scholarly journals Cyclophilin B mediates cyclosporin A incorporation in human blood T-lymphocytes through the specific binding of complexed drug to the cell surface

1996 ◽  
Vol 317 (2) ◽  
pp. 565-570 ◽  
Author(s):  
Fabrice ALLAIN ◽  
Agnès DENYS ◽  
Geneviève SPIK
Keyword(s):  
T Lymphocytes ◽  
Cell Surface ◽  
Cyclosporin A ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Binding Capacity ◽  
Specific Binding ◽  
Biological Fluids ◽  
Cell Surface Receptor ◽  
Cyclophilin B

Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein located within intracellular vesicles and released in biological fluids. We recently reported the specific binding of this protein to T-cell surface receptor which is internalized even in the presence of CsA. These results suggest that CyPB might target the drug to lymphocytes and consequently modify its activity. To verify this hypothesis, we have first investigated the binding capacity and internalization of the CsA–CyPB complex in human peripheral blood T-lymphocytes and secondly compared the inhibitory effect of both free and CyPB-complexed CsA on the CD3-induced activation and proliferation of T-cells. Here, we present evidence that both the CsA–CyPB complex and free CyPB bind to the T-lymphocyte surface, with similar values of Kd and number of sites. At 37 °C, the complex is internalized but, in contrast to the protein, the drug is accumulated within the cell. Moreover, CyPB receptors are internalized together with the ligand and rapidly recycled to the cell surface. Finally, we demonstrate that CyPB-complexed CsA remains as efficient as uncomplexed CsA and that CyPB enhances the immunosuppressive activity of the drug. Taken together, our results support the hypothesis that surface CyPB receptors may be related to the selective and variable action of CsA, through specific binding and targeting of the CyPB–CsA complex to peripheral blood T-lymphocytes.

scholarly journals Involvement of two classes of binding sites in the interactions of cyclophilin B with peripheral blood T-lymphocytes

1998 ◽  
Vol 336 (3) ◽  
pp. 689-697 ◽  
Author(s):  
Agnès DENYS ◽  
Fabrice ALLAIN ◽  
Mathieu CARPENTIER ◽  
Geneviève SPIK
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Cell Surface ◽  
Peripheral Blood ◽  
Binding Sites ◽  
Secretory Pathway ◽  
Biological Fluids ◽  
Type I ◽  
Cyclophilin B ◽  
Sulphated Glycosaminoglycans

Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway, and is released in biological fluids. We recently reported that CyPB specifically binds to T-lymphocytes and promotes enhanced incorporation of CsA. The interactions with cellular binding sites involved, at least in part, the specific N-terminal extension of the protein. In this study, we intended to specify further the nature of the CyPB-binding sites on peripheral blood T-lymphocytes. We first provide evidence that the CyPB binding to heparin-Sepharose is prevented by soluble sulphated glycosaminoglycans (GAG), raising the interesting possibility that such interactions may occur on the T-cell surface. We then characterized CyPB binding to T-cell surface GAG and found that these interactions involved the N-terminal extension of CyPB, but not its conserved CsA-binding domain. In addition, we determined the presence of a second CyPB binding site, which we termed a type I site, in contrast with type II for GAG interactions. The two binding sites exhibit a similar affinity but the expression of the type I site was 3-fold lower. The conclusion that CyPB binding to the type I site is distinct from the interactions with GAG was based on the findings that it was (1) resistant to NaCl wash and GAG-degrading enzyme treatments, (2) reduced in the presence of CsA or cyclophilin C, and (3) unmodified in the presence of either the N-terminal peptide of CyPB or protamine. Finally, we showed that the type I binding sites were involved in an endocytosis process, supporting the hypothesis that they may correspond to a functional receptor for CyPB.

scholarly journals Diacylglycerol activates the influx of extracellular cations in T-lymphocytes independently of intracellular calcium-store depletion and possibly involving endogenous TRP6 gene products

2002 ◽  
Vol 364 (1) ◽  
pp. 245-254 ◽  
Author(s):  
Alessandra GAMBERUCCI ◽  
Emanuele GIURISATO ◽  
Paola PIZZO ◽  
Maristella TASSI ◽  
Roberta GIUNTI ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
T Lymphocytes ◽  
Peripheral Blood ◽  
Transient Receptor Potential ◽  
Human Peripheral Blood ◽  
Receptor Potential ◽  
Animal Cells ◽  
Bivalent Cation ◽  
Intracellular Calcium Store ◽  
Transient Receptor

In Jurkat and human peripheral blood T-lymphocytes, 1-oleoyl-2-acetyl-sn-glycerol (OAG), a membrane-permeant analogue of diacylglycerol, activated the influx of Ca2+, Ba2+ and Sr2+. OAG also caused plasma-membrane depolarization in Ca2+-free media that was recovered by the addition of bivalent cation, indicating the activation of Na+ influx. OAG-induced cation influx was (i) mimicked by the natural dacylglycerol 1-stearoyl-2-arachidonyl-sn-glycerol, (ii) not blocked by inhibiting protein kinase C or in the absence of phopholipase C activity and (iii) blocked by La3+ and Gd3+. Differently from OAG, both thapsigargin and phytohaemagglutinin activated a potent influx of Ca2+, but little influx of Ba2+ and Sr2+. Moreover, the influx of Ca2+ activated by thapsigargin and that activated by OAG were additive. Furthermore, several drugs (i.e. econazole, SKF96365, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, 2-aminoethoxy diphenylborate and calyculin-A), while inhibiting the influx of Ca2+ induced by both thapsigargin and phytohaemagglutinin, did not affect OAG-stimulated cation influx. Transient receptor potential (TRP) 3 and TRP6 proteins have been shown previously to be activated by diacylglycerol when expressed heterologously in animal cells [Hofmann, Obukhov, Schaefer, Harteneck, Gudermann and Schultz (1999) Nature (London) 397, 259–263]. In both Jurkat and peripheral blood T-lymphocytes, mRNA encoding TRP proteins 1, 3, 4 and 6 was detected by reverse transcriptase PCR, and the TRP6 protein was detected by Western blotting in a purified plasma-membrane fraction. We conclude that T-cells express a diacylglycerol-activated cation channel, unrelated to the channel involved in capacitative Ca2+ entry, and associated with the expression of TRP6 protein.

Differential chemokine expression profiles in human peripheral blood T lymphocytes: dependence on T-cell coreceptor and calcineurin signaling

Blood ◽  
2003 ◽  
Vol 101 (1) ◽  
pp. 216-225 ◽  
Author(s):  
Anthony D. Cristillo ◽  
Mirtha J. Macri ◽  
Barbara E. Bierer
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Signaling Pathways ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Expression Profiles ◽  
Lymphoid Tissues ◽  
Rnase Protection ◽  
Chemokine Expression ◽  
Calcineurin Activity

Abstract The chemokine superfamily consists of small (8-10 kDa) molecules that function to attract, selectively, different subsets of leukocytes. Binding of chemokines to their appropriate G-protein–coupled receptors is necessary for primary immune responses and for homing of leukocytes to lymphoid tissues. Here, we have characterized the signaling pathways in primary T lymphocytes that regulate chemokine gene induction using an RNase protection assay. Dependence on stimulation through the coreceptor CD28 and sensitivity to the calcineurin inhibitors cyclosporine and tacrolimus were studied using purified human peripheral blood lymphocytes. Lymphotactin (Ltn), macrophage inflammatory protein (MIP)–1α, and MIP-1β were all rapidly induced and sensitive to cyclosporine treatment. At later time points, the expression of MIP-1α and MIP-1β, but not of Ltn, was restored despite the inhibition of calcineurin activity. By contrast, the induction of interleukin-8 was delayed and was found to be cyclosporine insensitive. Calcineurin activity of IP-10 mRNA induction was contingent on the specific T-cell stimulation conditions, suggesting that IP-10 expression is modulated by calcineurin-dependent and -independent signaling pathways. Differential chemokine expression profiles result from the engagement of T-cell coreceptors and the requirement for, and the dependence on, calcineurin phosphatase activity.

scholarly journals Expression of IκBα in the nucleus of human peripheral blood T lymphocytes

Oncogene ◽  
1999 ◽  
Vol 18 (8) ◽  
pp. 1581-1588 ◽  
Author(s):  
Teresa Laín de Lera ◽  
Lola Folgueira ◽  
Angel G Martín ◽  
Catherine Dargemont ◽  
María-Antonia Pedraza ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Peripheral Blood ◽  
Human Peripheral Blood

scholarly journals Derivation of Induced Pluripotent Stem Cells from Human Peripheral Blood T Lymphocytes

PLoS ONE ◽  
2010 ◽  
Vol 5 (6) ◽  
pp. e11373 ◽  
Author(s):  
Matthew E. Brown ◽  
Elizabeth Rondon ◽  
Deepika Rajesh ◽  
Amanda Mack ◽  
Rachel Lewis ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Lymphocytes ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Induced Pluripotent

TCR γ Chain Expression on Human Peripheral Blood T Lymphocytes

1989 ◽  
pp. 551-553
Author(s):  
Frits Koning ◽  
Rafick P. Sekaly ◽  
Erwin Tschachler ◽  
Roberto Biassoni ◽  
Marvin S. Reitz ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Peripheral Blood ◽  
Human Peripheral Blood
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