摘要:

AbstractThe orientation (chemotaxis) and locomotion (chemokinesis) of human polymorphonuclear leukocytes (PMNs) are generated by an internal movement mechanism that involves active cytoplasmic movement; they are influenced by external environmental and ionic conditions. We have studied the degree to which the orientation and movement mechanisms of PMNs are self‐contained within the cell and the degree to which they are under membrane control.PMNs were partially and selectively demembranated by treatment with the non‐ionic detergent, octyl‐phenoxyl‐polyethoxyethanol (commercially known as Triton X‐100) under controlled conditions. The tritonated PMNs (referred to in the literature as models) were non‐motile and non‐locomotory. Addition of ATP/Mg+ +with a trace amount of Ca+ +to the medium was followed by reactivation of the tritonated PMN models to move again as motile cells. Although these reactivated PMN models actively locomoted, they could no longer orient to chemoattractants. Thus, the reactivation process restored the physical self‐contained movement parameters but could not reestablish the orientation capacity (chemotactic responsiveness) that was characteristic of live PMNs. The demembranation process apparently destroyed the chemotactic receptors and/or eradicated the coordination function of the membrane.Videotapes of normal (control) as well as reactivated PMN movement were analyzed for movement characteristics. These characteristics were objectively analyzed with a newly designed computer‐assisted micro‐image‐processing technique whereby the videotapes were digitized and quantified and the actual PMN movement printed out in computer‐graphics and tracings (Freeman codes) for confirmation of orientation and movement arising as a result of reactivation.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.203

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractA rapid method for mass isolation of functionally intact hepatocytes and reticuloendothelial cells from a single rat liver is described. The technique is based on collagenase perfusion of the liver, isopycnic sedimentation in Percoll, and selective adherence of the cells. The Kupffer cells (KC) attach and spread on glass or plastic in serum‐free medium 15 min following seeding. Cultures of KC are 90%–95% pure with about 5% liver endothelial cells (LEC),<1% parenchymal cells (PC) and a maximum of 5% stellate cells (SC). The LEC adhere and spread on fibronectin 60–120 min following seeding, forming cultures that are contaminated with 5–10% SC and<1% KC and PC. The yield of plated LEC is 50–60 × 106per 200‐g rat. Ultrastructural analysis shows that Percoll does not associate with the cells during the separation procedure.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.213

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractCyclosporin A (CsA) is a potent immunosuppressive agent that inhibits T‐cell proliferation and lymphokine production. There is less information on the direct effect of CsA on B‐cells. We investigated the proliferative responses of human tonsillar B‐lymphocytes to a “T dependent” mitogen, pokeweed mitogen (PWM), and to a “T independent” mitogen, Staphylococcus aureus (SA). Both responses were strongly inhibited by CsA. Nonspecific cytotoxicity was ruled out, and the inhibition was not reversed by adding IL1, IL2, or BCGF individually or in combination. Maximal inhibition of the PWM response occurred when CsA was added early in the culture period. Cyclosporin A added 18 hours after the start of culture was less effective, and adding CsA after 36 hours resulted in only minimal inhibition. However, with SA as mitogen, addition after 36 hours still affected substantial inhibition. These results, on the time of action and resistance to reversal by exogenous growth factors, suggest that CsA can directly inhibit human B‐cells by a mechanism similar to its action on T‐lymphocytes, blocking an early event critical to entry into cell cycle, but an additional mechanism of inhibition later in the cell cycle may also operate when the proliferative signal is provided by the T‐independent mitogen SA.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.231

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractThe numbers of resident peritoneal cells recovered from NZB and (NZB × NZW)F1hybrid mice, which develop systemic lupus erythematosus (SLE), increased strikingly with age as compared with cells recovered from normal mice. The rise paralleled the onset of anti‐DNA antibodies, occurring earlier in females than in males. The increased number of cells was due to an accumulation of medium‐sized cells with an indeterminate appearance, but with the functional characteristics and cell markers typical of a macrophage. The unusual cells were esterase, F4/80 and Mac‐1 positive, peroxidase, and Alcian blue negative, and were shown on sedimentation velocity separation to be phagocytic for EA and C3 (serum‐treated) zymosan; 47–48% of peritoneal cells were la positive.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.241

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractMacrophages are known to be present in the murine uterus and are known to be among those cells comprising the uterine decidual response to pregnancy. The extent of macrophage involvement in the decidual response has not been documented, and there are unresolved questions regarding expression of markers normally associated with macrophages on cells within the decidua.Using tissue immunohistology, macrophages were identified in virgin and pregnant murine uteri. A significant increase in macrophage density was noted during all stages of pregnancy. When uteri from virgin and pregnant mice were enzymatically digested, 10% of uterine cells from virgin and 22% from pregnant mice expressed macrophage markers (binding of rabbit antimouse macrophage serum, Fc γ receptor expression). Double labeling immunofluorescence demonstrated that the two markers were associated with the same cells. Those results were confirmed in “panning” experiments using a monoclonal antimouse macrophage reagent. In cell suspensions from pregnant murine (C3H/HeN) uteri, 50% of cells exhibiting macrophage markers were I‐Akpositive, and macrophages accounted for nearly all I‐Akpositive cells in uterine cell suspensions. The results of this study demonstrate that the murine decidual response to pregnancy includes an increase in Fc γ receptor‐bearing macrophages and that a relatively high percentage of those macrophages are la positive.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.255

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractHuman peritoneal macrophages (HPM) obtained via laparoscopy were examined for the presence and functional capacity of complement and Fc receptors. Between 5 and 20 ml of peritoneal fluid containing 1–2 × 106macrophages/ml was available for each study. Macrophages made up 80–95% of the cells in the fluid. Fc and C3 receptors on HPM were characterized by rosette formation with, and phagocytosis of, IgG‐ and C3‐coated sheep erythrocytes (E). EIgG were bound by 82% and ingested by 63% of HPM, with 4–15 E ingested/HPM. The HPM formed rosettes with EC3b (56%) and EC3bi (71%) but not EC3d,g or EC3d. Antibodies to complement receptors type 1 (CR1) and type 3 (CR3) inhibited rosette formation with EC3b and EC3bi, respectively, indicating that HPM possessed separate and distinct receptors for the C3b and iC3b ligands. In 60% of the samples studied, HPM demonstrated the ability to ingest both EC3b and EC3bi, as well as ElgG. Because of the heterogeneous nature of the cells obtained in peritoneal fluid, due to their progressive change from monocytelike cells into mature macrophages, HPM were separated by 1 g velocity sedimentation into fractions of increasing maturity. They were then examined for phagocytosis via Fc and complement receptors. Fc receptor mediated phagocytosis occurred throughout the monocyte‐to‐macrophage maturation sequence, while the ability of HPM to ingest via CR1and CR3was maturation dependent, with ingestion via CR3occurring before CR1, in a manner analogous to in vitro differentiation of monocyte‐derived macrophages.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.267

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractTwo reproducible and sensitive assays have been developed for the detection of cell‐free HLA‐DR antigen, an antibody‐mediated complement‐dependent cytotoxicity inhibition assay (CIA) and a competition enzyme‐linked immunosorbent assay (CELISA). One unit of cell‐free HLA‐DR antigen has been quantitated to be the equivalent of 27.5 ng pure DR antigen/ml. Human peripheral blood adherent mononuclear cells (PBAMC), as well as DR+monocytes and B lymphoblastoid cell lines but not T lymphocytes, were observed to shed DR‐bearing vesicles in vitro. Human recombinant IFN‐γ induces the expression of Ia/DR antigen by PBAMC by increasing both the number of cells expressing DR antigen and the density per cell. After incubation with IFN‐γ, PBAMC also shed significantly more DR antigen. The degree of DR expression and shedding is dependent on the dose of IFN‐γ. The shedding of DR antigen occurred concomitantly with the inductive phase of cell membrane antigen expression, and very little DR antigen appeared in culture supernatants subsequent to the removal of IFN‐γ. The possible physiologic significance of shed DR antigen is discussed.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.279

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractKupffer cells isolated from the liver of normal rats were checked for their antiviral activities. The intrinsic antiviral effect against vaccinia virus was high whether the cells were activated in vitro with endotoxin or not. The expression of the extrinsic antiviral activity measured by mixing isolated Kupffer cells with vaccinia‐virus‐infected target cells was remarkably enhanced by prior treatment of the Kupffer cells with LPS. Interferon was shown to be not responsible for that inhibitory activity. Different experimental data suggest that the virus replication in the target cells is blocked at a late stage in the replication cycle.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.293

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractBiological and immunological characteristics of the reticuloendothelial system of irradiation bone marrow chimeric mice and macrophages collected from various tissue sources of the mice were studied. The chimeras showed comparable activities in carbon clearance to those of normal donor or recipient mice. The macrophages from spleen, lymph node, bone marrow, peripheral blood, liver, peritoneal cavity, and lung were demonstrated to be of donor marrow origin. They showed almost the same enzyme activities and phagocytic capability of sheep erythrocytes (SRBC, E), SRBC sensitized with anti‐SRBC IgG (EA), and SRBC sensitized with anti‐SRBC IgM and coated with complement (EAC) as those of normal mice. Proportions of Fc receptor and complement receptor‐positive cells are also in normal range. In addition, the antigen‐presenting capability of the chimeric macrophages for in vitro primary antibody response to SRBC was intact. These observations suggest that the reticuloendothelial system and macrophages of allogeneic bone marrow chimeras where donor and recipient differ at the major histocompatibility complex have no defect so far as could be ascertained by the present study.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.305

出版商:Wiley    

年代:1985

数据来源: WILEY

摘要:

AbstractPreviously, we reported that one of the factors that determines whether or not an animal will be prepared for hapten help after priming is the type of adjuvant used. The present work was undertaken, therefore, to determine which of a diverse variety of adjuvants or biological response modifiers would be effective. They included Freund's complete (CFA) and incomplete (FICA) adjuvants, particulate glucan, muramyl dipeptide (MDP), and its L‐ala‐glycerol‐mycolate derivative. Help by the azobenzenearsonate (ABA) hapten was measured as the augmentation of the anti‐bovine γ‐globulin (BGG) plaque‐forming cell (PFC) response to ABA‐BGG of mice that had been hapten‐primed with ABA conjugated to ovalbumin (OVA). The results showed that FICA was ineffective. MDP was effective but only if administered with FICA during hapten‐priming. MDP‐L‐ala‐glycerol‐mycolate was effective without any adjuvant but only within a narrow dose range. Particulate glucan was as effective as CFA in preparing mice for hapten help.As the macrophage is the primary cellular target of those biological response modifiers that were effective, we conclude that it plays an important role in the cellular interaction involved in the mediation of hapten help.

ISSN:0741-5400    

DOI:10.1002/jlb.38.2.317

出版商:Wiley    

年代:1985

数据来源: WILEY