Novel monoclonal antibodies against proteolipid protein peptide 139–151 demonstrate demyelination and myelin uptake by macrophages in MS and marmoset EAE lesions
Copyright policy )Novel monoclonal antibodies against proteolipid protein peptide 139–151 demonstrate demyelination and myelin uptake by macrophages in MS and marmoset EAE lesions
Experimental autoimmune encephalomyelitis (EAE) induced by immunization of mice with epitopes of the proteolipid protein (PLP), a major myelin constituent, forms a useful model for the study of multiple sclerosis (MS). In addition, MS patients display PLP-specific T- and B-cell responses, suggesting that PLP reactivity is relevant to pathogenesis.Here, the generation and characterization of a panel of mouse monoclonal antibodies (Mab) against PLP139-151, the prominent encephalitogenic sequence in SJL/J mice is described. Five Mab were generated by conventional immunization of an SJL/J mouse and hybridoma generation. These Mab reacted well with the PLP139-151 peptide in ELISA and belonged to the IgG2a and IgG2b subclasses, consistent with CD4+ T helper 1-cell-supported antibody formation. The Mab also efficiently detected PLP peptide-BSA conjugates in Western blot, confirming their multi-assay applicability. The Mab were subsequently used to determine the occurrence of demyelination in brains of MS patients and marmoset monkeys with EAE. Immunohistochemistry on both paraffin and frozen sections demonstrated a homogeneous expression of PLP139-151 in normal myelin, and a complete absence in lesions containing demyelinated areas, confirming that the Mab can be used as a general myelin marker. In active demyelinating MS lesions, the Mab visualized the peptide in the cytoplasm of macrophages containing phagocytosed myelin. In conclusion, this panel of Mab against the encephalitogenic PLP139-151 epitope forms a useful tool for further study of autoantigen expression, demyelination/remyelination and the staging of lesional activity in MS patients, as well as in EAE models in distinct animal species.
- Vrije Universiteit Amsterdam Netherlands
- Erasmus University Rotterdam Netherlands
- Amsterdam UMC, location VUmc Netherlands
- Delft University of Technology Netherlands
- Biomedical Primate Research Centre Netherlands
EMC MM-02-72-02, Lesional staging, Axonal loss, multiple sclerosis, paraffin, Mice, allergic encephalomyelitis, Reference Values, Myelin Sheath, antibody production, epitope, Th1 cell, phagocytosis, Antibodies, Monoclonal, Callithrix, Immunohistochemistry, frozen section, unclassified drug, myelin, priority journal, immunohistochemistry, cytoplasm, Female, demyelination, Demyelination, monkey, CD4 antigen, immunoblotting, immunoreactivity, Encephalomyelitis, Autoimmune, Experimental, Multiple Sclerosis, proteolipid protein[139-151], animal experiment, brain region, Mice, Inbred Strains, macrophage, immunization, hybridoma, animal tissue, proteolipid protein, immunoglobulin G2a, Animals, Humans, Animal model, immunoglobulin G2b, controlled study, protein interaction, Myelin Proteolipid Protein, protein expression, mouse, nonhuman, animal model, Macrophages, Multiple sclerosis (MS), Peptide Fragments, enzyme linked immunosorbent assay, monoclonal antibody, antigen expression, Demyelinating Diseases
EMC MM-02-72-02, Lesional staging, Axonal loss, multiple sclerosis, paraffin, Mice, allergic encephalomyelitis, Reference Values, Myelin Sheath, antibody production, epitope, Th1 cell, phagocytosis, Antibodies, Monoclonal, Callithrix, Immunohistochemistry, frozen section, unclassified drug, myelin, priority journal, immunohistochemistry, cytoplasm, Female, demyelination, Demyelination, monkey, CD4 antigen, immunoblotting, immunoreactivity, Encephalomyelitis, Autoimmune, Experimental, Multiple Sclerosis, proteolipid protein[139-151], animal experiment, brain region, Mice, Inbred Strains, macrophage, immunization, hybridoma, animal tissue, proteolipid protein, immunoglobulin G2a, Animals, Humans, Animal model, immunoglobulin G2b, controlled study, protein interaction, Myelin Proteolipid Protein, protein expression, mouse, nonhuman, animal model, Macrophages, Multiple sclerosis (MS), Peptide Fragments, enzyme linked immunosorbent assay, monoclonal antibody, antigen expression, Demyelinating Diseases
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