bluuebunny/crossref_metadata_2025 huggingface dataset:
Human microglial cells have phenotypic and functional characteristics in common with both macrophages and dendritic antigen-presenting cells
Authors: Ulvestad Elling , Williams Kenneth , Bjerkvig Rolf , Tiekotter Kenneth , Antel Jack , Matre Roald
Published: January 2018

Resting microglia comprise up to 13% of the cells in human central nervous system (CNS) white matter. Their large number and dendritic morphology make them ideally suited to survey the CNS for noxious stimuli. Upon activation microglia gradually lose dendritic processes and transform into typical phagocytic macrophages. Microglia have been implicated as the main antigen presenting cell within the CNS, and appear to be of central importance as effectors and regulators of demyelination. To further characterize the capacity for immune reactivity within the human CNS, we have studied several characteristics of microglia, both in situ and in vitro. We find that human microglia have ultrastructural, phenotypic (CD11c, CD68, acid phosphatase), and functional (FcR and CR mediated phagocytosis) properties typical for cells of the monocyte lineage. Our data indicate that microglia also have properties in common with dendritic antigen-presenting cells. Electron microscopy studies show extended dendritic cell processes on cultured microglia, and microglia are, like dendritic cells, negative for the monocyte markers nonspecific esterase, endogenous peroxidase, CD14, and RFD7. Microglia constitutively express HLA-DR in situ, and express the dendritic cell marker RFD1 upon activation. Coculturing of microglia with CD4+ T cells results in clustering of T cells around microglia and initiation of a mixed lymphocyte reaction, both distinguishing features of dendritic cells. These functional properties of microglia may be of importance for the maintenance of an immunologic response in the CNS, an organ where dendritic cells, in contrast to other organs, have not previously been identified. J. Leukoc. Biol. 56: 732–740; 1994.
Modulation of the chemotactic responsiveness of guinea pig neutrophils to hrIL-8 and fMLP
Authors: Dai Yalei , Holgate Stephen T , Church Martin K , Shute Janis K
Published: January 2018

Neutrophils elicited in the peritoneal cavity of guinea pigs were purified on Percoll gradients and their chemotactic response to hrIL-8 and fMLP measured in vitro. hrIL-8 and fMLP were effective chemoattractants with optimal concentrations of 6 10-9 and 1 10-7 M, respectively. Scatchard analysis revealed approximately 205,000 IL-8 receptors/cell and 34,000 fMLP receptors/cell with K  d values of 4.1 10-9 and 3.3 10-8 M, respectively. At suboptimal concentrations of chemoattractants the response was inhibited by dibutyryl cyclic AMP, histamine, and adenosine in the presence of a phosphodiesterase inhibitor. IL-8 and fMLP induced an increase in cellular cyclic AMP and the response to optimal concentrations of chemoattractants was inhibited by Calphostin C and Ro 31-8220, inhibitors of protein kinase C (PKC). Our results indicate that the chemoattractants activate the same PKC-dependent pathway that is down-regulated by cyclic AMP-dependent mechanisms. J. Leukoc. Biol. 56: 776–783; 1994.
Glycosylphosphatidylinositol-anchored NAD glycohydrolase is released from peritoneal macrophages activated by interferon-γ and lipopolysaccharide
Authors: Han Myung Kwan , Yim Chang-Yeol , An Nyeon-Hyoung , Kim Hyung-Rho , Kim Uh-Hyun
Published: January 2018

We have previously shown that an ectoenzyme, NAD glycohydrolase (NADase) could be solubilized by treatment with bacterial phosphatidylinositol phospholipase C (PIPLC). However, it is unknown whether endogenous PIPLC can cleave this ectoenzyme. In this study, we used mouse peritoneal exudate macrophages which have been known to have relatively high activity of NADase. The results show that release of ecto-NADase was markedly increased when mouse peritoneal macrophages were costimulated with interferon-γ (IFN-γ) and bacterial lipopolysaccharide (LPS), compared to unstimulated cells. This increase was preceded by markedly enhanced activity of endogenous glycosylphosphatidy-linositol phospholipase C (GPIPLC). The cross-reacting determinant (CRD) of the glycosylphosphatidylinositol anchor in released NADase from activated macrophages was detected by immunoblotting with anti-CRD antibody. Taken together, ecto-NADase is released from peritoneal exudate macrophages during IFN-γ/LPS-induced activation and endogenous GPIPLC is involved in the NADase release from the activated macrophages. J. Leukoc. Biol. 56: 792–796; 1994.
Notes from the Incoming Editor
Authors: Oppenheim, M.D. Joost J
Published: January 2018

I have asked myself many times why I agreed to become the Editor-in-Chief of the Journal of Leukocyte Biology. It is a timeconsuming, at times contentious and hazardous position with the potential to lose friends and make enemies.
Ligation of CD23 activates soluble guanylate cyclase in human monocytes via an L-arginine–dependent mechanism
Authors: Paul-Eugène Nathalie , Kolb Jean-Pierre , Sarfati Marika , Arock Michel , Ouaaz Fateh , Debré Patrice , Mossalayi Djavad M , Dugas Bernard
Published: January 2018

Transduction through FcR2/CD23 was analyzed in normal human monocytes using immunoglobulin E (IgE)-anti-IgE immune complexes (IgE ICs) and monoclonal antibodies (mAbs) to CD23. Anti-CD23 mAb and IgE IC triggered a time-dependent increase in cGMP and cAMP in interleukin-4–preincubated (CD23+) but not in unstimulated (CD23-) monocytes. Maximal cGMP and cAMP accumulations were observed 10 and 20 min, respectively, after the onset of CD23 ligation. The increase in cGMP was inhibited with Nω-monomethyl-l-arginine (L-NMMA), which also partially affected cAMP accumulation. Addition of an anti-CD23 mAb Fab fragment inhibited the IgE IC– and the anti-CD23 mAb–induced cGMP and cAMP accumulation, confirming the engagement of CD23. In addition, IgE IC and anti-CD23 mAb induced, at least in some donors, a production of nitrite that was inhibited in the presence of L-NMMA. Taken together, these findings suggest a possible involvement of the nitric oxide synthase pathway in IgE IC–mediated activation of CD23+ monocytes. J. Leukoc. Biol. 57: 160–167; 1995.
Expression of both types of human interleukin-8 receptors on mature neutrophils, monocytes, and natural killer cells
Authors: Morohashi Hirohisa , Miyawaki Toshio , Nomura Hideki , Kuno Kouji , Murakami Seishi , Matsushima Kouji , Mukaida Naofumi
Published: January 2018

cDNA cloning revealed the presence of two related but distinct types of human interleukin-8 (IL-8) receptors, type I (type A) and type II (type B). By immunizing rabbits with glutathione-S-transferase fused with the NH2-terminal domain of each type of IL-8 receptor, we prepared polyclonal antibodies that specifically recognized the NH2-terminal domain of each type of IL-8 receptor. Immunofluorescence analysis of human peripheral blood leukocytes demonstrated that mature granulocytes except eosinophils express both types of IL-8 receptors. A majority of monocytes and CD16+ natural killer (NK) cells in peripheral blood were stained with both antibodies, whereas CD3+ T or CD20+ B lymphocytes in peripheral blood or CD34+ cells in cord blood were not stained with either antibody. These results suggest that both types of human IL-8 receptors were coordinately and selectively expressed in mature granulocytes, monocytes, and CD16+ NK cells. J. Leukoc. Biol. 57: 180–187; 1995.
Distinct granzyme expression in human CD3- CD56+ large granular- and CD3- CD56+ small high density-lymphocytes displaying non-MHC-restricted cytolytic activity
Authors: Smyth Mark J , Browne Kylie A , Kinnear Beverley F , Trapani Joseph A , Warren Hilary S
Published: January 2018

Cultured natural killer (NK) cells derived from CD3- CD56+ high-density small lymphocytes (HDLs) exhibit similar morphology and high levels of non-major histocompatibility complex-restricted (NK) cytotoxicity equivalent to those of cultured NK cells from CD3- CD56+ low-density large granular lymphocytes (LGLs). To examine the similarities and differences between NK cells from HDLs and NK cells from LGLs, we investigated the expression of three distinct members of the granule serine protease (granzyme) family within cultured CD3- CD56+ LGLs and HDLs. CD3- subpopulations of nonadherent peripheral blood mononuclear cells, LGLs (density < 1.063 g/ml), and HDLs (density >1.063 g/ml) were stimulated to proliferate in culture. The cultured cells from each population were entirely CD3- CD56+ and were indistinguishable in terms of their increased granularity and size once activated. All cultured CD3- CD56+ LGLs and HDLs displayed cytolytic activity against K562 and immunoglobulin-coated P815. Western analysis detected perforin in both cultured LGL and HDL populations. Cultured HDLs and LGLs both expressed BLT-esterase activity and human granzyme A mRNA. Granzyme B mRNA and protein and Asp-ase activity were detected in unstimulated and cultured LGLs and cultured HDLs. By contrast, unstimulated HDLs did not express significant levels of granzyme B. High levels of Hu-Met-1 granzyme mRNA and Met-ase activity were detected only in cultured LGLs. Thus, despite the development of large granular morphology during proliferation, interleukin-2 cultured CD3- CD56+ HDLs display a different pattern of granzyme expression from CD3- CD56+ LGLs. These data also further suggest an unusually restricted expression of the Hu-Met-1 granzyme in LGLs. J. Leukoc. Biol. 57: 88–93; 1995.
Temporal and spatial changes of quinolinic acid immunoreactivity in the immune system of lipopolysaccharide-stimulated mice
Authors: Espey Michael Graham , Moffett John Rodney , Namboodiri M A Aryan
Published: January 2018

Quinolinic acid (Quin), a metabolite of tryptophan, is a neurotoxin that has been implicated in a variety of neuropathologic disorders that have immune components. The goal of this study was to characterize the changes in the cellular localization of Quin immunoreactivity in a paradigm of immune stimulation with lipopolysaccharide (LPS) in vivo to provide a basis for further studies on the physiological role of Quin in the immune system. Intraperitoneal LPS injection significantly increased Quin immunoreactivity (IR) in lymphoid tissues within 24 h. Spatial changes in splenic Quin-IR demonstrated a shift from the periarterial lymphoid sheaths to the follicles before returning to control levels by 72 h post-LPS. The strongly Quin-IR cells were tentatively identified as interdigitating dendritic cells and macrophages. Only minimal Quin-IR was detected in liver and lung, even under conditions of LPS stimulation combined with tryptophan loading. These data emphasize the temporally and spatially specific nature of Quin-IR changes in lymphoid tissues under conditions of immune stimulation and raise the possibility that Quin may have an immunomodulatory function. J. Leukoc. Biol. 57: 199–206; 1995.
Effects of oxo and dihydro metabolites of 12-hydroxy-5,8,10,14-eicosatetraenoic acid on chemotaxis and cytosolic calcium levels in human neutrophils
Authors: Powell William S , Hashefi Mandana , Falck J R , Chauhan Kamlesh , Rokach Joshua , Wang Steven S , Mills Elaine , MacLeod R John
Published: January 2018

One of the pathways of metabolism of leukotriene B4 (LTB4) and 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) in leukocytes is oxidation of the 12-hydroxyl group, followed by reduction of the 10,11-double bond. In the case of 12R-HETE and 12S-HETE, this results in the formation of 12-oxo-ETE, 10,11-dihydro-12-oxo-ETE, and the 12R and 12S isomers of 10,11-dihydro-12-HETE (i.e., 12R-HETrE and 12S-HETrE). We investigated the effects of metabolites of 12-HETE formed by this pathway on cytosolic calcium levels and chemotaxis in human neutrophils. Of the above series of metabolites, 12S-HETrE (which has the same absolute stereochemistry at C-12 as 12R-HETE) was the most potent in stimulating both cytosolic calcium levels and chemotaxis. It was slightly less potent than 12R-HETE, consistent with the concept that reduction of the 10,11-double bond results in a loss of biological activity on neutrophils. The effect of 12S-HETrE on calcium levels was blocked by preincubation of these cells with LTB4, suggesting that it acted by stimulating the LTB4 receptor. 12R-HETrE was about 20 times less potent than its 12S isomer in stimulating cytosolic calcium in neutrophils and was also less active as a chemotactic agent. Oxidation of the 12-hydroxyl group to an oxo group resulted in a further loss of biological activity. 12-OxoETE, 8-trans-12-oxo-ETE, and 12-oxo-ETrE had only modest effects on cytosolic calcium levels at concentrations as high as 10 μM and did not display detectable chemotactic activity. However, 12-oxo-ETE and its 8-trans isomer inhibited calcium responses to LTB4 by about 40%. It is concluded that reduction of the 10,11-double bond of 12-HETE results in a slight loss of biological activity on neutrophils, whereas oxidation of the 12-hydroxyl group results in a considerably greater loss of activity. J. Leukoc. Biol. 57: 257–263; 1995.
Modulation of IL-8 receptor expression on purified human T lymphocytes is associated with changed chemotactic responses to IL-8
Authors: Xu Luoling , Kelvin David J , Ye George Q , Taub Dennis D , Ben-Baruch Adit , Oppenheim Joost J , Wang Ji Ming
Published: January 2018

Interleukin-8 is a member of the chemokine superfamily and is a major mediator of acute inflammation. Although IL-8 has been reported by some laboratories also to be a chemoattractant for T lymphocytes, this has been difficult to confirm and remains a controversial issue. By using freshly purified human T cells (90–95% CD3+), we could demonstrate consistent directional migration of T cells to recombinant human IL-8. IL-8 was as potent as RANTES, MIP1α, and M1P1β in inducing T cell chemotaxis. Highly purified T cells, however, incubated at 37° for more than 12 h or cultured over-night with anti-CD3 antibody cross-linked to plastic dishes, showed a markedly reduced capacity to migrate in response to IL-8. This was associated with a decrease in binding of radioiodinated IL-8 to T cells. Northern blot and polymerase chain reaction analyses showed that freshly purified T cells expressed mRNA for both IL-8 receptor type A and type B. Steady-state levels of mRNA for the IL-8RA and IL-8RB genes were also reduced by incubation of the cells with or without anti-CD3 for 12 h at 37°. These results indicate that T cells are indeed one of the target cell populations for IL-8. The regulation of IL-8 receptor expression on T lymphocytes may contribute to the pathophysiological role of IL-8 in inducing the homing and infiltration of T cells. J. Leukoc. Biol. 57: 335–342; 1995.
Monoclonal antibody EG2 reactivity cannot be used as an immunohistochemical activation marker for eosinophils
Authors: Jahnsen Frode L , Brandtzaeg Per
Published: January 2018

To the Editor: Rosenberg and Tiffany recently described [1] that the presumed “activation-specific” antibody EG2 recognized only one (the smallest) of three glycosylated forms (18 kDa, 20 kDa, and 22 kDa) of eosinophil cationic protein (ECP). On the basis of this finding they proposed two alternative ways to explain how the epitope for EG2 was accessible only in activated eosinophils. They suggested that deglycosylation of ECP could occur in conjunction with activation and secretion; this was unlikely though because the 18-kDa form constitutes a substantial fraction of ECP (by Western blotting) even in resting eosinophils [1, 2]. Then they speculated that the 18-kDa form of ECP might be functionally undetectable prior to secretion.1 We have recently shown, however, that cellular EG2 reactivity does not reliably discriminate between resting and activated eosinophils by immunocytochemistry and immunohistochemistry [3]. All blood eosinophils from healthy individuals were strongly EG2-positive when fixed in formalin, and a substantial fraction reacted in the unfixed or acetone-fixed state as well, although with variable intensity. Even after maximum activation (phorbol ester) EG2 could not discriminate eosinophils from their resting counterparts [4].
Monoclonal antibody EG2 reactivity: the need for clarification
Authors: Jahnsen Frode L , Brandtzaeg Per
Published: January 2018

A final comment: We regret that Rosenberg and Tiffany in their response (1) apparently considered our letter (2) about immunohistochemical reactivity of the monoclonal antibody EG2 as criticism of their excellent work (3). Our intention was simply to help explain their biochemical findings in terms of immunocytochemical and immunohistochemical staining performance of EG2 in the light of recent information (4).
Induction of macrophage binding to virally infected cells by low levels of lipopolysaccharide
Authors: Derrick Steven C , LeBlanc Paul A
Published: January 2018

Bone marrow culture-derived macrophages (BMCM) and vesicular stomatitis virus-infected BALB/e-3T3 cells (3T3-VSV) were used to determine whether macrophages could be activated to bind virally infccted cells. Although lipopolysaccharide (LPS)-activated BMCM bound some uninfected BALB/c-3T3 cells, the number of targets that were bound increased with increasing times between infection and assay. Fur thermore, LPS-activated BMCM bound more 3T3-VSV cells than did control macrophages. As more targets were added, the number of targets bound by the unactivated macrophagcs remained relatively level. However, the number of targets bound by the activated macrophages increased with increasing concentrations of added targets until they reached a plateau that was eight times greater than that bound by the unactivated BMCM. When BMCM were exposed to LPS for 24 h before assay, they lost both their ability to bind to 3T3-VSV and their cyto lytic activity against those targets. However, as when using P815, a standard tumor target, the acquisition of binding of 3T3-VSV could be separated from macrophage cytolytic activity against those targets. The amount of LPS required to activated BMCM for increased binding of 3T3-VSV cells was 10–100 times lower than that needed to induce cytolytic activity for 3T3-VSV cells. Each of these values was approximately 100-fold lower than the amount of LPS required to induce the corresponding ac tivity (binding or cytotoxicity) by using P815 targets. J. Lcukoc. Biol 57: 569–573; 1995.
Intracellular multiplication of Legionella pneumophila in HL-60 cells functionally differentiated in response to 22-oxacalcitriol
Authors: Matsuzaki Miwako , Shimamoto Yoshinori , Watanabe Masayuki , Kukita Akiko , Yoshida Shin-ichi , Tadano Jutaro
Published: January 2018

The agent 1,25-dihydroxy vitamin D3 (D3) induces the differentiation of HL-60 human leukemia cells into functional monocyte-like cells that can support the intracellular multiplication of Legionella pneumophila. 22-Oxacalcitriol (OCT), a synthetic analogue of D3, ex hibits greater differentiation-inducing activity than D3 in WEHI-3 mouse leukemia cells and has been suggested to be clinically more useful because of its lower hypercalcemic activity. The abilities of OCT and D3 to induce the functional differentiation of human leukemia HL-60 cells have now been investigated. OCT induced the differentiation of HL-60 cells into monocyte-like cells to a similar extent as D3. Thus, both OCT and D3 increased (1) the surface expression of CD11b, CD11c, CD14, and CD35; (2) nonspecific esterase staining; and (3) phagocytic ac tivity toward fluorescent beads. HL-60 cells differentiated in response to OCT also supported the intracellular mul tiplication of L. pneumophila. Activation of both OCT-and D3-treated HL-60 cells with human recombinant interferon-γ (IFN-γ) for 24 h before infection markedly inhibited L. pneumophila multiplication. IFN-γ activation enhanced superoxide anion generation by D3-treated HL-60 cells but not by OCT-treated HL-60 cells, suggesting that the inhibition of L. pneumophila multiplication in IFN-γ-activated cells is independent of superoxide generation. Finally, D3, but not OCT, markedly stimu lated the formation of osteoclast-like multinudeated cells from mouse bone marrow cells, consistent with the lower hypercalcemic activity of OCT. J. Leukoc. Biol. 57: 574–580; 1995.
Effects of endotoxin on expression of VLA integrins by human bronchoalveolar lavage macrophages
Authors: Kang Yuan-Hsu , Lee Che-Hung , Brummel Sara E , Newball Harold H , Forrester Joe
Published: January 2018

Endotoxin (lipopolysaccharide, LPS) is known to induce inflammatory responses, such as mono cyte/macrophage adherence, migration, and accumulation. Recruitment and accumulation of macrophages during infection and inflammation are regulated by integrin-mediated cell-extracellular matrix interactions. In the present report, we studied the effects of LPS on the expression of VLA-5 (α  5  β  1), VLA-3 (α  3  β  1), and VLA-2 (α  2  β  1  ) integrins and fibronectin (FN) by human alveolar macrophages in an attempt to understand the mechanism by which LPS regulates macrophage adhesion to matrix proteins. Bronchoalveolar lavage macrophages were treated with varying concentrations of Escherichia coli LPS for different times and evaluated for expression of the integrins and FN by immunofluorescence, immunoelectron microscopy, autoradiography, and radioim munoassay. Immunofluorescent and immunoelectron microscopic observations showed that VLA integrins were constitutively expressed on the cell surface and con centrated on the microvilli and pseudopodia of the mac rophages. The effects of LPS on expression of the inte grins were dose and time related. VLA-5 expression was increased after 30 min of stimulation by LPS, suggesting that LPS may induce rapid secretion of the integrin. However, incubations with LPS longer than 30 min decreased VLA-5 expression in a dose-dependent pattern. LPS also caused dose-related decreases in the expression of VLA-3 and VLA-2 integrins and increases of intracel lular FN 24 h after stimulation. The results suggest that a prolonged exposure to LPS may impede VLA inte grin-mediated migration and result in local accumulation of macrophages in the lung. J. Leukoc. Biol. 57: 624–634; 1995.
Immune complexes increase nitric oxide production by interferon-γ-stimulated murine macrophage-like J774.16 cells
Authors: Mozaffarian Neelufar , Berman Joan W , Casadevall Arturo
Published: January 2018

Murine macrophage-like J774.16 cells were tested for changes in nitric oxide production upon incu bation with immune complexes. Cryptococcus neoformans capsular polysaccharide and polysaccharidespecific monoclonal antibodies were added to J774.16 cells in the presence and absence of recombinant murine interferon-γ (IFN-γ). The effect of immune complexes on nitrite synthesis was both concentration dependent and isotype dependent. In the presence of IFN-γ, immune complexes of IgGl, IgG2a, IgG2b, or IgG3 isotype in creased nitrite levels, whereas complexes of IgM isotype did not. Immune complexes did not alter nitrite production by unstimulated macrophages. Antibody alone, anti gen alone, and antigen with irrelevant IgGl antibody did not augment nitrite formation, either in the presence or absence of IFN-γ. indicating a requirement for FCγR cross-linking. These results suggest that IgG isotypes may offer additional protection against pathogens by enhancing macrophage nitric oxide production, J. Lcukoc. Biol. 57: 657–662; 1995.
Thrombopoietin, the c-mpl ligand, is a major regulator of platelet production
Authors: Broudy Virginia C , Kaushansky Kenneth
Published: January 2018

Hematopoiesis is regulated by a family of glycoproteins, the hematopoietic growth factors. Although the cytokines that influence the late stages of granulopoiesis (granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor) or erythropoiesis (erythropoietin) have been identified, characterized, and cloned, and have been in clinical use since that late 1980’s, the cytokine that stimulates thrombopoiesis had remained elusive. By using strategies based on the c-mpl receptor, several groups have recently succeeded in purifying and cloning thrombopoietin [1–6]. This study outlines the cloning and biological characterization in vitro and in vivo of thrombopoietin, a major regulator of platelet production. J. Leukoc. Biol. 57: 719–725; 1995.
Cytokine production profile of early thymocytes and the characterization of a new class of chemokine
Authors: Kelner Gregory S , Zlotnik Albert
Published: January 2018

By using a variety of molecular techniques, we have characterized the cytokine profile of pro-T cells. During this characterization we cloned a novel cytokine that has chemotactic activities. We have designated this cytokine lymphotactin. The structural analysis, chromosomal location, and biological properties of lymphotactin provide strong evidence that this cytokine represents a new class of chemokine. J. Leukoc. Biol. 57: 778–781; 1995.
Receptors, signal transduction, and spectrum of action of monocyte chemotactic protein-1 and related chemokines
Authors: Sozzani Silvano , Locati Massimo , Zhou Dan , Rieppi Monica , Luini Walter , Lamorte Giuseppe , Bianchi Giancario , Polentarutti Nadia , Allavena Paola , Mantovani Alberto
Published: January 2018

Chemokines are a bipartite family of chemotactic proteins that bear the structural hallmark of four cysteine residues, the first two of which are in tandem. The spectrum of action of C-C chemokines, monocyte chemotactic protein-1 (MCP-1), MCP-2, and MCP-3, in particular, encompasses, in addition to monocytes, other leukocyte populations. Evidence is presented that MCP-1, MCP-2, and MCP-3 are active on natural killer cells. Available information on receptor usage by MCP-1 and related chemokines and signal transduction pathways is reviewed. A better understanding of signaling mechanisms will provide a new basis for therapeutic strategies. J. Leukoc. Biol. 57: 788–794; 1995.
Neutrophils are essential for resolution of primary and secondary infection with Listeria monocytogenes
Authors: Rakhmilevich Alexander L
Published: January 2018

The role for neutrophils in the resolution of primary and secondary infection with Listeria monocytogenes was studied. The results show that although control mice started to clear Listeria from their spleens and livers between days 2 and 4 of sublethal primary infection and eradicated bacteria in 2 weeks, mice given a specific granulocyte-depleting antibody (RB6-8C5) on days 4 or 6 of infection developed lethal listeriosis. Likewise, treatment of immunized mice with RB6-8C5 monoclonal antibody abolished their acquired ability to resolve a lethal challenge infection. The results demonstrate that neutrophils are necessary for the resolution of secondary and primary Listeria infection. J. Leukoc. Biol. 57: 827–831; 1995.
Slow Ca2+ waves in large myeloid cells as a result of a diffusible cytosolic factor
Authors: Roberts G M , Davies E V , Hallett M B
Published: January 2018

In the work reported here evidence is provided that shows the slow wave of Ca2+ large neonatal myeloid cells provoked by formyl-Met-Leu-Phe was generated by spatially delayed Ca2+ influx. Evidence is provided that the delay in Ca2+ influx was the result of diffusion of a factor from the Ca2+ storage site, which is responsible for Ca2+ channel opening. The location of the Ca2+ release site was correlated with a region near the nucleus, probably a specialized region of endoplasmic reticulum. It is proposed that similar mechanisms of Ca2+ signaling occur in mature myeloid cells, such as neutrophils, but on a shorter time scale as a consequence of their smaller size. J. Leukoc. Biol. 57: 837–841; 1995.
Profile of immunoglobulin heavy chain variable gene repertoires and highly selective detection of malignant clonotypes in acute lymphoblastic leukemia
Authors: Greenberg Steven J , Choi Youngnim , Ballow Mark , Du Tian-Long , Ward Pamela M , Rickert Michael H , Frankel Stanley , Bernstein Steven H , Brecher Martin L
Published: January 2018

The predominant B cell immunoglobulin heavy chain variable gene (IgH-V) usage and the uniquely rearranged, clonotype-specific variable-diversity-joining region gene (VDJ) sequences were identified in patients with B cell acute lymphoblastic leukemia (B-ALL) using a novel DNA-based gene amplification strategy. The approach allows a thorough and sensitive determination of the number of clonal leukemic IgH rearrangements and their precise V gene usage. This strategy may be applied in the detection of minimal residual disease, in surveillance after induction of disease-free states, and in analyzing the effectiveness of purging autologous bone marrow of malignant clones. An initial primary polymerase chain reaction (PCR), directed by an IgH-J generic primer and a complement of family-specific IgH-V primers, defined the major B cell IgH-V gene usage. Use of an IgH-J generic primer supplanted the use of a constant region primer anchor and thus eliminated the need to target mRNA by the traditional RNA reverse transcription–PCR amplification method. Monoclonality of rearranged VDJ bands was further substantiated by high-resolution denaturant gel electrophoretic analysis. The predominant amplified bands were subcloned and sequenced. By sequencing through VDJ juxtaposed regions, that is, the third complementarity-determining region, clonotype-specific primers were developed and used in a secondary clonotype primer-directed PCR (CPD-PCR) to detect, with extreme sensitivity and specificity, a unique B cell clone. Analysis of the products of the CPD-PCR permitted the detection of a single malignant cell among 1 million polyclonal cells and superseded the constraints of prior studies that have provided a limited evaluation of family variable gene repertoire usage. Leukemic clonal rearrangements were detected in 100% of the eight cases of pediatric and two cases of adult B-ALL studied. Two or more clonal IgH-VDJ amplified sequences were observed in 50% of the B-ALL bone marrows analyzed. In two cases, clonotype-specific oligodeoxynucleotide primers, derived from B-ALL VDJ sequences, directed the secondary CPD-PCR, and disease activity was monitored after chemotherapy and allogeneic bone marrow transplantation. J. Leukoc. Biol. 57: 856–864; 1995.
Poxvirus-based vaccine candidates for cancer, AIDS, and other infectious diseases
Authors: Perkvs Marion E , Tartaglia James , Paoletti Enzo
Published: January 2018

Over the past 12 years, the poxvirus vector technology has provided scientists with valuable reagents to achieve high-level expression of proteins, to address questions of structure-function relationship of specific polypeptides, to investigate the immunobiology of specific pathogens, and to develop recombinant vaccine candidates. It is this last role that has drawn enthusiasm from the medical community because of the potential this technology has to provide novel approaches for addressing urgent needs in human and veterinary medicine. From one perspective, the safety issues surrounding the use of vaccinia-based vaccine candidates have been addressed with the development of the NYVAC and ALVAC vectors. Evaluation of these novel poxvirus vectors are in progress to determine their potential impact on cancer and infectious disease. J. Leukoc. Biol. 58: 1–13; 1995.
Binding of Gc globulin (vitamin D binding protein) to C5a or C5a des Arg is not necessary for co-chemotactic activity
Authors: Kew Richard R , Mollison Karl W , Webster Robert O
Published: January 2018

Gc globulin (vitamin D binding protein) has been shown to augment significantly die leukocyte chemotactic activity of the activated complement peptides C5a and C5a des Arg. However, the mechanism of chemotaxis enhancement is not known. Natural C5-derived peptides contain a carbohydrate side chain that comprises approximately 25% of the mass of the 11-kDa peptides. Previous studies have demonstrated that Gc globulin binds to C5-derived peptides via sialic acid residues on this carbohydrate side chain. The necessity of this carbohydrate side chain for chemotaxis enhancement by Gc globulin was investigated by using both natural (glycosylated) and recombinant (carbohydrate-free) peptides. The dose-response curves of neutrophil chemotaxis to recombinant C5a or C5a des Arg plus Gc globulin were identical to those observed with the naturally derived peptides, despite the fact that natural C5a bound to Gc globulin while the recombinant C5a failed to bind this protein. Moreover, neutrophils pretreated with Gc globulin then washed before addition to the chemotaxis assay displayed significantly enhanced movement to C5a alone. These results indicate that the binding of C5a/C5a des Arg to Gc globulin is not necessary for their chemotactic activity to be augmented. Furthermore, these results demonstrate that the co-chemotactic activity of Gc globulin is generated on the cell surface, independent of C5a binding to its receptor. J. Leuhoc. Biol. 58: 55–58; 1995.
Murine Th1 and Th2 cell clones differentially regulate macrophage nitric oxide production
Authors: Taub Dennis D , Cox George W
Published: January 2018

Previous studies have demonstrated that the combination of the T helper cell 1 (Thl)-derived cytokines interleukin (IL)-2 and interferon (IFN)-γ induces nitric oxide (NO) production and tumor cytolysis by mouse peritoneal macrophages and the mouse macrophage cell line ANA-1 in vitro. Conversely, the Th2-derived cytokine IL-4 inhibits IL-2 and IFN-γ-induced NO production and tumor cytolysis by ANA-1 macrophages. To examine the paracrine regulatory effects of Thl and Th2 cells on macrophages, various mouse T cell clones were tested for their ability to regulate NO production by mouse peritoneal macrophages or ANA-1 macrophages. Antigen, superantigen, and mitogen stimulated Th1 cells but not Th2 cells induced NO production by macrophages. Supernatants from these activated Th1 clones also induced NO production by peritoneal macrophages and ANA-1 macrophages. Neutralization analysis using monoclonal anticytokine antibodies revealed that both IL-2 and IFN-γ production by activated Th1 cells were required for the production of NO by macrophages. Co-culture studies using a panel of Th2 cell clones that share the same antigen specificity revealed that these cells suppressed Th1-mediated macrophage activation. The Th2-mediated impairment of Th1-induced NO production was primarily due to the secretion of IL-4. IL-4 appeared to have a direct effect on macrophage activation because neither mitogen-induced proliferation of Th1 cells nor cytokine production by Th1 cells were affected by IL-4. Overall, these results suggest that a potent paracrine regulatory network involving Th1 cells and Th2 cells may control the activation of macrophages for NO production and antitumor cytotoxicity. J. Leukoc. Biol. 58: 80—89; 1995.
Gamma/delta T lymphocytes in viral Infections
Authors: Wallace Marianne , Malkovsky Miroslav , Carding Simon R
Published: January 2018

T lymphocyte progenitors differentiate into two distinct T cell lineages. Although the αβ and γδ T cell lineages resemble each other phenotypically and functionally, there are some striking differences. Some γδ T cells recognize, similarly to αβ T cells, peptides presented by major histocompatibility complex (MHC) proteins or MHC-like molecules. However, there are γδ T cells that recognize MHC molecules in a fundamentally different manner in comparison with αβ T cells. Also in contrast to aLβ T lymphocytes, many γδ T cells are capable of recognizing nonpeptide antigens. Most responses of γδ T cells appear to be directed against microbial pathogenic agents including bacteria, parasites, and viruses. In particular, the potent cytotoxic responses of γδ T cells against cells infected with, for example, herpesviruses or lentiviruses may be essential for the overall antiviral defense of vertebrates. The analysis of antiviral immunosurveillance by γδ T cells is crucial for understanding the unique biological role of this lymphocyte subset.
Costimulation of CD4+ and CD8+ T cells through CD26: The ADA-binding epitope is not essential for complete signaling
Authors: De Meester Ingrid A , Kostens Luc L , Vanham Guido L , Vanhoof Greet C , Vingerhoets Johan H , Gigase Paul L , Scharpé Simon L
Published: January 2018

It was previously shown that CD26 (DPP IV, EC 3.4.14.5) is a binding site for adenosine deaminase (ADA, EC 3.5.4.4) on T cells and that costimulation by some anti-CD26 monoclonal antibodies (mAbs) and anti-CD3 induces CD4+ T cell proliferation. The CD26 epitopes involved in costimulation, the precise sequence of the events preceding proliferation, and the response of CD8+ compared with CD4+ T cells to CD26 were not extensively studied. We therefore compared the effects of the novel TA5.9 anti-CD26 mAb, recognizing an ADA-binding epitope, and the clearly distinct anti-Ta1 reference anti-CD26 mAb for their costimulatory properties in various T cell subsets. Both purified CD4+ and CD8+ T cells proliferated upon costimulation with anti-CD3 and either anti-CD26 mAb, but anti-TA5.9 mAb induced a more potent response than anti-Tal. Either anti-CD26 mAb, together with anti-CD3, caused a similar sequential up-regulation of CD69, CD25 (IL-2Rα), and CD71 (transferrin receptor) expression on CD4+ and CD8+ T cells. The activation markers appeared faster on the CD45R0+ than on the CD45R0− subsets. After costimulation, CD4+ T cell cultures contained significant amounts of the Th1 cytokines EL-2, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α). In CD8+ T cell cultures relatively more IFN-γ and TNF-α but almost no IL-2 was measured after triggering of CD3 and CD26. Our data demonstrate that the recognition of the ADA-binding epitope is not a prerequisite for the costimulatory capacity of anti-CD26 mAbs. Both CD4+ and CD8+ T cells and their CD45R0−and CD45R0+ subsets are sensitive to various aspects of activation via CD26, but the magnitude and/or kinetics differ according to the anti-CD26 used and the T cell subset studied.
Involvement of nitric oxide in the regulation of peripheral blood leukocyte counts
Authors: Geffner Jorge R , Trevani Analía S , de D’Elia Isabel , Diament Miriam , Klein Danka , Giordano Mirta
Published: January 2018

A role for nitric oxide (NO) in the regulation of blood leukocyte numbers was examined in BALB/c mice by employing the NO synthase inhibitor NG-nitro L-arginine methyl ester (L-NAME). Treatment of animals with a single dose of 50 mg/kg body wt caused a dramatic increase in the number of circulating neutrophils and a moderate decrease in the number of circulating lymphocytes. These effects were partially reversed by the simultaneous inoculation of L-arginine (250 mg/kg body wt.) but not by D-arginine. A second NO synthase inhibitor, NG-nitro L-arginine, induced changes comparable to those elicited by L-NAME. Because catecholamines and glucocorticoids are well-known modulators of blood leukocyte counts, experiments were carried out in adrenalectomized mice. It was found that adrenalectomy did not modify the increase in the number of circulating neutrophils induced by L-NAME but completely prevented the decrease of circulating lymphocytes. Taken together, these findings support the hypothesis that NO plays an important role in the regulation of the peripheral blood number of neutrophils and lymphocytes, and that this function involves, in each case, the participation of different mechanisms.
Expression of leukocyte differentiation antigens during the differentiation of HL-60 cells induced by 1,25-dihydroxyvitamin D3: comparison with the maturation of normal monocytic and granulocytic bone marrow cells
Authors: Brackman Damien , Lund-Johansen Fridtjof , Aarskog Dagfinn
Published: January 2018

1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] induces monocytic differentiation of the HL-60 leukemic cell line. The present study investigated whether and to what extent this differentiation resembles the normal maturation of monocytic cells in the bone marrow. Multidimensional flow cytometry was used to identify changes in antigen expression that occur in normal bone marrow cells at distinct stages of monocytic and granulocytic maturation. HL-60 cells were analyzed in the same manner after exposure to 1,25-(OH)2D3 to determine whether the hormone induces a similar sequence of phenotypic changes. In the leukemic cells, monocytic features were sequentially induced and several maturational steps could be resolved. CD14, CD32, CD53, CD15, CDw65, CD29, CD16, and CD66b were modulated in 1,25-(OH)2D3-induced HL-60 cells as in the normal monocytic maturational pathway. Differences were observed for CD15s and CDw17. The expression pattern of CD44 during differentiation of HL-60 cells resembled that in granulocytic cells. The results therefore suggest that 1,25-(OH)2D3 induces a differentiation program in HL-60 cells that in many ways resembles that of normal monocytic cells in the bone marrow but also carries elements of the granulocytic pathway.
Autocrine regulation of collagenase gene expression by TNF-α in U937 cells
Authors: Callaghan Máire M , Lovis Rosa M , Rammohan Chidambaram , Lu Yang , Pope Richard M
Published: January 2018

Tumor necrosis factor α (TNF-α) has been shown to induce the production of interstitial collagenase by fibroblasts and chondrocytes. We investigated the role of TNF-α in collagenase gene expression by U937 monocyte/macrophage cells. Transcription of the TNF-α gene was observed after 0.5 h of phorbol myristate acetate (PMA) stimulation. Collagenase mRNA expression was not observed until 5–7 h of activation with PMA. TNF-α was detected in the culture supernatants 2–3 h before transcription of the collagenase gene. Neutralization of TNF-α protein with anti-TNF-α antibodies significantly reduced collagenase mRNA expression. Protein kinase C (PKC) and protein tyrosine kinase (PTK) inhibition essentially abolished both PMA-induced TNF-α protein secretion and collagenase mRNA expression. Collagenase gene expression induced by exogenous TNF-α in U937 cells stimulated with a suboptimal concentration of PMA was sup-pressed by PTK, but not PKC, inhibition. The pyrrolidine derivative of dithiocarbamate, a potent inhibitor of nuclear factor-κB (NF-κB) activation, resulted in a marked reduction in collagenase gene transcription, however, no reduction of TNF-α secretion was noted. Anti-TNF-α antibodies inhibited PMA-induced NF-κB activation. These observations demonstrate an important role for TNF-α in the autocrine regulation of collagenase gene expression by U937 cells. Additionally, TNF-α-induced PTK and NF-κB activation were important in collagenase gene expression in this cell line.
Human monocyte chemotactic proteins-2 and -3: structural and functional comparison with MCP-1
Authors: Proost Paul , Wuyts Anja , van Damme Jo
Published: January 2018

Structurally, the monocyte chemotactic proteins MCP-1, -2, and -3 form a subfamily of the C-C or β-chemokines. Like other chemokines, MCPs are produced by a variety of cells on stimulation with cytokines (interleukin-1, tumor necrosis factor-α, interferon-γ), bacterial and viral products or mitogens. MCP-1 levels are enhanced during infection and inflammation, which are characterized by leukocyte infiltration. In vitro, MCPs are chemotactic for a distinct spectrum of target cells and show different specific biological activities depending on the cell type and the chemokine tested. MCP-3 has the broadest range in that it activates monocytes, dendritic cells, lymphocytes, natural killer cells, eosinophils, basophils, and neutrophils. The most sensitive cells to all three MCPs are lymphocytes and monocytes. MCP-1 is a potent basophil activator but does not attract eosinophils, whereas, at higher concentrations, MCP-2 also stimulates both eosinophils and basophils. The signal transduction of MCPs on monocytes involves at least two G protein-linked C-C chemokine receptors: C-C CKR-1 binds MCP-3 and C-C CKR-2 binds MCP-1 and MCP-3 but not MCP-2. Receptor binding leads to enhanced [Ca2+]i for all chemokines except for MCP-2.
Novel insight into molecular mechanism of endotoxin shock: biochemical analysis of LPS receptor signaling in a cell-free system targeting NF-κB and regulation of cytokine production/action through β2 integrin in vivo
Authors: Mukaida Naofumi , Ishikawa Yuji , Lkeda Naoki , Fujioka Nakaba , Watanabe Shun-ichi , Kuno Kouji , Matsushima Kouji
Published: January 2018

Lipopolysaccharide (LPS), a constituent of gram-negative bacteria cell wall, plays an essential role in the pathogenesis of septic shock by generating endogenous mediators such as cytokines, nitrous oxide, superoxide anions, and lipid mediators. In vitro, LPS induces the transcription of a set of genes involved in inflammatory reactions by activating several types of transcription factors, particularly nuclear factor-κB (NF-κB). An analysis of NF-κB activation using a cell-free system demonstrated that two pathways converge to activate NF-κB; one is staurosporine-sensitive, the other is staurosporine-insensitive and tyrosine kinase-dependent. Furthermore, the latter pathway culminates in IκBaL phosphorylation at serine/threonine residues in its carboxyl-terminal acidic region with dissociation of IκBα from NF-κB, thereby activating NF-κB. The requirement for the phosphorylation at this site was confirmed by the specific inhibition of NF-κB activation in a cell-free system by the synthetic peptide corresponding to this site. The in vivo administration of an anti-CD18 antibody prevented elevation of plasma tumor necrosis factor (TNF) levels and acute lethality induced by injection of a low dose of LPS into Propionibacterium acnes-primed rabbits or by the administration of a single high dose of LPS into animals. Anti-CD18 also prevented acute lethality induced by one of the main mediators of endotoxin shock, TNF-α. Furthermore, an antibody to a ligand for CD18, intercellular adhesion molecule-1, also prevented TNF-induced shock as well as endotoxin shock in rabbits. These observations suggest that the interaction between leukoytes and endothelium through β2. integrin adhesion molecules may be of primary importance in mediating LPS signals in vivo.
Role of CSBP/p38/RK stress response kinase in LPS and cytokine signaling mechanisms
Authors: Lee John C , Young Peter R
Published: January 2018

A new member of the mitogen-activated protein kinase family, alternatively termed CSBP, p38, or RK, has been identified independently by several laboratories recently. Activation of this novel protein kinase via dual phosphorylation has been observed in different cell systems upon stimulation by a wide spectrum of stimuli, such as physicochemical stress and treatment with lipopolysaccharide or proinflammatory cytokines such as interleukin-1 and tumor necrosis factor. Furthermore, CSAID™ cytokine biosynthesis inhibitors have now been determined to be potent and selective inhibitors of CSBP/p38/RK kinase activity. These inhibitors will help to dissect signaling pathways involved in inflammatory responses. In particular, for the first time a definitive signal transduction pathway can be prescribed to the action of lipopolysaccharide in cytokine production in macrophages.