTH) reactions to histoplasmin, but had deficient NK cell cytotoxic activity against YAC‐1 target cells. Studies of macrophage killing ofH. capsulatumin vitro showed that T lymphocytes of eitherbg/+ orbg/bgmice were able to activate fungal killing bybg/+ but not bybg/bgmacrophages. These studies, while not excluding a role for the NK cell, suggest that macrophage dysfunction may be critical in the greater susceptibility of thebglbgmouse and, by extension, that macrophage function is of major importance in host defense againstH. capsulatum.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.228

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe purpose of this study was to determine whether endotoxin decreased hepatic glucocorticoid binding by the action of mediator(s). Steroid binding in liver cytosol, plasma glucose levels, and plasma corticosterone levels were assayed in C3HeB/FeJ LPS normoresponsive and C3H/HeJ LPS hyporesponsive mice. In C3HeB/FeJ mice, endotoxin significantly depressed the maximum number of steroid binding sites (Bmax) to 30% of control. Plasma glucose levels were decreased to 50% of control, and plasma corticosterone levels increased 4‐fold. No changes in these parameters were seen in C3H/HeJ mice given endotoxin, except for decreased plasma glucose levels at the highest dose of endotoxin. Decreased steroid binding was observed in C3H/HeJ mice 4‐6 hours after receiving C3HeB/FeJ peritoneal exudate cells (elicited with thiogiycollate) and endotoxin. No change in steroid binding was observed in C3H/HeJ mice that received C3H/HeJ peritoneal exudate cells and endotoxin. Mediator‐rich plasma was produced in CF‐1 mice by infecting them with 1 x 107BCG and by challenging them with endotoxin (2 μg) 2 weeks later for 2 h. Transfer of BCG‐endotoxin plasma to C3H/HeJ mice also resulted in decreased steroid binding and plasma glucose. These results indicate that perturbation of glucocorticoid action during endotoxin shock is mediated by soluble factor(s) other than endotoxin. A likely source of mediator(s) is the mononuclear phagocyte.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.236

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe subcellular localization of lysozyme (LZ) has been investigated by immunogold electron microscopic cytochemistry in human neutrophils from bone marrow and blood. Intact cells or subcellular granule fractions were fixed in glutaraldehyde and embedded in glycol methacrylate. Thin sections were incubated with monospecific antibodies followed by antiglobulins coupled to colloidal gold. LZ was detected within both elliptical and spherical primary granules in bone marrow neutrophil promyelocytes. In myelocytes and more mature neutrophils immunolabeling for LZ was observed within the primary granules, although fainter than in promyelocytes. However secondary granules from bone marrow and blood neutrophils were not consistently labeled by gold particles. Immunogold staining was then performed on sections of subcellular fractions of secondary granules: immunogold staining of lactoferrin demonstrated 95% of secondary granules in this fraction. Labeling for LZ of this granule fraction was intense, and except that the few primary granules were also labeled, looked similar to that of lactoferrin.In conclusion, this study is the first to utilize electron microscopic cytochemistry to show that LZ is present in both the primary and secondary granules of blood and bone marrow neutrophils. This technique has the advantage of allowing LZ distribution to be studied within a single organelle and/or in relation to the rest of the cell structure.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.242

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe effect of selected monosaccharides on the random migration of normal adult rabbit alveolar macrophages (AM) was investigated. It was observed that 10 mM of L‐fucose, L‐galactose, or D‐mannose stimulated AM migration 1.5‐2.0 times. In addition, derivatives of L‐fucose and D‐mannose occupying the carbon‐6 position such as L‐fucosyl‐lactose, D‐mannose‐6‐phosphate, D‐mannitol, and mannan enhanced the migration of AM, whereas derivatives of L‐fucose and D‐mannose in the carbon‐1 position produced no migration enhancement. Macrophage migration enhancement activity that was produced spontaneously by spleen cell cultures from normal young rabbits was destroyed by treatment with L‐fucosidase. Accordingly, the migration enhancement factor (MEF) found in spleen cell culture supernatants appeared to depend on L‐fucose conjugated to some protein carrier because MEF was non‐dialyzable. When normal adult AM were treated with L‐fucosidase, they lost their responsiveness to migration inhibitory factor (MIF) but retained their responsiveness to MEF. We have interpreted this to mean that the MIF and MEF receptors are distinct. Synthetic MEFs were prepared by conjugating L‐fucose, D‐mannose, of L‐galactose to bovine serum albumin (BSA). It was noted that these sugar‐BSA conjugates were about 200 times more effective than the corresponding free sugars in producing migration enhancement. In addition, these sugar‐BSA conjugates neutralized MIF activity in a migration inhibition test.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.248

出版商:Wiley    

年代:1987

数据来源: WILEY

摘要:

AbstractThe suppressive effect of IFN‐alpha and IFN‐beta on the induction of tumoricidal activity in mouse bone marrow‐derived macrophages was investigated. Macrophages incubated for 24 hr with IFN‐beta developed lower levels of cytolytic activity when stimulated with IFN‐gamma and LPS, in comparison with macrophages pretreated with medium. The suppressive effect was dependent on the pretreatment dose of IFN‐beta over a concentration range of 1 to 1,000 U/ml. Analysis of IFN‐gamma dose response curves of IFN‐beta treated macrophages showed that these cells were less sensitive to IFN‐gamma. The suppressive effects were fully neutralized by an antiserum to IFN‐alpha/beta. Prostaglandins were apparently not involved in this process since the addition of indomethacin to IFN‐beta treated macrophages did not prevent the loss of responsiveness to activating stimuli. In contrast to the results obtained with IFN‐alpha and IFN‐beta, macrophages pretreated with IFN‐gamma did not develop lower levels of cytolytic activity when again stimulated with IFN‐gamma and LPS. These observations provide evidence for a potentially important negative regulatory role for IFN‐alpha and IFN‐beta in macrophage activation for tumor cell killing.

ISSN:0741-5400    

DOI:10.1002/jlb.41.3.257

出版商:Wiley    

年代:1987

数据来源: WILEY