Whole-Proteome Peptide Microarrays for Profiling Autoantibody Repertoires within Multiple Sclerosis and Narcolepsy

The underlying molecular mechanisms of autoimmune diseases are poorly understood. To unravel the autoimmune processes across diseases, comprehensive and unbiased analyses of proteins targets recognized by the adaptive immune system are needed. Here we present an approach starting from high-density p...

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Vydáno v:Journal of proteome research Ročník 16; číslo 3; s. 1300 - 1314
Hlavní autoři: Zandian, Arash, Forsström, Björn, Häggmark-Månberg, Anna, Schwenk, Jochen M, Uhlén, Mathias, Nilsson, Peter, Ayoglu, Burcu
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 03.03.2017
Témata:
Adolescent
Adult
Aged
Autoantibodies - blood
Autoantigens - blood
Cell Adhesion Molecules, Neuronal - blood
Child
Female
Humans
Male
MAP Kinase Kinase Kinases - blood
Middle Aged
Multiple Sclerosis - blood
Narcolepsy - blood
Nerve Tissue Proteins - blood
Protein Array Analysis - methods
Proteome - analysis
Young Adult
autoantibody profiling
peptide microarrays
narcolepsy
multiple sclerosis
epitope mapping
ISSN:1535-3907
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Abstract The underlying molecular mechanisms of autoimmune diseases are poorly understood. To unravel the autoimmune processes across diseases, comprehensive and unbiased analyses of proteins targets recognized by the adaptive immune system are needed. Here we present an approach starting from high-density peptide arrays to characterize autoantibody repertoires and to identify new autoantigens. A set of ten plasma and serum samples from subjects with multiple sclerosis, narcolepsy, and without any disease diagnosis were profiled on a peptide array representing the whole proteome, hosting 2.2 million 12-mer peptides with a six amino acid lateral shift. On the basis of the IgG reactivities found on these whole-proteome peptide microarrays, a set of 23 samples was then studied on a targeted array with 174 000 12-mer peptides of single amino acid lateral shift. Finally, verification of IgG reactivities was conducted with a larger sample set (n = 448) using the bead-based peptide microarrays. The presented workflow employed three different peptide microarray formats to discover and resolve the epitopes of human autoantibodies and revealed two potentially new autoantigens: MAP3K7 in multiple sclerosis and NRXN1 in narcolepsy. The presented strategy provides insights into antibody repertoire reactivity at a peptide level and may accelerate the discovery and validation of autoantigens in human diseases.
AbstractList The underlying molecular mechanisms of autoimmune diseases are poorly understood. To unravel the autoimmune processes across diseases, comprehensive and unbiased analyses of proteins targets recognized by the adaptive immune system are needed. Here we present an approach starting from high-density peptide arrays to characterize autoantibody repertoires and to identify new autoantigens. A set of ten plasma and serum samples from subjects with multiple sclerosis, narcolepsy, and without any disease diagnosis were profiled on a peptide array representing the whole proteome, hosting 2.2 million 12-mer peptides with a six amino acid lateral shift. On the basis of the IgG reactivities found on these whole-proteome peptide microarrays, a set of 23 samples was then studied on a targeted array with 174 000 12-mer peptides of single amino acid lateral shift. Finally, verification of IgG reactivities was conducted with a larger sample set (n = 448) using the bead-based peptide microarrays. The presented workflow employed three different peptide microarray formats to discover and resolve the epitopes of human autoantibodies and revealed two potentially new autoantigens: MAP3K7 in multiple sclerosis and NRXN1 in narcolepsy. The presented strategy provides insights into antibody repertoire reactivity at a peptide level and may accelerate the discovery and validation of autoantigens in human diseases.
Author Schwenk, Jochen M
Ayoglu, Burcu
Zandian, Arash
Forsström, Björn
Uhlén, Mathias
Nilsson, Peter
Häggmark-Månberg, Anna
Author_xml – sequence: 1
  givenname: Arash
  orcidid: 0000-0003-1242-0873
  surname: Zandian
  fullname: Zandian, Arash
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  givenname: Björn
  surname: Forsström
  fullname: Forsström, Björn
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  givenname: Anna
  surname: Häggmark-Månberg
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  givenname: Jochen M
  surname: Schwenk
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  surname: Nilsson
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  organization: Affinity Proteomics, SciLifeLab, School of Biotechnology, KTH - Royal Institute of Technology , SE-171 21 Solna, Sweden
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  givenname: Burcu
  orcidid: 0000-0001-7843-2960
  surname: Ayoglu
  fullname: Ayoglu, Burcu
  organization: Affinity Proteomics, SciLifeLab, School of Biotechnology, KTH - Royal Institute of Technology , SE-171 21 Solna, Sweden
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Keywords autoantibody profiling
peptide microarrays
narcolepsy
multiple sclerosis
epitope mapping
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Snippet The underlying molecular mechanisms of autoimmune diseases are poorly understood. To unravel the autoimmune processes across diseases, comprehensive and...
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SubjectTerms Adolescent
Adult
Aged
Autoantibodies - blood
Autoantigens - blood
Cell Adhesion Molecules, Neuronal - blood
Child
Female
Humans
Male
MAP Kinase Kinase Kinases - blood
Middle Aged
Multiple Sclerosis - blood
Narcolepsy - blood
Nerve Tissue Proteins - blood
Protein Array Analysis - methods
Proteome - analysis
Young Adult
Title Whole-Proteome Peptide Microarrays for Profiling Autoantibody Repertoires within Multiple Sclerosis and Narcolepsy
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