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  • 1
    ISSN: 1432-1440
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Human blood lymphocytes were separated by density centrifugation in a discontinuous albumin gradient. Prior to centrifugation the lymphocytes were purified by passage through a nylon wool column. A characteristic pattern of density distribution of lymphocytes was found for individual donors. The majority of cells was consistently recovered from the interfaces representing the regions of higher density of the gradient (〉1,068 g/cm3). The separated cell fractions were cultured in vitro and stimulated by phytohemagglutinin (PHA) and streptolysin 0 antigen (SLO). Most of the PHA-reacting cells were found in the region of low density (〈1060 g/cm3). The optimal response to SLO antigen was observed in cell cultures recovered from the same region of low density. Cell fractions 1–5 of the low density region comprised not more than 12% of the total cells. Unstimulated lymphocytes, too, from the low density region consistently showed a higher uptake of uridine and thymidine compared to the original lymphocyte suspension. The significance of these findings is discussed with respect to enrichment of antigen-reactive cells and to the mechanism of transformation of lymphocytes in vitro.
    Notes: Zusammenfassung Humane Lymphocyten aus dem peripheren Blut wurden durch Zentrifugation im diskontinuierlichen Albumingradienten nach ihrer Dichte in verschiedene Populationen getrennt. Vor der Zentrifugation wurden die Lymphocyten von haftenden Zellen durch Filtration an Nylonwolle gereinigt. Für jeden Lymphocytenspender fand sich ein charakteristisches Verteilungsmuster. Der größte Teil der Zellen reicherte sich in den dichteren Zonen des Gradienten an (〉1,068 g/cm3). Die getrennten Zellfraktionen wurden in vitro kultiviert und mit Phytohämagglutinin und Streptolysin 0 Antigen stimuliert. Eine Anreicherung der PHA-reaktiven Zellen fand sich in den Schichten geringerer Dichte (〈1060 g/cm3). In den gleichen Zonen befanden sich die auf SLO Antigen optimal reagierenden Zellen. In den Zellschichten 1–5, den Zonen geringerer Dichte, ließen sich nie mehr als 12% aller Zellen gewinnen. Auch unstimulierte Lymphocyten aus den Zonen geringerer Dichte wiesen in allen Fällen einen höheren Einbau von Uridin und Thymidin auf als die entsprechenden Originalzellen. Die Bedeutung dieser Befunde für die Anreicherung von antigenreaktiven Zellen und für den Mechanismus der Transformation von Lymphocyten in vitro wird diskutiert.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 53 (1975), S. 1155-1159 
    ISSN: 1432-1440
    Keywords: Lymphocytes ; membrane-bound immunoglobulin ; phytohaemagglutinin ; mixed-lymphocyte reaction ; appendix ; appendicitis ; Lymphocyten ; membrangebundenes Immunglobulin ; Phytohämagglutinin-Stimulation ; Lymphocytenmischkultur ; Appendix ; Appendicitis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Aus der Submukosa von frisch opierten humanen Appendices wurden die Lymphocyten isoliert und die Verteilung der T- und B-Lymphocyten untersucht. Der Anteil der T-Lymphocyten aus 34 Appendices, bestimmt durch Bildung von spontanen E-Rosetten, betrug 50,0%. Durchschnittlich ließen sich 46,8% der Lymphocyten aus 10 Appendices mit dem immunautoradiographischen Nachweis von Oberflächenimmunglobulin als B-Lymphocyten charakterisieren. Von den B-Lymphocyten waren 17,7% IgM-positiv, 24,4% IgG-positiv und 4,7% IgA-positiv. Die Lymphocyten ließen sich mit Phytohaemagglutinin (PHA) stimulieren und zeigten eine positive Reaktion in der Lymphocytenmischkultur. Die Befunde zeigen, daß Suspensionen von Appendixlymphocyten einen hohen Anteil an T-Lymphocyten besitzen, der auch funktionell als solcher zu charakterisieren ist. Eine deutliche Korrelation zwischen dem histologischen Entzündungsgrad der Appendices und der Verteilung sowie der Reaktivität ihrer Lymphocyten konnte nicht gefunden werden.
    Notes: Summary The isolation of viable lymphocytes from fresh human appendices is described. Under standardized experimental conditions of cell preparation no selective enrichment of either T- or B-lymphocytes was observed. The mean percentage of T-lymphocytes as judged by spontaneous rosette formation found in 34 appendices examined was 50.0%. After overnight incubation surface-Ig was detected by immunoradioautography on 46.8% of the recovered lymphocytes from 10 appendices. For the detection of surface-Ig specifically purified antibodies to heavy chains µ, γ and α were used. Of the 46.8% surface-Ig positive lymphocytes the class distribution was as follows: IgM 17.7%, IgG 24.4% and IgA 4.7%. The isolated lymphocytes showed a variable mitotic response to PHA (stimulation index ranging from 3.3 to 200.9) as well as to allogeneic lymphocytes in a one-way mixed lymphocyte culture. So far, a distinct correlation could not be found between the histological degree of inflammation and the class or mitotic response of appendix lymphocytes.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1440
    Keywords: Tγ lymphocytes ; Monocytic antigens ; Natural cytotoxicity ; Antibody-dependent cellular cytotoxicity ; Tγ-Lymphozyten ; monozytäre Antigene ; spontane Zytotoxizität ; antikörper-abhängige zellvermittelte Zytotoxizität
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Zwei Patienten, bei denen wir eine Proliferation von Tγ Lymphozyten nachweisen konnten, boten ein ungewöhnliches klinisches Bild (disseminierte intravasale Gerinnung im Anschluß an eine niedrigdosierte Milzbestrahlung, isolierte absolute Neutropenie). Während bei einem Patienten ohne Zweifel eine maligne Erkrankung vorlag, ist bei dem anderen Patienten eher eine reaktive Zellproliferation anzunehmen. Neben T-Zell-Oberflächeneigenschaften wiesen die proliferierenden Zellen ein monozytäres Antigen auf. Sie zeigten keine Suppression der Differenzierung von B-Lymphozyten in Plasmazellen. Im Gegensatz dazu entwickelten die proliferierenden Zellen, besonders bei einem Patienten, deutlich spontane und antikörpervermittelte Zytotoxizität mit Melanom-und MOLT 4-Zielzellen. Bei dem zweiten Patienten konnten wir phagozytäre Eigenschaften der Tγ-Zellen nachweisen. Die Ergebnisse lassen vermuten, daß Untereinheiten der Tγ-Zellen Beziehungen zu monozytären Zellen aufweisen und daß diese Zellen sowohl spontane als auch antikörpervermittelte Zytotoxizität und teilweise phagozytäre Eigenschaften entwickeln können.
    Notes: Summary Two patients suffering from proliferation of Tγ cells exhibited uncommon clinical features, such as activation of intravascular coagulation after low dose irradiation of the enlarged spleen in one patient and isolated neutropenia in the other patient. While the malignant nature of the disease was doubtless in one patient, cell proliferation in the other patient was more likely reactive. In addition to T cell determinants the proliferating cells expressed a monocytic antigen. They did not suppress B-lymphocyte differentiation into plasma cells. In contrast the proliferating cells, especially in one patient, acted as potent effectors in NK and ADCC using melanoma and MOLT 4 target cells. Erythrophagocytosis by Tγ cells was seen in one patient. The data suggest that subsets of Tγ cells are related to the monocytic lineage and that these cells can mediate both NKA and ADCC and partly can develop phagocytic activity.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 64 (1986), S. 1119-1123 
    ISSN: 1432-1440
    Keywords: Transferrin receptors ; Tumor cells ; Bone marrow cells ; Natural killer cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Two different experimental approaches based on the specificity of monoclonal antibodies (mAbs) have been taken to verify the hypothesis that the transferrin receptor (TfR) on proliferating cells serves as a common target structure for natural killer (NK) cells. Thus, by the lysostrip technique the TfR was removed from the surface of K562 and Molt4 tumor cells by incubation with two different anti-TfR mAbs. The effect of removal of the TfR was controlled by uptake of radiolabeled transferrin, and by binding of non-cross-reacting monoclonal anti-TfR receptor antibodies. Though the modulation of TfR on the membrane of viable cells was nearly complete, the cells remained fully susceptible to NK lysis. The second approach consisted in removal of TfR-bearing cells from bone marrow cell suspensions by an indirect rosetting technique. Using mAbs bound to ox erythrocytes the rosetted TfR-bearing cells could be removed from bone marrow cell suspension by density centrifugation with an efficiency of 〉99%. It could be shown that both fractions, TfR+ and TfR− cells, could be lysed to the same degree by NK cells. Thus, the evidence obtained speaks against a role of TfR as a recognition structure for NK cells.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 54 (1976), S. 1011-1019 
    ISSN: 1432-1440
    Keywords: Hairy cell leukemia ; Hairy cells ; Cytochemistry ; Surface immunoglobulins ; Fc-receptors ; Phagocytosis ; Mitogen stimulation ; E-rosettes ; Haarzell-Leukämie ; Haarzellen ; Cytochemie ; Oberflächenimmunglobuline ; Fc-Rezeptoren ; Phagocytose ; Mitogen-Stimulation ; E-Rosetten
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Bei vier Patienten mit Haarzell-Leukämie wurden die leukämischen Zellen auf das Vorkommen von Oberflächenimmunglobulinen und Fc-Rezeptoren sowie die Stimulierbarkeit mit verschiedenen Mitogenen hin untersucht. Die Analyse dieser Eigenschaften erfolgte mit Hilfe einer kombinierten zytochemisch-radioautographischen Technik, bei der die radioaktiv markierten Zellen durch den Nachweis des Tartrat-resistenten Isoenzyms der sauren Phosphatase als Haarzellen identifiziert werden konnten. Für den Nachweis der Oberflächenimmunglobuline wurden125I-markierte (Fab')2-Fragmente von monospezifischen Antikörpern gegen schwere Immunoglobulinketten verwendet. Bei zwei Patienten wurden auf der Oberfläche der isoenzymhaltigen Zellen μ- und δ-Immunglobulinketten nachgewiesen, die bei einem großen Teil dieser Zellen gleichzeitig auf der Zelloberfläche ausgedrückt waren. Bei einem Patienten zeigten die isoenzymhaltigen Zellen γ- und δ-Ketten und bei einem vierten Patienten nur γ-Ketten. Bei einer Patientin konnten μ- und δ-Ketten nach einer in vitro-Kultur von 14 Tagen in Humanserum-freiem Medium in gleicher Weise nachgewiesen werden wie auf frischen Zellen, was für eine Synthese der Oberflächenimmunglobuline durch die Haarzellen selbst spricht. In der indirekten Immunfluoreszenztechnik wurden die Oberflächenimmunglobuline sehr schnell über einem Zellareal konzentriert, in dem auch Latexpartikel nach der Phagozytose lokalisiert waren. Mit Hilfe von125I-markierten Aggregaten von humanem IgG konnten bei allen vier Patienten Fc-Rezeptoren auf fast 100% der isoenzymhaltigen Zellen nachgewiesen werden. Eine deutliche, wenn auch geringe Stimulation der isoenzymhaltigen Zellen konnte beobachtet werden, wenn die Haarzellen mit Pokeweed-Mitogen oder Lima-Bohnen-Lektin (B- und T-Zell-Mitogene) kultiviert wurden, nicht jedoch, wenn die Stimulation mit Phytohämagglutinin oder Concanavalin-A (T-Zell-Mitogene) erfolgte. Bei keinem der drei untersuchten Patienten bildeten die Haarzellen Spontanrosetten mit Schaferythrozyten.
    Notes: Summary In four patients with hairy cell leukemia the expression of surface immunoglobulins and Fc-receptors on the leukemic cells as well as the stimulation of the leukemic cells by various T- and B-cell mitogens was studied. Using a combined cytochemical radioautographic method the tartrate resistant isoenzyme of the acid phosphatase and immunological markers could be demonstrated simultaneously on single cells. Surface immunoglobulins were detected by125I-labelled (Fab')2-fragments of monospecific anti-heavy-chain-antibodies. In two patients only μ- and δ-determinants were found on the isoenzyme-positive hairy cells; in one patient 86% and in the other 44% of these cells carried both heavy chains on the same cell. Another patient showed both γ-and δ-chains on the isoenzyme-positive cells and the fourth patient γ-chains only. When the hairy cells of one patient were cultured in vitro in human serum-free medium for 14 days, μ- and δ-determinants were found just as on freshly isolated cells suggesting that hairy cells synthesize immunoglobulins. By indirect immunofluorescence the surface-Ig on hairy cells was shown to be capped very rapidly at room temperature. The concentrated surface-Ig was found over that cytoplasmic area where most of the ingested latex particles were located. In addition, using125I-labelled aggregated human IgG (M.W. 5−15×106), Fc-receptors were found on virtually all of the isoenzyme-containing hairy cells in all four patients. Furthermore, a distinct, low-degree stimulation of the isoenzyme-positive hairy cells could be demonstrated when the cells were cultured in the presence of pokeweed mitogen or Lima-bean lectin (B- and T-cell-mitogens), whereas no stimulation was observed by phytohem-agglutinin and concanavalin A (T-cell-mitogens). In three patients studied, isoenzyme-positive hairy cells were negative with regard to rosette formation with sheep erythrocytes.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1440
    Keywords: Type I diabetes ; Autoimmunity ; Ia-antigen bearing cells ; ICA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Peripheral blood lymphocytes have been investigated in 20 newly diagnosed type-I diabetics and 10 healthy subjects using monoclonal antibodies. Mononuclear cells were marked with anti-T-lymphocytes (Leu2, 3, 4, 12) and anti-Ia-antibodies (K14, L243) using indirect immunofluorescence. The percentage of circulating K14- and L243-positive cells was significantly higher in all diabetics than in normal controls. An increase in the number of K14-bearing cells was found in newly diagnosed patients with duration of less than 7 days (n=10) compared with diabetics of longer duration (1 to 8 months;n=10). Using dual-color immunofluorescence with fluorescein-conjugated anti-T-lymphocytes and rhodamin-conjugated anti-Ia-antibodies it was not possible to identify Ia-antigen bearing cells (Ia cells) as helper or suppressor lymphocytes. In addition, there was no significant difference in the number of Ia cells in diabetics with and without islet cell antibodies. It is concluded that there is evidence of activation of cellular immune response in type I diabetes, particularly in the early days of manifestation. However, previous assumptions that Ia cells represent T-cell activation have to be questioned.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 265 (1977), S. 158-160 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The data were obtained with an ADCC system5,6 consisting of cultured melanoma cells as target cells, different preparations of human peripheral blood lymphocytes as effector cells, and anti-melanoma cell antisera obtained from melanoma patients. The cytolytic activity of lymphocytes was measured by ...
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: To investigate the mechanisms of action underlying the therapeutic effect of CD4 monoclonal antibody therapy in rheumatoid arthritis (RA), clinical responses were compared with several laboratory parameters. Twenty-nine RA patients received either 10 mg, 50 mg or 100 mg of cM-T412, a chimeric CD4 MoAb, for 7 days. The CD4 binding sites on circulating lymphocytes were saturated directly with cM-T412 and serum levels of unbound cM-T412 accumulated towards day 7 of treatment only in the patients treated with 50 and 100 mg. The treatment induced an instant and prolonged depression of the number of circulating CD4+ cells, similar for all dosages. Clinical improvement was observed predominantly in the patients treated with 50 or 100mgcM-T412daily and did not correlate with changes in counts of circulating leucocyte subsets nor with changes in serum cytokine levels. An antiglobulin response against cM-T412 developed in a majority of the patients. Side effects on the first day of treatment were correlated with an increase of serum IL-6 levels. This study indicates that a favourable clinical effect of cM-T412 administration was associated with the presence of unbound cM-T412 in the circulation of RA patients. Therefore penetration of unbound cM-T412 into the site of inflammation might determine the therapeutic effect in RA.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 6 (1977), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: This paper describes the successful use of 125I-labeled IgG aggregate to detect Ia-type alloantibodies in pregnancy sera. The blockade of aggregate uptake of a variety of normal mononuclear and leukemic cell types by anti-Ia alloantibodies is analyzed. Fc receptors and Ia alloantigens are clearly two distinct molecular entities. The association between Fc receptors and Ia alloantigens on a quantitative level seems to depend on a ligand-binding mechanism to control their interaction rather than the presence of a topographical molecular tandem arrangement.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 5 (1976), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: An in vitro lymphocyte-mediated cytotoxicity assay using [3H]proline-labeled target cells is described. The assay, modified from an original procedure of Bean et al., assesses the release of [3H]proline by filtering the total culture fluid containing both trypsinized tumor cells and effector cells. Filtration is performed with a semiautomatic harvesting device using low suction pressure and large-diameter glass filters. Pretreatment of filters with whole serum diminishes adsorption of cell-free radioactive material considerably and thus increases the sensitivity of the assay. Nearly 100% of the radioactivity could be recovered with this harvesting device. The technique allowed the detection of cytolytic activities of lymphocytes after 6 h of incubation. Lymphocytes from patients with primary malignant melanoma showed a significantly higher cytolytic reactivity (P 〈 0.001) than normal donors’ lymphocytes against three different melanoma cell lines. In a series of parallel experiments on 36 patients and IS normal donors, this modification of the [3H]proline test was compared with three different assays: the conventional microcytotoxicity test of Takasugi and Klein, the original [3H]proline microcytotoxicity test of Bean et al., and the viability count of tumor cells.
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