Visualization of myelin basic protein (MBP) T cell epitopes in multiple sclerosis lesions using a monoclonal antibody specific for the human histocompatibility leukocyte antigen (HLA)-DR2-MBP 85-99 complex.

Journal article

Susceptibility to multiple sclerosis (MS) is associated with the human histocompatibility leukocyte antigen (HLA)-DR2 haplotype, suggesting that major histocompatibility complex class II-restricted presentation of central nervous system-derived antigens is important in the disease process. Antibodies specific for defined HLA-DR2-peptide complexes may therefore be valuable tools for studying antigen presentation in MS. We have used phage display technology to select HLA-DR2-peptide-specific antibodies from HLA-DR2-transgenic mice immunized with HLA-DR2 molecules complexed with an immunodominant myelin basic protein (MBP) peptide (residues 85-99). Detailed characterization of one clone (MK16) demonstrated that both DR2 and the MBP peptide were required for recognition. Furthermore, MK16 labeled intra- and extracellular HLA-DR2-MBP peptide complexes when antigen-presenting cells (APCs) were pulsed with recombinant MBP. In addition, MK16 inhibited interleukin 2 secretion by two transfectants that expressed human MBP-specific T cell receptors. Analysis of the structural requirement for MK16 binding demonstrated that the two major HLA-DR2 anchor residues of MBP 85-99 and the COOH-terminal part of the peptide, in particular residues Val-96, Pro-98, and Arg-99, were important for binding. Based on these results, the antibody was used to determine if the HLA-DR2-MBP peptide complex is presented in MS lesions. The antibody stained APCs in MS lesions, in particular microglia/macrophages but also in some cases hypertrophic astrocytes. Staining of APCs was only observed in MS cases with the HLA-DR2 haplotype but not in cases that carried other haplotypes. These results demonstrate that HLA-DR2 molecules in MS lesions present a myelin-derived self-peptide and suggest that microglia/macrophages rather than astrocytes are the predominant APCs in these lesions.

Authors: Krogsgaard, M; Wucherpfennig, K; Cannella, B; Hansen, B; Svejgaard, A

• Journal: Journal of experimental medicine
• Volume: 191
• Issue: 8
• Pages: 1395-1412
• Publication date: 2000-04-01
• DOI: 10.1084/jem.191.8.1395
• EISSN: 1540-9538
• ISSN: 0022-1007
• Language: English
• Keywords: Humans; Mice, Inbred DBA; Animals; Recombinant Proteins; HLA-DR2 Antigen; Mice; Drosophila melanogaster; Multiple Sclerosis; Molecular Sequence Data; Myelin Basic Protein; Peptide Fragments; Immunohistochemistry; Immunodominant Epitopes; Mice, Transgenic; Cell Line; Antibody Specificity; Amino Acid Sequence; Binding Sites; Antibodies, Monoclonal; Mice, Inbred BALB C