ImmunoMethods最新文献_第5页

High-Resolution Proton NMR Studies of Lymphocyte Extracts 淋巴细胞提取物的高分辨率质子核磁共振研究

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1013

Sze Daniel Y., Jardetzky Oleg

引用次数: 18

On-Line Studies of Activation Events in Primary Human T Lymphocytes 人类原代T淋巴细胞激活事件的在线研究

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1016

Bental Michal, Deutsch Carol

{"title":"On-Line Studies of Activation Events in Primary Human T Lymphocytes","authors":"Bental Michal, Deutsch Carol","doi":"10.1006/immu.1994.1016","DOIUrl":"10.1006/immu.1994.1016","url":null,"abstract":"<div><p>In this paper, we review our NMR studies of human peripheral blood T lymphocytes. These studies focus on the physiological and biochemical alterations accompanying cell cycle progression. In particular, we have characterized phosphorus metabolism, glucose utilization and lactate production, and pH regulation using <sup>31</sup>P, <sup>13</sup>C, and <sup>19</sup>F NMR, respectively. These studies required developing new methods for monitoring on-line stimulation of quiescent T cells under sterile, physiological conditions (i.e., CO<sub>2</sub>/HCO<sup>−</sup><sub>3</sub> buffer, 37°C) for prolonged periods of time. A perfusion system optimized for T lymphocytes inside agarose beads is described. In addition, custom-designed <sup>19</sup>F NMR pH indicators were synthesized, characterized, and used to determine intracellular pH in quiescent lymphocytes, stimulated lymphocytes, and lymphocytes undergoing the G<sub>0</sub>-G<sub>1</sub>, transition. These unique molecular probes are described in detail. Finally, the physiological relevance of our findings regarding carbon metabolism and pH regulation is considered in the context of mitogenesis .</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 148-162"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

31P and 13C Nuclear Magnetic Resonance Studies of Macrophages 巨噬细胞的31P和13C核磁共振研究

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1018

Seguin François, Le Pape Alain

{"title":"31P and 13C Nuclear Magnetic Resonance Studies of Macrophages","authors":"Seguin François, Le Pape Alain","doi":"10.1006/immu.1994.1018","DOIUrl":"10.1006/immu.1994.1018","url":null,"abstract":"<div><p>Biochemical events associated with differentiation and activation of monocyte-macrophage cell lines are of major interest in the understanding of pathophysiological processes as well as in research on immunopharmacological modulation of these cells. Nuclear magnetic resonance is the technique of choice for kinetic studies of metabolic events under such experimental conditions. This approach was used with the P388-D1 model of mature macrophages. Cells primed <em>in vivo</em> were triggered <em>in vitro</em> during NMR analysis and the results were compared to those from chemiluminescence tests performed simultaneously. Three preliminary phases were achieved: (i) <sup>31</sup>P and <sup>13</sup>C NMR spectroscopy of perchloric acid extracts, (ii) optimization of culture and perfusion conditions with validation of macrophage viability and functionality, and (iii) development of a data processing technique to improve the time resolution of kinetic studies. Based on their phosphocreatine content, cells primed <em>in vivo</em> exhibited greater maturation than control cells. After the respiratory burst of primed macrophages was triggered by concanavalin A, <sup>31</sup>P NMR spectra reflected both a transient increase in ADP phosphorylation and intracellular acidification. <sup>13</sup>C NMR studies indicated an acceleration of metabolism following <em>in vitro</em> triggering. The phenomenon was associated with an increased glucose consumption, implicating the hexose monophosphate shunt. These occurred concomitantly with the appearance of new peaks attributed to phosphorylated sugars.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 179-187"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

1H Magnetic Resonance Spectroscopy of Primary Human and Murine Cells of the Myeloid Lineage 人和小鼠原代髓系细胞的1H磁共振波谱分析

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1019

King Nicholas J.C., Delikatny Edward J., Holmes Kerry T.

{"title":"1H Magnetic Resonance Spectroscopy of Primary Human and Murine Cells of the Myeloid Lineage","authors":"King Nicholas J.C., Delikatny Edward J., Holmes Kerry T.","doi":"10.1006/immu.1994.1019","DOIUrl":"10.1006/immu.1994.1019","url":null,"abstract":"<div><p>Murine and human cells of the myeloid lineage can exhibit proton spectra arising primarily from neutral lipid such as triglyceride, similar to other activated and transformed cell types. These spectral characteristics are independent of proliferation or proliferative ability and can be induced <em>in vivo</em> and <em>in vitro</em> in murine peritoneal macrophages by <em>Listeria</em> and interferon-γ stimulation, respectively. In human peripheral monocytes, this spectrum can develop in the apparent absence of external activating stimuli such as interferon or endotoxin and also occurs in the absence of proliferation. Furthermore, 16-h culture of mixed peripheral blood cells causes concomitant and similar spectral changes in lymphocytes, which do not occur in pure lymphocyte cultures. The lipid spectra induced in mixed cultures develop to the same extent regardless of cell density or the ability to adhere or form aggregates and regardless of whether serum supplementation is of fetal bovine or adult human origin. Unlike peritoneal macrophages, the addition of interferon-γ causes no additional spectral changes in adherent or nonadherent cultures. However, reduction of serum concentrations in culture medium causes a dose-dependent decrease in the acquired lipid signal. The observed spectral differences between human and murine myeloid populations could be species-dependent but could also be due to their differentiation status.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 188-198"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 29

Proton Magnetic Resonance Spectroscopy of Lymphocytes: An Historical Perspective 淋巴细胞的质子磁共振波谱:一个历史的视角

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1012

Mountford Carolyn E., Mackinnon Wanda B.

{"title":"Proton Magnetic Resonance Spectroscopy of Lymphocytes: An Historical Perspective","authors":"Mountford Carolyn E., Mackinnon Wanda B.","doi":"10.1006/immu.1994.1012","DOIUrl":"10.1006/immu.1994.1012","url":null,"abstract":"<div><p>Proton magnetic resonance spectroscopy can be used to elucidate alterations to cellular chemistry associated with specific biological functions. It became apparent that this type of information was present in the magnetic resonance spectra from intact viable lymphocytes in the late 1970s. It was not until the 1980s, however, that one-dimensional multipulse sequences were used to filter the large signal contributions from water and fat, which had until then masked weaker signals from other molecules. When this technology was combined with two-dimensional spectroscopy, unambiguous assignment of the biologically relevant chemical species became possible. <em>In vitro</em> activated, stimulated, transformed, and malignant lymphocytes, as well as embryonic fibroblasts and malignant cells of epithelial origin, all gave rise to a strong triglyceride spectrum and resonances from a multitude of cellular metabolites. Two-dimensional spectroscopy and the analyses of highly purified membranes determined that the triglyceride signals originated, at least in part, from the plasma membrane. Based on physicochemical data, a new model for the structural arrangement of plasma membrane lipid in these cells was proposed. While differences exist between the proton magnetic resonance spectra of stimulated lymphocytes and malignant cells <em>in vitro</em>, they share a high-resolution lipid spectrum. In tissue, however, the presence of activated lymphocytes does not always produce the lipid spectrum, particularly in the vicinity of tumors.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 98-112"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Two-Dimensional 1H NMR Studies of Membrane Changes during the Activation of Primary T Lymphocytes 原发性T淋巴细胞活化过程中膜变化的二维1H NMR研究

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1014

Dingley Andrew J., Veale Margaret F., King Nicholas J.C., King Glenn F.

{"title":"Two-Dimensional 1H NMR Studies of Membrane Changes during the Activation of Primary T Lymphocytes","authors":"Dingley Andrew J., Veale Margaret F., King Nicholas J.C., King Glenn F.","doi":"10.1006/immu.1994.1014","DOIUrl":"10.1006/immu.1994.1014","url":null,"abstract":"<div><p>Two-dimensional <sup>1</sup>H NMR spectroscopy was used to quantify the level of \"mobile\" plasma membrane triglyceride and the intracellular concentrations of water-soluble phospholipid precursors during the activation of both mature and immature primary T lymphocytes. The concentration of mobile triglyceride in the plasma membrane was seen to increase ∼35-fold during 72 h of activation of murine thymic and splenic T lymphocytes with ionomycin and phorbol 12-myristate 13-acetate. This dramatic increase in mobile plasma membrane triglyceride during the activation of both mature and immature T-lymphocyte populations supports the hypothesis that immune cell activation is associated with increased plasma membrane fluidity. The intracellular concentrations of various phospholipid precursors were shown to increase during the early stages of T-lymphocyte activation and then remain at levels above those in resting cells. This may facilitate <em>de novo</em> phospholipid biosynthesis, which is presumably necessary since cell volume, and hence the plasma membrane surface area, was demonstrated to increase significantly during T-lymphocyte activation. Various models that might explain the origin of the NMR-visible plasma membrane triglyceride that is observed during immune cell activation and malignant transformation are examined</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 127-138"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 18

Immobilization Methods for NMR Studies of Cellular Metabolism - A Practical Guide 细胞代谢核磁共振研究的固定化方法-实用指南

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1017

Lundberg Peter, Roy Sushmita, Kuchel Philip W

引用次数: 12

Theoretical and Practical Aspects of NMR Studies of Cells 细胞核磁共振研究的理论和实践方面

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1011

King Glenn F., Kuchel Philip W.

引用次数: 9

Lymphocyte Activation-31P Magnetic Resonance Studies of Energy Metabolism and Phospholipid Pathways 淋巴细胞活化- 31p磁共振研究能量代谢和磷脂途径

ImmunoMethods Pub Date : 1994-04-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1015

Kaplan Ofer, Cohen Jack S.

{"title":"Lymphocyte Activation-31P Magnetic Resonance Studies of Energy Metabolism and Phospholipid Pathways","authors":"Kaplan Ofer, Cohen Jack S.","doi":"10.1006/immu.1994.1015","DOIUrl":"10.1006/immu.1994.1015","url":null,"abstract":"<div><p><sup>31</sup>P NMR spectra of perfused lymphocytes embedded in alginate capsules and activated by interleukin-2 (IL-2) are remarkably different from those of control lymphocytes. The main differences are the appearance and gradual increase of phosphodiester signals, glycerophosphocholine and glycerophosphoethanolamine. These metabolic changes also occur following perfusion with phorbol ester and after incubation with phytohemagglutinin (PHA) and are not dependent on a special growth medium. Nifedipine, a calcium channel blocking drug, inhibits the effects of PHA, but not of IL-2. There are no NMR spectral differences between peripheral lymphocytes, stimulated for 3 weeks, and tumor-infiltrating lymphocytes. Thus, sustained accelerated turnover of phosphatidylcholine (PC) and phosphatidylethanolamine is an inherent feature of the activation process. <sup>31</sup>P NMR spectra of lymphocytes are characterized by a low phosphocholine signal. Perfusion studies with high concentrations of choline and the use of dapsone, an inhibitor of phosphocholine cytidyltransferase, indicate that choline kinase plays a key role in regulating PC synthesis in human lymphocytes.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 2","pages":"Pages 139-147"},"PeriodicalIF":0.0,"publicationDate":"1994-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Natural and Recombinant Soluble Low-Affinity FcγR: Detection, Purification, and Functional Activities 天然和重组可溶性低亲和力fc - γ - r:检测、纯化和功能活性

ImmunoMethods Pub Date : 1994-02-01 Epub Date: 2002-05-25 DOI: 10.1006/immu.1994.1007

Teillaud J.L., Bouchard C., Astier A., Teillaud C., Tartour E., Michon J., Galinha A., Moncuit J., Mazières N., Spagnoli R., Fridman W.H., Sautès C.

{"title":"Natural and Recombinant Soluble Low-Affinity FcγR: Detection, Purification, and Functional Activities","authors":"Teillaud J.L., Bouchard C., Astier A., Teillaud C., Tartour E., Michon J., Galinha A., Moncuit J., Mazières N., Spagnoli R., Fridman W.H., Sautès C.","doi":"10.1006/immu.1994.1007","DOIUrl":"10.1006/immu.1994.1007","url":null,"abstract":"<div><p>Studies on the identification, cloning, and biochemical characterization of natural and recombinant human and mouse low-affinity soluble FcγR (sFcγR) have been developed using various methods. RT-PCR and/or biochemical analyses have demonstrated that low-affinity sFcγR (i) are generated by enzymatic cleavage of membrane-associated receptors or by an alternative splicing of the transmembrane region encoding exon and (ii) comprise only the extracellular domains or these domains plus the intracellular region of the membrane-associated molecules, respectively. Functional studies indicated that recombinant sFcγR bind mouse and human IgG subclasses with a binding profile identical to that of their membrane counterparts and inhibit FcγR-mediated functions such as immune complex binding or ADCC. In addition, it has been demonstrated that a mouse recombinant truncated sFcγRII inhibits antibody responses to T-dependent antigens as well as B-cell proliferation and that a human recombinant truncated sFcγRIIIB blocks the Ig production by activated human peripheral blood mononuclear cells. Finally, different immunoassays devised to detect and quantitate circulating sFcγR showed that sFcγR serum levels vary in circumstances such as injections of protein antigens, in parasitic infections, in tumor-bearing mice, in patients with multiple myeloma (MM), or upon infusions of IgG or Fcγ fragments in MM or immune thrombocytopenic purpura patients. The use of recombinant sFcγR, as well as the availability of monoclonal and polyclonal antibodies directed against different regions of these molecules, makes it possible to characterize further the biological effects of sFcγR and their biochemical and immunochemical characteristics, as well as to define their putative ligands on cell membranes.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"4 1","pages":"Pages 48-64"},"PeriodicalIF":0.0,"publicationDate":"1994-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19062883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 41