PMID- 24416182
OWN - NLM
STAT- MEDLINE
DCOM- 20140908
LR  - 20240313
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 9
IP  - 1
DP  - 2014
TI  - Stable expression of human muscle-specific kinase in HEp-2 M4 cells for automatic 
      immunofluorescence diagnostics of myasthenia gravis.
PG  - e83924
LID - 10.1371/journal.pone.0083924 [doi]
LID - e83924
AB  - Muscle-specific kinase (MuSK) belongs to the nicotinic acetylcholine receptor 
      complex which is targeted by pathogenic autoantibodies causing Myasthenia gravis. 
      While up to 95% of patients with generalized Myasthenia gravis were shown to be 
      positive for acetylcholine receptor-specific autoantibodies, up to 70% of the 
      remaining patients develop autoantibodies against MuSK. Discrimination of the 
      autoantibody specificity is important for therapy of Myasthenia gravis. Recently, 
      the new automatic fluorescence assessment platform AKLIDES has been developed for 
      immunofluorescence-based diagnostics of autoimmune diseases. In order to 
      establish an AKLIDES procedure for the detection of MuSK-specific autoantibodies 
      (anti-MuSK), we developed a recombinant HEp-2 cell clone expressing the human 
      MuSK cDNA. Here we show at the mRNA and protein level that the cell clone HEp-2 
      M4 stably expresses human MuSK. We provide evidence for a localization of MuSK at 
      the cell membrane. Using cell clone HEp-2 M4 on the AKLIDES system, we 
      investigated 34 patient sera that were previously tested anti-MuSK positive by 
      radioimmunoassay as positive controls. As negative controls, we tested 29 
      acetylcholine receptor-positive but MuSK-negative patient sera, 30 amytrophic 
      lateral sclerosis (ALS) patient sera and 45 blood donors. HEp-2 M4 cells revealed 
      a high specificity for the detection of MuSK autoantibodies from 25 patient sera 
      assessed by a specific pattern on HEp-2 M4 cells. By using appropriate cell 
      culture additives, the fraction of cells stained positive with anti-MuSK 
      containing sera can be increased from 2-16% to 10-48%, depending on the serum. In 
      conclusion, we provide data showing that the novel recombinant cell line HEp-2 M4 
      can be used to screen for anti-MuSK with the automatic AKLIDES system.
FAU - George, Sandra
AU  - George S
AD  - Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, 
      Senftenberg, Germany ; Institute of Immunology, Technical University Dresden, 
      Dresden, Germany.
FAU - Paulick, Silvia
AU  - Paulick S
AD  - Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, 
      Senftenberg, Germany.
FAU - Knutter, Ilka
AU  - Knutter I
AD  - GA Generic Assays GmbH, Dahlewitz/Berlin, Germany.
FAU - Rober, Nadja
AU  - Rober N
AD  - Institute of Immunology, Technical University Dresden, Dresden, Germany.
FAU - Hiemann, Rico
AU  - Hiemann R
AD  - GA Generic Assays GmbH, Dahlewitz/Berlin, Germany.
FAU - Roggenbuck, Dirk
AU  - Roggenbuck D
AD  - Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, 
      Senftenberg, Germany ; GA Generic Assays GmbH, Dahlewitz/Berlin, Germany.
FAU - Conrad, Karsten
AU  - Conrad K
AD  - Institute of Immunology, Technical University Dresden, Dresden, Germany.
FAU - Kupper, Jan-Heiner
AU  - Kupper JH
AD  - Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, 
      Senftenberg, Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140109
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Autoantibodies)
RN  - 0 (Receptors, Cholinergic)
RN  - EC 2.7.10.1 (MUSK protein, human)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
SB  - IM
MH  - Autoantibodies/blood/immunology
MH  - Automation
MH  - Cell Line
MH  - Cell Proliferation
MH  - Cell Shape
MH  - Fluorescent Antibody Technique
MH  - Humans
MH  - Myasthenia Gravis/blood/*diagnosis/*enzymology/immunology
MH  - Organ Specificity
MH  - Receptor Protein-Tyrosine Kinases/*metabolism
MH  - Receptors, Cholinergic/*metabolism
PMC - PMC3886972
COIS- Competing Interests: Dirk Roggenbuck is a shareholder of GA Generic Assays GmbH 
      and Medipan GmbH. Ilka Knutter and Rico Hiemann are employees of GA Generic 
      Assays GmbH. This does not alter the authors' adherence to all the PLOS ONE 
      policies on sharing data and material. The remaining authors declare that they 
      have no competing financial or other interests.
EDAT- 2014/01/15 06:00
MHDA- 2014/09/10 06:00
PMCR- 2014/01/09
CRDT- 2014/01/14 06:00
PHST- 2013/08/09 00:00 [received]
PHST- 2013/11/11 00:00 [accepted]
PHST- 2014/01/14 06:00 [entrez]
PHST- 2014/01/15 06:00 [pubmed]
PHST- 2014/09/10 06:00 [medline]
PHST- 2014/01/09 00:00 [pmc-release]
AID - PONE-D-13-32944 [pii]
AID - 10.1371/journal.pone.0083924 [doi]
PST - epublish
SO  - PLoS One. 2014 Jan 9;9(1):e83924. doi: 10.1371/journal.pone.0083924. eCollection 
      2014.