Antigen-specific therapy of experimental allergic encephalomyelitis by soluble class II major histocompatibility complex-peptide complexes.

Abstract

Experimental allergic encephalomyelitis is a T-cell-mediated, major histocompatibility complex (MHC) MATERIALS AND METHODS Mice. Female SJL/J mice were purchased from The Jackson Laboratory and were used between 8 and 12 weeks of age.Antigens. Peptides of MBP and PLP corresponding to amino acids 1-14, 91-103, and 139-151 were synthesized by solid-phase 9-fluorenylmethoxycarbonyl (FMOC) procedures. Peptides were purified by reverse-phase HPLC and were characterized by both HPLC and mass spectroscopy.Purification of dass H MHC. I-Al protein was purified from a Nonidet P-40 extract of spleen cell membranes from SJL/J mice by affinity chromatography using the monoclonal antibody 10-2.16 (specific for I-Ak and I-Al), coupled to Sepharose 4B beads. Extracted lysate from the high-speed (100,000x g) membrane fraction in a buffer of 10 mM Tris HCI, pH 8.3/0.5% Nonidet P-40/0.1 M sodium chloride/5 mM EDTA/0.02% sodium azide/1 mM phenylmethylsulfonyl fluoride was recycled over the preequilibrated column at 40C for 16 hr. The column was washed first with 10 bed volumes of deoxycholate buffer/10 mM Tris-HCl, pH 8.3/0.5% deoxycholate/0.1 M sodium chloride/5 mM EDTA/0.02% sodium azide/1 mM phenylmethylsulfonyl fluoride and then by 5 bed volumes of phosphate-buffered saline (PBS)/1% 1-octyl P-Dglycopyranoside buffer. Finally, I-AS was eluted with 20 mM phosphate buffer, pH 11/0.1 M sodium chloride/1% octyl glucoside/0.02% sodium azide/1 mM phenylmethylsulfonyl fluoride. Fractions were neutralized with acetic acid to a final Abbreviations: EAE, experimental allergic encephalomyelitis; MBP, myelin basic protein; PLP, proteolipoprotein; MHC, major histocompatibility complex. tTo whom reprint requests should be addressed. 11465The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

Interaction of papain-digested HLA class I molecules with human alloreactive cytotoxic T lymphocytes (CTL)

“…In a recent paper, experitnental allergic encephaiomyelitis was prevented by soluble complexes of MHC class II molecules and antigetiic peptides. Additionally, specific T cell suppression by these cotnplexes //( riiro was also reported, underlining the suggestion of functional T cell inhibition by binding of soluble MHC molecules to the T cell receptor [25]. In previous reports we and others described specific inhibition of alloreactive T cells by purified intact M HC class I molecules [9,1 ()].…”

Expression of Recombinant HLA-DR2 Molecules

“…Soluble MHC⅐peptide complexes may also be useful for the treatment of autoimmune diseases. Studies in the experimental autoimmune encephalomyelitis model have demonstrated that an autoimmune disease can be treated by the administration of soluble MHC⅐peptide complexes (Sharma et al, 1991).…”

A Single-Chain Class II MHC-IgG3 Fusion Protein Inhibits Autoimmune Arthritis by Induction of Antigen-Specific Hyporesponsiveness

“…Soluble class I (12)(13)(14)(15)(16) and class II (17-21) MHC molecules have been produced that can be loaded with specific peptides and are able to signal T cells in vitro. Soluble class II have been reported to inhibit experimental allergic encephalomyelitis (22,23). However, in general, soluble MHC molecules have not been effective at delivering inhibitory T cell signals in vivo.…”

“…A dimeric class I MHC-IgG fusion protein was recently shown to induce Ag-specific blockade in vivo (26). A few studies demonstrating signaling by soluble class II may have used aggregated MHC molecules (20,22,23).…”