Enhancement of immunogenicity of Jeg3 cells by ectopic expression of HLA-A*0201 and CD80

PROBLEM: The choriocarcinoma cell line Jeg3 suppresses immunity in vitro by secretion of soluble factors like leukemia inhibitory factor suppressing leukocyte activation. The cells lack expression of classical human leukocyte antigen (HLA)-A and -B alleles but express some HLA-C, and non-classical HLA-G and -E. Upon binding to killing inhibitory receptor on natural killer (NK) cells, HLA-G prevents activation of cytolytic activity. We investigated whether Jeg3 cells are capable of immune stimulation after complementation with classical HLA and T cell costimulatory signal CD80. METHOD OF STUDY: Jeg3 cells were transduced to express HLA-A*0201 and/or CD80. Parental Jeg3 or transfectants Jeg3-A2, Jeg3-CD80 or Jeg3-CD80-A2 were used to stimulate allogeneic resting and activated peripheral blood lymphocytes (PBL). The different cell lines were loaded with a HLA-A2-restricted Epstein-Barr virus (EBV) recall antigen peptide epitope and antigen presenting ability was examined. T cell lines specific for Jeg3 and transfectants were generated from HLA-A2 matched and nonmatched donors and compared for expansion, phenotypes and cytolytic activity. RESULTS: While all Jeg3 cell lines induced only marginal proliferation of resting T cells, phytohemagglutinin (PHA)-activated T cells were stimulated by CD80 or CD80-A2 expressing Jeg3. Only the transfectant Jeg3-CD80-A2 was capable of specific T cell stimulation by EBV recall antigen presentation. T cell lines of HLA-A2 non-matched donors stimulated with the Jeg3 transfectants showed significant expansion only when HLA-A2 and the costimulus CD80 were present. T cells from HLA-A2 positive donors did not expand significantly or differentially. No NK cells grew under any condition. In Jeg3-CD80-A2 stimulated T cells lines CD8+ cells expanded preferentially. These T cells exerted cytolytic activity toward all Jeg3 cell lines. CONCLUSION: Our data suggest that, in spite of immunosuppressive mechanisms, proliferative and cytolytic T cell responses are induced by Jeg3 cells when classical HLA- and/or costimulatory signals are present on the cells.     

Einige Befunde zur Sinnesphysiologie der Kaltblüterzecke Amblyomma testudinis

Zusammenfassung Aus eigener Zucht stammende Larven, Nymphen und Imagines von Amblyomma testudinis wurden auf ihre Sinnesleistungen hinsichtlich Phototaxis, Thermotaxis, Geotaxis und Chemotaxis untersucht. Es ergab sich ein entwicklungsabhängiger Wandel im phototaktischen Verhalten von anfangs positiver Phototaxis bei den Larven über indifferente Phototaxis bei vollgesogenen Larven und nüchternen Nymphen zu negativer Phototaxis bei vollgesogenen Nympen sowie - und -Imagines. Sämtliche Entwicklungsstadien mit Ausnahme der -Imagines verhalten sich temperaturindifferent vor dem Kontakt mit einem Wirt; vollgesogene Larven und Nymphen dagegen bevorzugen niedrige Temperaturen. Nymphen und Imagines, die noch nicht gesogen haben, reagieren negativ geotaktisch. Sie erklettern vermutlich in der Natur die Spitzen von Pflanzen und Steinen, um dort das Vorbeikriechen eines Wirtes abzuwarten. Imagines von Amblyomma testudinis reagieren mit einer positiven Chemotaxis auf Schlangen stärker als auf Kröten. Auch Substrat aus Schlangenbehältern enthält mindestens für 30 Std nach der Entfernung der Schlangen chemotaktisch wirksame Stoffe.

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Contributions to sensory physiology of the tick Amblyomma testudinis

Summary Phototaxis, thermotaxis, geotaxis and chemotaxis of larvae, nymphs and adults of Amblyomma testudinis from own breedings were studied. The phototactic behaviour changed according to the different stages of development: unfed larvae were positively phototactic, fed larvae and unfed nymphs were indifferent, fed nymphs but also the adults showed a negative phototaxis.—All stages of development besides the females reacted without any preference for a distinct temperature, whereas larvae and nymphs after feeding prefered lower temperatures.—Unfed nymphs and adults were negatively geotactic which corresponds to their natural behaviour of climbing plants for catching hosts.—Adults of Amblyomma testudinis showed a higher rate of chemotaxis for snakes than for toads; substrate from snake cages containes chemotaxis inducing substances for more than 30 hours after elimination of the snakes.

     

Looking after yourself

People with HIV may face a wide array of medical choices, often when they are very ill and unable to make rational decisions on their own behalf. Healthcare proxies allow patients to appoint someone to make those decisions for them. The agent's power only starts when a doctor certifies that the patient can no longer make decisions. Medical providers should be notified that there is an agent who will make decisions on behalf of the patient, should the need arise. A number of issues relating to this topic are discussed, including medical directives, Do Not Resuscitate orders (DNR), and powers of attorney.     

Application of reflectance spectroscopy to the estimation of uric acid, urea and glucose: an evaluation of the Ames Seralyzer

An original approach to the measurement of analytes in clinical chemistry has now become available, in which dry reagent strip technology is linked to measurement by reflectance spectroscopy. The present studies have evaluated the performance of the first of these test systems—for uric acid, urea and glucose, in serum—by comparison with conventional liquid chemistry methods. Satisfactory performance in terms of both precision and accuracy was obtained for all three test systems, the current “state-of-the-art” performance criteria being met; the Seralyzer system proved reliable and easy to use in the hands of trained operators. It should find a place as a “Stat” analyser in the laboratory, in specified wards and in Health Centres.