Multiplexed phospho-protein analysis of immune cells in multiple sclerosis
Personalized immunogenicity testing in multiple sclerosis
Many drugs used in therapy will be recognized as foreign by the immune system. In some cases the efficacy of the drug can be completely inhibited by the formation of antibodies that neutralize the drug. It is therefore essential to develop tests to monitor the possible adverse activation of a patient’s immune system to the drug itself.
Many drugs used in medicine are identical or derivatives of substances that a human body can produce itself. However, no matter how sophisticated the technology of production of such drugs is, the immune system can and will recognize the drug as foreign in a considerable number of patients. When such a situation arises during treatment the immune system eventually produces antibodies that impair or neutralize the efficacy of the drug. In such a case the patient no longer benefits from taking the drug. It is therefore essential to develop tests to monitor the possible adverse activation of a patient’s immune system to the drug itself.
In multiple sclerosis, a debilitating disease of the central nervous system, patients are treated with interferon-beta, a compound that in humans is induced by viral infections. The interferon-beta preparations are produced in cell lines and are known to trigger an adverse immune response to the compound in many treated patients. Currently adverse effects are monitored in patients by laborious laboratory methods which are satisfactory in identifying patients with a severe immunogenic response to the drug. However, the question of when the efficacy of the drug is offset by the immunogenic response in a specific patient is still debated. We have developed a new method to address this problem that uses patient blood cells to evaluated adverse immune effects to the drug, This approach potentially allows a personalized test to identify patients that no longer or sub optimally benefit from the drug.
The method is based on analysis of the potential activation of the interferon-beta signaling pathway in primary immune cells from treated patients. Highly specific monoclonal antibodies that only recognize activated forms of molecules in this pathway are combined with single cell analysis by flow cytometry. This multiplexed approach identified immune cell subtypes in patients that are particularly promising as biomarkers for an adverse immune response to the drug in treated patients.
The idea and method and a detailed protocol have been published in previous years (see below) and the recent manuscript has been accepted for publication. Further data that has been generated in 2011 will additionally be submitted for publication in 2012: Individual phenotypic evaluation of the in vivo effect of recombinant interferon-beta in patients with multiple sclerosis.
The thesis will be submitted in March/April 2012.
1: Gavasso S, BT Gjertsen, E Anderssen, KM Myhr, C Vedeler. Immunogenic effects of recombinant interferon-beta therapy disrupt the JAK/STAT pathway in primary immune cells from patients with multiple sclerosis. Multiple Sclerosis Journal, accepted December 2011.
2: Gavasso S. Flow cytometry and cell activation. Methods Mol Biol.
2009;514:35-46. PubMed PMID: 19048212.
3: Gavasso S, Myhr KM, Vedeler C. Multiplexed phosphoprotein analysis in immune
cells. Acta Neurol Scand Suppl. 2006;183:58-60. Review. PubMed PMID: 16637932.
Monitoring drug response in patients with signaling pathway specific flow cytometry
ECTRIMS, Amsterdam, 19-22 October 2011
Phospho-specific flow cytometry and immunogenicity evaluation in primary cells from MS patients treated with interferon-beta
Danish Society for Flow cytometry in collaboration with the Norwegian Society. Halmstad, 4-5 April 2011