Activation of Interferon-γ Inducing Factor Mediated by Interleukin-1β Converting Enzyme
10.1126/science.275.5297.206
Crossref journal-article
American Association for the Advancement of Science (AAAS)
Science (221)
Abstract

The interleukin-1β (IL-1β) converting enzyme (ICE) processes the inactive IL-1β precursor to the proinflammatory cytokine. ICE was also shown to cleave the precursor of interferon-γ inducing factor (IGIF) at the authentic processing site with high efficiency, thereby activating IGIF and facilitating its export. Lipopolysaccharide-activated ICE-deficient (ICE −/− ) Kupffer cells synthesized the IGIF precursor but failed to process it into the active form. Interferon-γ and IGIF were diminished in the sera of ICE −/− mice exposed to Propionibacterium acnes and lipopolysaccharide. The lack of multiple proinflammatory cytokines in ICE −/− mice may account for their protection from septic shock.

Bibliography

Gu, Y., Kuida, K., Tsutsui, H., Ku, G., Hsiao, K., Fleming, M. A., Hayashi, N., Higashino, K., Okamura, H., Nakanishi, K., Kurimoto, M., Tanimoto, T., Flavell, R. A., Sato, V., Harding, M. W., Livingston, D. J., & Su, M. S.-S. (1997). Activation of Interferon-γ Inducing Factor Mediated by Interleukin-1β Converting Enzyme. Science, 275(5297), 206–209.

Authors 17
  1. Yong Gu (first)
  2. Keisuke Kuida (additional)
  3. Hiroko Tsutsui (additional)
  4. George Ku (additional)
  5. Kathy Hsiao (additional)
  6. Mark A. Fleming (additional)
  7. Nobuki Hayashi (additional)
  8. Kazuya Higashino (additional)
  9. Haruki Okamura (additional)
  10. Kenji Nakanishi (additional)
  11. Masashi Kurimoto (additional)
  12. Tadao Tanimoto (additional)
  13. Richard A. Flavell (additional)
  14. Vicki Sato (additional)
  15. Matthew W. Harding (additional)
  16. David J. Livingston (additional)
  17. Michael S.-S. Su (additional)
References 43 Referenced 964
  1. 10.1016/S0092-8674(00)81334-3
  2. 10.1126/science.1373520
  3. 10.1038/356768a0
  4. 10.1126/science.7535475
  5. 10.1016/0092-8674(95)90490-5
  6. Zheng H., et al., Immunity 3, 9 (1995). (10.1016/1074-7613(95)90154-X) / Immunity by Zheng H. (1995)
  7. Okamura H., et al., Infect. Immun. 63, 3966 (1995). (10.1128/iai.63.10.3966-3972.1995) / Infect. Immun. by Okamura H. (1995)
  8. Okamura H., et al., Nature 378, 88 (1995); (10.1038/378088a0) / Nature by Okamura H. (1995)
  9. Ushio S., et al., J. Immunol. 156, 4274 (1996). (10.4049/jimmunol.156.11.4274) / J. Immunol. by Ushio S. (1996)
  10. Heusel J. W., Wesselschmidt R. L., Shresta S., Russell J. H., Ley T. J., Cell 76, 977 (1994); (10.1016/0092-8674(94)90376-X) / Cell by Heusel J. W. (1994)
  11. Darmon A. J. , Nicholson D. W. , Bleackley R. C., Nature 377, 446 (1995); (10.1038/377446a0) / Nature by Darmon A. J. (1995)
  12. Quan L. T., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 1972 (1996). (10.1073/pnas.93.5.1972) / Proc. Natl. Acad. Sci. U.S.A. by Quan L. T. (1996)
  13. Gu Y., et al., J. Biol. Chem. 271, 10816 (1996). (10.1074/jbc.271.18.10816) / J. Biol. Chem. by Gu Y. (1996)
  14. A 0.6-kb cDNA encoding full-length murine proIGIF (6) was ligated into the mammalian expression vector pCDLSRα (28). Plasmids for the expression of active human ICE (11) TX (10) and CMH-1 (13) lacking the prosequence were as described. Expression plasmid for the active form of CPP32 lacking the prosequence (12) was constructed similarly in the pCDLSRα vector. Plasmids (3 μg) were transfected into COS cells in 35-mm dishes by the DEAE-dextran method (11). Twenty-four hours later cells were lysed and the lysates were subjected to SDS-PAGE and immunoblotting with an antiserum to IGIF (6).
  15. 10.1002/j.1460-2075.1995.tb07183.x
  16. Gu Y., et al., ibid.1923. / ibid. by Gu Y.
  17. Fernandes-Alnemri T., Litwack G., Alnemri E. S., J. Biol. Chem. 269, 30761 (1994); (10.1016/S0021-9258(18)47344-9) / J. Biol. Chem. by Fernandes-Alnemri T. (1994)
  18. 10.1038/376037a0
  19. 10.1016/0092-8674(95)90541-3
  20. Lippke J. A., Gu Y., Sarnecki C., Caron P. R., S.-S. Su M., J. Biol. Chem. 271, 1825 (1996). (10.1074/jbc.271.4.1825) / J. Biol. Chem. by Lippke J. A. (1996)
  21. Expression plasmid for (His) 6 -proIGIF was created by introducing Nde I sites at the ends of murine proIGIF cDNA coding sequence (6) and ligating into Escherichia coli expression vector pET-15B (Novagen). The E. coli strain BL21(DE3) carrying the plasmid was induced with isopropyl-1-thio-β-D-galactopyranoside. (His) 6 -proIGIF protein was purified from soluble fractions by Ni-nitrilotriacetic acid-agarose (Qiagen) chromatography according to the manufacturer's instructions. In vitro cleavage reactions by ICE and ICE-like proteases were carried out as in (29). Conditions for cleavage by granzyme B were as in (8). Cleavage products were analyzed by SDS-PAGE on 16% gels and Coomassie blue staining and were subjected to NH 2 -terminal amino acid sequencing with an ABI automated peptide sequencer.
  22. [ 35 S]methionine-labeled proIGIF [∼3000 cpm prepared by in vitro transcription and translation with the TNT T7 coupled reticulocyte lysate system (Promega) and proIGIF cDNA in pSP73 vector as template] was incubated in reaction mixtures of 60 μl containing 0.1 to 1 nM recombinant ICE and 190 nM to 12 μM unlabeled proIGIF for 8 to 10 min at 37°C. Cleavage product concentrations were determined by SDS-PAGE and PhosphorImager analysis. The kinetic parameters were calculated by nonlinear regression fitting of the rate versus concentration data to the Michaelis-Menten equation by means of the program Enzfitter (Biosoft).
  23. Dolle R. E., et al., J. Med. Chem. 37, 563 (1994). (10.1021/jm00031a003) / J. Med. Chem. by Dolle R. E. (1994)
  24. The T H 1 A. E7 cells (30) (1.3 × 10 5 cells in 0.15 ml) or nonadherent splenic T cells (8 × 10 5 cells) in 96-well plates were treated with IGIF or conditioned medium for 18 to 20 hours and the culture supernatants were assayed for IFN-γ by ELISA (Endogen Cambridge MA).
  25. COS cells (3.5 × 10 5 cells in a 35-mm dish) were labeled for 7 hours with 1 ml of methionine-free Dulbecco's modified Eagle's medium (DMEM) containing 2.5% normal DMEM 1% dialyzed fetal bovine serum (FBS) and [ 35 S]methionine (300 μCi/ml; [ 35 S]Express Protein Labeling Mix New England Nuclear). Cell lysates [prepared in 20 mM Hepes (pH 7.2) 150 mM NaCl 0.1% Triton X-100 5 mM N -ethylmaleimide 1 mM phenylmethylsulfonyl fluoride and leupeptin (2.5 μg/ml)] or conditioned medium were immunoprecipitated with the antiserum to IGIF (6).
  26. COS cells (3.5 × 10 5 cells in a 35-mm dish) were transfected and grown in 1 ml of medium for 18 hours. Medium was harvested and used at 1→10 final dilution in the IFN-γ induction assay (17); COS cell pellets from the same transfection were lysed in 100 μl of 20 mM Hepes (pH 7.0) by three cycles of freezing and thawing. Lysates were cleared by centrifugation and were used at 1→10 dilution in the assay. On the basis of our analysis that 10% mature IGIF was exported out of the cells we estimated that the mature IGIF concentration in lysates is ∼90 times that of the conditioned medium.
  27. Wild-type or ICE-deficient mice were injected intraperitoneally with 1 mg of heat-killed P. acnes (5). Kupffer cells were prepared 7 days later (31) except that nycodenz gradient was used instead of metrizamide. For each experiment Kupffer cells (1 × 10 6 cells per milliliter) from two or three animals were pooled and cultured in RPMI 1640 supplemented with 10% FBS and LPS (1 μg/ml
  28. Difco E. coli strain 055:B5). Cell lysates and conditioned media were prepared 3 hours later. IGIF was determined by ELISA with protein G-purified rabbit polyclonal antibody to murine IGIF (6). Metabolic labeling and immunoprecipitation experiments were carried out as for COS cells (18) except that methionine-free RPMI 1640 was used in place of DMEM. To prepare conditioned medium of LPS-stimulated adherent splenocytes we cultured total splenic cells (6 × 10 7 cells in 1 ml) from wild-type or ICE −/− mice for 1 hour. Adherent cells were then stimulated with LPS (50 ng/ml) for 5 hours. Conditioned media were harvested and used at 1→4 dilution in the IFN-γ assay (17) in the presence or absence of anti-IGIF (25 μg/ml) (6).
  29. Wild-type or ICE-deficient mice were primed with P. acnes (20). Seven days later mice were exposed to LPS (1 μg intravenously). In some experiments recombinant mature IGIF (1 μg) or protein G-purified anti-IGIF (250 μg) was coinjected with LPS; sera were collected 3 hours after LPS exposure.
  30. Reduced IFN-γ was also observed in Listeria -infected (N. M. Tsuji et al. in preparation) and LPS-exposed (G. Ku et al. in preparation) ICE −/− mice.
  31. Trinchieri G., Annu. Rev. Immunol. 13, 251 (1995). (10.1146/annurev.iy.13.040195.001343) / Annu. Rev. Immunol. by Trinchieri G. (1995)
  32. Belardelli F., APMIS 103, 161 (1995); (10.1111/j.1699-0463.1995.tb01092.x) / APMIS by Belardelli F. (1995)
  33. Dinarello C. A.,Blood 87, 2095 (1996). (10.1182/blood.V87.6.2095.bloodjournal8762095) / Blood by Dinarello C. A. (1996)
  34. Margolis N. Dinarello C. unpublished data.
  35. Ku G. Harding M. W. unpublished data.
  36. 10.1126/science.8456300
  37. Huang S., et al., ibid., p. 1742; / ibid. by Huang S.
  38. Car B. D., et al., J. Exp. Med. 179, 1437 (1994). (10.1084/jem.179.5.1437) / J. Exp. Med. by Car B. D. (1994)
  39. Takebe Y., et al., Mol. Cell. Biol. 8, 466 (1988). / Mol. Cell. Biol. by Takebe Y. (1988)
  40. Gu Y., Sarnecki C., Aldape R. A., Livingston D. J., S.-S. Su M., J. Biol. Chem. 270, 18715 (1995). (10.1074/jbc.270.32.18715) / J. Biol. Chem. by Gu Y. (1995)
  41. Quill H., Schwartz R. H., J. Immunol. 138, 3704 (1987). (10.4049/jimmunol.138.11.3704) / J. Immunol. by Quill H. (1987)
  42. Tsutsui H., Mizoguchi Y., Morisawa S., Hepato-Gastroenterology 39, 553 (1992). / Hepato-Gastroenterology by Tsutsui H. (1992)
  43. We thank T. Fox and W. Chen for ICE and TX protein; A. Diu C. Faucheu and J.-L. Lalanne for TX cDNA; M. Rincon for A. E7 cells; J. Lippke for CPP32 and CMH-1 cDNA; B. O'Hare for oligonucleotide synthesis and DNA sequencing; T. Faust for ELISA; A. Heiser for animal surgery; and J. Boger for critical reading and discussion of the manuscript. R.A.F. is an HHMI Investigator.
Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 5:45 a.m.)
Deposited 1 year, 7 months ago (Jan. 13, 2024, 1:09 a.m.)
Indexed 1 week, 4 days ago (Aug. 21, 2025, 1:06 p.m.)
Issued 28 years, 7 months ago (Jan. 10, 1997)
Published 28 years, 7 months ago (Jan. 10, 1997)
Published Print 28 years, 7 months ago (Jan. 10, 1997)
Funders 0

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@article{Gu_1997, title={Activation of Interferon-γ Inducing Factor Mediated by Interleukin-1β Converting Enzyme}, volume={275}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.275.5297.206}, DOI={10.1126/science.275.5297.206}, number={5297}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Gu, Yong and Kuida, Keisuke and Tsutsui, Hiroko and Ku, George and Hsiao, Kathy and Fleming, Mark A. and Hayashi, Nobuki and Higashino, Kazuya and Okamura, Haruki and Nakanishi, Kenji and Kurimoto, Masashi and Tanimoto, Tadao and Flavell, Richard A. and Sato, Vicki and Harding, Matthew W. and Livingston, David J. and Su, Michael S.-S.}, year={1997}, month=jan, pages={206–209} }