Method of detecting the presence or absence of autoantibodies
The invention relates to methods of detecting the presence or absence of autoantibodies in an individual and to related methods and kits. In particular, the disclosure concerns measuring from one or more samples from an individual suspected of having a disease with an autoimmune component for binding of one or more autoantibodies to membrane vesicles comprising corresponding one or more antigens.
1. A method of measuring from a sample from an individual the presence or absence of one or more autoantibodies each directed against an antigen, the method comprising:
transforming or transfecting a cell with polynucleotide sequences that encode the one or more antigens,
expressing said one or more antigens in the cell;
collecting membrane vesicles from said cell to obtain a preparation of membrane vesicles comprising said one or more antigens;
drying said membrane vesicles;
contacting the sample from the individual with said preparation of membrane vesicles comprising the one or more antigens; and
measuring for the presence or absence of the one or more autoantibodies by detecting in the sample from the individual whether or not one or more autoantibodies in the sample binds to said one or more antigens,
wherein said one or more antigens are selected from the group consisting of aquaporin-4 (AQP4), myelin-oligodendrocyte glycoprotein (MOG), acetylcholine receptor, muscle specific tyrosine kinase (MuSK), N-methyl-D-aspartate (NMDA) receptor, leucine-rich glioma inactivated protein 1 (LGI1), contactin associated protein-like 2 (CASPR2), glycine receptor alpha 1, gamma-aminobutyric acid type B (GABA-B) receptor and a combination thereof.
2. A method according to claim 1 , wherein the membrane vesicles are exosomes.
3. A method according to claim 1 , wherein the membrane vesicles are:
(a) substantially isolated from cells;
(b) isolated from cells; or
(c) present in a ratio of more than 5,000 membrane vesicles per cell.
4. A method according to claim 1 , wherein the membrane vesicles are:
(a) dried;
(b) attached to a solid support; or
(c) derived from a cell transfected or transformed with the antigen.
5. A method according to claim 1 , wherein the presence or absence of the one or more autoantibodies are detected by an enzyme- linked immunosorbent assay (ELISA), radioimmunoassay, competition assay, inhibition assay, sandwich assay, fluorescent microscopy, microarray or fluorescence-activated cell sorting (FACS) analysis.
6. A method according to claim 1 , wherein when the presence of the one or more autoantibodies in the individual is identified, the individual is administered a therapeutically effective amount of an immunotherapy.
7. The method of claim 1 , wherein the individual is a mother having an unborn baby and when the presence of the one or more autoantibodies in the mother is identified, the mother is administered a therapeutically effective amount of an immunotherapy.
8. The method of claim 1 , wherein said antigen is acetylcholine receptor and said membrane vesicles comprising the acetylcholine receptor co-express rapsyn.
9. The method of claim 1 , wherein said antigen is NMDA receptor and said membrane vesicles comprising NMDA receptor co-express postsynaptic density protein (PSD).