MxA Gene Expression Analysis as an Interferon-β Bioactivity Measurement in Patients with Multiple Sclerosis and the Identification of Antibody-Mediated Decreased Bioactivity
Tóm tắt
Background: Interferon-β (IFNβ) has proven to be an important advance in the therapy of multiple sclerosis (MS), but optimal markers for bioactivity have not been identified. To accurately measure bioactivity in MS patients treated with IFNβ, we developed and tested a real-time reverse transcriptase (RT)-PCR assay for gene expression of MxA, an IFNβ-induced gene in the peripheral blood of patients treated with IFNβ.
Methods: We compared IFNβ-treated patients with MS to controls in expression of MxA relative to the house-keeping gene, GAPDH. 2′-5′oligoadenylate synthetase (OAS) gene expression was also tested by real-time RT-PCR on RNA from the same patient specimens. Anti-IFNβ antibody was measured by ELISA and a cytopathic effect assay.
Results: Seven of 54 patients were found to have complete loss of bioactivity. MxA expression correlated well with OAS expression. All patients with lost bioactivity had high levels of binding antibodies or neutralizing antibodies.
Conclusions: This is the first demonstration that a real-time RT-PCR assay can be used to monitor therapy with interferons. These data identify MxA mRNA as an excellent biomarker for INFβ action on the IFN receptor, and clarify the relationship between anti-IFNβ antibodies and bioactivity in patients with MS treated with IFNβ.
Tài liệu tham khảo
The IFNB Multiple Sclerosis Study Group. Interferon beta-1b is effective in relapsing-remitting multiple sclerosis. Neurology 1993; 43: 665–1
Runkel L, De Dios C, Karpusas M, et al. Mapping of IFN-beta epitopes important for receptor binding and biologic activation: comparison of results achieved using antibody-based methods and alanine substitution mutagenesis. J Interferon Cytokine Res 2001; 21(11): 931–41
Kochs G, Haener M, Aebi U, et al. Self-assembly of human MxA GTPase into highly ordered dynamin-like oligomers. J Biol Chem 2002; 277(16): 14172–6
von Wussow P, Jakschies D, Hochkeppel HK, et al. The human intracellular Mx-homologous protein is specifically induced by type I interferons. Eur J Immunol 1990; 20: 2015–9
Deisenhammer F, Reindl M, Harvey J, et al. Bioavailability of interferon beta 1b in MS patients with and without neutralizing antibodies. Neurology 1999; 52(6): 1239–43
Deisenhammer F, Mayringer I, Harvey J, et al. A comparative study of the relative bioavailability of different interferon beta preparations. Neurology 2000; 54(11): 2055–60
Bertolotto A, Gilli F, Sala A, et al. Evaluation of bioavailability of three types of IFNb in multiple sclerosis patients by a new quantitative-competitive-PCR method for MxA quantification. J Immunol Methods 2001; 256: 141–52
Szuchet S, Plachetzki DC, Karialukas R. Oligodendrocytes express an alpha/beta-interferon-susceptible Mx gene: molecular characterization of the encoded protein. Glia 2002; 37(2): 183–9
Bertolotto A, Gilli F, Sala A, et al. Persistent neutralizing antibodies abolish the interferon beta bioavailability in MS patients. Neurology 2003; 60(4): 634–9
Pachner AR. Anti-cytokine antibodies in IFNβ-treated MS patients and need for testing: the plight of the practicing neurologist. Neurology 1997; 49: 647–50
Pachner AR. Measurement of antibodies to interferon beta in patients with multiple sclerosis. Arch Neurol 2001; 58(8): 1299–300
Giovannoni G, Munschauer III FE, Deisenhammer F. Neutralising antibodies to interferon beta during the treatment of multiple sclerosis. J Neurol Neurosurg Psychiatry 2002; 73(5): 465–9
Whitaker JN. Commentary on neutralizing antibodies to interferon beta in patients with multiple sclerosis. Arch Neurol 2001; 58: 1301
Rice G. The significance of neutralizing antibodies in patients with multiple sclerosis treated with interferon beta. Arch Neurol 2001; 58: 1297–300
Schellekens H. Bioequivalence and the immunogenicity of biopharmaceuticals. Nat Rev Drug Discov 2002; 1: 457–62
PRISMS Study Group. PRISMS-4: long-term efficacy of interferon-b-1a in relapsing MS. Neurology 2001; 56: 1628–36
The IFNB Multiple Sclerosis Study Group. Neutralizing antibodies during treatment of multiple sclerosis with interferon beta-1b: experience during the first three years. Neurology 1996; 47: 889–94
Rudick RA, Simonian NA, Alam JA, et al. Incidence and significance of neutralizing antibodies to interferon beta-1a in multiple sclerosis: Multiple Sclerosis Collaborative Research Group (MSCRG). Neurology 1998; 50(5): 1206–8
Richert N, Zierak MC, Bash C, et al. MRI evaluation of multiple sclerosis patients after cessation of interferonβ-1b [abstract]. Ann Neurol 1998 Sep; 44(3): 505A
Richert ND, Zierak MC, Bash CN, et al. MRI and clinical activity in MS patients after terminating treatment with interferonβ. Mult Scler 2000; 6(2): 86–90
Brickelmaier M, Hochman PS, Baciu R, et al. ELISA methods for the analysis of antibody responses induced in multiple sclerosis patients treated with recombinant interferon-β. J Immunol Methods 1999; 227: 121–35
PAXgene blood RNA handbook. Franklin Lakes (NJ): Becton-Dickinson, 2001
Kurtzke JF. On the evaluation of disability in multiple sclerosis. Neurology 1961; 11: 686–93
Grossberg SE, Kawade Y. The expression of potency of neutralizing antibodies for interferons and other cytokines. Biotherapy 1997; 10(1): 93–8
Grossberg SE, Kawade Y, Kohase M, et al. The neutralization of interferons by antibody: I. Quantitative and theoretical analyses of the neutralization reaction in different bioassay systems. J Interferon Cytokine Res 2001; 21(9): 729–42
Grossberg SE, Kawade Y, Kohase M, et al. The neutralization of interferons by antibody: II. Neutralizing antibody unitage and its relationship to bioassay sensitivity: the tenfold reduction unit. J Interferon Cytokine Res 2001; 21(9): 743–55
Kracke A, von Wussow P, Al-Masri AN, et al. Mx proteins in blood leukocytes for monitoring interferon beta-1b therapy in patients with MS. Neurology 2000; 54(11): 193–9
Vallittu A-M, Halminen M, Peltoniemi J, et al. Neutralizing antibodies reduce MxA protein induction in interferon-beta-1a-treated MS patients. Neurology 2002 Jun; 58: 1786–90
Rainen L, Oelmueller U, Jurgensen S, et al. Stabilization of mRNA expression in whole blood samples. Clin Chem 2002; 48(11): 1883–90
Sachs AB. Messenger RNA degradation in eukaryotes. Cell 1993; 74: 413–21
Beelman CA, Parker R. Degradation of mRNA in eukaryotes. Cell 1995; 81: 179–83
Muller MC, Merx K, Wieber A, et al. Improvement of molecular monitoring of residual disease in leukemias by bedside RNA stabilization. Leukemia 2002; 16: 2395–9
Russell-Harde D, Wagner TC, Perez HD, et al. Formation of a uniquely stable type I interferon receptor complex by interferon beta is dependent upon particular interactions between interferon beta and its receptor and independent of tyrosine phosphorylation. Biochem Biophys Res Commun 1999; 255(2): 539–44
Wandinger KP, Sturzebecher CS, Bielekova B, et al. Complex immunomodulatory effects of interferon-beta in multiple sclerosis include the upregulation of T helper 1-associated marker genes. Ann Neurol 2001; 50(3): 349–57
Floris S, Ruuls SR, Wierinckx A, et al. Interferon-beta directly influences monocyte infiltration into the central nervous system. J Neuroimmunol 2002; 127(1–2): 69–79
Van Weyenbergh J, Wietzerbin J, Rouillard D, et al. Treatment of multiple sclerosis patients with interferon-beta primes monocyte-derived macrophages for apoptotic cell death. J Leukoc Biol 2001; 70(5): 745–8