Abstract
β-Chemokines induce the directional migration of monocytes and T lymphocytes that are implicated in the pathogenesis of multiple sclerosis (MS) lesions. RANTES is a member of the β-chemokine family that has been detected in the lesions of MS patients. However, the cellular sources of RANTES message and the molecular basis for the regulation of its production in MS lesions are not well understood. Glial cells may be a major source of RANTES in vivo and have been shown to produce RANTES in vitro. Thus, the objective of this study was to establish a model system for studying the regulation of RANTES expression by cytokines in cultured human glial cells, and to determine the mechanism involved in the process. We show that the Th1 cytokines TNF-α and IL-1β independently induce RANTES mRNA and chemokine levels in human U-251 MG astroglial cells, and that these effects are time- and concentration-dependent. In addition, we demonstrate that both cytokines increased the rate of transcription of the RANTES gene, as estimated by in vitro nuclear transcript elongation assays. The transcriptional activity in TNF-α-treated U-251 MG cells started to increase at 2 h and peaked at 8 h, with levels more than 14 times greater than controls. We further show that NF-κB may play a critical role in the up-regulation of human RANTES gene expression in this system. Gel shift assays revealed an induction of in vitro nuclear extract binding activity to the NF-κB element of RANTES in cells incubated with the Th1 cytokines. These observations suggest that human astroglia, within diseased brain, may be stimulated to produce RANTES chemokine in response to TNF-α and IL-1β, and that this effect of the Th1 cytokines is attributed to increase of transcription.
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REFERENCES
Trapp, B. D., Peterson, J., Ransohoff, R. M., Rudick, R., Mork, S., and Bo, L. 1998. Axonal transection in the lesions of multiple sclerosis. N. Engl. J. Med. 338:278–285.
McFarlin, D. E., and McFarland, H. F. 1982. Multiple sclerosis. N. Engl. J. Med. 307:1183–1188 and 1246-1251.
Prineas, J. W., and Wright, R. G. 1978. Macrophages, lymphocytes, and plasma cells in the perivascular compartment in chronic multiple sclerosis. Lab. Invest. 38:409–418.
Prineas, J. W. 1985. The neuropathology of MS. Handb. Clin. Neurol. 3:213–257.
Traugott, U., Shevach, E., Chiba, J., and Raine. C. S. 1981. Autoimmune encephalomyelitis: Simultaneous identification of T-and B-cells in the target organ. Science 214:1251–1253.
Traugott, U., Reinherz, E. L., and Raine, C. S. 1983. Multiple sclerosis. Distribution of T-cell subsets within active chronic lesions. Science 219:308–310.
Lindholm, D., Castren, E., and Keifer, R. 1992. Transforming growth factor-β1 in the rat brain: increase after injury and inhibition of astrocyte proliferation. J. Cell Biol. 117:395–400.
Traugott, U. and Lebon, P. 1988. Multiple sclerosis: involvement of interferons in lesion pathogenesis. Ann. Neurol. 24:243–250.
Schmidt, B., Stoll, G., Toyka, K. V., and Hartung, H.-P. 1990. Rat astrocytes express interferon-γ immunoreactivity in normal optic nerve and after nerve transection. Brain Res. 515:347–355.
Eizenberg, O., Faberelman, A., Lotan, A., and Schwartz, M. 1995. Interleukin-2 transcripts in human and rodent brains-possible expression by astrocytes. J. Neurochem. 64:1928–1936.
Fontana, A., Kristensen, F., and Dubs, R. 1982. Production of prostaglandin E and an interleukin-1 like factor by cultured astrocytes and C6 glioma cells. J. Immunol. 129:2413–2419.
Lieberman, A. P., Pitha, P. M., Shin, H. S., and Shin, M. L. 1989. Production of tumor necrosis factor and other cytokines by astrocytes stimulated with lipopolysaccharide or a neurotropic virus. Proc. Natl. Acad. Sci. USA 86:6348–6352.
Frei, K., Bodmer, S., Schwerdel, C., and Fontana, A. 1985. Astrocytes of the brain synthesize interleukin 3-like factors. J. Immunol. 135:4044–4047.
Frei, K., Malipiero, U. V., and Leist, T. P. 1989. On the cellular source and function of interleukin 6 produced in the central nervous system in viral diseases. Eur. J. Immunol. 19:689–694.
Benveniste, E. N., Sparacio, S. M., and Norris, J. G. 1990. Induction and regulation of interleukin-6 gene expression in rat astrocytes. J. Neuroimmunol. 30:201–212.
Hao, C., Guilbert, L. J., and Fedoroff, S. 1990. Production of colony-stimulating factor-1 (CSF-1) by mouse astroglia in vitro. J. Neurosci. Res. 27:314–323.
Tedeschi, B., Barrett, J. N., and Keane, R. W. 1986. Astrocytes produce interferon that enhances the expression of H-2 antigens on a subpopulation of brain cells. J. Cell Biol. 102:2244–2253.
da Cunha, A. and Vitkovic, L. 1992. Transforming growth factor-β1 (TGF-β1) expression and regulation in rat cortical astrocytes. J. Neuroimmunol. 36:157–169.
Aloisi, F., Care, A., and Borsellino, G. 1992. Production of hemolymphopoietic cytokines (IL-6, IL-8, colony stimulating factors) by normal human astrocytes in response to IL-1β and tumor necrosis factor-α. J. Immunol. 149:2358–2366.
Barnes, D. A., Huston, M., Holmes, R., Benveniste, E. N., Yong, V. W., and Perez, D. H. 1996. Induction of RANTES expression by astrocytes and astrocytoma cell lines. J. Neuroimmunol. 71:207–214.
Janabi, N., Stefano, M. D., Wallon, C., Hery, C., Chiodi, F., and Tardieu, M. 1998. Induction of human immunodeficiency virus type 1 replication in human glial cells after proinflammatory cytokines stimulation: Effect of IFN-γ, IL-1β, and TNF-α on differentiation and chemokine production in glial cells. Glia 23:304–315.
Schall, T. J., Bacon, K., Toy, K. J., and Goddel, D. V. 1990. Selective attraction of monocytes and T lymphocytes of memory phenotype by cytokine RANTES. Nature 347:669–671.
Schall, T. J. 1991. Biology of the RANTES/SIS cytokine family. Cytokine 3:165–183.
Bigner, D. D., Bigner, S. H., Ponten, J., Westermark, B., Eny, L. F., and Wikstrand, C. F. 1981. Heterogeneity of genotypic and phenotypic characteristics of fifteen permanent cell lines derived from human gliomas. J. Neuropathol. Exp. Neurol. 40:210–218.
Noe, K. H., Cenciarelli, C., Moyer, S. A., Rota, P. A., and Shin, M. L. 1999. Requirements for measles virus induction of RANTES chemokine in human astrocytoma-derived U373 cells. J. Virol. 73:3117–3124.
Chomczynski, P. and Sacchi, N. 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159.
Li, Q., Falkler, W. A., and Bever, C. T. 1997. Endotoxin induces increased intracellular cathepsin B activity in THP-1 cells. Immunopharmacol. Immunotoxicol. 19:215–237.
Li, Q. and Bever, C. T. 1997. Interferon-γ induced increases in intracellular cathepsin B activity in THP-1 cells are dependent on RNA transcription. J. Neuroimmunol. 74:77–84.
Li, Q., Bostick-Bruton, F., and Reed, E. 1998. Effect of interleukin-1α and tumor necrosis factor-α on cisplatin-induced ERCC-1 mRNA expression in a human ovarian carcinoma cell line. Anticancer Res. 18:2283–2288.
Li, Q., Tsang, B., Bostick-Bruton, F., and Reed, E. 1999. Modulation of ERCC-1 messenger RNA expression by pharmacological agents in human ovarian carcinoma cells. Biochem. Pharmacol. 57:347–353.
Li, Q., Yu, J. J., Mu, C., Yunmbam, M. K., Slavsky, D., Cross, C. L., Bostick-Bruton, F., and Reed, E. 2000. Association between the level of ERCC-1 expression and the repair of cisplatin-induced DNA damage in human ovarian cancer cells. Anticancer Res. 20:645–652.
Sambrook, J., Fritsch, E. F., and Maniatis, T. 1989. Molecular Cloning: A laboratory Manual. Cold Spring Harbor, New York: Cold Spring Harbor.
Raj, N. B. K. and Pitha, P. M. 1983. Two levels of regulation of beta-interferon gene expression in human cells. Proc. Natl. Acad. Sci. USA 80:3923–3927.
Li, Q., Gardner, K., Zhang, L., Tsang, B., Bostick-Bruton, F., and Reed, E. 1998. Cisplatin induction of ERCC-1 mRNA expression in A2780/CP70 human ovarian cancer cells. J. Biol. Chem. 273:23419–23425.
Li, Q., Ding, L., and Bever, C. T. 1998. Interferon-γ induces cathepsin B expression in a human macrophage-like cell line by increasing both transcription and mRNA stability. Int. J. Mol. Med. 2:181–186.
Li, Q., Zhang, L., Tsang, B., Gardner, K., Bostick-Bruton, F., and E. Reed. 1999. Phorbol ester exposure activates an AP-1-mediated increase in ERCC-1 messenger RNA expression in human ovarian tumor cells. Cell. Mol. Life Sci. 55:456–466.
Zhang, L., Zhan, S., Li, Q., Kim, M., Seth, P., and Helman, L. J. 1998. AP-2 may contribute to IGF-II overexpression in rhabdomyosarcoma. Oncogene 17:1261–1270.
Nelson, P. J., Kim, H. T., Manning, W. C., Goralski, T. J., and Krensky, A. M. 1993. Genomic organization and transcriptional regulation of the RANTES chemokine gene. J. Immunol. 151:2601–2612.
Snedecor, G. W. 1956. Statistical Methods. Ames, Iowa: Iowa State University.
Snedecor, G. W. and Cochran, W. G. 1967. Statistical methods. 6th ed. Ames, IA: The Iowa State University Press, Pages 1–593.
Moriuchi, H., Moriuchi, M., and Fauci, A. S. 1997. Nuclear factor-κB potently up-regulates the promoter activity of RANTES, a chemokine that blocks HIV infection. J. Immunol. 158:3483–3491.
Thomas, L. H., Friedland, J. S., Sharland, M., and Becker, S. 1998. Respiratory syncytial virus-induced RANTES production from human bronchial epithelial cells is dependent on nuclear factor-κB nuclear binding and is inhibited by adenovirus-mediated expression of inhibitor of κBα. J. Immunol. 161:1007–1016.
Ayad, O., Stark, J. M., Fiedler, M. M., Menendez, I. Y., Ryan, M. A., and Wong, H. R. 1998. The heat shock response inhibits RANTES gene expression in cultured human lung epithelium. J. Immunol. 161:2594–2599.
Zoja, C., Donadelli, R., Colleoni, S., Figliuzzi, M., Morigi, M., and Remuzzi, G. 1998. Protein overload stimulates RANTES production by proximal tubular cells depending on NF-κB activation. Kidney Int. 53:1608–1615.
Wang, X., Jobin, C., Allen, J. B., Roberts, W. L., and Jaffe, G. J. 1999. Suppression of NF-κB-dependent proinflammatory gene expression in human RPE cells by a proteasome inhibitor. Invest. Ophthalmol. Vis. Sci. 40:477–486.
Hu, S., Chao, C. C., Sheng, W. S., Sutton, R. L., Rockswold, G. L., and Peterson, P. K. 1999. Inhibition of microglial cell RANTES production by IL-10 and TGF-β. J. Leukoc. Biol. 65:815–821.
Oppenheim, J. J., Zachariae, C. O., Mukaida, N., and Matsushima, K. 1991. Properties of the novel proinflammatory supergene & quot; intercrine & quot; cytokine family. Annu. Rev. Immunol. 9:617–648.
Van Damme, J. 1994. Interleukin-8 and related molecules. Pages 185–208. In: A. Thompson, ed. The Cytokine Handbook. New York: Academic Press.
Karpus, W. J., Lukacs, N. W., McRae, B. L., Strieter, R. M., Kunkel, S. L., and Miller, S. D. 1995. An important role for the chemokine macrophage inflammatory protein-1α in the pathogenesis of the T cell-mediated autoimmune disease, experimental autoimmune encephalomyelitis. J. Immunol. 155:5003–5010.
Karpus, W. J., and Ransohoff, R. M. 1998. Chemokine regulation of experimental autoimmune encephalomyelitis: Temporal and spatial expression patterns govern disease pathogenesis. J. Immunol. 161:2667–2671.
Glabinski, A. R., Tuohy, V. K., and Ransohoff, R. M. 1998. Expression of chemokines RANTES, MIP-1α and GRO-α correlates with inflammation in acute experimental autoimmune encephalomyelitis. Neuroimmunomodulation 5:166–171.
Sun, D., Hu, X., Liu, X., Whitaker, J. N., and Walker, W. S. 1997. Expression of chemokine genes in rat glial cells: The effect of myelin basic protein-reactive encephalitogenic T cells. J. Neuroscience Res. 48:192–200.
Schall, T. J., Jongstra, J., Bradley, J. D., Jorgensen, J., Davis, M. M., and Krensky, A. M. 1988. A human T cell-specific molecule is a member of a new gene family. J. Immunol. 141:1018–1025.
Kuna, P., Reddigari, S. R., Schall, T. J., Rucinski, D., Sadick, M., and Kaplan, A. P. 1993. Characterization of the human basophil response to cytokines, growth factors, and histamine releasing factors of the intercrine/chemokine family. J. Immunol. 150:1932–1943.
Godiska, R., Chantry, D., Dietsch, G. N., and Gray, P. W. 1995. Chemokine expression in murine experimental allergic encephalomyelitis. J. Neuroimmunol. 58:167–176.
Noe, K. H., Fisher, S. N., Dhib-Jalbut, S. S., and Shin, M. L. 1996. Induction of cytokine RANTES by virus in astrocytes. J. Neurochem. 66:S71D.
Marfaing-Koka, A., Devergne, O., Gorgone, G., Portier, A., Schall, T. J. and Emilie, D. 1995. Regulation of the production of the RANTES chemokine by endothelial cells: Synergistic induction by IFN plus TNF-α and inhibition by IL-4 and IL-13. J. Immunol. 154:1870–1877.
Selmaj, K., Raine, C. S., Cannella, B., and Brosnan, C. F. 1991. Identification of lymphotoxin and tumor necrosis factor in multiple sclerosis brain lesions. J. Clin. Invest. 87:949–954.
Hvas, J., McLean, C., Justesen, J., Kannourakis, G., Steinman, L., and Bernard, C. C. A. 1997. Perivascular T cells express the pro-inflammatory chemokine RANTES mRNA in multiple sclerosis lesions. Scand. J. Immunol. 46:195–203.
Hvas, J. and Bernard, C. C. A. 1998. Molecular detection and quantitation of the chemokine RANTES in mRNA neurological brain. APMIS 106:598–604.
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Li, Q.Q., Bever, C.T. Th1 Cytokines Stimulate RANTES Chemokine Secretion by Human Astroglial Cells Depending on De Novo Transcription. Neurochem Res 26, 125–133 (2001). https://doi.org/10.1023/A:1011042711631
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DOI: https://doi.org/10.1023/A:1011042711631