Search PubMed⌕ Search

Biomedical subjects

C Aussel

Publications and source records attributed to C Aussel.

At least 73 records · Page 4Linked to original sources
PubMed1992

Changes in phospholipid metabolism induced by quinine, 4-aminopyridine and tetraethylammonium in the monocytic cell line THP1.

Quinine, 4-aminopyridine and tetraethylammonium, three compounds generally used as effectors of K+ channels, strongly modify phospholipid metabolism. In the human monocytic cell line THP1, the three drugs enhanced the incorporation of [3H]serine into phosphatidylserine and that of [3H]inositol into phosphatidylinositol in the absence of significant changes in the uptake of the 3H labels. On the contrary, the biosynthesis of both phosphatidylcholine and phosphatidylethanolamine was strongly inhibited. This inhibition appeared to be mainly due to the inhibition of both [3H]choline and [3H]ethanolamine uptake by the cells, by impairment of choline transport in a competitive mode.

4-Aminopyridine↗
PubMed1992

A diacylglycerol kinase is involved in the regulation of interleukin-2 synthesis in Jurkat T cells.

Diacylglycerol (DAG) plays a prominent role in several activation processes because of its ability, in the presence of calcium ions and phosphatidylserine, to activate C kinase. In T lymphocytes, however, DAG, produced in response to activating mAb or lectins, is rapidly metabolized into phosphatidic acid (PA) which may participate in further steps of activation. We thus investigated the involvement of a DAG kinase in several events subsequent to the activation of Jurkat T cells by CD3 mAb or phytohemagglutinin (PHA). We showed that three inhibitors of DAG kinase abrogated PA production and impaired calcium release from intracellular compartment, restored phosphatidylserine synthesis, and finally blocked IL-2 production in CD3- and PHA-stimulated cells. Postactivation DAG levels were not modified and DAG kinase inhibitors lowered IL-2 secretion even in the presence of phorbol ester that directly activates the C kinase. These results clearly demonstrate that, beside the effect of DAG on C kinase, DAG kinase dependent production of PA is crucial for further steps of T cell activation.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

Inhibition of phosphatidylserine synthesis induced by a CD4 mAb, B66.6 in Jurkat T cells.

A monoclonal antibody, B66.6, previously classified in the cluster of differentiation 4 (CD4), has been studied and compared with another CD4 monoclonal antibody, IOT4. It was found that B66.6 but not IOT4 was able to mobilize Ca2+ from intracellular stores in the Jurkat T cell line. Ca2+ mobilization was followed by a decrease in the extent of phosphatidylserine synthesis. In the presence of the phorbol ester, phorbol 12,13-dibutyrate, B66.6 induced interleukin-2 synthesis. Altogether, the results indicate that the CD4 monoclonal antibody, B66.6, mimics other T cell activators such as CD3 and confirm that the inhibition of phosphatidylserine synthesis in activated T cells follows the mobilization of Ca2+ from intracellular stores and is independent of the activation of the Ca(2+)-and phospholipid-dependent protein kinase C.

Antibodies, Monoclonal↗
PubMed1991

Diacylglycerol production in Jurkat T-cells: differences between CD3, CD2 and PHA activation pathways.

Diacylglycerol (DAG) production induced after stimulation with either CD3 mAb, a pair of CD2 mAbs or phytohaemagglutinin has been monitored in Jurkat T-cells prelabelled to isotopic equilibrium with seven [3H]- or [14C] fatty acids. It was found that CD3 induced a high production of arachidonic acid-labelled DAG and a modest production of oleic acid-DAG. The reverse was observed when using CD2 as activator. Phytohaemagglutinin induced a high production of these two DAG subspecies and in addition induced the production of linolenic acid-labelled DAG. Whatever the activator used no changes were observed in DAG production when cellular phospholipids were prelabelled with either myristic, palmitic, stearic or linoleic acids. All together our results strongly suggest that the three activation pathways previously described in T-lymphocytes might differ either at the level of the transduction mechanism or the phospholipid pools solicited during the activation process.

Antibodies, Monoclonal↗
PubMed1991

A phosphatidic acid-sensitive intracellular pool of calcium is released by anti-CD3 in Jurkat T cells.

Jurkat T cells, loaded with the fluorescent calcium probe Indo 1, responded to exogenous phosphatidic acid (PA) by transiently increasing their cytosolic Ca2+ concentration. This effect was dose-dependent, remained unmodified when external Ca2+ was chelated with EGTA, and was totally inhibited when cells were first exposed to CD3 monoclonal antibodies, indicating that it was solely due to the release of an intracellular pool, which is also mobilized during a stimulation via the CD3 T-cell receptor (TcR) molecular complex. CD3- and phytohaemagglutinin (PHA)-stimulated Jurkat cells also produced PA, the dose-responses and kinetics of which were consistent with those of calcium release. Moreover, diacylglycerol (DAG) kinase inhibitors abrogated PA production and lowered calcium release by CD3-stimulated cells. PA did not induce any apparent increase in inositol triphosphates (IP3), nor did it modify the increase entailed by activation of the CD3 pathway, pointing out that IP3 can be supplemented in mobilizing calcium from intracellular stores. Conversely, a first exposure to PA only partially inhibited the CD3- or ionomycin-induced internal release of calcium, suggesting either a rapid restoration of the PA-sensitive stores, or a contribution of other mediators, such as IP3, in the CD3 activation pathway.

Antibodies, Monoclonal↗
PubMed1991

Rheumatoid arthritis modifies basal and insulin-mediated glucose uptake by human synoviocytes.

Basal and insulin-mediated glucose uptake were studied in human synovial cells. Cell cultures were established from samples of synovium obtained at synovectomy from patients with osteoarthritis (non-rheumatoid synovial cells, NRSC) or rheumatoid arthritis (rheumatoid synovial cells, RSC). Basal glucose uptake was significantly higher in RSC than in NRSC. NRSC were sensitive to insulin at near-physiological concentrations (10(-8)M), with maximal transport occurring after a 30-min. association time (27.2 +/- 2.0%, mean +/- SEM). Insulin did not stimulate significantly 2-déoxy-D-glucose uptake in RSC, regardless of the association time (up to 120 min.) or the insulin concentration (10(-10) to 10(-6) M). Treatment of NRSC with human recombinant interleukin-1 beta (IL-1 beta) enhanced glucose uptake to a level similar to basal uptake by RSC. These results suggest that autocrine production of IL-1 beta by RSC could be responsible for the higher basal glucose uptake by these cells.

Arthritis, Rheumatoid↗
PubMed1990

Action of ornithine alpha-ketoglutarate, ornithine hydrochloride, and calcium alpha-ketoglutarate on plasma amino acid and hormonal patterns in healthy subjects.

Ornithine alpha-ketoglutarate (OKG) has been useful as an adjuvant of enteral and parenteral nutrition. However, its metabolism and mechanism of action remain unclear although it is known that alpha-ketoglutarate (alpha KG) and ornithine (ORN) follow, in part, common metabolic pathways. Six fasting healthy male subjects underwent three separate oral load tests: (i) they received 10 g of OKG (i.e., 3.6 g of alpha KG and 6.4 g of ORN); (ii) 6.4 g of ORN as ornithine hydrochloride, and (iii) 3.6 g of alpha KG as calcium alpha-ketoglutarate. Blood was drawn 15 times over a five-hour period for measurements of plasma amino acids, alpha KG, insulin, and glucagon. After OKG and ORN administration, plasma ORN peaked at 60-75 min (494 +/- 91 and 541 +/- 85 mumol/L). The increase in plasma alpha KG was very small. OKG, alpha KG, and ORN all increased glutamate concentrations at 60 min (mean: +43%, +68%, +68%, respectively, p less than 0.05 compared to basal values). However, only OKG increased proline and arginine levels at 60 min (mean: +35%, p less than 0.01 and mean: +41%, p less than 0.05). Furthermore, glutamate, proline, and arginine concentrations correlated linearly with ornithine levels at 60 min. Finally, OKG increased insulinemia and glucagonemia (mean: +24% at 15 min, p less than 0.05 and +30% at 60 min, p less than 0.01, respectively). These data provide evidence that the combination of ORN and alpha KG modifies amino acid metabolism in a way which is not observed when they are administered separately. In addition, the OKG-mediated increase in insulin levels probably does not appear to result from a direct action of ORN on pancreatic secretion.

Administration, Oral↗
PubMed1989

Modulation of T cell activation by differential regulation of the phosphorylation of two cytosolic proteins. Implication of both Ca2+ and cyclic AMP-dependent protein kinases.

Interleukin 2 production by activated Jurkat T cells is markedly decreased by prostaglandin E2 (PGE2). The target of PGE2 action has been investigated in the present study. Among the biochemical events occurring after CD3.TCR triggering by anti-CD3 monoclonal antibody, phosphorylation of two cytosolic proteins, pp21 and pp23, was strongly inhibited by PGE2, forskolin, and 8-bromo-cAMP, whereas anti-CD3 monoclonal antibody-induced CD3.TCR modulation and Ca2+ influx were not affected. The inhibition of both pp21 and pp23 phosphorylation and interleukin 2 synthesis by PGE2 can be largely reversed by the cAMP-dependent protein kinase inhibitor, N-[2-(methylamino)-ethyl-1]-5-isoquinoline sulfonamide. Together with the demonstration of a cAMP-dependent protein kinase activity in Jurkat T cells, these results are consistent with the participation of the cAMP-dependent protein kinase mediating the inhibitory action of PGE2, probably through the inhibition of pp21 and pp23 phosphorylation. Thus, it appears that the modulation of the phosphorylation of these cytosolic proteins represents an essential step in the regulation of T lymphocyte activation.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1989

Phosphorylation of two cytosolic proteins. An early event of T-cell activation.

T lymphocytes can be activated to proliferate by triggering the T-cell antigen-receptor complex (CD3-Ti) with anti-CD3 (Cluster of Differentiation 3) monoclonal antibody (mAb) or with the mitogenic lectin phytohaemagglutinin A (PHA). We have investigated the relationship between lymphocyte activation and protein phosphorylation in the human leukaemic T-cell line Jurkat. Incubation of 32P-labelled Jurkat cells with anti-CD3 mAb or PHA induced the phosphorylation of two cytosolic proteins that migrate with apparent Mr values of 21,000 (pp21) and 23,000 (pp23) and pI values of 5.1 and 5.0 respectively. Peptide mapping of the two proteins produced the same phosphopeptides pattern, suggesting that pp21 and pp23 are closely related. The phosphorylation of pp21 and pp23 induced by anti-CD3 mAb appeared to be transient, since it was already detected 2 min after the addition of the mAb, reached a maximum at 10 min and recovered its basal level after 1 h. Phosphorylation of pp21 and pp23 could also be elicited by the Ca2+ ionophore A23187 and sodium orthovanadate (Na3VO4), two agents that bypass the T-cell-receptor complex and produced an increase in cytosolic Ca2+ concentration. In addition, we found that vanadate, like the Ca2+ ionophore, induced the secretion of interleukin-2 (IL-2) when used in combination with a submitogenic concentration of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate. These results show that the Ca2+-dependent phosphorylation of pp21 and pp23 represents an early event in the process of signal transduction through the CD3-Ti receptor complex.

Calcimycin↗
PubMed1989

Chymotryptic-type protease inhibitors block the increase in Ca2+ and Il-2 production in activated Jurkat T cells.

Several chymotryptic-type protease inhibitors were found to inhibit both anti-CD3 mAb- and PHA-induced rise in Ca2+ and IL-2 production in Jurkat T cells. The magnitude of inhibition was a function of the effectors used to stimulate Ca2+ entry and depended on the concentration of the inhibitors. Neither tryptic-type protease inhibitors nor an elastase substrate prevented anti-CD3 mAb- or PHA-induced Ca2+ rise in Jurkat cells. The inhibitory effect of N-alpha-p-tosyl-L-phenylalanine chloromethyl-ketone on anti-CD3 mAb- and PHA-induced rise in Ca2+ resulted from a rapid increase in Ca2+ efflux. The inhibitors which were effective on Ca2+ mobilization also inhibited IL-2 production initiated by an anti-CD3 mAb in the presence of 12-O-tetradecanoylphorbol-13-acetate, and to a lesser extent by PHA or the calcium ionophore A23187. No inhibition of IL-2 production was observed when tryptic-type protease inhibitors or the elastase inhibitor were used. In addition, membrane preparations from Jurkat cells were found to hydrolyze the chymotryptic substrate Suc-Ala-Ala-Phe-paranitroaniline, an effect markedly inhibited by N-alpha-p-tosyl-L-phenylalanine chloromethylketone. Moreover, this inhibitor protected one potential endogenous substrate (Mr 38 kDa) from proteolysis. Taken together, these observations show that chymotryptic-type protease inhibitors block the responses generated by the binding of anti-CD3 mAb to Jurkat cells, and suggest that a chymotryptic-like membrane protease contributes to T cell activation.

Antibodies, Monoclonal↗
PubMed1989

Neutral amino acid transport in human synovial cells: substrate specificity of adaptative regulation and transinhibition.

Neutral amino acid transport was characterized in human synovial cells. The amino acids tested are transported by all three major neutral amino acid transport systems, that is, A, L, and ASC. The model amino acid 2-aminoisobutyric acid (AIB) was found to be a strong specific substrate for system A in synovial cells. When cells were starved of amino acids, the activity of AIB transport increased, reaching a maximum within 1 h. The stimulation of transport activity was not blocked by cycloheximide and would thus appear to be related to a release from transinhibition. Similarly, the decrease in the activity of AIB transport observed after the addition of alpha-methyl-aminoisobutyric acid (meAIB) appeared to be related to transinhibition. However, using a different approach, that is, amino acid starvation followed by incubation with 10 mM meAIB and transfer to an amino acid-free medium with or without cycloheximide supplementation, a clear increase in AIB uptake, due both to derepression and a release from transinhibition, was observed. Unlike human fibroblasts, the depression of system A in these synovial cells was not serum-dependent. The process of derepression was observed only after preloading with meAIB. Neither AIB nor alanine produced this phenomenon. Moreover, alanine preloading led to a large increase in AIB transport activity due to a release from transinhibition. These observations indicate that the process of derepression and release from transinhibition are specific to the substrates present in the culture medium prior to amino acid starvation.

Alanine↗
PubMed1989

Inhibition of phosphatidylserine synthesis induced by triggering CD2 or the CD3-TCR complex in a human T cell line. Relationships with G proteins and receptors modulation.

Activation of Jurkat T cells with phytohemagglutinin, CD3 or CD2 mAbs results in a marked inhibition of phosphatidylserine (PS) synthesis. Monitoring PS synthesis in T cells shows that: (i) after modulation of CD3 molecules the cells become refractory to further treatment with CD3 mAbs as well as to a further challenge with CD2 mAbs; and (ii) treatment of T cells with fluoride ions and cholera toxin, two known effectors of guanosine triphosphate-binding proteins, also resulted in a strong inhibition of the synthesis of this phospholipid. The inhibition of PS synthesis thus appears to be regulated similarly to the other activation events, suggesting that transmembrane signalling mechanisms leading to PS inhibition are the same as those previously proposed for increasing phosphatidylinositides turnover and subsequent rise in the intracellular calcium concn in lymphocytes.

Antibodies, Monoclonal↗
PubMed1989

Phospholipid metabolism and T cell activation: receptor triggering is associated with the inhibition of phosphatidylserine synthesis.

Activation of Jurkat T lymphocytes to produce IL2 is accompanied by a strong inhibition of phosphatidylserine (PS) synthesis. This inhibition was obtained either with the mitogenic lectin PHA, anti-CD3 monoclonal antibodies (mAb), anti-CD2 mAb or anti-Ti mAb. Bypassing membrane receptor signalling, by using a Ca2+ ionophore or a protein phosphatase inhibitor, sodium ortho-vanadate, also results in a marked inhibition of PS synthesis. Activators of phospholipid -Ca2+ dependent protein kinase C (PKC) did not significantly modify PS synthesis, suggesting that the observed changes only involve the transduction of the first activation signal. PS being a necessary cofactor for PKC, our results strongly suggest that the inhibition of PS synthesis induced by receptor triggering exerts a feed back control on PKC therefore leading to a transient activation of the enzyme upon full lymphocyte activation.

Antibodies, Monoclonal↗
PubMed1989

A chymotryptic-type protease inhibitor decreases interleukin 2 synthesis and induces prostaglandin production in Jurkat T cells.

TPCK (N-alpha-p-tosyl-L-phenylalanine chloromethylketone), a potent inhibitor of chymotryptic-type serine proteases, was found to decrease IL2 synthesis in Jurkat T cells. Conversely, the tryptic-type protease inhibitor, TLCK (N-alpha-p-tosyl-lysine chloromethylketone), which structurally is very similar to TPCK, had no effect on IL2 synthesis. Prostaglandin synthesis, a process that is known to reduce IL2 production in T cells, was increased by TPCK but not by TLCK, suggesting that this process could be, at least in part, responsible for the inhibition of IL2 production. Our results imply that a chymotryptic-type serine protease plays an active role in the regulation of IL2 synthesis and thus in the whole process of T-lymphocyte activation.

Amino Acid Chloromethyl Ketones↗
PubMed1988

IL-1 signaling for IL-2 production in T cells involves a rise in phosphatidylserine synthesis.

Activation of Jurkat T cells with anti-TCR, anti-CD3, anti-CD2, or PHA is accompanied by a strong inhibition of phosphatidylserine (PS) synthesis. The inhibition of the synthesis of this phospholipid could be partially reversed by IL-1. In Jurkat cells, IL-1 did not activate phosphodiesterases as demonstrated by the lack of change of inositol triphosphate and diacylglycerol levels as well as the lack of change in cytosolic Ca2+ concentration. Furthermore, IL-1 did not modify the intracellular level of cGMP and cAMP, suggesting that the observed rise of PS synthesis could play the role of mediator IL-1 action. As PS is a necessary cofactor for the activation of protein kinase C, our results suggest strongly that IL-1 modulate protein kinase C activity in the activated lymphocyte through its action on PS synthesis.

Cell Line↗
PubMed1988

Inhibition and activation of interleukin 2 synthesis by direct modification of guanosine triphosphate-binding proteins.

To investigate whether guanosine triphosphate-binding proteins (G proteins) are involved in T cell activation, tests were made of the effect of pertussis toxin, cholera toxin, guanosine 5'-(3-O-thio)-triphosphate, and fluoride ions on interleukin 2 (IL-2) synthesis in Jurkat cells. It was found: 1) that pertussis toxin interferes with the first pathway of T cell activation insofar as it can substitute for phytohemagglutinin or monoclonal antibodies directed against the CD3 surface proteins, suggesting that a G protein serves as transducer for signals via the T cell receptor-CD3 complex; and 2) that fluoride ions induce the release of diacylglycerol (DAG) from [3H] arachidonic acid or [3H]oleic acid-prelabeled cells. In [3H]inositol or 32P-prelabeled cells, the increase in DAG production was also found to be accompanied by a 280% increase of intracellular inositol phosphate (IP), without significant modification of IP2 and IP3. These results suggest that a G protein controls the activity of a phospholipase C in Jurkat cells that upon stimulation releases DAG but not IP3. Inasmuch as DAG, like the phorbol ester tetradecanoyl phorbol acetate, activates protein kinase C, it suggests that a G protein is also involved in the transduction of the second signal for lymphocyte activation. Fluoride ions were found to be as effective as tetradecanoyl phorbol acetate to stimulate IL-2 synthesis in Jurkat cells when used in combination with phytohemagglutinin. Finally, cholera toxin and guanosine 5'-(3-O-thio)-triphosphate were found to increase intracellular cyclic adenosine triphosphate and to inhibit IL-2 synthesis. All together these results suggest that several G proteins are involved in the transduction of the two signals necessary for T cell activation as well as in the negative regulation of IL-2 synthesis.

Antigens, Differentiation, T-Lymphocyte↗