Flow cytometry model for the detection of neutralizing antibodies against of IFN-β
Abstract
Introduction. Interferon beta (IFN-β) is a treatment for relapsing remitting multiple sclerosis. However, the therapeutic use of recombinant proteins induces a humoral immunologic response resulting in the induction of binding (BAb) or neutralizing (NAb) antibodies against the biological product. The presence of neutralizing antibodies has been associated with decreased IFN-β treatment efficacy.
Materials and methods. Two tumor cell lines (K562 and U937) were cultivated with human recombinant IFN-β1a at different concentrations and lengths of time in order to measure the expression of intracellular ISG15, an inducible molecule in the IFN-β1a signaling cascade. Blood was obtained from non-immunized and IFN-β1a immunized (100,000 IU) New Zealand rabbits. The presence of BAbwas evaluated by ELISA. For NAb detection, sera 1:20 dilution were added to the IFN-β1a-stimulated cell lines, and ISG15 expression was evaluated by flow cytometry.
Results. K562 cells provided the better cell line for the assay, stimulated with a dose of 1,000 IU ofIFN-β1a, and a 1:100 dilution for the primary antibody and a 1:200 dilution for the secondary antibody. ISG15 expression was compared between cells alone or cultivated with IFN-β1a. Mean fluorescence intensity (MFI) for ISG-15 expression median was 198 arbitrary units (AU) with interquartile ranges of173-231 AU for non-stimulated cells and 430 AU with interquartile ranges of 316-611.5 AU for IFN-β1a stimulated cells (p<0.01). Immunized rabbit sera decreased the expression of ISG-15 in K562 cells stimulated with IFN-β1a, whereas non-immunized rabbit sera did not.
Conclusions. This rabbit model demonstrates that ISG15 expression evaluated with flow cytometry can be used as a detection assay for NAb.
doi: http://dx.doi.org/10.7705/biomedica.v32i4.637
Downloads
References
Compston A, Coles A. Multiple sclerosis. Lancet. 2008;372:1502-17.
Ascherio A, Munger K. Epidemiology of MS: From risk factors to prevention. Semin Neurol. 2008;1:17-28.
Toro J, Sarmiento OL, Díaz del Castillo A, Satizabal CL, Ramírez, JD, Montenegro AC, et al. Prevalence of multiple sclerosis in Bogotá, Colombia. Neuroepidemiology. 2007;28:33-8.
Coyle PK. Existing therapies for multiple sclerosis offer proven efficacy and safety. Curr Opin Neurol. 2009;22:S4-9.
Yong VW, Chabot S, Stuve O, Williams G. Interferon beta in the treatment of multiple sclerosis: Mechanisms of action. Neurology. 1998;51:682-9.
Polman CH, Bertolotto A, Deisenhammer F, Giovannoni G, Hartung HP, Hemmer B, et al. Recommendations for clinical use of data on neutralizing antibodies to interferonbeta therapy in multiple sclerosis. Lancet Neurol. 2010;9:740-50.
Sorensen PS, Ross C, Clemmesen KM, Bendtzen K, Frederiksen JL, Kristensen O, el at. Clinical importance of neutralizing antibodies against interferon beta in patients with relapsing-remitting multiple sclerosis. Lancet. 2003;362:1184-91.
Goodin DS, Frohman EM, Hurwitz B, O’Connor PW, Oger JJ, Reger AT, et al. Neutralizing antibodies to interferon beta: Assessment of their clinical and radiographic impact. An evidence report - Report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. Neurology. 2007;68:977-84.
PRISMS (Prevention of Relapses and Disability by Interferon beta-1a Subcutaneously in Multiple Sclerosis) Study Group. Randomized double-blind placebo-controlled study of interferon beta-1a in relapsing/remitting multiple sclerosis. Lancet. 1998;352:1498-504.
PRISMS-4 Study Group. Long-term efficacy of interferonbeta-1a in relapsing MS. Neurology. 2001;56:1628-36.
Soelberg Sorensen PS, Koch-Henriksen N, Ross C, Clemmense KM, Danish Multiple Sclerosis Study Group. Appearance and disappearance of neutralizing antibodies during interferon-beta therapy. Neurology. 2005;65:33-9.
Farrell RA, Giovannoni G. Measuring and management of anti-interferon beta antibodies in subjec 13. Oliver B, Órpez T, Mayorga C. Neutralizing antibodies against IFN beta in patients with multiple sclerosis: A comparative study of two cytopathic effect tests (CPE) for their detection. J Immunol Methods. 2009;351:41-5.
Massart C, Gibassier J, de Seze J, Debouviere M, Moreau T, Pelletier J, et al.. Determination of interferon beta neutralizing antibodies in multiple sclerosis: Improvement of clinical sensitivity of a cytopathic effect assay. Clin Chim Acta. 2008;391:98-101.
Serrano-Fernández P, Möller S, Goertsches R, Fiedler H, Koczan D, Thiesen HJ, et al. Time course transcriptomics of IFNB1b drug therapy in multiple sclerosis. Autoimmunity. 2010;43:172-8.
Platanias, L. Mechanisms of type-I- and type-II-interferonmediated signaling. Nat Rev Immunol. 2005;5:375-86.
Some similar items:
- María Clara Echeverry, Nubia Catalina Tovar, Guillermo Mora, Presence of antibodies to cardiac neuroreceptors in patients with Chagas disease , Biomedica: Vol. 29 No. 3 (2009)
- Nina Paola Lamprea, Lina María Ortega, Gerardo Santamaría, Ladys Sarmiento, Orlando Torres-Fernández, Production and evaluation of an antiserum for immunohistochemical detection of rabies virus in aldehyde fixed brain tissues , Biomedica: Vol. 30 No. 1 (2010)
- Vanihamín Domínguez, Itzen Aguiñiga, Leticia Moreno, Beatriz Torres, Edelmiro Santiago-Osorio, Sodium caseinate increases the number of B lymphocytes in mouse , Biomedica: Vol. 37 No. 4 (2017)
- Yenny M. Montenegro-Medina, Luz Aída Rey-Caro, Jurg Niederbacher, Ruth Aralí Martínez-Vega, Fredi Alexander Díaz-Quijano, Luis Ángel Villar-Centeno, Roll of antibodies antiplatelets in viral infection: a systematic review of literature , Biomedica: Vol. 31 No. 1 (2011)
- Sofía Duque, Rubén Santiago Nicholls, Adriana Arévalo, Rafael Guerrero, Serodiagnosis of giardiasis: identiflcation of immunoglobulin G anti-Giardia duodenalis in sera by ELlSA , Biomedica: Vol. 21 No. 3 (2001)
- Viviana Marcela Rodríguez, Adriana Cuéllar, Lyda Marcela Cuspoca, Carmen Lucía Contreras, Marcela Mercado, Alberto Gómez, Phenotypical determinants of stem cell subpopulations derived from human umbilical cord blood. , Biomedica: Vol. 26 No. 1 (2006)
- Piedad Agudelo Flórez, Marcos Restrepo, María Amparo Lotero, Evaluation of indirect immunofluorescence assay for diagnosis of human leptospirosis. , Biomedica: Vol. 26 No. 2 (2006)
- Beatriz Eugenia Ferro, Ana Lucía Rodríguez, Mauricio Pérez, Bruno Luis Travi, Seroprevalence of Leptospira infection in habitants of peripheral neighborhoods in Cali, Colombia. , Biomedica: Vol. 26 No. 2 (2006)
- Yenny Alviarez, María Lares, Mercedes Viettri, Cruz M. Aguilar, Leidi Herrera, Elizabeth Ferrer, Standardization of a direct agglutination test for the immunodiagnosis of Chagas disease , Biomedica: Vol. 34 No. 2 (2014)
- Luis Alfonso Zarco, Sonia Patricia Millán, Darío Londoño, Ludy Parada, Alejandra Taborda, Miguel Germán Borda, The cost-effectiveness of interferon beta treatment in patients with a clinically isolated syndrome in Colombia , Biomedica: Vol. 34 No. 1 (2014)
| Article metrics | |
|---|---|
| Abstract views | |
| Galley vies | |
| PDF Views | |
| HTML views | |
| Other views | |
