Abstract
An experimental therapy for improvement of macrophage dysfunction caused by transforming growth factor‐β (TGF‐β) was tried in EL4 tumor‐bearing mice. TGF‐β was detected in cell‐free ascitic fluid from EL4‐bearers, but not in tbat from normal mice, by western blot analysis. The ascites also showed growth‐suppressive activity against MvlLn cells, and the suppressive activity was potentiated by transient acidification. To investigate whether the functions of peritoneal macrophagcs were suppressed in EL4‐bearers, the abilities to produce nitric oxide and tumor necrosis factor‐α (TNF‐α) upon lipopolysaccharide (LPS) stimulation were measured. Both abilities of macrophages in EL4‐bearing mice were suppressed remarkably on day 9, and decreased further by day 14, compared with non‐tumor‐bearing controls. TGF‐β activity was abrogated by administration of anti‐TGF‐α antibody to EL4‐bearing mice. While a large amount of TGF‐β was detected in ascitic fluid from control EL4‐bearers, little TGF‐β was detectable in ascites from EL4‐bearers given anti‐TGF‐β antibody. Furthermore, while control macrophages exhibited little or no production of nitric oxide and TNF‐α on LPS stimulation in vitro, macrophages from EL4‐bearers administered with anti‐TGF‐β antibody showed the same ability as normal macrophages. These results clearly indicate that TGF‐β contributes to macrophage dysfunction and that the administration of specific antibody for TGF‐β reverses macrophage dysfunction in EL4‐bearing hosts.
Keywords: TGF‐β, Ascitic fluid, EL4, Anti‐TGF‐β antibody, Immunosuppression
Full Text
The Full Text of this article is available as a PDF (473.0 KB).
REFERENCES
- 1. ) Fujiwara , H. and Ellner , J. J.Spontaneous production of a suppressor factor by the human macrophage‐like cell line U937 . J. Immunol , 136 , 181 – 185 ( 1986. ). [PubMed] [Google Scholar]
- 2. ) Riley , L. B. , Pellis , N. R. , Schantz , S. P. , Freedman , R. S. , Balch , C. M. and Itoh , K.Humoral modulation of lymphokine‐activated killer (LAK)‐cell induction in humans: IgG‐related and non‐IgG inhibitors in sera from cancer patients . Int. J. Cancer , 46 , 785 – 791 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Fu , Y. X. , Watson , G. A. , Kasahara , M. and Lopez , D. M.The role of tumor‐derived cytokines on the immune system of mice bearing a mammary adenocarcinoma. I. Induction of regulatory macrophages in normal mice by the in vivo administration of rGM‐CSF. J . Immunol , 146 , 783 – 789 ( 1991. ). [PubMed] [Google Scholar]
- 4. ) MacCubbin , D. L. , Mace , K. F. , Ehrke , M. J. and Mihich , E.Modification of host antitumor defense mechanism in mice by progressively growing tumor . Cancer Res. , 49 , 4216 – 4224 ( 1989. ). [PubMed] [Google Scholar]
- 5. ) Roberts , A. B. , Anzano , M. A. , Lamb , L. C. , Smith , J. M. and Sporn , M. B.New class of transforming growth factors potentiated by epidermal growth factor: isolation from nonspecific tissues . Proc. Natl. Acad. Sci. USA , 78 , 5339 – 5343 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6. ) Ristow , H. J.BSC‐1 growth inhibitor/type β transforming growth factor is a strong inhibitor of thymocyte proliferation . Proc. Natl. Acad Sci. USA , 83 , 5531 – 5533 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. ) Inge , T. H. , Hoover , S. K. , Susskind , B. M. , Barrett , S. K. and Bear , H. D.Inhibition of tumor‐specific cytotoxic T‐lymphocyte responses by transforming growth factorβ 1 . Cancer Res. , 52 , 1386 – 1392 ( 1992. ). [PubMed] [Google Scholar]
- 8. ) Kehrl , J. H. , Wakefield , L. M. , Roberts , A. B.Jakowlew , S. , Alvarez‐Mon , M. , Derynck , R. , Sporn , M. B. and Fauci , A. S , Production of transforming growth factor β by human lymphocytes and its potential role in the regulation of T cell growth . J. Exp. Med. , 163 , 1037 – 1050 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. ) Tada , T. , Ohzeki , S. , Utsumi , K. , Takiguchi , H. , Muramatsu , M. , Li , X.‐F. , Shimizu , J. , Fujiwara , H. and Hamaoka , T.Transforming growth factors‐induced inhibition of T cell function: susceptibility difference in T cells of various phenotypes and functions and its relevance to immunosuppression in tumor‐bearing state. J . Immunol. , 146 , 1077 – 1082 ( 1991. ). [PubMed] [Google Scholar]
- 10. ) Zou , J. P. , Shimozu , J. , Ikegame , K. , Yamamoto , N. , Ono , S. , Fujiwara , H , and Hamaoka , T.Tumor‐bearing mice exhibit a progressive increase in tumor antigen‐responsive CD4+ T cell activity . J. Immunol , 148 , 648 – 655 ( 1992. ). [PubMed] [Google Scholar]
- 11. ) Kehrl , J. H. , Roberts , A. B. , Wakefield , L. M. , Jakowlew , S. , Sporn , M. B. and Fauci , A. S.Transforming growth factor β is an important immunomodulatory protein for human B lymphocytes . J. Immunol , 137 , 3855 – 3860 ( 1986. ). [PubMed] [Google Scholar]
- 12. ) Rook , A. H , Kehrl , J. H. , Wakefield , L. M. , Roberts , A. B. , Sporn , M. B. , Burlington , B. , Lane , H. C. and Fauci , A. S.Effects of transforming growth factor β on the functions of natural killer cells: depressed cytolytic activity and blunting of interferon responsiveness. J . Immunol. , 136 , 3916 – 3920 ( 1986. ). [PubMed] [Google Scholar]
- 13. ) Geller , R. L. , Smyth , M. L , Stribl , S. L. , Bach , F. H. , Ruscetti , F. W. , Longo , D. L. and Ochoa , A. C.Generation of lymphokine‐activated killer activity in T cells. J . Immunol. , 146 , 3280 – 3288 ( 1991. ). [PubMed] [Google Scholar]
- 14. ) Wahl , S. M. , Hunt , D. A. , Wakefield , L. M. , McCartney‐Francis , N. , Wahl , L. M. , Roberts , A. B. and Sporn , M. B.Transforming growth factor type β induces monocyte chemotaxis and growth factor production . Proc. Natl. Acad. Sci. USA , 84 , 5788 – 5792 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. ) Tsunawaki , S. , Sporn , M. , Ding , A. and Nathan , C.Deactivation of macrophages by transforming growth factor‐β . Nature , 334 , 260 – 262 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 16. ) Ding , A. , Nathan , C. F. , Graycar , J. , Derynck , R. , Stuehr , D. J. and Srimal , S.Macrophage deactivating factor and transforming growth factors‐β 1, ‐β2 and ‐β 3 inhibit induction of macrophage nitrogen oxide synthesis by IFN‐η . J, Immunol , 145 , 940 – 944 ( 1990. ). [PubMed] [Google Scholar]
- 17. ) Vodovitz , Y. , Bogdan , C. , Paik , J. , Xie , Q.‐W. and Nathan , C.Mechanisms of suppression of macrophage nitric oxide release by transforming growth factor‐β . J. Exp. Med. , 178 , 605 – 613 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18. ) Chantry , D. , Turner , M. , Abney , E. and Feldmann , M.Modulation of cytokine production by transforming growth factor‐β. J . Immunol , 142 , 4295 – 4300 ( 1989. ). [PubMed] [Google Scholar]
- 19. ) Roberts , A. B. and Sporn , M. B.The transforming growth factor‐β . Handb. Exp. Pharmacol , 95 , 419 – 472 ( 1990. ). [Google Scholar]
- 20. ) Like , B. and Massague , J.The antiproliferative effect of type β transforming growth factor occurs at a level distal from receptors for growth‐activating factors . J, Biol. Chem. , 261 , 13426 – 13429 ( 1986. ). [PubMed] [Google Scholar]
- 21. ) Green , L. C. , De Luzuriaga , K. R. , Wagner , D. A. , Rand , W. , Istfan , N. , Young , V. R. and Tannenbaum , S. R.Nitrate biosynthesis in man . Proc. Natl. Acad. Sci. USA , 78 , 7764 – 7768 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. ) Young , M. R. , Wright , M. A. , Coogan , M. , Young , M. E. and Bagash , J.Tumor‐derived cytokines induce bone marrow suppressor cells that mediate immunosuppression through transforming growth factor beta . Cancer Immunol Immunother. , 35 , 14 – 18 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23. ) Guillermo , T. A. , Beauchamp , R. D. , Koeppen , H. , Park , B. H. , Schreiber , H. , Moses , H. L. and Rowley , D. A.A highly immunogenic tumor transfected with a murine transforming growth factor type‐β 1 cDNA escapes immune surveillance . Proc. Natl. Acad. Sci. USA , 87 , 1486 – 1490 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 24. ) Nycum , L. M. and Fishman , M.The effect of phorbol esters on tumor cell sensitivity to macrophage‐mediated cytostasis . Cell Immunol , 118 , 147 – 156 ( 1989. ). [DOI] [PubMed] [Google Scholar]
- 25. ) Haak‐Frendscho , M. , Wynn , T. A. , Czuprynski , C. J , and Paulnock , D.Transforming growth factor‐β 1 inhibits activation of macrophage cell line RAW 264.7 for cell killing . Clin. Exp. Immunol , 82 , 404 – 410 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. ) Keller , R. and Van Der Meide , P. H.Modulation of tumoricidal activity, induced in bone‐marrow‐derived mononuclear phagocytes by interferon η or Corynebacterium parvum, by interferon β, tumor necrosis factor, prostaglandin E2, and transforming growth factor β . Int. J. Cancer , 49 , 796 – 800 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 27. ) Craznieski , C. W. , Chiu , H. H. , Wong , G. H. W. , McCabe , S. M. and Palladino , M. A.Transforming growth factor‐β 1 modulates the expression of class II histocompatibility antigens on human cells . J. Immunol , 140 , 4217 – 4223 ( 1988. ). [PubMed] [Google Scholar]
- 28. ) Espevik , T. , Figari , L S. , Shalaby , M. R. , Lackides , G. A. , Lewis , G. D. , Shepard , H. M. and Palladino , M. A.Inhibition of cytokine production by cyclosporin A and transforming growth factor β . J. Exp. Med. , 166 , 571 – 576 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 29. ) Saiki , I. , Maeda , H. , Sakurai , T. , Murata , J. , Lida , J. , Kiso , M. , Hasegawa , A. and Azuma , I.Induction of an endogenous tumor necrosis factor in mice by murine recombinant interferon‐η combined with a lipid A subunit analog (GLA‐60) of low toxicity . Cancer Immunol Immunother. , 29 , 101 – 108 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 30. ) Li , X.‐F. , Takiuchi , H. , Zou , J.‐P. , Katagiri , T. , Yamamoto , N. , Nagata , T. , Ono , S. , Fujiwara , H. and Hamaoka , T.Transforming growth factor‐β (TGF‐β)‐mediated immunosuppression in the tumor‐bearing state: enhanced production of TGF‐β and a progressive increase in TGF‐β susceptibility of anti‐tumor CD4+ T cell function . Jpn. J. Cancer Res. , 84 , 315 – 325 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31. ) Ueki , N. , Ohkawa , T. , Yokoyama , Y. , Maeda , J. , Kawai , Y. , Ikeda , T. , Amuro , Y. , Hada , T. and Higashino , K.Potentiation of metastatic capacity by transforming growth factor‐β 1 gene transfection . Jpn. J. Cancer Res. , 84 , 589 – 593 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32. ) Mooradian , D. L. , McCarthy , J. B. , Komanduri , K. V. and Furcht , L. T.Effects of transforming growth factor‐β 1 on human pulmonary adenocarcinoma cell adhesion, motility, and invasion in vitro . J. Natl. Cancer Inst. , 84 , 523 – 527 ( 1992. ). [DOI] [PubMed] [Google Scholar]
- 33. ) Arrick , B. A. , Lopez , A. R. , Elfman , F. , Ebner , R. , Damsky , C. H. and Derynck , R.Altered metabolic and adhesive properties and increased tumorigenesis associated with increased expression of transforming growth factor β 1 . J. Cell Biol. , 118 , 715 – 726 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 34. ) Wu , S. , Theodorescu , D. , Kerbel , R. S , Willson , J. K. V. , Mulder , K. M. , Humphrey , L. E. and Brattain , M. G.TGF‐β l is an autocrine‐negative growth regulator of human colon carcinoma FET cells in vitro as revealed by transfection of an antisense expression vector. J . Cell Biol. , 116 , 187 – 196 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]