bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of acridine tracer dyes on neutrophil function
Authors: Hansell Peter , Berger Elaine , Chambers J David , Arfors Karl-E
Published: January 2018

A study was performed to elucidate the effect of two commonly used fluorescent dyes in in vivo microscopic studies, acridine orange (AO) and acridine red (AR), on the ability of phorbol myristate acetate (PMA)-or formyl peptide (fMLP)-stimulated human neutrophils to adhere to a bovine serum albumin matrix and to generate superoxide anions (SOX). Unlabeled stimulated human neutrophils showed 36 ± 9% (PMA, 10-7 M) and 11 ± 7% (fMLP, 10-7 M) adherence to the matrix. This adhesion was CD18 dependent as evidenced by 98% and 92% reduction, respectively, when the anti-CD18 antibody IB4 was included. A dose-dependent inhibition of stimulated human neutrophil adhesion was evident after 30 min of in vitro dye labeling and the EC50 was approximately 70 μg/ml (AR) and 145 μg/ml (AO). SOX generation by PMA-stimulated neutrophils was unaffected up to 100 μg/ml AR and AO but was reduced by 40–60% at higher doses. Rabbit neutrophils labeled in vivo or in vitro with 100 μg/ml AR exhibited 41% and 61% lower SOX generation, respectively. The study indicates that neutrophil function, in terms of ability to adhere to a BSA matrix using CD11/CD18 integrins and to generate SOX upon stimulation, is reduced depending on the dose and choice of fluorescent dye. Caution should be exercised when using these compounds at high concentrations in studies of PMN function. J. Leukoc. Biol. 56: 464–468; 1994.
Interleukin-4 suppresses inflammatory cytokine gene transcription in porcine macrophages
Authors: Zhou Yaling , Lin Gaofeng , Baarsch Mary Jo , Scamurra Ronald W , Murtaugh Michael P
Published: January 2018

Inflammatory cytokines, including interleukin (IL)-1α, IL-1β, IL-8, and tumor necrosis factor α (TNF-α) are produced by macrophages in response to a variety of pathogenic stimuli. We show here that the expression of inflammatory cytokines is suppressed by IL-4 at the transcriptional level. Interleukin-4, when added together with bacterial lipopolysaccharide (LPS), suppressed LPS-induced increases in mRNA levels of IL-1α, IL-1β, IL-8, and TNF-α in alveolar macrophages. The level of suppression was dependent on dose and time of exposure and reached a maximum of 75–80% of uninduced values for IL-1α, IL-8, and TNF. Interleukin-1β expression was completely inhibited by IL-4. The amount of secreted protein, as determined by TNF-α bioassay, was also suppressed by IL-4. Half-maximal suppression occurred at IL-4 concentrations between 0.02 and 0.1 ng/ml for all inflammatory cytokines. Nuclear run-on assays showed that IL-4 suppressed transcriptional activity of all inflammatory cytokines. Messenger RNA stability was not changed by IL-4. The data suggest that IL-4 plays an important transcriptional role in the regulation of alveolar macrophage inflammatory activities in respiratory disease and raise the possibility that IL-4 may function in vivo as a coordinator of inflammatory and immune responses J. Leukoc. Biol. 56: 507–513; 1994.
Cytokine regulation of TNF-α mRNA and protein production by unprimed macrophages from C57BI/6 and NZW mice
Authors: Schook Lawrence B , Albrecht Hugo , Gallay Philippe , Jongeneel C Victor
Published: January 2018

It is well known that bacterial lipopolysaccharide (LPS) induces the synthesis of tumor necrosis factor α (TNF-α) and other inflammatory cytokines by primary monocytes and macrophages and that the Thl lymphokines, interleukin-2 (IL-2) and interferon-γ (IFN-γ), augment this response. We investigated the ability of IL-2 and IFN-γ to induce the production of TNF-α mRNA and protein independently of LPS and the modulation of this response by macrophage colony-stimulating factor (M-CSF) and IL-10. We found that IL-2 and IFN-γ were both able to induce the accumulation of TNF-α mRNA, albeit with slower kinetics than LPS, and that they acted synergistically. However, very little TNF bioactivity was secreted by lymphokine-stimulated macrophages unless LPS was also added. This finding underscores the importance of translational effects in the control of TNF production. M-CSF and IL-10 strongly inhibited TNF production at the level of both mRNA and bioactivity but had no effect on the production of IL-6. Bone marrow-derived or thiogycollate-elicited macrophages from the NZW mouse strain, which have been reported to be deficient in their ability to produce TNF, were at least as responsive to LPS or lymphokines as those taken from the C57B1/6 strain and were similarly affected by M-CSF and IL-10. Therefore, the genetic defect of NZW mice is not a primary deficiency in TNF production J. Leukoc. Biol. 56: 514–520; 1994.
IgE and eosinophil regulation in a murine model of allergic aspergillosis
Authors: Kurup Viswanath P , Choi Hongyung , Murali Pazhayannur S , Coffman Robert L
Published: January 2018

Exposure of BALB/c mice to Aspergillus fumigatus (Af), the antigen responsible for causing allergic bronchopulmonary aspergillosis in humans, caused elevated levels of serum immunoglobulin E (IgE) and peripheral blood and lung eosinophilia similar to that observed in the human disease. We have investigated the role of interleukin-4 (IL-4), IL-5 and interferon-γ in regulating IgE and eosinophilia in the mouse model. Animals were immunized by intraperitoneal injections of soluble Af antigens adsorbed to alum. These animals developed elevated IgE and Af specific IgG1 and were then treated with anticytokine monoclonal antibodies before the final exposure to particulate Af antigens by the intranasal route. The results showed that anti-IL-5 abrogated eosinophilia in mice, while those treated with anti-IL-4 retained the same or reduced IgE levels compared to pretreatment levels. All anti–IL-5, anti–IFN-γ, and control antibody–treated animals showed enhanced IgE levels. Anti–IFN-γ treatment of mice resulted in marked enhancement of eosinophilia compared to all other groups. Eosinophil numbers observed in the histological sections of the lungs confirmed the eosinophilia detected in the peripheral blood. These results indicate that the increase in IgE and eosinophils after exposure to Af antigens in BALB/c mice are due to Af-induced production of IL-4 and IL-5 and that both IgE and eosinophilia are independently regulated. J. Leukoc. Biol. 56: 593–598; 1994.
Dietary polyunsaturated fatty acid effects on immune cells of the porcine lung
Authors: Turek J J , Schoenlein I A , Clark L K , Van Alstine W G
Published: January 2018

The effects of various dietary polyunsaturated fatty acids (PUFAs) on the function of immune cells of the porcine lung was studied. Groups of six pigs were fed diets containing 10.5% corn oil [CO; enriched in linoleic acid (18:2, n-6)], linseed oil (LO; enriched in α-linolenic acid (18:3, n-3)], menhaden oil (MO; enriched in eicosapentaenoic (20:5; n-3) and docosahexaenoic (22:6; n-3) acids], linseed + corn oil (1:1; LC), and menhaden + corn oil (1:1; MC) for 28–30 days. Basal levels of alveolar macrophage (mφ) tumor necrosis factor-α (TNF-α) production were higher (P < .05) for LC- and MC-fed pigs than for CO- and LO-fed pigs. Lipopolysaccharide (LPS)-stimulated LC and MC mφs produced more TNF than mφs from pigs fed CO, LO, and MO diets. Macrophages from pigs receiving the CO and LC diets had higher (P < .05) levels of leucine aminopeptidase than mfs from the other dietary groups. Lipopolysaccharide did not increase mφ nitrite production over basal levels except in the MO diet group. However, LPS-stimulated mφs from the CO, MO, and LC dietary groups produced more nitrite than mfs from MC-fed pigs. Alveolar lymphocytes from pigs receiving the MC diet produced more T cell growth factors than LO and MO mφs. Alveolar mφs from the different dietary groups did not differ in their capacity for non-immune-mediated phagocytosis of fluorescent latex beads. These results indicate that dietary PUFAs can modulate some functions of porcine alveolar immune cells and that this may prove significant for host response to respiratory disease agents. J. Leukoc. Biol. 56: 599–604; 1994.
Hyaluronan receptor (CD44) expression and function in human peripheral blood monocytes and alveolar macrophages
Authors: Culty Martine , O’Mara Thomas E , Underhill Charles B , Yeager Henry , Swartz Rodney P
Published: January 2018

CD44 glycoproteins are present on the surfaces of many hematopoietic cells and in some cases can bind hyaluronan, a major component of the extracellular matrix. In the present study, we have found that newly explanted human peripheral blood monocytes (PBMs) exhibit a major CD44 band of 85 kDa, whereas autologous alveolar macrophages (AMφ) express multiple isoforms ranging from 85 to 200 kDa. Within 4 h in culture, PBMs began expressing new CD44 isoforms of 120, 150, and 180 kDa. Newly explanted AMφ specifically bound [3H]hyaluronan (135 cpm/μg protein), but newly explanted PBMs did not. However, in vitro cultured PBM progressively acquired the ability to bind [3H]hyaluronan and exhibited specific binding of hyaluronan similar to that of AMφ (113 cpm/μg protein) after 4 days in culture. In both cases, the binding of [3H]hyaluronan was specifically inhibited by the addition of monoclonal antibody directed against CD44. AMφ readily degraded [3H]hyaluronan and reached a plateau after 4 days in culture (115 cpm/μg protein). Newly explanted PBM exhibit no hyaluronan degradation and only a small degradative activity after 4 days in culture (6 to 11 cpm/μg protein). Thus, CD44 expression and function appear to change as PBM mature in vitro resembling more that found in AMφ. J. Leukoc. Biol. 56: 605–611; 1994.
Role for oxygen radicals in self-sustained HIV-1 replication in monocyte-derived macrophages: enhanced HIV-1 replication by N-acetyl-L-cysteine
Authors: Nottet Hans S L M , van Asbeck B Sweder , de Graaf Loek , de Vos N Machiel , Visser Maarten R , Verhoef Jan
Published: January 2018

N-acetyl-l-cysteine (NAC) has been proposed as a therapeutic agent for AIDS patients because it reduces human immunodeficiency virus type 1 (HIV-1) replication in stimulated T cells. However, NAC and glutathione enhanced acute HIV-1 replication in monocyte-derived macrophages. Buthionine sulfoximine did not affect NAC-mediated enhanced HIV-1 replication, indicating that the NAC-mediated effects are glutathione-independent. Superoxide dismutase and the hydroxyl radical scavengers dimethylthiourea and thiourea, but not urea, inhibited acute HIV-1 replication in macrophages. NAC reduced ferricytochrome c and increased dose-dependently Fe(III)-citrate and Fe(III)-EDTA-catalyzed hydroxyl radical formation in a system using glucose and glucose oxidase. Dimethylthiourea and thiourea, but not urea and superoxide dismutase, dose-dependently inhibited NAC-mediated enhancement of HIV-1 replication. These data suggest that oxygen radicals play an important role in self-sustained HIV-1 replication in macrophages and that oxygen radical scavengers other than NAC should be considered as therapeutic agents for AIDS patients. J. Leukoc. Biol. 56: 702–707; 1994.
Characterization of nitric oxide–stimulated ADP-ribosylation of various proteins from the mouse macrophage cell line ANA-1 using sodium nitroprusside and the novel nitric oxide-donating compound diethylamine dinitric oxide
Authors: Sheffler Lynn A , Wink David A , Melillo Giovanni , Cox George W
Published: January 2018

We examined the ability of nitric oxide (NO) to stimulate the ADP-ribosylation of proteins from the mouse macrophage cell line ANA-1. To demonstrate a specific effect of NO, we used a novel compound named diethylamine dinitric oxide (DEA/NO; l,l-diethyl-2-hydroxy-2-nitrosohydrazine, sodium salt; [Et2NN(O)NO]Na), which releases NO in aqueous solution at neutral pH. DEA/NO stimulated the ADP-ribosylation of at least three cytosolic proteins (Mr = 28,000, 33,000, and 39,000) from ANA-1 macrophages. The effect of DEA/NO on the ADP-ribosylation of the predominant target p39 was dose dependent (EC50 = 80 μM). Moreover, the effect of DEA/NO was attributed specifically to released NO rather than diethylamine or nitrite. Sodium nitroprusside (SNP) also stimulated the ADP-ribosylation of cytosolic proteins from ANA-1 mouse macrophages. However, SNP exhibited different time- and dose-dependent effects on the modification of p39. NO synthesized via the activity of interferon-γ plus lipopolysaccharide-induced NO synthase also enhanced the ADP-ribosylation of p39, confirming that the effects of DEA/NO and SNP could be attributed to NO or reactive nitrogen oxide species. Neither pertussis toxin nor cholera toxin stimulated the ADP-ribosylation of p39; however, cholera toxin stimulated the ADP-ribosylation of proteins with approximate molecular weights of 28,000 and 33,000. These data suggest that the induced expression of NO synthase in tumoricidal macrophages may be associated with autocrine and paracrine effects of NO that include the ADP-ribosylation of various proteins. Moreover, these results indicate that DEA/NO and related compounds may be useful as pharmacologic tools for investigating the effects of NO and reactive nitrogen oxide species on macrophages. J. Leukoc. Biol. 57: 152–159; 1995.
Differential effects of the anti-inflammatory compounds heparin, mannose-6-phosphate, and castanospermine on degradation of the vascular basement membrane by leukocytes, endothelial cells, and platelets
Authors: Bartlett Mark R , Cowden William B , Parish Christopher R
Published: January 2018

Recent studies suggest that heparin, mannose-6-phosphate (M6P), and castanospermine (CS) may mediate their anti-inflammatory effects by inhibiting the passage of leukocytes through the subendothelial basement membrane (BM). In order to test this hypothesis, heparin, M6P, and CS were examined for their ability to prevent the in vitro degradation of a 35SO4-labeled extracellular matrix (ECM) by neutrophils, lymphocytes, endothelial cells (ECs), and platelets, the labeled ECM degradation products being analyzed by gel filtration chromatography. All three compounds inhibited 35SO4-labeled ECM degradation, but M6P and CS were cell-type specific in their effects. Heparin inhibited the heparanase activity of all cell types examined, confirming the results of previous studies using similar in vitro techniques. M6P selectively inhibited lymphocyte heparanase but not that of platelets, neutrophils, or ECs. CS selectively inhibited phorbol myristate acetate (PMA)-induced EC heparanase and sulfatase activity but did not affect the constitutive expression of degradative enzymes by non-stimulated ECs. These findings provide important clues to the mode of action of these compounds and the characteristic inflammatory pathology associated with the use of each anti-inflammatory agent. In particular, the data support the view that leukocytes markedly differ in the mechanisms they use to degrade BM/ECM to enable extravasation and that some degree of cooperation with EC is required in this process. J. Leukoc. Biol. 57: 207–213; 1995.
Functional deficiencies in two distinct interferon α-producing cell populations in peripheral blood mononuclear cells from human immunodeficiency virus seropositive patients
Authors: Feldman Stephen B , Milone Michael C , Kloser Patricia , Fitzgerald-Bocarsly Patricia
Published: January 2018

Two populations of IFN-α producing cells (IPC) were examined to determine whether they are coordinately dysregulated in human immunodeficiency virus (HIV) disease. IFN-α produced in response to herpes simplex virus (HSV) and Sendai virus (SV) was measured and the frequencies of the IPC were obtained by ELISpot assay. IPC that respond to HSV (natural IFN-α producing cells) and those responding to SV (predominantly monocytes) were present, on average, at 7.6 and 138 per 104 PBMC in healthy controls, respectively. More patients had a reduced IFN-α response to HSV than to SV, and individual patients did not show a decreased response to SV without a decreased response to HSV. Neither IPC function was correlated with CD4+ cell levels. We conclude that the defects in IFN-α production in these two cell populations arise independently, possibly due to differences in susceptibility to HIV infection or molecular regulation. J. Leukoc. Biol. 57: 214–220; 1995.
Effects of bafilomycin A1 on functional capabilities of LPS-activated alveolar macrophages
Authors: Bidani Akhil , Heming Thomas A
Published: January 2018

Resident alveolar macrophages (mφ) possess plasmalemmal vacuolar-type H+-ATPase (V-ATPase) that plays a crucial role in regulation of intracellular pH (pHi). To assess the importance of this V-ATPase to mφ effector functions, resident alveolar mφ from rabbits were activated with E. coli-derived lipopolysaccharide (LPS) and exposed to bafilomycin A1 a specific inhibitor of V-ATPase. Bafilomycin caused a significant cytosolic acidification in both the absence and presence of CO2-HCO3-, and in both unstimulated and activated mφ. Superoxide production and Fc receptor-mediated phagocytosis also were reduced in bafilomycin-treated mφ. Similar effects were elicited by acidifying the cytoplasm in the absence of bafilomycin, by lowering extracellular pH (pHo) from 7.4 to 6.5–6.6. Thus, the effects of bafilomycin on phagocytosis and superoxide production probably were related to cytosolic acidification, secondary to blockade of V-ATPase-mediated H+ extrusion across the plasma membrane. Conversely, bafilomycin significantly increased TNF-α release. This effect cannot be explained by a bafilomycin-induced acidosis because acidic pHo significantly reduced TNF-α release. The results demonstrate that V-ATPase activity is an important determinant of the effector functions of LPS-activated mφ. J. Leukoc. Biol. 57: 275–281; 1995.
Interleukin-9 and its receptor: involvement in mast cell differentiation and T cell oncogenesis
Authors: Renauld Jean-Christophe , Kermouni Abdenaim , Vink Anne , Louahed Jamila , Van Snick Jacques
Published: January 2018

Intcrleukin-9 (IL-9) is a multifunctional cytokine produced by activated TH2 clones in vitro and during TH2-like T cell responses in vivo. The IL-9 receptor is a member of the hemopoietin receptor superfamily and interacts with the γ chain of the IL-2 receptor for signal transduction. Various observations indicate that IL-9 is actively involved in mast cell responses by inducing the proliferation and differentiation of these cells. The role of IL-9 in T cell responses is less clear. Although freshly isolated normal T cells do not respond to IL-9, this cytokine induces the proliferation of murine T cell lymphomas in vitro and in vivo overexpression of IL-9 results in the development of thymic lymphomas. In the human, the existence of an IL-9-mediated autocrine loop has been suggested for some malignancies such as Hodgkin’s disease. Other potential biological targets for IL-9 include B lymphocytes, hematopoietic progenitors, and immature neuronal cell lines. J. Leukoc. Biol. 57: 353–360; 1995.
Immunocytochemical detection of lipid peroxidation in phagosomes of human neutrophils: correlation with expression of flavocytochrome b
Authors: Quinn Mark T , Linner John G , Siemsen Daniel , Dratz Edward A , Buescher E Stephen , Jesaitis Algirdas J
Published: January 2018

Oxidants generated by the NADPH oxidase of activated neutrophils can react with a number of tissue targets to form toxic metabolites such as 4-hydroxynonenal (4-HNE). 4-HNE is a lipid peroxidation product generated by free radical attack on ω-6 polyunsaturated fatty acids and is a marker for membrane lipid peroxidation. In this study, we examined the accumulation of 4-HNE-protein adducts in phagosomes of neutrophils obtained from a male patient with homozygous X-linked, flavocytochrome b-deficient chronic granulomatous disease (CGD), his heterozygous mother, and his normal father. Specific polyclonal antibodies recognizing 4-HNE-protein adducts and gp91-phox (flavocytochrome b large subunit) were prepared and used to immunocytochemically detect these antigens in cryofixed, molecular distillation-dried neutrophils. No 4-HNE-protein adducts were detected in flavocytochrome b-deficient cells from the homozygous patient or from the heterozygous CGD carrier. However, in gp91-phox-positive cells from both the normal and heterozygous CGD carrier, significant 4-HNE-protein adduct labeling was observed, primarily in the phagosomes. When data from single- and doublelabeled cells were combined, the frequency distribution of the labels in phagosomes supported this observation, showing that neutrophils from the heterozygous CGD carrier were 71% 4-HNE-protein adduct-positive and 56% gp91-phox-positive, while cells from the normal father were >97% positive for both 4-HNE-protein adducts and gp91-phox. These results confirmed the nitroblue tetrazolium tests of 100%, 60 ± 2%, and 0% positive for the father’s, mother’s, and son’s cells, respectively, and demonstrated that 4-HNE-protein adduct antibodies are useful and accurate probes of the occurrence of lipid peroxidation in vivo. We conclude that 4-HNE and resulting 4-HNE-protein adducts are generated as a result of NADPH oxidase activity in the phagosomes of human neutrophils and that these lipid peroxidation products may contribute to microbial killing and/or damage of neutrophil phagolysosomal proteins. J. Leukoc. Biol. 57: 415–421; 1995.
Distinct actions of benzene and its metabolites on nitric oxide production by bone marrow leukocytes
Authors: Laskin Jeffrey D , Rao N R , Punjabi Chitra J , Laskin Debra L , Snyder Robert
Published: January 2018

Benzene is a widely used industrial solvent known to cause bone marrow depression. This is associated with increased production of reactive oxygen metabolites and nitric oxide by bone marrow phagocytes, which have been implicated in hematotoxicity. Benzene metabolism to phenolic intermediates appears to be an important factor in bone marrow toxicity. In the present studies, we compared the effects of benzene and several of its metabolites on nitric oxide production by murine bone marrow leukocytes. Bone marrow cells readily produced nitric oxide in response to the inflammatory mediators lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Treatment of mice with benzene (800 mg/kg), or its metabolites hydroquinone (100 mg/kg), 1,2,4-benzenetriol (25 mg/kg), or p-benzoquinone (2 mg/kg), at doses that impair hematopoiesis, sensitized bone marrow leukocytes to produce increased amounts of nitric oxide in response to LPS and IFN-γ. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) augmented bone marrow leukocyte production of nitric oxide induced by inflammatory mediators. Benzene, as well as its metabolites, markedly increased the sensitivity of the cells to both GM-CSF and M-CSF. Cells from hydroquinone- or 1,2,4-benzenetriol–treated mice were significantly more responsive to the inflammatory cytokines and growth factors than cells isolated from benzene- or p-benzoquinone–treated mice, suggesting that the phenolic metabolites of benzene are important biological reactive intermediates. Because nitric oxide suppresses cell growth and can be metabolized to mutagens and carcinogens, the ability of benzene and its metabolites to modulate its production in the bone marrow may be important in their mechanism of action. J. Leukoc. Biol. 57: 422–426; 1995.
Normal human sweat contains interleukin-8
Authors: Jones Anne P , Webb Louise M C , Anderson Arthur O , Leonardo Edward J , Rot Antal
Published: January 2018

Sweating in humans is induced by physical or emotional stress, which raises the possibility that sweating may relate to host defense. We therefore asked whether human eccrine sweat attracts leukocytes and found that it is chemotactic for human neutrophils. This activity was due to several chemoattractants, one of which was interleukin-8 (IL-8). Using immunohistochemistry and in situ hybridization IL-8 and its mRNA have been detected in sweat gland epithelium, indicating that IL-8 is produced in situ. This establishes a pattern of physiological IL-8 secretion by exocrine glands and suggests that, in addition to its role as a major inflammatory mediator, IL-8 also has physiological homeostatic functions. J. Leukoc. Biol. 57: 434–437; 1995.
LPS directly induces oxygen radical production in human monocytes via LPS binding protein and CD14
Authors: Landmann Regine , Scherer Franziska , Schumann Ralf , Link Susanne , Sansano Sebastiano , Zimmerli Werner
Published: January 2018

In human monocytes, superoxide (O2-) generation accompanies phagocytosis and is important for bactericidal activity. It also contributes to tissue damage in inflammation. In the present study we investigated, whether lipopolysaccharide (LPS) directly stimulates monocyte O2- production with kinetics known for other LPS effects and, if so, by which mechanism. LPS caused a time- and dose-dependent O2- release in nonadherent purified monocytes. The effect appeared after 5 min, peaked at 30 min, and disappeared after 2 h. It was maximal with 10 ng/ml lipid A (+148 ± 22%, P < .001), 1 ng/ml LPS Escherichia coli Re (+226 ± 68%, P < .001), and 100 ng/ml LPS Salmonella abortus equi sm (+272 ± 52%, P < .001), respectively. The effect was not observed in buffer, even when using 10 üg/ml LPS. It was dependent on the presence of heat-inactivated AB serum, with a maximal effect at ≥;0.5%. Serum could be replaced by LPS-binding protein (LBP). Polymyxin B and anti-LBP antiserum, respectively, blocked the LPS effect. LPS-induced O2- generation was also completely blocked by anti-CD14 antibodies (3C10 and 63D3) and by their corresponding F(ab’)2 fragments. Monocytes treated with phosphoinositol-specific phospholipase C and monocytes from patients with paroxysmal nocturnal hemoglobinuria, lacking the phosphatidylinositol-anchored CD14, did not respond to LPS stimulation with O2- production. Similarly to LPS, E. coli caused stronger O2- production with heat-inactivated serum than without, and this effect was blocked by anti-CD14 antibodies. In conclusion, these data indicate that LPS directly stimulates O2- production in human monocytes via CD14 depending on LBP. J. Leukoc. Biol. 57: 440–449; 1995.
A mouse monoclonal antibody specific for the Vβ5.3 chain of the human TcR recognizes a subgroup of the mouse TcR Vβ8.2 chains
Authors: Bleux C , Lacoste-Eleaume A S , Cabaniols J P , Carrière D , Poncelet P , Kanellopoulos-Langevin C
Published: January 2018

We have analyzed the reactivity of a mouse monoclonal antibody directed against the human T cell receptor for antigen (TCR). This antibody (111-427) of immunoglobulin G1 isotype has been produced in a BALB/c mouse immunized with HPB-ALL cells and normal human peripheral blood leukocytes. It reacts specifically with the HPB-ALL lymphoma and 2 to 7% of normal human blood lymphocytes, on which it has a mitogenic effect in vitro. We have shown that it immunoprecipitates the αβ TCR of HPB-ALL and that it is specific for the Vβ5.3 chain of the human TCR. In addition, we have observed that this antibody stains a minor fraction of T lymphocytes in different strains of mice. We have screened a number of murine T cell clones or hybridomas and have found that the T cell hybrid line D0.11.10.S4.4 is positive. We have been unable to immunoprecipitate reproducibly the molecule recognized by 111-427 after 125I cell surface labeling and cell lysis in NP-40 or digitonin, probably because of low-affinity binding. On Western blotting, 111-427 revealed one band that has an apparent molecular mass of 89 kDa in nonreducing conditions and disappears after reduction. Similar results were obtained in parallel with the F23.1 and F23.2 antibodies. Thus, this antibody appears to recognize an epitope present primarily on the Vβ8.2 chain of the mouse TCR. We have assayed its capacity to stimulate splenic T lymphocytes in vitro. We have observed that it is capable of triggering, to a minor degree in soluble form and very effectively when coupled to Sepharose beads, the proliferation of spleen T lymphocytes from mice chronically infected with the blood parasite Trypanosoma cruzi. J. Leukoc. Biol. 57: 491–499; 1995.
Phenotypic and functional heterogeneity of the murine alveolar macrophage-derived cell line MH-S
Authors: Sankaran Kumaresan , Herscowitz Herbert B
Published: January 2018

We have previously reported that MH-S, an established murine alveolar macrophage-derived cell line, mediated profound inhibition of in vitro antibody production, as did their freshly isolated alveolar macro phage (AM) counterparts. In this communication we show that like freshly recovered AMs, the MH-S cell line also displays phenotypic and functional heterogeneity. Sorting of parental MH-S cells by flow cytometry based on reactivity with anti-Mac-1 antibody yielded two sub sets. Further analysis by staining with monoclonal anti bodies against well-characterized murine macrophage cell surface markers revealed that both Mac-1+ and Mac-1-subsets expressed the mature murine macrophage antigen (F4/80) and class II major histocompatibility complex molecules, but with different intensity. In contrast, the two subsets stained equivalently with antibody against the FCγII receptor, whereas neither subset stained with anti-CD4 antibody. Examination by light microscopy revealed pleomorphism in the Mac-1+ population with many of the cells appearing spindle shaped and having elongated processes, whereas a majority of the cells in the Mac-1- population were spherical in shape. Functionally, cells from the Mac-1+ population were less inhibitory of in vitro antibody production and produced significantly more nitric oxide in response to stimulation with lipopolysaccharide than were cells in the Mac-1- population. Essentially similar results were obtained using cloned Mac-1+ and Mac-1- MH-S cells. The finding of het erogeneity in an established cell line that displays functions similar to those of freshly recovered AMs suggests that distinct subsets of AMs may be involved in the pathogenesis of disease processes in the lung. J. Leukoc. Biol. 57: 562–568; 1995.
Effects of myristate phorbol ester on V-ATPase activity and Na+-H+ exchange in alveolar macrophages
Authors: Heming Thomas A , Bidani Akhil
Published: January 2018

The roles of protein kinase G (PKC) in regulation of the plasmalemmal vacuolar-type H+-ATPase (V-ATPase) and Na+-H+ exchanger (NHE) of rabbit alveolar macro phages (mφ) were investigated using phorbol 12-myristate 13-acetate (PMA). At an extracellular pH (pHo) of 7.4 (nominal absence of CO2-HCO-  3), PMA caused a dose-dependent increase in the rate of cellular H+ extrusion with little change in intracellular pH (pHi). PMA caused a prolonged cytosolic acidification at pHo < 6.8. PMA-induced changes in pHi were sensitive to bafilomycin A1, but were insensitive to amiloride. Studies of pHi recovery following intracellular acid challenge showed that both V-ATPase and the NHE were up-regulated by PMA. An inactive analog, 4α-phorbol, had no detectable effects on pHi homeostasis. These data indicate that (a) PKC is involved in regulation of V-ATPase and the NHE of resident alveolar mφ and (b) V-ATPase is the predominant mechanism for pHi homeostasis in unstimulated and PMA-activated mφ. J. Leukoc. Biol. 57: 600–608; 1995.
IFN-γ increases cathepsin H mRNA levels in mouse macrophages
Authors: Lafuse William P , Brown David , Castle Lynn , Zwilling Bruce S
Published: January 2018

Expression of major histocompatibility complex (MHC) class II molecules and ability to present anti gen to T lymphocytes is acquired upon activation of the macrophage by intcrfcron-γ (IFN-γ). Little information is available concerning immune regulation of protease gene expression in mouse macrophages. We have isolated a eDNA clone for cathepsin H, a lysosomal cysteine pro teinase from a cDNA subtraction library of mouse macro phage genes induced by IFN-γ, and have characterized its expression. The level of cathepsin H mRNA increased in mouse peritoneal macrophages following addition of IFN-γ. Cathepsin H mRNA levels began to increase 8 h after the addition of IFN-γ and was maximal at 24–48 h. This increase was concordant in time with appearance of MHC class II Eβ mRNA and Ia invariant chain mRNA. The increase in cathepsin H mRNA levels by IFN-γ was dose dependent. Cycloheximide treatment of peritoneal macrophages inhibited the increase in cathepsin H mRNA levels induced by IFN-γ, suggesting that the in crease in cathepsin mRNA levels requires de novo protein synthesis. Lipopolysaccharide and cytokines interleukin-2 (IL-2), IL-4, IL-10, and tumor necrosis factor α were found to have no effect on cathepsin H mRNA levels in mouse peritoneal macrophages. J. Leukoc. Biol. 57: 663–669; 1995.
Contribution of mannose receptor to signal transduction in Fcγ receptor-mediated phagocytosis of mouse peritoneal macrophages induced by liposomes
Authors: Murai Makoto , Aramaki Yukihiko , Tsuchiya Seishi
Published: January 2018

The contribution of mannose receptors on the cell surface of mouse peritoneal macrophages to the pro cess of liposome-induced phagocytosis of immunoglobu lin G-opsonized sheep red blood cells (SRBCs) through Fcγ receptor has been investigated. Fcγ receptor-mediated phagocytosis of opsonized SRBCs was activated by modified α  2-macroglobulin, which was produced in the incubation mixture of α  2-macroglobulin and liposometreated splenic B cells. The phagocytosis was specifically inhibited by the addition of D-mannose, and the inhibition was dependent on the D-mannose concentration. The binding of modified α  2-macroglobulin to macro phages was also reduced by the addition of D-mannose. The activation effect of modified α  2-macroglobulin was not inhibited when in the presence of α  2-macroglobulintrypsin and -methylamine complexes. In the presence of cydoheximide, activated phagocytosis was reduced to the control level. By Scatchard plot analysis of IgG binding studies, the number of Fcγ receptors of a macrophage had been increased to 4.6-fold that of a control macrophage by treatment with modified α  2-macroglobulin. These findings suggest that macrophage mannose receptors are involved in activating the process of FCγ receptor-mcdiatcd phagocytosis of opsonized SRBCs induced by modified a α  2-macro globulin. Lectins may participate in a signal transduction in macrophage activation by lipo somes, J, Lcukoc, Biol. 57: 687–691; 1995.
CD4 T cell development and cytokine polarization: an overview
Authors: Swain Susan L
Published: January 2018

We have studied two key stages of CD4 development that serve as important checkpoints in determining whether a response will occur, how big it will be, and how long it will last. The response of naive CD4 cells is strictly regulated by the requirement for interaction of the potential precursor with an antigen-presenting cell-expressing multiple costimulatory molecules such as B7 family and intercellular adhesion molecule-1. The extent of naive cell proliferation is critically dependent on interleukin-2, whose autocrine production is similarly regulated. We show that effector CD4 cells can either die, expand, and/or generate memory. Antigen stimulation accelerates death or promotes expansion dependent on the presence of cytokines. Interleukin-2 and transforming growth factor-β synergize to block apoptosis and favor extensive effector expansion. J. Leukoc. Biol. 57: 795–798; 1995.
Regulatory effect of aminopeptidase inhibitor (bestatin) on the cervix during induction of ripening by interleukin-8
Authors: El Maradny Emad , Kanayama Naohiro , Halim Abdul , Maehara Kayoko , Sumimoto Kazuhiro , Terao Toshihiko
Published: January 2018

Bestatin is an immunomodulatory peptide that stimulates the humoral and cell-mediated immune system. It also has an inhibitory effect on multiple aminopeptidases. Recently we found that aminopeptidase N inactivates interleukin-8 in vitro. Bestatin successfully suppresses the effect of aminopeptidase N on interleukin-8. During cervical maturation many biochemical changes occur including decrease in collagen concentration and increase in collagenase and elastase activities. Interleukin-8, which has a potent neutrophil chemotactic effect, was found to induce cervical ripening in rabbits. The combination of interleukin-8 with bestatin also induced cervical ripening by providing approximately regular levels of neutrophil numbers, collagenase, and elastase activities. We therefore suggest that this regulatory mechanism also takes place in vivo through the inhibitory effect of bestatin on aminopeptidase N. J. Leukoc. Biol. 57: 832–836; 1995.
Ultraviolet radiation reduces phagocytosis and intracellular killing of mycobacteria and inhibits nitric oxide production by macrophages in mice
Authors: Jeevan Amminikutty , Bucana Corazon D , Dong Zhongyun , Dizon Victor V , Thomas Stephanne L , Lloyd Thomas E , Kripke Margaret L
Published: January 2018

Exposure of mice to a single or multiple low doses of ultraviolet radiation (UVR) decreases the induction of the delayed-type hypersensitivity (DTH) response to Mycobacterium bovis BCG and Mycobacterium lepraemurium (MLM) and impairs the clearance of bacteria from the lymphoid organs. This study is an attempt to address the mechanism by which UV radiation impairs the clearance of bacteria from the lymphoid organs by determining whether alterations in macrophage function such as ingestion and intracellular killing of mycobacteria or production of reactive nitrogen intermediates might be responsible for these effects. BALB/c or C3H/HeN mice were exposed to a single dose of UVB (280–320 nm) radiation ranging from 0.35 to 45 kJ/m2, and at regular intervals after irradiation, the peritoneal and splenic macrophages were collected, cultured, and infected with live BCG or MLM. Phagocytosis was assessed at 6 h by counting the number of acid-fast bacteria per macrophage after Ziehl-Neelsen staining. The rate of intracellular killing was assessed by lysing the macrophages at 6, 12, 24, and 48 h after BCG infection, plating the suspension on 7H11 agar, and counting the number of colony-forming units 21 days later. Similarly, the nitric oxide production, as measured by nitrite, by macrophages obtained from UVB-irradiated and nonirradiated mice in response to BCG was assessed. There was a significant reduction in the uptake of organisms by both peritoneal and splenic macrophages collected from UV-irradiated mice. The intracellular killing of organisms was also significantly reduced, as was the production of nitric oxide by peritoneal macrophages infected with BCG in vitro. These results indicate that UVR affects macrophage functions and are consistent with our hypothesis that impaired clearance of bacteria in vivo results from an alteration in macrophage function. J. Leukoc. Biol. 57: 883–890; 1995.
Influence of serum on zinc, toxic shock syndrome toxin-1, and lipopolysaccharide-induced production of IFN-γ and IL-1β by human mononuclear cells
Authors: Driessen Christoph , Hirv Kaimo , Wellinghausen Nele , Kirchner Holger , Rink Lothar
Published: January 2018

Measurement of cytokine secretion in vitro is usually performed in culture medium supplemented with human serum. We compared the secretion of interferon-γ and interleukin-1β as a parameter for lymphocyte and monocyte activation in RPMI 1640 medium supplemented with fetal calf or autologous serum in serum-free medium and protein-free medium. Four different stimulatory mechanisms were tested: phytohemagglutinin, toxic shock syndrome toxin-1 (TSST-1), lipopolysaccharide (LPS), and zinc ions. We found that the optimal stimulatory zinc concentration depended on the total protein content of the medium, whereas the monokine levels were dependent on the concentration of transport proteins such as transferrin. Monokine induction by LPS or TSST-1 were each influenced by the protein and serum composition in a specific manner. Our results show that the differing mechanisms of cytokine induction are influenced by the medium and serum composition in a diverse but specific manner. Serum- or protein-free medium are especially suitable after superantigen challenge when LPS activity needs to be ruled out or after activation by agents with only a weak stimulatory capacity. J. Leukoc. Biol. 57: 904–908; 1995.
Role of YopH in the suppression of tyrosine phosphorylation and respiratory burst activity in murine macrophages infected with Yersinia enterocolitica
Authors: Green Simon P , Hartland Elizabeth L , Robins-Browne Roy M , Phillips Wayne A
Published: January 2018

Tyrosine phosphorylation is an important component of the signaling pathways responsible for the activation of the macrophage respiratory burst. Because the virulence plasmid of Yersinia enterocolitica encodes a phosphotyrosine phosphatase, YopH, it is possible that the pathogenic strategy of Y. enterocolitica involves the disruption of tyrosine phosphorylation in the macrophage leading to inhibition of respiratory burst activity. We have investigated the effects of Yersinia infection on tyrosine phosphorylation and respiratory burst activity in murine bone marrow-derived macrophages. Infection of macrophages with virulent [Ye(pYV+)] but not avirulent [Ye(pYV-)] strains of Y. enterocolitica was found to suppress both tyrosine phosphorylation and respiratory burst activity in response to zymosan. Mutational inactivation of YopH reversed the suppressive effect of Ye(pYV+) on zymosan-induced tyrosine phosphorylation, indicating that YopH is responsible for the dephosphorylation of macrophage phosphotyrosine-containing proteins observed in macrophages infected with the virulent strain of Y. enterocolitica. In contrast, mutational loss of YopH failed to reverse the inhibitory effect of Ye(pYV+) on the zymosan-triggered respiratory burst. We conclude that the inhibition of the macrophage respiratory burst by Y. enterocolitica involves a plasmid-encoded virulence protein(s) other than, or in addition to, YopH J. Leukoc. Biol. 57: 972–977; 1995.
Dissociation of Rac translocation from p47phox/p67phox movements in human neutrophils by tyrosine kinase inhibitors
Authors: Dorseuil Olivier , Quinn Mark T , Bokoch Gary M
Published: January 2018

Cytosolic components of the phagocyte NADPH oxidase (p47phox, p67phox, and Rac2) translocate to the plasma membrane on cell activation where they interact with a membrane-bound cytochrome b to generate superoxide anion. Phosphorylation reactions are known to be important for activity of NADPH oxidase. Translocation of Rac2, p47phox, and p67phox were all enhanced in formyl-Met-Leu-Phe-stimulated neutrophils treated with 50 nM of the protein phosphatase 1/2A inhibitor calyculin A. Rac translocation was blocked by the tyrosine kinase inhibitors genistein (50 μM) and herbimycin (17 μM), whereas movement of p47phox and p67phox were not inhibited. Cell-free analysis of Rac translocation also demonstrated that translocation of p47phox and p67phox were not linked to the movement or availability of Rac2. Thus, Rac2 does not appear to regulate NADPH oxidase by controlling movements of the cytosolic components to the membrane-associated enzyme but may exert its effect at the level of the assembled complex. Tyrosine kinase activity is required for translocation of Rac in the chemoattractant-stimulated human neutrophil. J. Leukoc. Biol. 58: 108–113; 1995.
Hyperglycosylation of eosinophil ribonucleases in a promyelocytic leukemia cell line and in differentiated peripheral blood progenitor cells
Authors: Lee Tiffany H , Li Fei , Rosenberg Helene F
Published: January 2018

We evaluated two independent models of eosinophil differentiation for their ability to synthesize the ribonuclease toxins eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP). Cells from the done 15 subline of HL-60 (human promyelocytic leukemia) produced both EDN and ECP; production of EDN increased in response to butyric acid (BA). CD34+ peripheral blood progenitor cells (PBPCs) grown with cytokines promoting eosinophil differentiation also produced EDN. EDN from both the done 15 and PBPCs was more heterogeneous and heavily glycosylated (~22–45 kDa) than EDN from mature peripheral blood eosinophils (18–25 kDa). The heterogeneity of EDN from the done 15 cells was not altered by endogiycosidase Hf, whereas treatment with peptide-N-giycosidase F (PNGase F) produced a single-band immunoreactive band (~15 kDa). In contrast, only the highest molecular weight forms of EDN from differentiated PBPCs were eliminated by PNGase F (reduced to 22–35 kDa), suggesting the presence of uncharacteristic forms of posttranslational modification. Synthesis of hyperglycosylated proteins has not been previously reported in PBPCs and is a feature shared with tumor cells and cell lines. J. Leukoc. Biol. 58: 49–54; 1995.
Modulation of human microglial cell superoxide production by cytokines
Authors: Chao Chun C , Hu Shuxian , Peterson Phillip K
Published: January 2018

Reactive oxygen intermediates (e.g., superoxide [O2-]) generated by microglia may play a role in host defense and injury within the central nervous system. We investigated the effect of cytokines on human microglial cell O2- production on stimulation with phorbol myristate acetate. Priming of microglial cell cultures with interferon-γ or tumor necrosis factor-α resulted in a dose- and time-dependent enhancement of (O2- production. The priming effects of these cytokines were mediated through a protein kinase C signal transduction pathway. In contrast, astrocytes did not generate detectable O2- on phorbol myristate acetate stimulation. Treatment of microglia with transforming growth factor-β, interleukin-4, or interleukin-10 suppressed in a dose-dependent manner the priming effects of tumor necrosis factor-α and interferon-γ. The results of this study have implications for understanding the mechanisms by which cytokines and microglia contribute to processes of host defense and neurodegeneiration via generation of reactive oxygen intermediates. J. Leukoc. Biol. 58: 65–70; 1995.