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Multiple Sclerosis, Vol. 12, No. 1, 39-46 (2006)
DOI: 10.1191/1352458506ms1272oa

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

C Ross

Institute for Inflammation Research, Rigshospitalet University Hospital, Copenhagen

K M Clemmesen

Institute for Inflammation Research, Rigshospitalet University Hospital, Copenhagen

P S Sørensen

Department of Neurology, Rigshospitalet University Hospital, Copenhagen

N Koch-Henriksen

Aalborg Hospital, Ålborg, Department of Neurology, Denmark

K Bendtzen

Institute for Inflammation Research, Rigshospitalet University Hospital, Copenhagen

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 IFNß-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.

Key Words: binding antibodies • clinical validation • multiple sclerosis • neutralizing antibodies • radioimmunoassay • screening methods • therapeutic use • type 1 interferon


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