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. 1997 Aug;91(4):609–617. doi: 10.1046/j.1365-2567.1997.00296.x

Distribution of cyclophilin B-binding sites in the subsets of human peripheral blood lymphocytes.

A Denys 1, F Allain 1, B Foxwell 1, G Spik 1
PMCID: PMC1363883  PMID: 9378502

Abstract

Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway and released in biological fluids. We have recently demonstrated that both free CyPB and CyPB-CsA complex specifically bind to peripheral blood T lymphocytes and are internalized. These results suggest that CyPB might promote the targeting of the drug into sensitive cells. Peripheral blood lymphocytes are subdivided in several populations according to their biological functions and sensitivity to CsA. We have investigated the binding of CyPB to these different subsets using a CyPB derivatized by fluorescein through its single cysteine which retains its binding properties. We have confirmed that only T cells were involved in the interaction with CyPB. The ligand binding was found to be heterogeneously distributed on the different T-cell subsets and surface-bound CyPB was mainly associated with the CD4-positive cells. No significant difference was noted between the CD45RA and CD45RO subsets, demonstrating that CyPB-binding sites were equally distributed between native and memory T cells. CD3 stimulation of T lymphocytes led to a decrease in the CyPB-binding capacity, that may be explained by a down-regulation of the CyPB-receptor expression upon T-cell activation. Finally, we demonstrated that CyPB-receptor-positive cells, isolated on CyPB sulphydryl-coupled affinity matrices, are more sensitive to CyPB-complexed CsA than mixed peripheral blood lymphocytes, suggesting that CyPB potentiates CsA activity through the binding of the complex. Taken together, our results demonstrate that CyPB-binding sites are mainly associated with resting cells of the helper T lymphocyte, and that CyPB might modulate the distribution of CsA through the drug targeting to sensitive cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akbar A. N., Salmon M., Janossy G. The synergy between naive and memory T cells during activation. Immunol Today. 1991 Jun;12(6):184–188. doi: 10.1016/0167-5699(91)90050-4. [DOI] [PubMed] [Google Scholar]
  2. Allain F., Boutillon C., Mariller C., Spik G. Selective assay for CyPA and CyPB in human blood using highly specific anti-peptide antibodies. J Immunol Methods. 1995 Jan 13;178(1):113–120. doi: 10.1016/0022-1759(94)00249-v. [DOI] [PubMed] [Google Scholar]
  3. Allain F., Denys A., Spik G. Cyclophilin B mediates cyclosporin A incorporation in human blood T-lymphocytes through the specific binding of complexed drug to the cell surface. Biochem J. 1996 Jul 15;317(Pt 2):565–570. doi: 10.1042/bj3170565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arber S., Krause K. H., Caroni P. s-cyclophilin is retained intracellularly via a unique COOH-terminal sequence and colocalizes with the calcium storage protein calreticulin. J Cell Biol. 1992 Jan;116(1):113–125. doi: 10.1083/jcb.116.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Borel J. F., Feurer C., Gubler H. U., Stähelin H. Biological effects of cyclosporin A: a new antilymphocytic agent. Agents Actions. 1976 Jul;6(4):468–475. doi: 10.1007/BF01973261. [DOI] [PubMed] [Google Scholar]
  6. Bunjes D., Hardt C., Röllinghoff M., Wagner H. Cyclosporin A mediates immunosuppression of primary cytotoxic T cell responses by impairing the release of interleukin 1 and interleukin 2. Eur J Immunol. 1981 Aug;11(8):657–661. doi: 10.1002/eji.1830110812. [DOI] [PubMed] [Google Scholar]
  7. Davis T. R., Tabatabai L., Bruns K., Hamilton R. T., Nilsen-Hamilton M. Basic fibroblast growth factor induces 3T3 fibroblasts to synthesize and secrete a cyclophilin-like protein and beta 2-microglobulin. Biochim Biophys Acta. 1991 Oct 26;1095(2):145–152. doi: 10.1016/0167-4889(91)90077-b. [DOI] [PubMed] [Google Scholar]
  8. Fischer G., Wittmann-Liebold B., Lang K., Kiefhaber T., Schmid F. X. Cyclophilin and peptidyl-prolyl cis-trans isomerase are probably identical proteins. Nature. 1989 Feb 2;337(6206):476–478. doi: 10.1038/337476a0. [DOI] [PubMed] [Google Scholar]
  9. Galat A. Peptidylproline cis-trans-isomerases: immunophilins. Eur J Biochem. 1993 Sep 15;216(3):689–707. doi: 10.1111/j.1432-1033.1993.tb18189.x. [DOI] [PubMed] [Google Scholar]
  10. Granelli-Piperno A., Inaba K., Steinman R. M. Stimulation of lymphokine release from T lymphoblasts. Requirement for mRNA synthesis and inhibition by cyclosporin A. J Exp Med. 1984 Dec 1;160(6):1792–1802. doi: 10.1084/jem.160.6.1792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Handschumacher R. E., Harding M. W., Rice J., Drugge R. J., Speicher D. W. Cyclophilin: a specific cytosolic binding protein for cyclosporin A. Science. 1984 Nov 2;226(4674):544–547. doi: 10.1126/science.6238408. [DOI] [PubMed] [Google Scholar]
  12. Hannet I., Erkeller-Yuksel F., Lydyard P., Deneys V., DeBruyère M. Developmental and maturational changes in human blood lymphocyte subpopulations. Immunol Today. 1992 Jun;13(6):215–218. doi: 10.1016/0167-5699(92)90157-3. [DOI] [PubMed] [Google Scholar]
  13. Hess A. D., Tutschka P. J. Effect of cyclosporin A on human lymphocyte responses in vitro. I. CsA allows for the expression of alloantigen-activated suppressor cells while preferentially inhibiting the induction of cytolytic effector lymphocytes in MLR. J Immunol. 1980 Jun;124(6):2601–2608. [PubMed] [Google Scholar]
  14. Jobbágy A., Király K. Chemical characterization of fluorescein isothiocyanate-protein conjugates. Biochim Biophys Acta. 1966 Jul 27;124(1):166–175. doi: 10.1016/0304-4165(66)90325-4. [DOI] [PubMed] [Google Scholar]
  15. June C. H., Ledbetter J. A., Linsley P. S., Thompson C. B. Role of the CD28 receptor in T-cell activation. Immunol Today. 1990 Jun;11(6):211–216. doi: 10.1016/0167-5699(90)90085-n. [DOI] [PubMed] [Google Scholar]
  16. Liu J., Albers M. W., Wandless T. J., Luan S., Alberg D. G., Belshaw P. J., Cohen P., MacKintosh C., Klee C. B., Schreiber S. L. Inhibition of T cell signaling by immunophilin-ligand complexes correlates with loss of calcineurin phosphatase activity. Biochemistry. 1992 Apr 28;31(16):3896–3901. doi: 10.1021/bi00131a002. [DOI] [PubMed] [Google Scholar]
  17. Liu J., Farmer J. D., Jr, Lane W. S., Friedman J., Weissman I., Schreiber S. L. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807–815. doi: 10.1016/0092-8674(91)90124-h. [DOI] [PubMed] [Google Scholar]
  18. Mariller C., Haendler B., Allain F., Denys A., Spik G. Involvement of the N-terminal part of cyclophilin B in the interaction with specific Jurkat T-cell binding sites. Biochem J. 1996 Jul 15;317(Pt 2):571–576. doi: 10.1042/bj3170571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Motta I., Colle J. H., Shidani B., Truffa-Bachi P. Interleukin 2/interleukin 4-independent T helper cell generation during an in vitro antigenic stimulation of mouse spleen cells in the presence of cyclosporin A. Eur J Immunol. 1991 Mar;21(3):551–557. doi: 10.1002/eji.1830210304. [DOI] [PubMed] [Google Scholar]
  20. Schreiber S. L., Crabtree G. R. The mechanism of action of cyclosporin A and FK506. Immunol Today. 1992 Apr;13(4):136–142. doi: 10.1016/0167-5699(92)90111-J. [DOI] [PubMed] [Google Scholar]
  21. Sherry B., Yarlett N., Strupp A., Cerami A. Identification of cyclophilin as a proinflammatory secretory product of lipopolysaccharide-activated macrophages. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3511–3515. doi: 10.1073/pnas.89.8.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Spik G., Haendler B., Delmas O., Mariller C., Chamoux M., Maes P., Tartar A., Montreuil J., Stedman K., Kocher H. P. A novel secreted cyclophilin-like protein (SCYLP). J Biol Chem. 1991 Jun 15;266(17):10735–10738. [PubMed] [Google Scholar]
  23. Takahashi N., Hayano T., Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporin A-binding protein cyclophilin. Nature. 1989 Feb 2;337(6206):473–475. doi: 10.1038/337473a0. [DOI] [PubMed] [Google Scholar]

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