Expression, Purification, and Encephalitogenicity of Recombinant Human Myelin Oligodendrocyte Glycoprotein

Abstract

Myelin oligodendrocyte glycoprotein (MaG), a putative autoantigen in multiple sclerosis (MS), is a quantitatively minor component of the CNS. In view of the difficulties associated with the purification of MOG from brain tissues, the extracellular domain of human MOG corresponding to the N-terminal 121 amino acids was expressed in Escherichia co/i as a glutathione sulfotransferase fusion protein. The expressed protein was localized to inclusion bodies, and varying the growth parameters resulted in the solubilization of small amounts of GST-MOG that could be affinity purified on glutathione agarose columns. The fusion protein found in the inclusion bodies could be solubilized with urea. The solubilized fusion protein was cleaved with thrombin, and the extracellular domain was purified by CM Sephadex 50 chromatography to homogeneity. Injection of recombinant human MOG into different strains of mice resulted in the induction of an MS-like disease, characterized by severe neurological impairment and extensive CNS demyelinated lesions. Recombinant MOG produced in E. co/i should prove to be useful as a highly purified biological reagent for immunological, pathological, functional, and structural studies.

Rat and Human Myelin Oligodendrocyte Glycoproteins Induce Experimental Autoimmune Encephalomyelitis by Different Mechanisms in C57BL/6 Mice

“…Previous analyses suggested that the CNS infiltrate in mice immunized with human MOG protein was predominantly mononuclear (3,21), but it is difficult to determine from the published data whether PMN were present in these lesions. In the experiments presented in this study with mice immunized with human MOG protein, it is likely that PMN were attracted to the CNS by as yet unidentified chemotactic factors.…”

T-cell reactivity to multiple myelin antigens in multiple sclerosis patients and healthy controls

“…Human MBP was purified from white matter of human brain, as described (Deibler et al, 1972). Endotoxin-free extracellular domain (rMOG) was prepared as described previously (Bettadapura et al, 1998) (184)(185)(186)(187)(188)(189)(190)(191)(192)(193)(194)(195)(196)(197)(198)(199) were synthesized and HPLC purified (Ͼ 95% purity) by Severn Biotech Ltd (Worcester, UK). The amino acid sequences of the peptides are shown in Table II.…”

B cell recognition of myelin oligodendrocyte glycoprotein autoantigen depends on immunization with protein rather than short peptide, while B cell invasion of the CNS in autoimmunity does not

“…It is also important to note that all expression systems that we have identified produce fusion proteins consisting of the MOG extracellular domain, various purification tags, and often additional sequences that we cannot identify. Most incorporate either human or rat MOG 1-125 (Bettadapura et al, 1998), and therefore when used in mice represent a cross-reactive response not ideal for some investigations of autoimmune-specific cell interactions.…”