Sage Journals: Discover world-class research

Abstract

Author for correspondence: Klaus Bendtzen, Institute for Inflammation Research IIR 7521, Rigshospitalet University Hospital, 9 Blegdamsvej, DK-2100 Copenhagen, Denmark. E-mail: address: kben@mail.dk Administration of interferons (IFNs) may induce antibodies that interfere with therapeutic efficacy. We have optimized and validated methods for large-scale economic screening. Sera from patients with relapsing-remitting multiple sclerosis (MS) were investigated for binding antibody (BAb) by protein-G affinity-chromatography radioimmunoassay and a commercially available enzyme immunoassay (EIA). Neutralizing antibody (NAb) was investigated by cytopathic effect assays (CEA) using both fixed amount and serially diluted sera. BAb correlated with log10-transformed titres obtained by EIA (r=0.70, p<0.0001); the latter, however, failed to demonstrate low-level BAb. Comparison of clinical significance of NAb-positivity measured by biological assays with different sensitivities demonstrated an optimal odds ratio for relapse rate using 10 laboratory units (LU)/mL. Purification of IgG prior to CEA removed toxicity from toxic sera. The neutralizing capacity data correlated linearly with log10-transformed titres obtained by a Kawade 10-to-1 LU/mL CEA (r=0.77, p<0.0001). In conclusion, neutralizing capacity CEA utilizing a fixed amount of serum predicts differences in relapse rates in IFNβ-treated MS patients and correlates with NAb titres of the 10-to- 1 LU/mL CEA. Neutralizing capacity CEA is less laborious and more economical than titre-based NAb assays and suitable for large-scale screenings of MS patients.

Keywords

binding antibodies clinical validation multiple sclerosis neutralizing antibodies radioimmunoassay screening methods therapeutic use type 1 interferon

Get full access to this article

View all access options for this article.

References

Vilcek J , Feldmann M. Historical review: Cytokines as therapeutics and targets of therapeutics . Trends Pharmacol Sci 2004; 25: 201-209 .

Antonelli G , Currenti M , Turriziani O , Dianzani F. Neutralizing antibodies to interferon-a: Relative frequency in patients treated with different interferon preparations . J Infect Dis 1991; 163: 882-885 .

The IFNβ Multiple Sclerosis Study Group and the University of British Columbia MS/MRI Analysis Group . Neutralizing antibodies during treatment of multiple sclerosis with interferon b-1b: experience during the first three years . Neurology 1996; 47: 889-894 .

Abdul-Ahad A , Shah S , Galazka A. Incidence of antibodies to interferon-b in patients treated with recombinant human interferon-b1a from mammalian cells . Cytokines Cell Mol Ther 1997; 3: 27-32 .

Antonelli G , Bagnato F , Pozzilli C , Simeoni E , Bastianelli S , Currenti M et al. Development of neutralizing antibodies in patients with relapsing-remitting multiple sclerosis treated with IFNβ1a . J Interferon Cytokine Res 1998; 18: 345-350 .

Sørensen PS , Ross C , Clemmesen KM , Bendtzen K , Frederiksen JL , Jensen K et al. Clinical importance of neutralising antibodies against interferon beta in patients with relapsing-remitting multiple sclerosis . Lancet 2003; 362: 1184-1191 .

Files JG , Gray JL , Do LT , Foley WP , Gabe JD , Nestaas E et al. A novel sensitive and selective bioassay for human type I interferons . J Interferon Cytokine Res 1998; 18: 1019-1024 .

Pungor E, Jr , Files JG , Gabe JD , Do LT , Foley WP , Gray JL et al. A novel bioassay for the determination of neutralizing antibodies to IFN-beta1b . J Interferon Cytokine Res 1998; 18: 1025-1030 .

Brickelmaier M , Hochman PS , Baciu R , Chao B , Cuervo JH , Whitty A. ELISA methods for the analysis of antibody responses induced in multiple sclerosis patients treated with recombinant interferon-beta . J Immunol Methods 1999; 227: 121-135 .

10.

Grossberg SE , Kawade Y , Kohase M , Klein JP. 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: 729-742 .

11.

Grossberg SE , Kawade Y , Kohase M , Klein JP. 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: 743-755 .

12.

Kawade Y. An analysis of neutralization reaction of interferon by antibody: a proposal on the expression of neutralization titre . J Interferon Res 1980; 1: 61-70 .

13.

Bendtzen K. Anti-IFN BAb and NAb antibodies: A minireview . Neurology 2003; 61(Suppl. 5): S6-S10 .

14.

The IFNβ Multiple Sclerosis Study Group . Inter-feron beta-1b is effective in relapsing-remitting multiple sclerosis. I. Clinical results of a multicenter, randomized, double-blind, placebo-controlled trial . Neurology 1993; 43: 655-661 .

15.

The IFNβ Multiple Sclerosis Study Group and The University of British Columbia MS/MRI Analysis Group . Interferon b-1b in the treatment of multiple sclerosis: final outcome of the randomized controlled trial . Neurology 1995; 45: 1277-1285 .

16.

Kivisäkk P , Alm GV , Tian WZ , Matusevicius D , Fredrikson S , Link H. Neutralising and binding anti-interferon-b-1 b (IFN-b-1b) antibodies during IFN-b-1b treatment of multiple sclerosis . Mult Scler 1997; 3: 184-190 .

17.

Rudick RA , Simonian NA , Alam JA , Campion M , Scaramucci JO , Jones W et al. Incidence and significance of neutralizing antibodies to interferon beta-1a in multiple sclerosis . Neurology 1998; 50: 1266-1272 .

18.

Ross C , Clemmesen KM , Svenson M , Sørensen PS , Koch-Henriksen N , Skovgaard GL et al. Immunogenicity of interferon-b in multiple sclerosis patients: influence of preparation, dosage, dose frequency, and route of administration. Danish Multiple Sclerosis Study Group . Ann Neurology 2000; 48: 706-712 .

19.

Bendtzen K , Ross C , Hansen MB , Svenson M. Natural and induced anti-cytokine antibodies. In Ciliberto G , Savino R , eds. Cytokine Inhibitors. New York: Marcel Dekker , 2000: 53-95.

20.

Ross C , Engler CB , Sander B , Bendtzen K. IFN-a antibodies in patients with age-related macular degeneration treated with recombinant human IFN-a2a . J Interferon Cytokine Res 2002; 22: 421-426 .

21.

Bendtzen K , Hansen MB , Ross C , Svenson M. Detection of autoantibodies to cytokines . Mol Biotechnol 2000; 14: 251-261 .

22.

Giovannoni G , Munschauser FE 3rd , Deisenhammer F. Neutralising antibodies to interferon beta during the treatment of multiple sclerosis . J Neurol Neurosurg PS 2002; 73: 465-469 .

23.

Pachner AR. An improved ELISA for screening for neutralizing anti-IFN-beta antibodies in MS patients . Neurology 2003; 61: 1444-1446 .

24.

Giovannoni G. Strategies to treat and prevent the development of neutralizing anti-interferon-b antibodies . Neurology 2003; 61(Suppl. 5): S13-S17 .

Measuring and evaluating interferon b-induced antibodies in patients with multiple sclerosis

Abstract

Keywords

Get full access to this article

References