bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of 8-Aminoguanosine on the Toxicity of Guanosine and Deoxyguanosine for Malignant and Normal Lymphoid Cells
Authors: van der Kraan Peter M , van Zandvoort Peter M , De Abreu Ronney A , Bakkeren Jan A J M 
Published: January 2018

Abstract
The toxicity of guanosine and deoxyguanosine in the presence or absence of the purine nucleoside phosphorylase inhibitor, 8-aminoguanosine, for malignant lymphoid cell lines and mitogen-stimulated peripheral blood lymphocytes has been studied. Deoxyguanosine inhibited the proliferation of lymphoid cells more strongly than guanosine. Addition of 100 μM 8-aminoguanosine neither enhanced nor diminished the toxicity of guanosine to the lymphoid cells. Only the toxicity of deoxyguanosine for the leukemic T cell line, MOLT 4, and the leukemic nonBnonT cell line, KM-3, was enhanced by the addition of 100 μM 8-aminoguanosine. These data suggest a possible role of purine nucleoside phosphorylase inhibitors in the treatment of lymphoproliferative disorders of the T-acute lymphoblastic leukemia (ALL) as well as the nonBnonT-ALL subclass.
Long-Term Cultivation of Functional Human Macrophages in Teflon Dishes With Serum-Free Media
Authors: Helinski Ernest H , Bielat Kenneth L , Ovak Geraldine M , Pauly John L 
Published: January 2018

Abstract
Reported 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 (&gt;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.
SHORT COMMUNICATION: Use of Serum-Free, Compositionally Defined Medium for Analysis of Macrophage Differentiation In Vitro
Authors: Vogel Stefanie N , Perera Pin Yu , Hogan M Michele , Majde Jeannine A 
Published: January 2018

Abstract
Macrophage 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.
Myotonic Dystrophy: Defective Oxidative Burst of Polymorphonuclear Leukocytes
Authors: Mege J L , Pouget J , Capo C , Andre P , Benoliel A M , Serratrice G , Bongrand P 
Published: January 2018

Abstract
Because myotonic dystrophy (MD) is an autosomal dominant multisystemic disorder affecting plasma membrane, we have studied the oxidative burst of PMNs. The PMA and fMet-Leu-Phe-stimulated superoxide generation is defective in the patient group as compared to controls: the response is both delayed and low. The kinetic parameters of the NADPH oxidase complex are not affected. We have not found any abnormalities in the membrane potential changes. In addition, the cytosolic protein kinase C (PKC) activity of resting PMNs is similar in MD patients and controls, and the translocation of protein kinase C in response to PMA is not impaired. The decrease of the oxidative response of PMNs from MD patients may be related to an abnormality of the environment of the NADPH oxidase.
Establishment and Characterization of Long-Term Cultured Cell Lines of Murine Resident Macrophages
Authors: Lombard Yves , Bartholeyns Jacques , Chokri Mohamed , Illinger Dominique , Hartmann Dominique , Dumont Serge , Kaufmann Stefan H E , Landmann Regine , Loor Francis , Poindron Phillippe 
Published: January 2018

Abstract
Murine resident macrophages can proliferate in vitro when they are grown in coculture on a layer of mesothelial or endothelial type feeder cells. Resident macrophages were obtained from lung expiants of C57BI/6 Ipr/lpr mice and from spleen expiants or peritoneal washing of Balb/c mice; the cells were seeded without further washing. After 3-4 weeks of culture, the macrophages began to proliferate on a confluent layer of feeder cells. The macrophages then could be collected in the fluid phase and reseeded for permanent culture after generation of a new feeder layer. These cells were characterized as macrophages by the following criteria: 1) their morphology, ultrastructure, and adherence properties; 2) more than 90% of the macrophages phagocytized yeasts compared with less than 1% of the feeder cells; 3) the presence of functional Fc and mannose receptors, nonspecific cytoplasmic esterases, and membrane ectoenzymes such as nicotinamide adenine dinucleotide (NAD) glycohydrolase and nucleotide pyrophosphatase; 4) by cytofluorographic phenotype analysis with monoclonal antibodies, characterizing a normal macrophage population (MAC1+, Fcrec +, H-2K+, THY1 -, LYT2 -, L3T4 -). 5) by functional studies proving that the expanded macrophages could function as accessory cells in the induction of lymphocyte proliferation in response to concanavalin A (Con A), that they generated reactive oxygen radicals and that they were cytotoxic for tumor cells. During coculture, growth or activating factors such as macrophage colony-stimulating factor or gamma-interferon were released in the medium. Long-term cultured macrophages had chromosomal abnormalities. Our study suggests that tissue macrophages can proliferate in vitro and hence that it is possible to establish long-term cultured cell lines of macrophages of defined and reproducible characteristics.
Abnormal Macrophage Development in Hodgkin’s Disease: A Morphometric Study
Authors: Sokol R J , Hudson G , James N T , Wales J 
Published: January 2018

Abstract
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.
Comparison of Thymocyte and T Lymphocyte Gangliosides From C3H/HeN and C3H/HeJ Mice
Authors: Yohe Herbert C , Cuny Carole L , Berenson Charles S , Ryan John L 
Published: January 2018

Abstract
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.
Interactions Between Endogenous Glucocorticoids and Inflammatory Responses in Normal and Tumor-Bearing Mice: Role of T Cells
Authors: Normann S , Besedovsky H , Schardt M , del Ray A 
Published: January 2018

Abstract
Appropriately stimulated lymphocytes and macrophages produce factors in vitro that increase serum corticosterone levels when injected in vivo. In this study, we used euthymic and congenially athymic mice on a BALB/c background to explore the role of T cells in controlling corticosterone levels and the leukocyte response to inflammation. Adult athymic mice had more intense inflammatory reactions than euthymic mice despite higher basal corticosterone levels. This latter condition may be due to interleukin-1 (IL-1) since macrophages from athymic mice when stimulated in vitro by lipopolysaccharide produced more IL-1 than macrophages from euthymic mice. In response to mitogen stimulation, however, splenocytes from athymic mice produced a factor (not IL-1), which, upon injection, increased corticosterone levels and suppressed inflammation. Production of this factor was enhanced by T cells since splenocyte supernatants from euthymic mice were more potent in eliciting both effects. Evidence for in vivo participation of T cells in regulating corticosterone levels was obtained by tumor transplantation. Injection of syngeneic tumor cells or cell-free tumorous ascites rapidly increased corticosterone levels in euthymic but not athymic mice. Anti-inflammation correlated with increased corticosterone levels but was observed also in athymic mice receiving syngeneic tumor transplants. These studies demonstrate that T cells enhance production of a lymphokine that increases corticosterone levels and are required for the corticosterone response to tumor transplantation. In addition, the data suggest two pathways of anti-inflammation in tumor-bearing hosts: a corticosterone-independent, T cell-independent mechanism and a T cell-dependent mechanism that involves a lymphokine-mediated increase in corticosterone blood levels.
Effect of Fibronectin on Antigen-Induced Lymphoproliferation and Antibody Synthesis in Rats
Authors: Rybski James A , Lause David B , Reese Andy C 
Published: January 2018

Abstract
Macrophages are important positive and negative regulators of both primary and secondary antibody responses, and their activity may, in turn, be controlled by soluble mediators secreted by other cells. Fibronectin is a 440,000 dalton normal constituent of plasma and extracellular membranes that acts through macrophages to inhibit mitogen-and alloantigen-stimulated lymphoproliferation. We examined the effect of Fn on the antigen-stimulated lymphoproliferative and antibody responses in cells from trinitrophenol-derivitized keyhole limpet hemocyanin (TNP-KHL) primed rats. Fn in concentrations equivalent to normal plasma levels inhibited TNP-KLH-stimulated lymphoproliferation by unseparated lymph node leukocytes. When the experiment was repeated using purified lymph node T cells and added thioglycollate-induced peritoneal exudate macrophages or splenic adherent macrophages, Fn alone and TNP-KLH alone stimulated lymphoproliferation, but in combination they were strongly inhibitory. The effect was not due to decreased lymphocyte viability in the presence of both TNP-KLH and Fn. Nor was it due to complexes between TNP-KLH and Fn or to a simple alteration in the kinetics of lymphoproliferation. Fn had to be present with the TNP-KLH within the 1st hour of incubation. If macrophages were coincubated with TNP-KLH and Fn for 24 h, washed, and added to enriched T cells, inhibition was equivalent to that seen with continuous coculture. Similarly, coculture of TNP-KLH and Fn inhibited both total immunoglobulin and TNP-KLH-specific antibody synthesis at optimal concentrations of splenic adherent cells. However, at suboptimal levels of splenic macrophages, the combination was synergistic, stimulating more total immunoglobulin synthesis than either TNP-KLH or Fn alone. These data suggest that the inhibitory effect was dependent upon the concentration and phenotype of macrophages present in culture.
Factors Mediating Lipopolysaccharide-Induced Ganglioside Expression in Murine Peritoneal Macrophages
Authors: Berenson Charles S , Yohe Herbert C , Ryan John L 
Published: January 2018

Abstract
The ganglioside composition of endotoxin-responsive C3H/HeN murine peritoneal macrophages is known to undergo dramatic changes in vivo in response to intraperitoneal lipopolysaccharides (LPS), unlike endotoxin-hyporesponsive C3H/HeJ macrophages. To better investigate the mechanism behind LPS-induced macrophage ganglioside changes, resident C3H/HeN peritoneal macrophages were treated in vitro with 0.1-1.0 μg/ml LPS for 6-96 hr, but showed no differences in membrane ganglioside patterns. Coincubation of macrophages with lymphocytes and treating with LPS again elicited no ganglioside changes. In contrast, interferon γ (IFN-γ)-primed macrophages showed a dramatic shift in intensity of one ganglioside when treated with LPS in vitro; an additional macrophage ganglioside appeared when IFN-γ-primed, LPS-treated macrophages were coincubated with lymphocytes. Ganglioside expression induced in vitro still did not approach the complex changes seen in vivo. However, transplanting C3H/HeN macrophages intraperitoneally into C3H/HeJ mice, followed by administration of intraperitoneal LPS, did reveal striking changes in ganglioside expression that resembled the pattern seen in vivo. Thus, LPS alone does not provide the necessary direct signal to promote macrophage ganglioside change even though it alters macrophage function. IFN-γ appears to be one important mediator; however, complex interactions involving other cytokines or migration of independent populations of mononuclear cells may be required for the full manifestation of LPS-induced ganglioside expression in macrophages.
Production and Characterization of Monoclonal Antibodies to Fcγ 2a-Binding Protein Isolated From the Detergent Lysate of a Murine Macrophagelike Cell Line, P388D1
Authors: Kagami Masaru , Funatsu Yoshiyuki , Suzuki Tsuneo 
Published: January 2018

Abstract
Mybridoma cell lines were produced by fusion of SP2/0 murine myeloma cell line with the spleen cells of Wister rats which were immunized with IgG2a-rbinding protein isolated from the detergent lysate of a murine macrophagelike cell line, P388D1, by affinity chromatography on IgG-Sepharose 4B. A monoclonal clone (designated as 3A2) out of a total of 13 different antibody-secreting cell lines was found to secrete IgG1 class antibodies, which inhibited more than 70% of the binding of radio-iodinated myeloma IgG2a protein to P38SD1 cells. The 3A2 Fab fragments bound specifically to P3S8D1 cells at 4° with a KD of 1.9 × 10-8 M and Bmax of 2.9 × 105 per cell. This Fab fragment also specifically bound to Fcγ 2a receptor (R)-positive T cell line (S49) with a KD of 4.4 × 10-9 M and a Bmax of 1.0 × 104 but did not bind to Fcγ 2a-negative S49 variant cell line, cyc-. The flow cytometric analysis with the use of fluorescein-isothiocyanate-tagged 3A2 F(ab’)2 also showed that this antibody binds to Fcγ 2aR-positive cells, P388D1 and S49, but not to Fcγ 2aR-negatlve cells, cyc-. Monomeric and heat-aggregated IgG2a (13-fold molar excess) inhibited the binding of the radioiodinated 3A2 F(ab’)2 to P388D1 cells by 70 and 49%, respectively, whereas the inhibition by monomelic and heat-aggregated IgG2b was 17 and 39%, respectively; 3A2 F(ab’)2 (100-fold molar excess) inhibited the binding of IgG2a and IgG2b to P38SD1 cells by 90 and 24%, respectively, whereas the inhibition of binding of these IgG to S49 cells was 79 and 49%, respectively. Western blotting analysis showed that 3A2 antibody recognizes a major protein (Mr = 100,000) and a minor component (Mr = 80,000) separated by SDS-PAGE of P388D1 or S49 cell lysates under nonreducing condition, whereas under reducing condition, this antibody recognized a major protein (Mr = 50,000) and two additional minor components (Mr = 40,000 and 35,000). Fcγ 2aR may thus exist at the cell surface as a disulfide linked dimer of a subunit of Mr of 50,000, which could be partially degraded during the isolation to smaller fragments of 40,000 and 35,000 Mr peptides which are still held together by interchain disulfide bond. Furthermore, 3A2 antibody-binding protein isolated from P388D1 cell membrane lysate was found to exhibit casein kinase activity, which was previously found to be associated with IgG2a- but not with lgG2b-binding protein isolated from a similar cell lysate.
Mononuclear Cells From Human Lung Parenchyma Support Antigen-Induced T Lymphocyte Proliferation
Authors: Nicod Laurent P , Lipscomb Mary F , Weissler Jonathan C , Toews Galen B 
Published: January 2018

Abstract
We have previously demonstrated that there is a subpopulation of loosely adherent pulmonary mononuclear cells that can be isolated from minced and enzyme-digested lung tissue with a potent capacity to stimulate allogeneic T lymphocyte proliferation. We now demonstrate that these cells are also capable of stimulating an autologous mixed leukocyte reaction (AMLR) and presenting antigen to autologous T lymphocytes. These loosely adherent mononuclear cells (LAM) were more effective than either alveolar macrophages or monocytes as antigen-presenting cells. Depletion of phagocytic or Fc receptor-positive cells from the LAM population enhanced the stimulation of an reaction AMLR while preserving antigen-induced T lymphocyte proliferation. These results indicate that there are nonphagocytic, Fc receptor-negative accessory cells in human lung parenchyma capable of activating resting T cells in an AMLR and supporting antigen-specific T lymphocyte proliferation. The identity of these cells is uncertain, but the data strongly suggest that the cell is not a classical monocyte-derived macrophage. These antigen-presenting cells may be critical in the initiation of immune responses within the lung.
Inhibition by Monoclonal Anticomplement Receptor Type 1 on Interactions Between Senescent Human Red Blood Cells and Monocytic-Macrophagic Cells
Authors: Gattegno L , Saffar L , Vaysse J 
Published: January 2018

Abstract
The effect of different murine monoclonal antibodies (Mab) specific for the glycoprotein complement receptor type 1 (CR1), type 2 (CR2), and type 3 (CR3) on the adhesion to and on the phagocytosis of human senescent red blood cells (S-RBC) by monocytes or by monocyte-derived macrophages (Mø) was investigated.
Murine Mab anti-CR3 (anti-Leu 15 and OKM1) were found to inhibit, in the same order of magnitude, on one hand, the Fc receptors (FcR)-dependent rosetting and phagocytosis, and, on the other hand, the S-RBC rosetting and phagocytosis by adherent monocytes. Thus, the specific involvement of the CR3 epitopes recognized by Mab anti-Leu 15 or by OKM1 in the interactions between S-RBC and monocyte/macrophage could not be demonstrated.
Murine Mab anti-CR1 was found to be a significant inhibitor of binding to and of phagocytosis of S-RBC (but not of young [Y] RBC) by monocytes or Mø, whereas Mab OKM5 carrying the same isotype as Mab anti-CR1, but a different specificity, was devoid of any significant inhibitory effect Furthermore, Y-RBC or S-RBC opsonized with Mab anti-CR1 did not form FcR-dependent rosettes and were not internalized by monocytes; in addition, preincubation of phagocytes with Mab anti-CR1 did not inhibit FcR-dependent resetting and phagocytosis. These results suggest that the effect of anti-CR1 is mediated through a specific binding to CR1 and not through an FcR blockade.
As the role of specifically bound IgG on phagocytosis of human S-RBC by macrophages has previously been demonstrated by several authors, the present study suggests that rnonocyte-macrophage complement receptor type 1 may act in synergy with Fc receptors in the recognition of S-RBC by macrophages. It is shown in addition that the tripeptide Arg-Gly-Asp, identical to the region of iC3b recognized by CR3 and by several adhesion-promoting receptors that are structurally similar to CR3, such as fibronectin or vitronectin, is a significant inhibitor of the binding to and the phagocytosis of S-RBC by monocytic-macrophagic cells.
Interleukin-2 Augmentation of Interleukin-1 and Prostaglandin E2 Production
Authors: Tilden Arabella B , Dunlap Nancy E 
Published: January 2018

Abstract
Some of the major side effects of interleukin-2 (IL-2) therapy in the treatment of malignancies may be related to increased interleukin-1 (IL-1) and/or prostaglandin E2 (PGE2) production. We examined the effect of recombinant (rIL-2) on the in vitro production of IL-1β and PGE2 by unstimulated and LPS-activated human blood mononuclear cells (PBMC). We also compared the effect of rIL-2 on IL-1β production by adherent and nonadherent blood mononuclear cell populations. Cultures of PBMC (5 × 106/ml) were incubated for 24 hr in media only (control), 1,000 U/ml rIL-2, 2 μg/ml LPS, or both LPS and rIL-2. Supematants obtained from these cultures were analyzed for levels of IL-1β and PGE2 by radioimmunoassays. The addition of rIL-2 caused an increase in IL-1β production in 13 of 13 control PBMC cultures and in 11 of 13 LPS-stimulated cultures, which were significant increases as determined by paired f tests. When PBMC were fractionated into plastic adherent and nonadherent populations, the rIL-2 induced increases in IL-1β production were more consistent in control (six of seven cases) and LPS (seven of seven cases) cultures of plastic nonadherent cells than in control (three of seven cases) and LPS (four of seven cases) cultures of plastic adherent cells. Recombinant IL-2 did not increase PGE2 production in control PBMC cultures (none of four cases), but did so in LPS-stimulated PBMC cultures (three of four cases)). These results suggest that rIL-2 may increase IL-1 production in vivo and thus possibly account for some of the side effects of this therapy.