Multiple sclerosis is an autoimmune disease of the central nervous system believed to be mediated by pathogenic T lymphocytes. We have developed a next-generation therapy in which cells secrete specific therapeutic molecules to silence these aberrant T cells. We have shown that fibroblasts, transduced to secrete a myelin basic protein-derived peptide, abrogate disease in the murine experimental autoimmune encephalomyelitis model of multiple sclerosis, which we hypothesized using a low-zone tolerance mechanism. To determine the efficacy (or not) of this therapy in humans, we must ensure that patients receive comparable doses of therapeutic peptide. To this end, we have used liquid chromatography coupled to tandem mass spectrometry to detect a tryptic peptide, derived from the secreted therapeutic product, at nanomolar concentrations. Success depended on growing the transduced fibroblasts in defined PC-1 medium in the presence of a cocktail of protease inhibitors.
This is a preview of subscription content, access via your institution
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
Hemmer B, Archelos JJ, Hartung HP . New concepts in the immunopathogenesis of multiple sclerosis. Nat Rev Neurosci 2002; 3: 291–301.
Goverman J . Autoimmune T cell responses in the central nervous system. Nat Rev Immunol 2009; 9: 393–407.
Hemmer B, Hartung HP . Toward the development of rational therapies in multiple sclerosis: what is on the horizon? Ann Neurol 2007; 62: 314–326.
Freedman MS, Hughes B, Mikol DD, Bennett R, Cuffel B, Divan V et al. Efficacy of disease-modifying therapies in relapsing remitting multiple sclerosis: a systematic comparison. Eur Neurol 2008; 60: 1–11.
Weiner LP, Louie KA, Atalla LR, Kochounian HH, Du J, Wei W et al. Gene therapy in a murine model for clinical application to multiple sclerosis. Ann Neurol 2004; 55: 390–399.
Hochweller K, Sweenie CH, Anderton SM . Immunological tolerance using synthetic peptides--basic mechanisms and clinical application. Curr Mol Med 2006; 6: 631–643.
Louie KA, Weiner LP, Du J, Kochounian HH, Fling SP, Wei W et al. Cell-based gene therapy experiments in murine experimental autoimmune encephalomyelitis. Gene Ther 2005; 12: 1145–1153.
Tamvakopoulos C . Mass spectrometry for the quantification of bioactive peptides in biological fluids. Mass Spectrom Rev 2007; 26: 389–402.
Larche M, Wraith DC . Peptide-based therapeutic vaccines for allergic and autoimmune diseases. Nat Med 2005; 11: S69–S76.
This study was supported by a grant from the National Multiple Sclerosis Society (PP1387, MM). We thank Dr Leslie P Weiner for constant encouragement and helpful discussions and Sarah Olivo for preparing the paper and for editorial support.
KAL and MM declare potential, but no actual, conflict of interest.
About this article
Cite this article
Dadgari, J., Moore, R., Louie, K. et al. Mass spectrometry measurement of a therapeutic peptide for use in multiple sclerosis. Gene Ther 17, 709–712 (2010). https://doi.org/10.1038/gt.2010.19