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Volumn 271, Issue 5253, 1996, Pages 1272-1276

Blocked signal transduction to the ERK and JNK protein kinases in anergic CD4+ T cells

Author keywords

[No Author keywords available]

Indexed keywords

CALCINEURIN; CD28 ANTIGEN; CD3 ANTIGEN; INTERLEUKIN 2; LYMPHOCYTE ANTIGEN RECEPTOR; NUCLEAR FACTOR; PHORBOL 13 ACETATE 12 MYRISTATE; PROTEIN KINASE;

EID: 0029975023     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.271.5253.1272     Document Type: Article
Times cited : (411)

References (59)
  • 4
    • 0027767762 scopus 로고
    • C D Gimmi et al., Proc. Natl. Acad. Sci. U S.A 88, 6575 (1991); G J. Freeman et al., Science 262, 909 (1993).
    • (1993) Science , vol.262 , pp. 909
    • Freeman, G.J.1
  • 7
    • 0024327899 scopus 로고
    • M K. Jenkins, D. M. Pardoll, J Mizuguchi, T. M. Chused, R H. Schwartz, Proc. Natl. Acad. Sci U.S A 84, 5409 (1987); D. L. Mueller, M. K. Jenkins, R. H. Schwartz, Annu Rev. Immunol. 7, 445 (1989); D L. Mueller and M. K Jenkins, Curr. Opin. Immunol. 7, 375 (1995).
    • (1989) Annu Rev. Immunol. , vol.7 , pp. 445
    • Mueller, D.L.1    Jenkins, M.K.2    Schwartz, R.H.3
  • 8
    • 0029032046 scopus 로고
    • M K. Jenkins, D. M. Pardoll, J Mizuguchi, T. M. Chused, R H. Schwartz, Proc. Natl. Acad. Sci U.S A 84, 5409 (1987); D. L. Mueller, M. K. Jenkins, R. H. Schwartz, Annu Rev. Immunol. 7, 445 (1989); D L. Mueller and M. K Jenkins, Curr. Opin. Immunol. 7, 375 (1995).
    • (1995) Curr. Opin. Immunol. , vol.7 , pp. 375
    • Mueller, D.L.1    Jenkins, M.K.2
  • 9
    • 0026669383 scopus 로고
    • H Quill et al., J. Immunol. 149, 2887 (1992); E A Cho, M. P. Riley, A. L. Sillman, H. Quill, ibid. 151, 20 (1993), T. F. Gajewski, D. Qian, P. Fields, F W. Fitch, Proc. Natl Acad. Sci. U.S.A. 91, 38 (1994).
    • (1992) J. Immunol. , vol.149 , pp. 2887
    • Quill, H.1
  • 10
    • 0027301903 scopus 로고
    • H Quill et al., J. Immunol. 149, 2887 (1992); E A Cho, M. P. Riley, A. L. Sillman, H. Quill, ibid. 151, 20 (1993), T. F. Gajewski, D. Qian, P. Fields, F W. Fitch, Proc. Natl Acad. Sci. U.S.A. 91, 38 (1994).
    • (1993) J. Immunol. , vol.151 , pp. 20
    • Cho, E.A.1    Riley, M.P.2    Sillman, A.L.3    Quill, H.4
  • 12
    • 0026667860 scopus 로고
    • S.-M. Kang et al., Science 257, 1134 (1992).
    • (1992) Science , vol.257 , pp. 1134
    • Kang, S.-M.1
  • 13
    • 84920296693 scopus 로고    scopus 로고
    • note
    • + munne helper T cell clones A.E7 and 16B.2 used in this study, and their cultivation conditions, have been previously described (9, 10). Each clone produces IL-2 in response to Ag stimulation.
  • 19
    • 0027259359 scopus 로고
    • P G McCaffrey, B. A Perrino, T. R. Soderling, A Rao, J Biol. Chem. 268, 3747 (1993), J Jain, Z. Miner, A Rao, J. Immunol. 151, 837 (1993); J. Jain et al., Nature 366, 352 (1993).
    • (1993) J. Immunol. , vol.151 , pp. 837
    • Jain, J.1    Miner, Z.2    Rao, A.3
  • 20
    • 0027491776 scopus 로고
    • P G McCaffrey, B. A Perrino, T. R. Soderling, A Rao, J Biol. Chem. 268, 3747 (1993), J Jain, Z. Miner, A Rao, J. Immunol. 151, 837 (1993); J. Jain et al., Nature 366, 352 (1993).
    • (1993) Nature , vol.366 , pp. 352
    • Jain, J.1
  • 21
    • 84920296692 scopus 로고    scopus 로고
    • in preparation
    • A. Mondino et al , in preparation
    • Mondino, A.1
  • 22
    • 84920296691 scopus 로고    scopus 로고
    • note
    • Plasmid pNFATZH contains a multimer of the human NFAT enhancer element flanked by Xho I restriction sites that has been inserted into a minimal IL-2 promoter (-319 to +47 base pairs of IL-2 gene DNA containing an internal deletion and replacement of base pairs -296 to -72 by Xho I linker DNA) and fused to a bacterial lacZ gene (15). This plasmid also contains a hygromycin resistance gene under the control of the herpes simplex virus tk promoter. Plasmid pSXNeo/AP-1 contains five tandem repeats of the sheep metallothionein MT-II gene AP-1 srte (TGACTCA) ligated within an identical minimal IL-2 promoter at the internal Xho I site. To construct the pAP-1ZH plasmid, an Xmn I-Hind Ill fragment of pSXNeo/AP-1 containing the AP-1 multimer within the minimal IL-2 promoter was gel purified. The pNFATZH plasmid was similarly digested with Xmn I and Hind llI to remove enhancer sequences, and then purified vector DNA was ligated to the AP-1 multimer-containing fragment. The pAP-1ZH construct thus has a bacterial lacZ gene fused to a 5x AP-1 multimer-containing minimal IL-2 promoter, as well as a hygromycin resistance gene. The pZH enhancerless plasmid was prepared by simple digestion of pNFATZH with Xho I and ligation of the empty vector The construction of both plasmids was confirmed by restriction analysis
  • 24
    • 84920296690 scopus 로고    scopus 로고
    • note
    • + helper T cells were stably transfected with reporter gene DNA essentially according to the method of Kang et al. (7), with the use of hygromycin (Calbiochem) selection. Transfectants were maintained as pools of cells (called ZH or AP-1ZH, respectively) rather than clones, to eliminate any effects related to the site of integration.
  • 25
    • 0026534209 scopus 로고
    • C. E Whitehurst, T. G. Boulton, M. H. Cobb, T. D. Geppert, J. Immunol. 148, 3230 (1992); H. Owaki, R. Makar, T. G. Boulton, M. H. Cobb, T. D. Geppert, Biochem. Biophys Res. Commun. 182, 1416 (1992); M Izquierdo, S J. Leevers, C. J. Marshall, D. Cantrell, J. Exp. Med. 178, 1199 (1993), A. E. Nel, S. Gupta, L. Lee, J. A. Ledbetter, S. B. Kanner, J. Biol. Chem 270, 18428 (1995)
    • (1992) J. Immunol. , vol.148 , pp. 3230
    • Whitehurst, C.E.1    Boulton, T.G.2    Cobb, M.H.3    Geppert, T.D.4
  • 26
    • 0026578183 scopus 로고
    • C. E Whitehurst, T. G. Boulton, M. H. Cobb, T. D. Geppert, J. Immunol. 148, 3230 (1992); H. Owaki, R. Makar, T. G. Boulton, M. H. Cobb, T. D. Geppert, Biochem. Biophys Res. Commun. 182, 1416 (1992); M Izquierdo, S J. Leevers, C. J. Marshall, D. Cantrell, J. Exp. Med. 178, 1199 (1993), A. E. Nel, S. Gupta, L. Lee, J. A. Ledbetter, S. B. Kanner, J. Biol. Chem 270, 18428 (1995)
    • (1992) Biochem. Biophys Res. Commun. , vol.182 , pp. 1416
    • Owaki, H.1    Makar, R.2    Boulton, T.G.3    Cobb, M.H.4    Geppert, T.D.5
  • 27
    • 0027337561 scopus 로고
    • C. E Whitehurst, T. G. Boulton, M. H. Cobb, T. D. Geppert, J. Immunol. 148, 3230 (1992); H. Owaki, R. Makar, T. G. Boulton, M. H. Cobb, T. D. Geppert, Biochem. Biophys Res. Commun. 182, 1416 (1992); M Izquierdo, S J. Leevers, C. J. Marshall, D. Cantrell, J. Exp. Med. 178, 1199 (1993), A. E. Nel, S. Gupta, L. Lee, J. A. Ledbetter, S. B. Kanner, J. Biol. Chem 270, 18428 (1995)
    • (1993) J. Exp. Med. , vol.178 , pp. 1199
    • Izquierdo, M.1    Leevers, S.J.2    Marshall, C.J.3    Cantrell, D.4
  • 28
    • 0029117434 scopus 로고
    • C. E Whitehurst, T. G. Boulton, M. H. Cobb, T. D. Geppert, J. Immunol. 148, 3230 (1992); H. Owaki, R. Makar, T. G. Boulton, M. H. Cobb, T. D. Geppert, Biochem. Biophys Res. Commun. 182, 1416 (1992); M Izquierdo, S J. Leevers, C. J. Marshall, D. Cantrell, J. Exp. Med. 178, 1199 (1993), A. E. Nel, S. Gupta, L. Lee, J. A. Ledbetter, S. B. Kanner, J. Biol. Chem 270, 18428 (1995)
    • (1995) J. Biol. Chem , vol.270 , pp. 18428
    • Nel, A.E.1    Gupta, S.2    Lee, L.3    Ledbetter, J.A.4    Kanner, S.B.5
  • 29
    • 0026641090 scopus 로고
    • P. Dent et al., Science 257, 1404 (1992); C. M. Crews, A. Alessandrini, R L Erikson, ibid. 258, 478 (1992); S. A. Moodie, B. M. Wiltumsen, M J. Weber, A. Wolfman, ibid 260, 1658 (1993); M. Izquierdo, K. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
    • (1992) Science , vol.257 , pp. 1404
    • Dent, P.1
  • 30
    • 0026539931 scopus 로고
    • P. Dent et al., Science 257, 1404 (1992); C. M. Crews, A. Alessandrini, R L Erikson, ibid. 258, 478 (1992); S. A. Moodie, B. M. Wiltumsen, M J. Weber, A. Wolfman, ibid 260, 1658 (1993); M. Izquierdo, K. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
    • (1992) Science , vol.258 , pp. 478
    • Crews, C.M.1    Alessandrini, A.2    Erikson, R.L.3
  • 31
    • 0027337519 scopus 로고
    • P. Dent et al., Science 257, 1404 (1992); C. M. Crews, A. Alessandrini, R L Erikson, ibid. 258, 478 (1992); S. A. Moodie, B. M. Wiltumsen, M J. Weber, A. Wolfman, ibid 260, 1658 (1993); M. Izquierdo, K. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
    • (1993) Science , vol.260 , pp. 1658
    • Moodie, S.A.1    Wiltumsen, B.M.2    Weber, M.J.3    Wolfman, A.4
  • 32
    • 0028926511 scopus 로고
    • P. Dent et al., Science 257, 1404 (1992); C. M. Crews, A. Alessandrini, R L Erikson, ibid. 258, 478 (1992); S. A. Moodie, B. M. Wiltumsen, M J. Weber, A. Wolfman, ibid 260, 1658 (1993); M. Izquierdo, K. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
    • (1995) Immunol. Today , vol.16 , pp. 159
    • Izquierdo, M.1    Reif, K.2    Cantrell, D.3
  • 34
    • 0028329953 scopus 로고
    • B. Derijard et al , Cell 76, 1025 (1994); B. Su et al., ibid 77, 727 (1994).
    • (1994) Cell , vol.76 , pp. 1025
    • Derijard, B.1
  • 35
    • 0028291525 scopus 로고
    • B. Derijard et al , Cell 76, 1025 (1994); B. Su et al., ibid 77, 727 (1994).
    • (1994) Cell , vol.77 , pp. 727
    • Su, B.1
  • 36
    • 0029055812 scopus 로고    scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • (1995) Cell , vol.81 , pp. 1137
    • Coso, O.A.1
  • 37
    • 0029055812 scopus 로고    scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • Cell , pp. 1147
    • Minden, A.1    Lin, A.2    Claret, F.-X.3    Abo, A.4    Kann, M.5
  • 38
    • 0028998794 scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • (1995) Science , vol.269 , pp. 221
    • Knaus, U.G.1    Moms, S.2    Dong, H.-J.3    Chemoff, J.4    Bokoch, G.M.5
  • 39
    • 0028568217 scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • (1994) Science , vol.266 , pp. 1719
    • Minden, A.1
  • 40
    • 0028670788 scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • (1994) Nature , vol.372 , pp. 798
    • Yan, M.1
  • 41
    • 0029055812 scopus 로고    scopus 로고
    • O A. Coso et al., ibid. 81, 1137 (1995); A. Minden, A. Lin, F.-X. Claret, A Abo, M. Kann, ibid., p. 1147; U. G Knaus, S. Moms, H.-J Dong, J. Chemoff, G. M. Bokoch, Science 269, 221 (1995), A Minden et al., ibid. 266, 1719 (1994), M. Yan et al., Nature 372, 798 (1994); I. Sanchez et al., ibid , p. 794
    • Nature , pp. 794
    • Sanchez, I.1
  • 42
    • 0027297647 scopus 로고
    • R. Marais, J. Wynne, R. Treisman, Cell 73, 381 (1993), A. J. Whitrnarsh, P. Shore, A. D. Sharrocks, R. J. Davis, Science 269, 403 (1995)
    • (1993) Cell , vol.73 , pp. 381
    • Marais, R.1    Wynne, J.2    Treisman, R.3
  • 44
    • 84920296688 scopus 로고    scopus 로고
    • W. Li, data not shown
    • W. Li, data not shown.
  • 46
    • 0028125887 scopus 로고
    • Y. Ward et al , ibid. 367, 651 (1994).
    • (1994) Nature , vol.367 , pp. 651
    • Ward, Y.1
  • 47
    • 84920296687 scopus 로고    scopus 로고
    • note
    • T cells were mixed with T cell depleted splenic ACs, and then incubated for 64 hours in vitro with either no stimulus or with 1 μM Ag DASP, or with PMA (1 ng/ml) plus ionomycin (0 5 μM), as previously described (28), Thymidine incorporation was determined during the last 16 hours of culture, and delta counts per minute (δcpm) were calculated as the difference between unstimulated and stimulated cultures. Percent response = (δcpm of anergic cells)/(δcpm of normal cells) × 100. Three independent experiments were performed and the results are reported as the mean ± SEM of the percent response. We compared percent response of anergic cells to Ag with percent response of anergic cells to PMA plus ionomycin stimulation using the paired Student's t test.
  • 53
    • 84920296685 scopus 로고    scopus 로고
    • C. D. Whaley, data not shown
    • C. D. Whaley, data not shown.
  • 54
    • 84920296684 scopus 로고    scopus 로고
    • note
    • 6 normal or anergic A.E7 or 16B.2 T cells were pelleted and washed once with ice-cold phosphate-buffered saline containing protease and phosphatase inhibitors, and the cell pellet was then extracted with Triton X-100, as previously described (30) Lysates were recovered from the insoluble nuclear fraction and analyzed by protein immunoblot (8.5% SDS-PAGE with transfer to nitrocellulose) with a 1:1000 dilution of R59 rabbit antiserum to NFATp, goat antiserum to rabbit immunoglobulin G (IgG)-horseradish peroxidase (Bio-Rad), and ECL (Amer-sham) chemiluminescence reagents.
  • 56
    • 84920296683 scopus 로고    scopus 로고
    • note
    • 8 cells per milliliter) in 250 mM tns-HCl (pH 7 5). Freeze-thaw lysis of cells was followed by centrifugation and collection of the lysate, and protein concentration was determined by the BCA protein assay (Pierce). Duplicate enzyme reactions were carried out, followed by measurement of optical absorbance at 405 nm with a microplate reader (BIO-TEK Instruments). Units of activity were calculated based on a standard curve with purified enzyme (Sigma), and were corrected for the protein concentration of the extract. Values obtained in this assay for unstimulated AP-1ZH T cells are essentially identical to those obtained with either resting or activated ZH cells or untransfected T cells (32) and result from background optical absorbance in the assay.
  • 57
    • 84920296682 scopus 로고    scopus 로고
    • note
    • A protein immunoblot analysis of cellular proteins was also used to examine ERK and JNK protein abundance. Approximately 20 μg of whole-cell extract (33) was mixed 1:1 with 2× SDS sample buffer, and then separated by 10% SDS-PAGE. Proteins were transferred to nitrocellulose, and blotted as described (34) with either polyclonal antibody 691 to ERK-1 and ERK-2 at a 1:1000 dilution or with polyclonal antibody to JNK-1 (1 μg/ml) (Santa Cruz Biotechnology).
  • 58
    • 84920296681 scopus 로고    scopus 로고
    • note
    • 32P into the fusion protein during the first 15 min was linear with respect to both time and kinase concentration The immobilized proteins were washed three times with kinase wash buffer, eluted in 1× SDS sample buffer, and then analyzed on a 10% SDS-polyacrylamide gel, followed by autoradiography to detect the presence of a phosphorylated GST-c-Jun substrate. The phosphorylated fusion protein electrophoresed through an SDS gel as several bands because of protein degradation
  • 59
    • 84920296680 scopus 로고    scopus 로고
    • note
    • We thank G. R Crabtree (plasmids pNFATZH and pSXNeo/AP-1), T. Geppert (rabbit antibodies A249 to ERK-2 and 691 to ERK-1 and ERK-2), and A Rao (rabbit antiserum R59 to recombinant NFATp) for generously providing reagents and expertise critical to the work; T. Behrens and M. Mescher for critically reviewing the manuscript; L. Chiodetti, S. Garcia, and S. Seiffert for expert technical assistance; and M. K. Jenkins for many valuable discussions. Supported by a grant from NIH (R29 AI31669), as well as funding from the Minnesota Arthritis Foundation. A.M. is a recipient of a fellowship from the American-Italian Foundation for Cancer Research. D L.M. is the recipient of an Investigator Award from the National Arthritis Foundation.


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