Neutralizing antibodies against IFN beta in patients with multiple sclerosis: A comparative study of two cytopathic effect tests (CPE) for their detection

Oliver, B.; Órpez, Teresa; Mayorga, Cristobalina; Pinto-Medel, María Jesús; Leyva, Laura; Lopez‐Gómez, Carlos; Marín, Clara; Luque, Gloria M.; Ortega-Pinazo, Jesús; Fernández, Óscar

doi:10.1016/j.jim.2009.09.005

Cited by 9 publications

(11 citation statements)

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“…Además, para estudiar el efecto citopático se utilizan diferentes tipos de virus (VSV, MCMV) y diferentes tipos de líneas celulares; la lectura de los resultados se hace observando efectos citopáticos o disminución de la capacidad metabólica celular medida por MTT (13,14).…”

Section: Discussionunclassified

“…Sin embargo, se sabe que el uso de productos recombinantes puede inducir la producción de anticuerpos (6)(7)(8)(9)(10)(11)(12)(13)(14). Se han descrito dos tipos de anticuerpos contra el IFN-β terapéutico, los cuales se han clasificado como anticuerpos de unión (binding antibodies, BAb) y anticuerpos neutralizantes (neutralizing antibodies, NAb).…”

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“…La presencia de NAb está asociada con disminución de la actividad biológica del IFN-β, pérdida de eficacia clínica de éste y progresión de la enfermedad (7,8,11,12). Las técnicas existentes para la detección de NAb presentan inconvenientes al involucrar cultivos virales, depender del observador y no estar estandarizadas en cuanto al tipo de células y el virus usado (13,14).…”

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Modelo de detección de anticuerpos neutralizantes contra IFN-β mediante citometría de flujo

2012

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: ambos autores participaron por igual en el estudio, realizando los experimentos, analizando los datos y participando en la escritura del artículo. John Mario González: diseñó y coordinó el estudio, analizó los datos y escribió el artículo. NOTA TÉCNICA Modelo de detección de anticuerpos neutralizantes contra IFN-β mediante citometría de flujoJuan Carlos Villa-Camacho, Juan Camilo Vargas-Zambrano, John Mario González Introducción. El interferón beta (IFN-β) se usa para tratar la forma recaída-remisión de la esclerosis múltiple. Sin embargo, el uso de proteínas recombinantes como medicamentos puede generar la producción de anticuerpos, disminuyendo así la efectividad del tratamiento. Objetivo. Estandarizar una técnica de detección de anticuerpos neutralizantes contra IFN-β, mediante citometría de flujo. Materiales y métodos. Se cultivaron dos líneas tumorales humanas (U937 y K562) con IFN-β1a humano recombinante y mediante citometría de flujo se determinó la expresión de la proteína ISG15 intracelular. Los sueros se obtuvieron de un conejo Nueva Zelanda antes y después de la inmunización con 100.000 UI de IFN-β1a en adyuvante de Freund. Para la detección de anticuerpos neutralizantes, se estimularon células K562 con IFN-β1a preincubado con sueros de conejos a una dilución 1:20. Conclusions. This rabbit model demonstrates that ISG15 expression evaluated with flow cytometry can be used as a detection assay for NAb.

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Modelo de detección de anticuerpos neutralizantes contra IFN-β mediante citometría de flujo

2012

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“…The presence of neutralizing antibodies (NABs) against IFNß in serum samples of treated patients was tested by the cytopathic effect test following the WHO recommendations, as previously described [22]. The NABs were tested in the serum samples collected at the same time point as the blood used for isolating PBMC to avoid any possible influence on the activation of the Jak-STAT signaling pathway.…”

Section: Methodsmentioning

confidence: 99%

Activation of the JAK-STAT Signaling Pathway after In Vitro Stimulation with IFNß in Multiple Sclerosis Patients According to the Therapeutic Response to IFNß

et al. 2017

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Interferon beta (IFNß) is a common treatment used for multiple sclerosis (MS) which acts through the activation of the JAK-STAT pathway. However, this therapy is not always effective and currently there are no reliable biomarkers to predict therapeutic response. We postulate that the heterogeneity in the response to IFNß therapy could be related to differential activation patterns of the JAK-STAT signaling pathway. Our aim was to evaluate the basal levels and the short term activation of this pathway after IFNß stimulation in untreated and IFNß treated patients, as well as according to therapeutic response. Therefore, cell surface levels of IFNAR subunits (IFNAR1 and IFNAR2) and the activated forms of STAT1 and STAT2 were assessed in peripheral blood mononuclear cells from MS patients by flow cytometry. Basal levels of each of the markers strongly correlated with the expression of the others in untreated patients, but many of these correlations lost significance in treated patients and after short term activation with IFNß. Patients who had undergone IFNß treatment showed higher basal levels of IFNAR1 and pSTAT1, but a reduced response to in vitro exposure to IFNß. Conversely, untreated patients, with lower basal levels, showed a greater ability of short term activation of this pathway. Monocytes from responder patients had lower IFNAR1 levels (p = 0.039) and higher IFNAR2 levels (p = 0.035) than non-responders just after IFNß stimulation. A cluster analysis showed that levels of IFNAR1, IFNAR2 and pSTAT1-2 in monocytes grouped 13 out of 19 responder patients with a similar expression pattern, showing an association of this pattern with the phenotype of good response to IFNß (p = 0.013). Our findings suggest that an activation pattern of the IFNß signaling pathway in monocytes could be associated with a clinical phenotype of good response to IFNß treatment and that a differential modulation of the IFNAR subunits in monocytes could be related with treatment effectiveness.

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“…The presence of NABs in serum samples was determined after 12 and 24 months of treatment by the interferon-induced inhibition of virus cytopathic effect on human cells in culture (murine encephalomyocarditis virus and A-549 cells), following the World Health Organization (WHO) recommendations, as previously described [41], [42]. The neutralization titre of a serum sample was calculated according to Kawade et al [43] and expressed in 10-fold reduction units per millilitre (TRU/mL).…”

Section: Methodsmentioning

confidence: 99%

Candidate Gene Study of TRAIL and TRAIL Receptors: Association with Response to Interferon Beta Therapy in Multiple Sclerosis Patients

et al. 2013

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TRAIL and TRAIL Receptor genes have been implicated in Multiple Sclerosis pathology as well as in the response to IFN beta therapy. The objective of our study was to evaluate the association of these genes in relation to the age at disease onset (AAO) and to the clinical response upon IFN beta treatment in Spanish MS patients. We carried out a candidate gene study of TRAIL, TRAILR-1, TRAILR-2, TRAILR-3 and TRAILR-4 genes. A total of 54 SNPs were analysed in 509 MS patients under IFN beta treatment, and an additional cohort of 226 MS patients was used to validate the results. Associations of rs1047275 in TRAILR-2 and rs7011559 in TRAILR-4 genes with AAO under an additive model did not withstand Bonferroni correction. In contrast, patients with the TRAILR-1 rs20576-CC genotype showed a better clinical response to IFN beta therapy compared with patients carrying the A-allele (recessive model: p = 8.88×10−4, pc = 0.048, OR = 0.30). This SNP resulted in a non synonymous substitution of Glutamic acid to Alanine in position 228 (E228A), a change previously associated with susceptibility to different cancer types and risk of metastases, suggesting a lack of functionality of TRAILR-1. In order to unravel how this amino acid change in TRAILR-1 would affect to death signal, we performed a molecular modelling with both alleles. Neither TRAIL binding sites in the receptor nor the expression levels of TRAILR-1 in peripheral blood mononuclear cell subsets (monocytes, CD4+ and CD8+ T cells) were modified, suggesting that this SNP may be altering the death signal by some other mechanism. These findings show a role for TRAILR-1 gene variations in the clinical outcome of IFN beta therapy that might have relevance as a biomarker to predict the response to IFN beta in MS.

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Neutralizing antibodies against IFN beta in patients with multiple sclerosis: A comparative study of two cytopathic effect tests (CPE) for their detection

Cited by 9 publications

References 20 publications

Modelo de detección de anticuerpos neutralizantes contra IFN-β mediante citometría de flujo

Modelo de detección de anticuerpos neutralizantes contra IFN-β mediante citometría de flujo

Activation of the JAK-STAT Signaling Pathway after In Vitro Stimulation with IFNß in Multiple Sclerosis Patients According to the Therapeutic Response to IFNß

Candidate Gene Study of TRAIL and TRAIL Receptors: Association with Response to Interferon Beta Therapy in Multiple Sclerosis Patients

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