New inhibitors for the blastgenation of human lymphocytes isolation from edible mushrooms (original) (raw)
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Activation of cytotoxic activity of human blood lymphocytes by tumor-promoting compounds
Cancer research, 1984
Three categories of tumor promoters and chemically related but inactive substances were tested for their effect on the cytotoxic activity of human blood lymphocytes against K562 and Daudi targets. Lymphocytes incubated overnight in the presence of phorbol esters 12-O-tetradecanoylphorbol-13-acetate and phorbol-12,13-dibutyrate [P(Bu)2] had enhanced function. Incubation with 4-alpha-phorbol-12,13-didecanoate was without effect. Enhancing activity was also exerted by the indole alkaloids, teleocidin and lyngbyatoxin A, and the polyacetates, aplysiatoxin and debromoaplysiatoxin, but not by dihydroteleocidin. Only the tumor-promoting compounds activated the cytotoxic potential. The substances acted in a dose-dependent manner with optimal activity at characteristic concentrations. Overnight incubation of lymphocytes at 4 degrees did not change their spontaneous cytotoxicity but abolished the enhancing effect of P(Bu)2. Thus, P(Bu)2-induced activation occurred only on metabolically active...
Biulleten' eksperimental'noĭ biologii i meditsiny, 1982
A rise of temperature affects the immune response in vivo and also potentiates the response of lymphoid cells to stimulation by lectins, antigens, and allogeneic cells in vitro [i, 4, 5, 8, i0]. Increased production of leukocyte migration inhibition factor when the temperature of lymphocytes in culture is raised has been described . Since stimulation of lymphocytes by antigens or mitogens takes place with the participation of soluble mediators giving rise to proliferation, it was decided to study the effect of a rise of temperature on production of mitogenic lymphokines (ML) in cultures of lymphocytes stimulated by phytohemagglutinin (PHA).
PHA-Induced Activation of Suppressor Cells in Normal Human Peripheral Blood Lymphocytes
Scandinavian Journal of Immunology, 1976
Normal human peripheral blood and tonsil lymphocytes can be stimulated to proliferate by phytohemagglutinin (PHA). When cells cultured with this mitogen for 3 days were transferred fo fresh autologous lymphocytes in fresh medium with PHA, the mitogen response of the fresh lymphocytes was suppressed. The suppression required the presence of viable cells, in that culture supernatants alone were not inhibitory and cell extracts showed only marginal inhibition. Approximately equivalent numbers of previously stimulated cells were required to produce optimal suppression of the PHA response of fresh cells. Cells irradiated after PHA stimulation were as effective as nonirradiated cells in causing suppression. PHA-stimulated cells also inhibited concanavalinA-induced proliferation and a mixed lymphocyte reaction. However, PHAstimulated cells only partially inhibited the response to pokeweed mitogen. The suppressive effects were fully retained by a nylon-wool-enriched T-cell fraction but not by a B-cell-enriched fraction.
International Immunopharmacology, 2011
Lymphocytes proliferation after antigen-driven activation leads to an increase in cell count, which could last some week, until apoptosis mechanisms allow the homeostatic control of the system. During the first days of this stimulation, activated lymphocytes display high resistance to apoptosis and to most immunosuppressive drugs. According to the literature, few compounds have been described to kill recently activated cells, by inhibiting metabolic processes fundamental to proliferation. The aim of our work was to evaluate comparatively these different compounds, in order to identify the best strategy to kill cells that have undergone proliferation, while sparing the repertoire of resting cells. After preliminary experiments, 3-HAA and bortezomib were selected as the most suitable compounds for our purposes. The possible synergic effect of 3-HAA with bortezomib or with manganese ions was also assessed. 3-HAA was confirmed to be the most reliable pharmacologic approach to inhibit proliferation with acceptable toxicity on resting cells. While in the case of PHA stimulation 3-HAA led to death of most lymphocytes, only a minor percentage of cells were killed after allo-stimulation, suggesting that the effect is proportional to the percentage of stimulated lymphocytes. Manganese ions further enhanced this effect, while results with bortezomib seemed to be less consistent. These results deserve further investigations to develop new procedures for targeting activated cells with pharmacological approaches.
Blotting methods applied to investigations of the mitogenic activity of phytohaemagglutinin (PHA
Journal of Immunological Methods, 1985
PHA polypeptides were separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose filters which were then cut into identical discs and used to perform blastogenic stimulation on cultured human peripheral blood lymphocytes. This method offers the following advantages: (1) the high resolution of SDS-polyacrylamide gel electrophoresis, (2) the sensitive immunochemical identification of the polypeptides involved in the mitogenic responses of lymphocytes, (3) the determination in the same experiment of the molecular weight, antigenicity and biological activities of the proteins.
Two Lymphocyte Models in Vitro
Cucurbitacin E (CuE), like other cucurbitacins, is a potential anti-neoplastic agent. The aim of the present study was to investigate the in vitro effects of CuE on phytohaemagglutinin (PHA)-activated and unstimulated lymphocytes. CuE did not produce any significant cytotoxic effects on the two models. On the contrary, it had a stimulatory effect, and was capable of inducing and maintaining high proliferation rates in lymphocytes. The stimulatory effect of CuE was concentration-dependent with a median stimulatory concentration (SC 50) of 1.166 µM and reaching maximal effect at a concentration of 10-20 µM. The stimulatory effect of CuE was about 25% lower than that of PHA, but a combination of CuE and PHA had an additive effect producing a greater response than that induced by the two substances used separately. Agarose gel electrophoresis for DNA fragmentation failed to show any significant apoptotic activity in the cells after 48 hours exposure to CuE.
Journal of Immunological Methods, 1988
Anti-/~ preparations differ greatly in their ability to stimulate mouse B cells to incorporate tritiated thymidine (TdR). We have found that these differences may be due in part to different levels of lipopolysaccharide (LPS) content. In this report we show that LPS concentrations as low as 0.025 ng/ml stimulate the proliferation of T-depleted (C57BL/6 × DBA/2)F 1 (B6DEF1) spleen cells, provided that 5 × 10-5 M 2-mercaptoethanol is also present. Each of six commercial anti-/~ preparations tested for LPS content contained more than this amount. We describe a technique that uses polymyxin B-agarose to remove nanogram quantities of LPS from anti-/x preparations. In B6DEF 1 B cells, LPS-depleted anti-# preparations induced much more uniform tritiated thymidine incorporation than did non-depleted preparations; but there was little difference between the two preparations when tested on B cells from C3H/HeJ (LPS hyporesponsive) mice.