Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1995 Jul;85(3):400–407.

Transforming growth factor-beta 1 primes macrophages for enhanced expression of the nitric oxide synthase gene for nitric oxide-dependent cytotoxicity against Entamoeba histolytica.

J Y Lin 1, R Seguin 1, K Keller 1, K Chadee 1
PMCID: PMC1383913  PMID: 7558128

Abstract

Nitric oxide (NO) produced by activated macrophages is the major cytotoxic molecule for in vitro cytotoxicity against Entamoeba histolytica trophozoites. Transforming growth factor-beta 1 (TGF-beta 1) is a potent negative regulator of several macrophage functions, including NO production. In this study, we investigated the effect of TGF-beta 1 on macrophage nitric oxide synthase (mac-NOS) mRNA expression and NO production for macrophage cytotoxicity against E. histolytica trophozoites. TGF-beta 1 by itself was incapable of inducing mouse bone marrow-derived macrophage (BMM) amoebicidal activity and NO production (as measured by nitrite). In contrast, TGF-beta 1 pretreatment (4 hr) primed BMM for an enhanced amoebicidal activity of 15% and 23% in response to (interferon-gamma) IFN-gamma+tumour necrosis factor-alpha (TNF-alpha) or IFN-gamma+lipopolysaccharide LPS, concomitant with increased NO production of 85% and 27%, respectively. TGF-beta 1 pretreatment increased NO production in response to IFN-gamma+TNF-alpha/LPS stimulation in a time- and dose-dependent manner. By Northern blot analysis, the increased NO production of TGF-beta 1-pretreated BMM was preceded by markedly enhanced expression of mac-NOS mRNA. The priming effect of TGF-beta 1 on NO production was critically dependent on both a TNF-alpha (> or = 100 U) and a LPS (> or = 100 ng) triggering dose in the presence of IFN-gamma. TGF-beta 1 pretreatment enhanced TNF-alpha mRNA expression, but had no effect on TNF-alpha production in culture supernatants after 4 hr of stimulation with IFN-gamma+TNF-alpha/LPS; however, at a later time-point (16-48 hr), even though the levels of TNF-alpha mRNA expression were unaffected, TNF-alpha production was reduced. These data demonstrate that TGF-beta 1 priming for increased mac-NOS mRNA expression for NO-dependent cytotoxicity against E. histolytica in response to IFN-gamma+TNF-alpha/LPS stimulation may be involved in the modulation of a TNF-alpha triggering signal by TGF-beta 1.

Full text

PDF
400

Images in this article

403Figure 3
on p.403
404Figure 5
on p.404
405Figure 7
on p.405

Selected References

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

  1. Balkwill F. Cytokines in health and disease. Immunol Today. 1993 Apr;14(4):149–150. doi: 10.1016/0167-5699(93)90228-D. [DOI] [PubMed] [Google Scholar]
  2. Barral-Netto M., Barral A., Brownell C. E., Skeiky Y. A., Ellingsworth L. R., Twardzik D. R., Reed S. G. Transforming growth factor-beta in leishmanial infection: a parasite escape mechanism. Science. 1992 Jul 24;257(5069):545–548. doi: 10.1126/science.1636092. [DOI] [PubMed] [Google Scholar]
  3. Bermudez L. E., Covaro G., Remington J. Infection of murine macrophages with Toxoplasma gondii is associated with release of transforming growth factor beta and downregulation of expression of tumor necrosis factor receptors. Infect Immun. 1993 Oct;61(10):4126–4130. doi: 10.1128/iai.61.10.4126-4130.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brockhaus M., Schoenfeld H. J., Schlaeger E. J., Hunziker W., Lesslauer W., Loetscher H. Identification of two types of tumor necrosis factor receptors on human cell lines by monoclonal antibodies. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3127–3131. doi: 10.1073/pnas.87.8.3127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Celada A., Maki R. A. Transforming growth factor-beta enhances the M-CSF and GM-CSF-stimulated proliferation of macrophages. J Immunol. 1992 Feb 15;148(4):1102–1105. [PubMed] [Google Scholar]
  6. Chantry D., Turner M., Abney E., Feldmann M. Modulation of cytokine production by transforming growth factor-beta. J Immunol. 1989 Jun 15;142(12):4295–4300. [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Corradin S. B., Buchmuller-Rouiller Y., Smith J., Suardet L., Mauël J. Transforming growth factor beta 1 regulation of macrophage activation depends on the triggering stimulus. J Leukoc Biol. 1993 Nov;54(5):423–429. doi: 10.1002/jlb.54.5.423. [DOI] [PubMed] [Google Scholar]
  9. Deng W., Thiel B., Tannenbaum C. S., Hamilton T. A., Stuehr D. J. Synergistic cooperation between T cell lymphokines for induction of the nitric oxide synthase gene in murine peritoneal macrophages. J Immunol. 1993 Jul 1;151(1):322–329. [PubMed] [Google Scholar]
  10. Denis M., Chadee K. Cytokine activation of murine macrophages for in vitro killing of Entamoeba histolytica trophozoites. Infect Immun. 1989 Jun;57(6):1750–1756. doi: 10.1128/iai.57.6.1750-1756.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ding A. H., Nathan C. F., Stuehr D. J. Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. J Immunol. 1988 Oct 1;141(7):2407–2412. [PubMed] [Google Scholar]
  12. Ding A., Nathan C. F., Graycar J., Derynck R., Stuehr D. J., Srimal S. Macrophage deactivating factor and transforming growth factors-beta 1 -beta 2 and -beta 3 inhibit induction of macrophage nitrogen oxide synthesis by IFN-gamma. J Immunol. 1990 Aug 1;145(3):940–944. [PubMed] [Google Scholar]
  13. Drake W. T., Issekutz A. C. Transforming growth factor-beta 1 enhances polymorphonuclear leucocyte accumulation in dermal inflammation and transendothelial migration by a priming action. Immunology. 1993 Feb;78(2):197–204. [PMC free article] [PubMed] [Google Scholar]
  14. Fan K., Ruan Q., Sensenbrenner L., Chen B. Transforming growth factor-beta 1 bifunctionally regulates murine macrophage proliferation. Blood. 1992 Apr 1;79(7):1679–1685. [PubMed] [Google Scholar]
  15. Gatanaga T., Hwang C. D., Gatanaga M., Cappuccini F., Yamamoto R. S., Granger G. A. The regulation of TNF receptor mRNA synthesis, membrane expression, and release by PMA- and LPS-stimulated human monocytic THP-1 cells in vitro. Cell Immunol. 1991 Nov;138(1):1–10. doi: 10.1016/0008-8749(91)90127-w. [DOI] [PubMed] [Google Scholar]
  16. Gazzinelli R. T., Oswald I. P., Hieny S., James S. L., Sher A. The microbicidal activity of interferon-gamma-treated macrophages against Trypanosoma cruzi involves an L-arginine-dependent, nitrogen oxide-mediated mechanism inhibitable by interleukin-10 and transforming growth factor-beta. Eur J Immunol. 1992 Oct;22(10):2501–2506. doi: 10.1002/eji.1830221006. [DOI] [PubMed] [Google Scholar]
  17. Gilbert R. S., Herschman H. R. Transforming growth factor beta differentially modulates the inducible nitric oxide synthase gene in distinct cell types. Biochem Biophys Res Commun. 1993 Aug 31;195(1):380–384. doi: 10.1006/bbrc.1993.2054. [DOI] [PubMed] [Google Scholar]
  18. Hohmann H. P., Remy R., Brockhaus M., van Loon A. P. Two different cell types have different major receptors for human tumor necrosis factor (TNF alpha). J Biol Chem. 1989 Sep 5;264(25):14927–14934. [PubMed] [Google Scholar]
  19. Jacobsen S. E., Ruscetti F. W., Roberts A. B., Keller J. R. TGF-beta is a bidirectional modulator of cytokine receptor expression on murine bone marrow cells. Differential effects of TGF-beta 1 and TGF-beta 3. J Immunol. 1993 Nov 1;151(9):4534–4544. [PubMed] [Google Scholar]
  20. Kondo S., Isobe K., Ishiguro N., Nakashima I., Miura T. Transforming growth factor-beta 1 enhances the generation of allospecific cytotoxic T lymphocytes. Immunology. 1993 Jul;79(3):459–464. [PMC free article] [PubMed] [Google Scholar]
  21. Lin J. Y., Chadee K. Macrophage cytotoxicity against Entamoeba histolytica trophozoites is mediated by nitric oxide from L-arginine. J Immunol. 1992 Jun 15;148(12):3999–4005. [PubMed] [Google Scholar]
  22. Lin J. Y., Seguin R., Keller K., Chadee K. Tumor necrosis factor alpha augments nitric oxide-dependent macrophage cytotoxicity against Entamoeba histolytica by enhanced expression of the nitric oxide synthase gene. Infect Immun. 1994 May;62(5):1534–1541. doi: 10.1128/iai.62.5.1534-1541.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McCartney-Francis N. L., Wahl S. M. Transforming growth factor beta: a matter of life and death. J Leukoc Biol. 1994 Mar;55(3):401–409. doi: 10.1002/jlb.55.3.401. [DOI] [PubMed] [Google Scholar]
  24. McGee D. W., Beagley K. W., Aicher W. K., McGhee J. R. Transforming growth factor-beta enhances interleukin-6 secretion by intestinal epithelial cells. Immunology. 1992 Sep;77(1):7–12. [PMC free article] [PubMed] [Google Scholar]
  25. Nacy C. A., Nelson B. J., Meltzer M. S., Green S. J. Cytokines that regulate macrophage production of nitrogen oxides and expression of antileishmanial activities. Res Immunol. 1991 Sep;142(7):573–576. doi: 10.1016/0923-2494(91)90105-r. [DOI] [PubMed] [Google Scholar]
  26. Nelson B. J., Ralph P., Green S. J., Nacy C. A. Differential susceptibility of activated macrophage cytotoxic effector reactions to the suppressive effects of transforming growth factor-beta 1. J Immunol. 1991 Mar 15;146(6):1849–1857. [PubMed] [Google Scholar]
  27. Noble P. W., Henson P. M., Lucas C., Mora-Worms M., Carré P. C., Riches D. W. Transforming growth factor-beta primes macrophages to express inflammatory gene products in response to particulate stimuli by an autocrine/paracrine mechanism. J Immunol. 1993 Jul 15;151(2):979–989. [PubMed] [Google Scholar]
  28. Nussler A. K., Billiar T. R. Inflammation, immunoregulation, and inducible nitric oxide synthase. J Leukoc Biol. 1993 Aug;54(2):171–178. [PubMed] [Google Scholar]
  29. Oswald I. P., Wynn T. A., Sher A., James S. L. Interleukin 10 inhibits macrophage microbicidal activity by blocking the endogenous production of tumor necrosis factor alpha required as a costimulatory factor for interferon gamma-induced activation. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8676–8680. doi: 10.1073/pnas.89.18.8676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ruggiero V., Tavernier J., Fiers W., Baglioni C. Induction of the synthesis of tumor necrosis factor receptors by interferon-gamma. J Immunol. 1986 Apr 1;136(7):2445–2450. [PubMed] [Google Scholar]
  31. Silva J. S., Twardzik D. R., Reed S. G. Regulation of Trypanosoma cruzi infections in vitro and in vivo by transforming growth factor beta (TGF-beta). J Exp Med. 1991 Sep 1;174(3):539–545. doi: 10.1084/jem.174.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sporn M. B., Roberts A. B., Wakefield L. M., Assoian R. K. Transforming growth factor-beta: biological function and chemical structure. Science. 1986 Aug 1;233(4763):532–534. doi: 10.1126/science.3487831. [DOI] [PubMed] [Google Scholar]
  33. Tachibana K., Chen G. J., Huang D. S., Scuderi P., Watson R. R. Production of tumor necrosis factor alpha by resident and activated murine macrophages. J Leukoc Biol. 1992 Mar;51(3):251–255. doi: 10.1002/jlb.51.3.251. [DOI] [PubMed] [Google Scholar]
  34. Tsunawaki S., Sporn M., Ding A., Nathan C. Deactivation of macrophages by transforming growth factor-beta. Nature. 1988 Jul 21;334(6179):260–262. doi: 10.1038/334260a0. [DOI] [PubMed] [Google Scholar]
  35. Vodovotz Y., Bogdan C., Paik J., Xie Q. W., Nathan C. Mechanisms of suppression of macrophage nitric oxide release by transforming growth factor beta. J Exp Med. 1993 Aug 1;178(2):605–613. doi: 10.1084/jem.178.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Welch G. R., Wong H. L., Wahl S. M. Selective induction of Fc gamma RIII on human monocytes by transforming growth factor-beta. J Immunol. 1990 May 1;144(9):3444–3448. [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES