bluuebunny/crossref_metadata_2025_split huggingface dataset:
Local versus systemic control of numbers of endometrial T cells during pregnancy in sheep
Authors: Majewski A. C. , Tekin S. , Hansen P. J. 
Published: March 2003

SummaryPregnancy in sheep is associated with changes in numbers of specific T‐lymphocyte populations in the uterine endometrium. These changes probably contribute to evasion by the conceptus of maternal immunological rejection and indicate a possible role for T cells in placental growth, parturition and post‐parturient uterine defence against infection. The purpose of the present experiment was to evaluate the relative importance of systemic signals (i.e. those present throughout the uterus or from the circulation, including conceptus hormones secreted into the maternal blood) versus locally acting conceptus signals for regulating changes in numbers of endometrial lymphocytes during pregnancy. The approach taken was to surgically confine pregnancy to one uterine horn and compare differences in lymphocyte numbers between the two uterine horns as well as between both horns of pregnant ewes with those of ovariectomized ewes. As compared with ovariectomized ewes, there was a decline in numbers of CD45R+ lymphocytes within glandular epithelium and an increase in γδ T‐cell number within the luminal epithelium. These changes occurred in both the pregnant and non‐pregnant uterine horns of unilaterally pregnant ewes. Moreover, there were no significant differences in lymphocyte numbers between the two uterine horns of unilaterally pregnant ewes. Expression of CD25 was absent in tissues from both uterine horns. In conclusion, changes in numbers of endometrial lymphocytes during pregnancy, rather than due to locally acting signals of conceptus origin, are the result of hormonal signals of maternal or conceptus origin that either act directly on endometrial lymphocytes or stimulate the uterine endometrium to induce synthesis of regulatory molecules that affect lymphocyte dynamics.
Functional and phenotypic characterization of distinct porcine dendritic cells derived from peripheral blood monocytes
Authors: Paillot R. , Laval F. , Audonnet J.‐C. , Andreoni C. , Juillard V. 
Published: March 2003

SummaryDendritic cells (DCs) are bone marrow‐derived antigen‐presenting cells that have an exquisite capacity to interact with T cells and modulate their responses. Little is known about porcine DCs despite the fact that they represent an important target in strategies that are aimed at modulating resistance to infection in pigs and may be of major importance in transplantation biology. We generated immature monocyte‐derived porcine dendritic cells (MoDCs) directly from adherent peripheral blood cells treated with porcine granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and interleukin‐4 (IL‐4). The cells were observed via electron microscopy and their phenotype was characterized using monoclonal antibodies. The functionality of the porcine MoDCs was demonstrated showing that the cells were capable of different specialized functions relevant to antigen capture and were potent stimulators in a primary allo‐mixed leucocyte reaction. Treatment of the MoDCs with porcine cell line‐derived necrotic factors resulted in the phenotypic and functional maturation of MoDCs. We confirmed also that monocyte‐derived DCs were differentially regulated by cytokines, showing that transforming growth factor‐β1 (TGF‐β1) is able to redirect monocytic precursors into the differentiation pathway of Langerhans' cells presenting typical Birbeck granules. Interestingly, and in contrast to the human and murine model, we showed that the monocyte‐derived porcine Langerhans'‐type cells (MoLCs) were much more potent activators of allogeneic T cells than MoDCs obtained without TGF‐β1.
Differential endocytotic characteristics of a novel human B/DC cell line HBM‐Noda: effective macropinocytic and phagocytic function rather than scavenging function
Authors: Torii Ikuko , Morikawa Shigeru , Nagasaki Makoto , Nokano Akinobu , Morikawa Keiko 
Published: March 2003

SummaryIn order to characterize a novel human B cell‐lineage dendritic cell line (B/DC line) as an antigen‐presenting cell (APC), we compared three types of endocytosis (micropinocytosis via a clathrin‐coated pit, macropinocytosis via membrane ruffling, and phagocytosis) among myeloid‐related, macrophage (Mφ) cell lines and a B/DC line. In the present examination, we used a unique human dendritic cell (DC) line, HBM‐Noda (Noda). Flow cytometric and immunocytochemical analyses revealed that Noda not only expresses some DC markers, but also it expresses some B‐cell associated markers. Noda shows strong capacities to stimulate allogenic T cells, to produce immunoglobulin G (IgG), and to perform immunoglobulin gene rearrangment. These data strongly suggest that Noda is a B‐cell lineage DC line. The endocytic differences among these cell lines were as follows. (1) The level of micropinocytosis of Noda was significantly less than that of conventional human Mφ cell lines, and the formation of a clathrin‐coated pit was not observed in Noda. (2) The level of macropinocytosis of Noda was also smaller than that of conventional Mφ cells indicating that the active membrane ruffling of Noda induces rapid recycling. (3) Phagocytosis of opsonized sheep red blood cells (SRBC) was performed more efficiently in Noda than in other Mφ cell lines. Collectively, these data suggest that in human bone marrow cells, we can identify a unique DC subtype, B/DC line, which develops through a lymphoid DC‐differentiation pathway, and DC in this lineage plays an important role in the host immune response because of its effective uptake of a variety of size of antigens by using the skilful membrane ruffling and surface receptors
Third complementarity‐determining region of mutated VH immunoglobulin genes contains shorter V, D, J, P, and N components than non‐mutated genes
Authors: Rosner Karli , Winter David B. , Tarone Robert E. , Skovgaard Gunhild Lange , Bohr Vilhelm A. , Gearhart Patricia J. 
Published: March 2003

SummaryThe third complementarity‐determining region (CDR3) of immunoglobulin variable genes for the heavy chain (VH) has been shown to be shorter in length in hypermutated antibodies than in non‐hypermutated antibodies. To determine which components of CDR3 contribute to the shorter length, and if there is an effect of age on the length, we analysed 235 cDNA clones from human peripheral blood of VH6 genes rearranged to immunoglobulin M (IgM) constant genes. There was similar use of diversity (D) and joining (JH) gene segments between clones from young and old donors, and there was similar use of D segments among the mutated and non‐mutated heavy chains. However, in the mutated heavy chains, there was increased use of shorter JH4 segments and decreased use of longer JH6 segments compared to the non‐mutated proteins. The overall length of CDR3 did not change with age within the mutated and non‐mutated categories, but was significantly shorter by three amino acids in the mutated clones compared to the non‐mutated clones. Analyses of the individual components that comprise CDR3 indicated that they were all shorter in the mutated clones. Thus, there were more nucleotides deleted from the ends of VH, D, and JH gene segments, and fewer P and N nucleotides added. The results suggest that B cells bearing immunoglobulin receptors with shorter CDR3s have been selected for binding to antigen. A smaller CDR3 may allow room in the antibody binding pocket for antigen to interact with CDRs 1 and 2 as well, so that as the VDJ gene undergoes hypermutation, substitutions in all three CDRs can further contribute to the binding energy.
The pan‐chemokine inhibitor NR58‐3.14.3 abolishes tumour necrosis factor‐α accumulation and leucocyte recruitment induced by lipopolysaccharide in vivo
Authors: Reckless Jill , Tatalick Lauren M. , Grainger David J. 
Published: March 2003

SummaryChemokines participate in the regulation of leucocyte recruitment in a wide variety of inflammatory processes, including host defence and diseases such as asthma, atherosclerosis and autoimmune disorders. We have previously described the properties of Peptide 3, the first broad‐specificity chemokine inhibitor in vitro. Here, we report the properties of NR58‐3.14.3, a retroinverso analogue of Peptide 3. NR58‐3.14.3 inhibited leucocyte migration induced by a range of chemokines, including monocyte chemoattractant protein‐1 (MCP‐1) (2·5 nm), macrophage inflammatory protein‐1α (MIP‐1α) (5 nm), regulated on activation, normal T‐cell expressed and presumably secreted (RANTES) (20 nm), stromal cell‐derived factor‐1α (SDF‐1α) (25 nm) and interleukin‐8 (IL‐8) (30 nm), but did not affect migration induced by N‐formyl‐methionyl‐leucyl‐phenylalanine (FMLP) or complement C5a (> 100 µm). NR58‐3.14.3 is therefore ≈ 1000‐fold more potent than Peptide 3 but retains the broad‐spectrum chemokine inhibitory activity of the parent peptide. In vivo, pretreatment with a systemic dose of 10 mg of NR58‐3.14.3, but not the inactive derivative NR58‐3.14.4, abolished leucocyte recruitment in response to intradermal injection of 500 ng of MCP‐1 into rat skin. This suggests that NR58‐3.14.3 is a functional chemokine inhibitor in vivo as well as in vitro. We utilized NR58‐3.14.3 as a tool to investigate the role of chemokine activity during leucocyte recruitment in response to lipopolysaccharide (LPS) in vivo. NR58‐3.14.3, but not NR58‐3.14.4, abolished leucocyte recruitment in response to intradermal injection of 50 ng of LPS into rat skin. Furthermore, NR58‐3.14.3 completely inhibited LPS‐induced accumulation of tumour necrosis factor‐α (TNF‐α). This data is consistent with a model in which multiple chemokines act in parallel upstream of TNF‐α. NR58‐3.14.3 is therefore a powerful anti‐inflammatory agent in vivo, suppressing proinflammatory cytokine production and leucocyte recruitment in response to endotoxin stimulus in rat skin.
Allergen‐induced murine upper airway inflammation: local and systemic changes in murine experimental allergic rhinitis
Authors: Saito Hiroko , Howie Karen , Wattie Jennifer , Denburg Avram , Ellis Russ , Inman Mark D. , Denburg Judah A. 
Published: March 2003

SummaryThe role of inflammatory effector cells in the pathogenesis of airway allergy has been the subject of much investigation. However, whether systemic factors are involved in the development of local responses in both upper and lower airways has not been fully clarified. The present study was performed to investigate aspects of the pathogenesis of isolated allergic rhinitis in a murine model sensitized to ovalbumin (OVA). Both upper‐ and lower‐airway physiological responsiveness and inflammatory changes were assessed, as well as bone marrow progenitor responses, by culture and immunohistological methods. Significant nasal symptoms and hyper‐responsiveness appeared after intranasal OVA challenge (P < 0·0001 and P < 0·01, respectively), accompanied with significant nasal mucosal changes in CD4+ cells (P < 0·001), interleukin (IL)‐4+ cells (P < 0·01), IL‐5+ cells (P < 0·01), basophilic cells (P < 0·02) and eosinophils (P < 0·001), in the complete absence of hyper‐responsiveness or inflammatory changes in the lower airway. In the bone marrow, there were significant increases in CD34+ cells, as well as in eosinophils and basophilic cells. In the presence in vitro of mouse recombinant IL‐5, IL‐3 or granulocyte–macrophage colony‐stimulating factor (GM‐CSF), the level of bone marrow eosinophil/basophil (Eo/Baso) colony‐forming cells increased significantly in the OVA‐sensitized group. We conclude that, in this murine model of allergic rhinitis, haemopoietic progenitors are upregulated, which is consistent with the involvement of bone marrow in the pathogenesis of nasal mucosal inflammation. Both local and systemic events, initiated in response to allergen provocation, may be required for the pathogenesis of allergic rhinitis. Understanding these events and their regulation could provide new therapeutic targets for rhinitis and asthma.
Interleukin‐10‐secreting Peyer's patch cells are responsible for active suppression in low‐dose oral tolerance
Authors: Tsuji Noriko M. , Mizumachi Koko , Kurisaki Jun‐ichi 
Published: March 2003

SummaryWe demonstrate the induction of antigen‐specific interleukin‐10 (IL‐10)‐secreting cells in murine Peyer's patches (PPs) after low‐dose β‐lactoglobulin (BLG) feeding. In addition, we show that PP cells can inhibit the T‐cell proliferative response in vitro as well as T‐cell‐mediated inflammation in vivo. The active suppression mediated by these regulatory cells was seen only within a narrow range of antigen dosage (feeding), with the most prominent effect at 5 × 1 mg BLG. On either side of this range, T‐helper 1‐like cytokine responses were observed when PP cells were stimulated with antigen in vitro. This result correlated with reduced production of regulatory cytokines as well as reduced activity of bystander suppression. We found that changes in IL‐10 production correlated inversely with changes in interferon‐γ production. Inhibitory effects mediated by CD4+ PP cells were partially neutralized by antibodies to IL‐10 and transforming growth factor‐β. Interestingly, the generation of such regulatory cells after low‐dose BLG feeding exhibited organ dependence. Among spleen, lymph node and PP cells derived from orally tolerized mice, PP cells were the most effective in promoting bystander suppression in the presence of BLG, indicating the significance of PPs as an inductive site for antigen‐specific regulatory cells upon induction of low‐dose oral tolerance. Moreover, PP cells from mice fed 5 × 1 mg BLG were shown to suppress a BLG‐specific delayed‐type hypersensitivity response induced in footpads, suggesting that IL‐10‐secreting PP cells regulate systemic inflammation.
Multiple cytokines sharing the common receptor γ chain can induce CD154/CD40 ligand expression by human CD4+ T lymphocytes via a cyclosporin A‐resistant pathway
Authors: Fayen John D. 
Published: March 2003

SummaryExpression of CD154/CD40 ligand (CD154/CD40L), an important molecular component of CD4+ T‐cell help, can be triggered by T‐cell receptor (TCR) stimulation. Dephosphorylation of the transcriptional element Nuclear Factor of Activated T cells‐1 (NFAT1) is a critical activation step in the TCR‐initiated signal transduction cascade which promotes CD154/CD40L expression. Cyclosporin A (CsA), which interferes with NFAT1 activation, has been shown to be an effective inhibitor of TCR‐triggered CD154/CD40L expression by resting T cells. We now report that recombinant interleukin‐2 (rIL‐2) is also capable of inducing CD154/CD40L on CD4+ T lymphoblasts via a pathway triggered independently of the CD3/TCR receptor complex. Recombinant IL‐2‐mediated CD154/CD40L expression, in contrast to that triggered by CD3/TCR stimulation, is only partially inhibited by CsA. The capacity of rIL‐2 to induce CD154/CD40L expression by T lymphoblasts also extends to a restricted number of cytokines sharing the cytokine receptor common γ chain, including IL‐15, and, to a lesser extent, IL‐7, but not IL‐4. A similar CsA‐resistant CD154/CD40L induction pathway can be triggered in primary T cells by the combination of anti‐CD3 stimulation and recombinant lymphokines. In contrast to T lymphoblasts, the CsA‐resistant CD154/CD40L induction in primary lymphocytes can be efficiently triggered by multiple cytokines which bind the common γ chain receptor family. The data outline a novel pathway of CD154/CD40L induction which is, at least in part, independent of NFAT1 and resistant to CsA. A more complete understanding of the mechanisms governing CD154/CD40L expression may facilitate the rational design of specifically targeted immunotherapeutic agents.
Allograft inflammatory factor‐1 augments production of interleukin‐6, ‐10and ‐12 by a mouse macrophage line
Authors: Watano Keiko , Iwabuchi Kazuya , Fujii Satoshi , Ishimori Naoki , Mitsuhashi Shinya , Ato Manabu , Kitabatake Akira , Onoé Kazunori 
Published: March 2003

SummaryMouse allograft inflammatory factor‐1 (AIF‐1) cDNA was cloned and the AIF‐1‐specific monoclonal antibodies were established to examine its tissue distribution. The mouse AIF‐1 was highly conserved among all reported AIF‐1 from a variety of species, from invertebrates to mammals, and the cloned cDNA was in good accordance with putative expressed regions of genomic sequences in the mouse major histocompatibility complex (MHC) class III region. The messages of mouse AIF‐1 were abundantly expressed in the testis, moderately in the spleen and lymph nodes and slightly in the liver and thymus of normal BALB/c mice. Immunohistological examination revealed that differentiating germ cells in the testis and presumably macrophages in the red pulp of the spleen were positive for AIF‐1. To analyse the function of the AIF‐1, a macrophage cell line, RAW 264.7, was transfected with mouse AIF‐1 cDNA. Upon stimulation with bacterial lipopolysaccharide, the transfectants that overexpressed AIF‐1 showed marked morphological changes and produced significantly large amounts of interleukin (IL)‐6, IL‐10 and IL‐12p40 but not IL‐12p70 compared with control cells. No difference was noted in production of tumour necrosis factor‐α, transforming growth factor‐β1 and IL‐1α. These results suggest that AIF‐1 plays an important role in cells of a monocyte/macrophage lineage upon stimulation with inflammatory stimuli by augmenting particular cytokine production.
Failure to induce enhanced protection against tuberculosis by increasing T‐cell‐dependent interferon‐γ generation
Authors: Leal Irene S. , Smedegård Birgitte , Andersen Peter , Appelberg Rui 
Published: March 2003

SummaryWe evaluated the use of recombinant human interleukin‐6 (rhIL‐6) and a monoclonal antibody specific for interferon‐γ (IFN‐γ) as co‐adjuvants in a subunit vaccine against tuberculosis consisting of the culture filtrate proteins of Mycobacterium tuberculosis (ST‐CF) emulsified in the adjuvant dimethyl‐dioctadecylammonium bromide (DDA). Both the addition of rhIL‐6 and the neutralization of IFN‐γ resulted in an increased T helper type 1 (Th1) response characterized by enhanced IFN‐γ production and cell proliferation. Nevertheless, this did not result in the enhancement of protection against either an intravenous or an aerosol M. tuberculosis challenge. Our data stress the need to identify further correlates of protection in addition to IFN‐γ production to screen vaccines against tuberculosis infection.
Human immunodeficiency virus type 1 Tat binds to Candida albicans, inducing hyphae but augmenting phagocytosis in vitro
Authors: Gruber Andreas , Lell Claudia P. , Speth Cornelia , Stoiber Heribert , Lass‐Flörl Cornelia , Sonneborn Anja , Ernst Joachim F. , Dierich Manfred P. , Würzner Reinhard 
Published: March 2003

SummaryTat, the human immunodeficiency virus type 1 (HIV‐1) transactivating protein, binds through its RGD‐motif to human integrin receptors. Candida albicans, the commonest cause of mucosal candidiasis in subjects infected with HIV‐1, also possesses RGD‐binding capacity. The present study reveals that Tat binds to C. albicans but not to C. tropicalis. Tat binding was markedly reduced by laminin and to a lesser extent by a complement C3 peptide containing the RGD motif, but not by a control peptide. The outgrowth of C. albicans was accelerated following binding of Tat, but phagocytosis of opsonized C. albicans was also increased after Tat binding. Thus, Tat binding promotes fungal virulence by inducing hyphae but may also reduce it by augmenting phagocytosis. The net effect of Tat in vivo is difficult to judge but in view of the many disease‐promoting effects of Tat we propose that accelerating the formation of hyphae dominates over the augmentation of phagocytosis.
Expression of β‐defensin 1 and 2 mRNA by human monocytes, macrophages and dendritic cells
Authors: Duits Louise A. , Ravensbergen Bep , Rademaker Mirjam , Hiemstra Pieter S. , Nibbering Peter H. 
Published: March 2003

SummaryHuman β‐defensins are broad‐spectrum antimicrobial peptides known to be produced by epithelial cells. It was recently shown that β‐defensins also display chemotactic activity for dendritic cells (DC) and T cells, and thus may serve to link innate and adaptive immunity. The aim of the present study was to explore expression of mRNA for these peptides in mononuclear phagocytes and DC. The results revealed that monocytes, monocyte‐derived‐macrophages (MDM), and monocyte‐derived‐dendritic cells (DC) all express human‐β‐defensin‐1 (hBD‐1) mRNA. hBD‐1 mRNA expression by monocytes and MDM was increased after activation with interferon‐γ (IFN‐γ) and/or lipopolysaccharide (LPS) in a dose‐ and time‐dependent fashion. Alveolar macrophages showed an intense hBD‐1 expression, which could not be further increased. Expression of hBD‐1 mRNA by immature DC was low, and increased considerably after maturation. Monocytes, MDM, alveolar macrophages and DC showed a limited expression of human β‐defensin‐2 (hBD‐2) mRNA, which could only be increased in monocytes and alveolar macrophages by IFN‐γ and/or LPS in a dose‐ and time‐dependent fashion. Immunocytochemical stainings demonstrated the expression of hBD‐2 peptide by freshly isolated blood monocytes and alveolar macrophages in cytospin preparations.
Human small intestinal mucosa harbours a small population of cytolytically active CD8+ αβ T lymphocytes
Authors: Melgar Silvia , Bas Anna , Hammarström Sten , Hammarström Marie‐Louise 
Published: March 2003

SummaryIntraepithelial lymphocytes (IEL) in normal human small intestine exhibit cytotoxicity. This study was undertaken to characterize the effector cells and their mode of action. Freshly isolated jejunal IEL and lamina propria lymphocytes (LPL), as well as IEL and LPL depleted of CD4+, CD8+ and T‐cell receptor (TCR)‐γδ+ cells were used as effector cells in anti‐CD3‐mediated redirected cytotoxicity against a murine FcγR‐expressing cell line. Effector cell frequencies were estimated by effector to target cell titration and limiting dilution. The capacity of IEL and LPL to kill a Fas‐expressing human T‐cell line was also analysed. T‐cell subsets were analysed for perforin, granzyme B, Fas‐ligand (FasL), tumour necrosis factor‐α (TNF‐α) and TNF‐related apoptosis inducing ligand (TRAIL) mRNA expression by reverse transcription–polymerase chain reaction (RT‐PCR). Frequencies of IEL expressing the perforin and FasL proteins were determined by immunomorphometry. Both IEL and LPL exhibited significant Ca2+‐dependent, anti‐CD3‐mediated cytotoxicity, ≈ 30% specific lysis at the effector to target cell ratio 100. The cytotoxic cells constituted, however, only a small fraction of IEL and LPL (≈ 0·01%). CD8+ TCR‐αβ+ cells accounted for virtually all the cytotoxicity and expressed mRNA for all five cytotoxic proteins. The frequency of granzyme B‐expressing samples was higher in CD8+ cells than in CD4+ cells (P<0·05 and <0·01 for IEL and LPL, respectively). In addition, both IEL and LPL exhibited significant spontaneous anti‐CD3‐independent cytotoxicity against Fas‐expressing human T cells. This killing was mediated by Fas–FasL interaction. On average, 2–3% of the IEL expressed perforin and FasL. We speculate that CD8+ memory cells accumulate in the jejunal mucosa and that the CD8+ TCR‐αβ+ lymphocytes executing TCR/CD3‐mediated, Ca2+‐dependent cytotoxicity are classical cytotoxic T lymphocytes ‘caught in the act’ of eliminating infected epithelial cells through perforin/granzyme exocytosis. The observed Fas/FasL‐mediated cytotoxicity may be a reflection of ongoing down‐regulation of local immune responses by ‘activation‐induced cell death’.