IFN-β reduces the number and severity of exacerbations of multiple sclerosis (MS), presumably by modifying immune regulation. We used semiquantitative polymerase chain reaction (RT-PCR) to measure mRNA levels for cytokines before and after IFN β-1b therapy. mRNA was extracted from mononuclear cells of nine healthy controls and 31 patients with MS. Before therapy, IL-10 and leukemia inhibitory factor (UF) mRNA levels were elevated in stable MS compared to active MS. Twenty four hours after IFN β-1b treatment, mRNA levels for IL-1, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IFN-γ, TNF-α and UF had not changed. At 1 week, TNF-α mRNA increased and IL-10 and UF mRNA rose in 75% of patients. IL-2, IL-4, IL-12, IL-13 and IFN-γ did not change. At 3 months, cytokine mRNA returned to baseline levels. mRNA for the IFN-induced antiviral enzyme, 2, 5-OAS, rose by 24 h, peaked at 1 week, and remained elevated thereafter. Serum triglycerides and liver enzymes rose after therapy. Increased SGPT at 3 months correlated with TNF-α mRNA levels, suggesting that cytokines may cause some side effects of IFN β-1b. Baseline cytokine mRNA levels reflect disease activity, but the therapeutic effect of IFN β-1b does not appear to be explained by changes in cytokine mRNA levels.
1 Reder AT, Arnason BGW. (1985) Immunology of multiple sclerosis. In: Vinken PJ, Bruyn GW, Klawans HL, Koetsier JC.Handbook of Clinical Neurology: Demyelinating Diseases, Elsevier Science Publishers: Amsterdam. pp. 337–395.
2.
2 Maimone D, et al. (1991) Cytokine levels in the cerebrospinal fluid and sera of patients with multiple sclerosis. J Neuroimmunol32: 67–74.
3.
3 Rieckmann P, et al. (1994) Cytokine mRNA levels in mononuclear blood cells from patients with multiple sclerosis. Neurology44: 1523–1526.
4.
4 Sharief MK, Hentges R. (1991) Association between tumor necrosis factor-α and disease progression in patients with multiple sclerosis. N Engl J Med325: 467–472.
5.
5 Link J, et al. (1994) Increased transforming growth factor-β, interleukin-4, and interferon-7 in multiple sclerosis. Ann Neurol36: 379–386.
6.
6 Kennedy MK, et al. (1992) Analysis of cytokine mRNA expression in the central nervous system of mice with experimental autoimmune encephalomyelitis reveals that IL-10 mRNA expression correlates with recovery. J Immunol149: 2496–2505.
7.
7 Khoury SJ, Hancock WW, Weiner HL. (1992) Oral tolerance to myeline basic protein and natural recovery from experimental autoimmune encephalomyelitis are associated with downregulation of inflammatory cytokines and differential upregulation of transforming growth factor β, interleukin 4, and prostaglandin E expression in the brain. J Exp Med176: 1355–1364.
8.
8 Karpus WJ, Gould KE, Swanborg RH. (1992) CD4+ suppressor cells of autoimmune encephalomyelitis respond to T cell receptor-associated determinants on effector cells by interleukin-4 secretion. Eur J Immunol22: 1757–1763.
9.
9 The IFN-Beta Multiple Sclerosis Study Group. (1993) Interferon beta-1b is effective in relapsing-remitting multiple sclerosis. I. Clinical results of a multicenter, randomized, double-blind placebo-controlled trial.Neurology43: 655–661.
10.
10 Jacobs L, et al. (1994) Results of a phase III trial of intramuscular recombinant beta interferon as treatment for multiple sclerosis. Ann Neurol36: 259.
11.
11 The IFN-Beta Multiple Sclerosis Study Group. (1993) Interferon beta-1b is effective in relapsing-emitting multiple sclerosis. II. MRI analysis results of a multicenter, randomized, double-blind, placebo-controlled trial.Neurology43: 662–667.
13 Panitch HS, Folus JS, Johnson KP. (1987) Recombinant beta interferon inhibits gamma interferon production in multiple sclerosis. Ann Neurol22: 139A.
14.
14 Noronha A, Toscas A, Jensen MA. (1993) Interferon β decreases T cell activation and interferon γ production in multiple sclerosis. J Neuroimmunol46: 145–154.
15.
15 Abu-Khabar KS, Armstrong JA, Ho M. (1992) Type I interferons (IFN-α and-β) suppress cytotoxin (tumor necrosis factor-α and lymphotoxin) production by mitogen-stimulated human peripheral blood mononuclear cells. J Leukoc Biol52: 165–172.
16.
16 Panitch HS, Folus JS, Johnson KP. (1989) Beta interferon prevents HLA class II antigen induction by gamma interferon in MS.Neurology39 (Suppl 1): 171.
17.
17 Barna BP, et al. (1989) Interferon-β impairs induction of HLA-DR antigen expression in cultured adult human astrocytes. J Neuroimmunol23: 45–53.
18.
18 Ransohoff RM, et al. (1991). Interferon-b specifically inhibits interferon-g-induced class II major histocompatibility complex gene transcription in a human astrocytoma cell line. J Neuroimmunol33: 103–112.
19.
19 Reder AT, et al. (1993) Astrocyte cytolysis by MHC class II-specific mouse T cell clones. Transplantation56: 393–399.
20.
20 Porrini AM, Gambi D, Reder AT. (1995) Interferon beta effects on interleukin-10 secretion. Mononuclear cell response to interleukin-10 is normal in multiple sclerosis patients. J Neuroimmunol61: 27–34.
22 Bongioanni MR, et al. (1994) High-dose recombinant-alpha-2a-interferon (IFNA) therapy modulates cytokine prof iles in MS. J Neuroimmunol54: 1–2.
23.
23 Noronha A, Toscas A, Jensen MA. (1990) Interferon beta augments suppressor cell function in multiple sclerosis. Ann Neurol27: 207–210.
24.
24 Antel JP, et al. (1986) Activated suppressor cell dysfunction in chronic progressive multiple sclerosis. J Immunol137: 137–141.
25.
25 Chomczynski P, Sacchi N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chlorof orm extraction. Anal Biochem162: 156–159.
26.
26 Xie W-Q, Rothblum LI. (1991) Rapid, small-scale RNA isolation from tissue culture cells. Bio Techniques11: 324–327.
27.
27 Karp CL, et al. (1993) In vivo cytokine prof iles in patients with kala-azar. Marked elevation of both interleukin-10 and interferon-gamma. J Clin Invest91: 1644–1648.
28.
28 Wang AM, et al. (1985) Molecular cloning of the complementary DNA for human tumor necrosis factor. Science228: 149–154.
29.
29 Nedwin GE, et al. (1985) Human lymphotoxin and tumor necrosis factor genes: structure, homology and chromosomal localization. Nucleic Acids Research13: 6361–6373.
30.
30 Yamaura M, et al. (1992) Cytokine patterns of immunologically medicated tissue damage. J Immunol149: 1470–1475.
31.
31 Vieira P, et al. (1991) Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: Homology to Epstein-Barr virus open reading frame BCRFI. Proc Natl Acad Sci USA88: 1172–1176.
32.
32 Saito S, et al. (1993) Cytokine production by CD16 CD56bright natural killer cells in the human early pregnancy decidua. Intl Immunol5: 559–563.
33.
33 Marie I, Hovanessian AG. (1992) The 69-kDa 2–5A synthetase is composed of two homologous and adjacent functional domains. J Biol Chem267: 9933–9939.
34.
34 Tse WT, Forget BG. (1990) Reverse transcription and direct amplification of cellular RNA transcripts by Taq polymerase.Gene88: 293–296.
35.
35 Horikoshi T, et al. (1992) Quantitation of thymidylate synthase, dihydrof olate reductase, and DT-diaphorase gene expression in human tumors using the polymerase chain reaction. Cancer Res52: 108–116.
36.
36 Lagoo-Deenadayalan S et al. (1993) A standardized approach to PCR-based semiquantitiation of multiple cytokine gene transcripts from small cell samples. Lymphokine Cytokine Res12: 59–67.
37.
37 Braga VMM, Gendler SJ. (1994) Co-amplification of two cDNAs in RT-PCR can alter the yield of both products. BioTechniques17: 228–230.
38.
38 Bruchelt G, et al. (1992) Determination of 2′, 5′-oligoade-nylate synthetase in serum and peripheral blood mononuclear cells before and after subcutaneous application of recombinant interferon beta and gamma. Eur J Clin Chem Clin Biochem30: 521–528.
39.
39 Anegon I, et al. (1990) Production of human interleukin for DA cells (HILDA)/leukemia inhibitory factor (LIF) by activated monocytes. Cell Immunol130: 50–65.
40.
40 Barres BA, et al. (1993) Multiple extracellular signals are required for long-term oligodendrocyte survival. Development118: 283–295.
41.
41 Gard AL, et al. (1995) Astroglial control of oligodendrocyte survival mediated by PDGF and leukemia inhibitory factor-like protein. Development121: 2197–2197.
42.
42 Gruss HJ, Brach MA, Hermann F. (1992) Involvement of nuclear factor-kappa B in induction of the interleukin-6 gene by leukemia inhibitory factor. Blood80: 2563–2570.
43.
43 Aderka D, Le J, Vilček J. (1989) IL-6 inhibits lipopolysaccharide-induced tumor necrosis factor production in cultured human monocytes, U937 cells, and in mice. J Immunol143: 3517–3523.
44.
44 Schindler R, et al. (1990) Correlations and interactions in the production of IL-6, IL-1 and TNF in human blood mononuclear cells: IL-6 suppresses IL-1 and TNF. Blood75: 40–47.
45.
45 Patterson PH, Fann MJ. (1992) Further studies of the distribution of CDF/LIF mRNA. Ciba Found Symp167: 125–135.
46.
46 Hof man FM, et al. (1989) Tumor necrosis factor identified in multiple sclerosis brain. J Exp Med170: 607–612.
47.
47 Selmaj KW, Raine CS. (1988) Tumor necrosis factor mediates myeline and oligodendrocyte damage in vitro. Ann Neurol23: 339–346.
48.
48 Soliven B, Szuchet S, Nelson DJ. (1991) Tumor necrosis factor inhibits K+ current expression in cultured oligodendrocytes. J Membr Biol124: 127–137.
49.
49 Selmaj KW, et al. (1990) Proliferation of astrocytes in vitro in response to cytokines: a primary role for tumor necrosis factor. J Immunol144: 129–135.
50.
50 Durelli L, et al. (1994) Chronic systemic high-dose recombinant interferon alfa-2a reduces exacerbation rate, MRI signs of disease activity, and lymphocyte interferon gamma production in relapsing-remitting multiple sclerosis. Neurology44: 406–413.
51.
51 Porrini AM, Reder AT. (1994) IFN-γ, IFN-β and PGE1 affect monokine secretion: relevance to monocyte activation in multiple sclerosis. Cell Immunol157: 428–438.
52.
52 Dayal AS, et al. (1995) Inteferon-gamma-secreting cells in multiple sclerosis patients treated with interferon beta-1b. Neurology45: 2173–2177.
53.
53 Brinkman V, et al. (1993) Interferon a increases the frequency of interferon γ-producing human CD4+ T cells. J Exp Med178: 1655–1663.
54.
54 Powrie F, Menon S, Coffman RL. (1993) Interleukin-4 and interleukin-10 synergize to inhibit cell-mediated immunity in vivo. Eur J Immunol23: 2223–2229.
55.
55 Finkelman FD, et al. (1991) Regulation by interferon a of immunoglobulin isotype selection and lymphokine production in mice. J Exp Med174: 1179–1188.
56.
56 Kimura K, et al. (1987) Phase I study of recombinant human tumor necrosis factor. Cancer Chemother Pharmacol20: 223–229.
57.
57 Imagawa DK, et al. (1990) The role of tumor necrosis factor in allograft rejection. Transplantation50: 219–225.
58.
58 Assoian RK, et al. (1987) Expression and secretion of type β transforming growth factor by activated human macrophages. Proc Natl Acad Sci USA84: 6020–6024.
59.
59 Wanidworanun C, Strober W. (1993) Predominant role of tumor necrosis factor-α in human monocyte IL-10 synthesis. J Immunol151: 6853–6861.
60.
60 Genç K, Reder AT. (1996) Interferon regulation of B7 in mononuclear cells: Increased B7-1+ B cells in active multiple sclerosis, and reversal by IFNβ-1b therapy. (Submitted).
