Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1996 Feb 1;183(2):413–420. doi: 10.1084/jem.183.2.413

T cell responses in calcineurin A alpha-deficient mice

PMCID: PMC2192457  PMID: 8627154

Abstract

We have created embryonic stem (ES) cells and mice lacking the predominant isoform (alpha) of the calcineurin A subunit (CNA alpha) to study the role of this serine/threonine phosphatase in the immune system. T and B cell maturation appeared to be normal in CNA alpha -/- mice. CNA alpha -/- T cells responded normally to mitogenic stimulation (i.e., PMA plus ionomycin, concanavalin A, and anti-CD3 epsilon antibody). However, CNA alpha -/- mice generated defective antigen- specific T cell responses in vivo. Mice produced from CNA alpha -/- ES cells injected into RAG-2-deficient blastocysts had a similar defective T cell response, indicating that CNA alpha is required for T cell function per se, rather than for an activity of other cell types involved in the immune response. CNA alpha -/- T cells remained sensitive to both cyclosporin A and FK506, suggesting that CNA beta or another CNA-like molecule can mediate the action of these immunosuppressive drugs. CNA alpha -/- mice provide an animal model for dissecting the physiologic functions of calcineurin as well as the effects of FK506 and CsA.

Full Text

The Full Text of this article is available as a PDF (774.5 KB).

Selected References

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

  1. Bierer B. E., Holländer G., Fruman D., Burakoff S. J. Cyclosporin A and FK506: molecular mechanisms of immunosuppression and probes for transplantation biology. Curr Opin Immunol. 1993 Oct;5(5):763–773. doi: 10.1016/0952-7915(93)90135-f. [DOI] [PubMed] [Google Scholar]
  2. Chen J., Lansford R., Stewart V., Young F., Alt F. W. RAG-2-deficient blastocyst complementation: an assay of gene function in lymphocyte development. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4528–4532. doi: 10.1073/pnas.90.10.4528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clipstone N. A., Crabtree G. R. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature. 1992 Jun 25;357(6380):695–697. doi: 10.1038/357695a0. [DOI] [PubMed] [Google Scholar]
  4. Cohen P. The structure and regulation of protein phosphatases. Annu Rev Biochem. 1989;58:453–508. doi: 10.1146/annurev.bi.58.070189.002321. [DOI] [PubMed] [Google Scholar]
  5. Frantz B., Nordby E. C., Bren G., Steffan N., Paya C. V., Kincaid R. L., Tocci M. J., O'Keefe S. J., O'Neill E. A. Calcineurin acts in synergy with PMA to inactivate I kappa B/MAD3, an inhibitor of NF-kappa B. EMBO J. 1994 Feb 15;13(4):861–870. doi: 10.1002/j.1460-2075.1994.tb06329.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Friedman J., Weissman I. Two cytoplasmic candidates for immunophilin action are revealed by affinity for a new cyclophilin: one in the presence and one in the absence of CsA. Cell. 1991 Aug 23;66(4):799–806. doi: 10.1016/0092-8674(91)90123-g. [DOI] [PubMed] [Google Scholar]
  7. Guerini D., Klee C. B. Cloning of human calcineurin A: evidence for two isozymes and identification of a polyproline structural domain. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9183–9187. doi: 10.1073/pnas.86.23.9183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ito A., Hashimoto T., Hirai M., Takeda T., Shuntoh H., Kuno T., Tanaka C. The complete primary structure of calcineurin A, a calmodulin binding protein homologous with protein phosphatases 1 and 2A. Biochem Biophys Res Commun. 1989 Sep 29;163(3):1492–1497. doi: 10.1016/0006-291x(89)91148-0. [DOI] [PubMed] [Google Scholar]
  9. Kincaid R. L., Giri P. R., Higuchi S., Tamura J., Dixon S. C., Marietta C. A., Amorese D. A., Martin B. M. Cloning and characterization of molecular isoforms of the catalytic subunit of calcineurin using nonisotopic methods. J Biol Chem. 1990 Jul 5;265(19):11312–11319. [PubMed] [Google Scholar]
  10. Kincaid R. L., Takayama H., Billingsley M. L., Sitkovsky M. V. Differential expression of calmodulin-binding proteins in B, T lymphocytes and thymocytes. Nature. 1987 Nov 12;330(6144):176–178. doi: 10.1038/330176a0. [DOI] [PubMed] [Google Scholar]
  11. Kuno T., Mukai H., Ito A., Chang C. D., Kishima K., Saito N., Tanaka C. Distinct cellular expression of calcineurin A alpha and A beta in rat brain. J Neurochem. 1992 May;58(5):1643–1651. doi: 10.1111/j.1471-4159.1992.tb10036.x. [DOI] [PubMed] [Google Scholar]
  12. Kuno T., Takeda T., Hirai M., Ito A., Mukai H., Tanaka C. Evidence for a second isoform of the catalytic subunit of calmodulin-dependent protein phosphatase (calcineurin A). Biochem Biophys Res Commun. 1989 Dec 29;165(3):1352–1358. doi: 10.1016/0006-291x(89)92752-6. [DOI] [PubMed] [Google Scholar]
  13. Li E., Bestor T. H., Jaenisch R. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell. 1992 Jun 12;69(6):915–926. doi: 10.1016/0092-8674(92)90611-f. [DOI] [PubMed] [Google Scholar]
  14. Lieberman S. A., Spain L. M., Wang L., Berg L. J. Enhanced T cell maturation and altered lineage commitment in T cell receptor/CD4-transgenic mice. Cell Immunol. 1995 Apr 15;162(1):56–67. doi: 10.1006/cimm.1995.1051. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Mansour S. L., Thomas K. R., Capecchi M. R. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988 Nov 24;336(6197):348–352. doi: 10.1038/336348a0. [DOI] [PubMed] [Google Scholar]
  18. Mathis D. J., Benoist C., Williams V. E., 2nd, Kanter M., McDevitt H. O. Several mechanisms can account for defective E alpha gene expression in different mouse haplotypes. Proc Natl Acad Sci U S A. 1983 Jan;80(1):273–277. doi: 10.1073/pnas.80.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McKnight A. J., Zimmer G. J., Fogelman I., Wolf S. F., Abbas A. K. Effects of IL-12 on helper T cell-dependent immune responses in vivo. J Immunol. 1994 Mar 1;152(5):2172–2179. [PubMed] [Google Scholar]
  20. Metcalfe S., Alexander D., Turner J. FK506 and cyclosporin A each inhibit antigen-specific signaling in the T cell line 171 in the absence of a calcium signal. Cell Immunol. 1994 Oct 1;158(1):46–58. doi: 10.1006/cimm.1994.1255. [DOI] [PubMed] [Google Scholar]
  21. Mortensen R. M., Conner D. A., Chao S., Geisterfer-Lowrance A. A., Seidman J. G. Production of homozygous mutant ES cells with a single targeting construct. Mol Cell Biol. 1992 May;12(5):2391–2395. doi: 10.1128/mcb.12.5.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. O'Keefe S. J., Tamura J., Kincaid R. L., Tocci M. J., O'Neill E. A. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin. Nature. 1992 Jun 25;357(6380):692–694. doi: 10.1038/357692a0. [DOI] [PubMed] [Google Scholar]
  23. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Schreiber S. L. Immunophilin-sensitive protein phosphatase action in cell signaling pathways. Cell. 1992 Aug 7;70(3):365–368. doi: 10.1016/0092-8674(92)90158-9. [DOI] [PubMed] [Google Scholar]
  26. Sewell T. J., Lam E., Martin M. M., Leszyk J., Weidner J., Calaycay J., Griffin P., Williams H., Hung S., Cryan J. Inhibition of calcineurin by a novel FK-506-binding protein. J Biol Chem. 1994 Aug 19;269(33):21094–21102. [PubMed] [Google Scholar]
  27. Shinkai Y., Rathbun G., Lam K. P., Oltz E. M., Stewart V., Mendelsohn M., Charron J., Datta M., Young F., Stall A. M. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell. 1992 Mar 6;68(5):855–867. doi: 10.1016/0092-8674(92)90029-c. [DOI] [PubMed] [Google Scholar]
  28. Siekierka J. J., Sigal N. H. FK-506 and cyclosporin A: immunosuppressive mechanism of action and beyond. Curr Opin Immunol. 1992 Oct;4(5):548–552. doi: 10.1016/0952-7915(92)90024-9. [DOI] [PubMed] [Google Scholar]
  29. Stemmer P. M., Klee C. B. Dual calcium ion regulation of calcineurin by calmodulin and calcineurin B. Biochemistry. 1994 Jun 7;33(22):6859–6866. doi: 10.1021/bi00188a015. [DOI] [PubMed] [Google Scholar]
  30. Swanson S. K., Born T., Zydowsky L. D., Cho H., Chang H. Y., Walsh C. T., Rusnak F. Cyclosporin-mediated inhibition of bovine calcineurin by cyclophilins A and B. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3741–3745. doi: 10.1073/pnas.89.9.3741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Takaishi T., Saito N., Kuno T., Tanaka C. Differential distribution of the mRNA encoding two isoforms of the catalytic subunit of calcineurin in the rat brain. Biochem Biophys Res Commun. 1991 Jan 15;174(1):393–398. doi: 10.1016/0006-291x(91)90533-d. [DOI] [PubMed] [Google Scholar]
  32. Thomas K. R., Capecchi M. R. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987 Nov 6;51(3):503–512. doi: 10.1016/0092-8674(87)90646-5. [DOI] [PubMed] [Google Scholar]
  33. Tsuboi A., Masuda E. S., Naito Y., Tokumitsu H., Arai K., Arai N. Calcineurin potentiates activation of the granulocyte-macrophage colony-stimulating factor gene in T cells: involvement of the conserved lymphokine element 0. Mol Biol Cell. 1994 Jan;5(1):119–128. doi: 10.1091/mbc.5.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES