Immunolabelling and flow cytometry as new tools to explore dysferlinopathies.
Dysferlinopathies are autosomal recessive muscular dystrophies caused by DYSF mutations, which lead to a reduced amount or a complete lack of dysferlin. One step in dysferlinopathies diagnosis consists in Western blot analysis of proteins extracted from muscle biopsy, or blood monocytes. We have taken advantage of dysferlin expression in monocytes to develop a whole blood flow cytometry (WBFC), using antibodies directed against dysferlin. Six patients were submitted to WBFC analysis and immunofluorescence analysis on monocytes. Results obtained are correlated to Western blot from monocytes and muscle biopsies. The possible usefulness of this flow cytometry analysis in routine diagnosis is presented.
Wein
N
N
From Inserm UMR_S 910 Génétique Médicale et Génomique Fonctionnelle, Université de la Méditerranée, Faculté de Médecine de Marseille, France.
Krahn
M
M
Courrier
S
S
Bartoli
M
M
Salort-Campana
E
E
Nguyen
K
K
Fernandez
C
C
Pouget
J
J
Fossat
C
C
Depetris
D
D
Leturcq
F
F
Cau
P
P
Levy
N
N
D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D000073939:Dysferlin; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism; D009124:Muscle Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism; D009136:Muscular Dystrophies / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation
D000906:Antibodies; C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins