KR960705038A - USES OF MYELIN OLIGODENDROCYTE GLYCOPROTEIN AND PEPTIDE PORTIONS THEREOF IN PROTOCOLS RELATED TO AUTOIMMUNE DESEASE - Google Patents

Abstract

The present invention provides nucleic acid molecules having a nucleotide sequence encoding human MOG which is a self-representative involved in demyelinized autoimmune disease. The invention also provides a recombinantly produced human MOG or antigen thereof. Antigen fragments of MOG that can be prepared synthetically represent T cell epitopes of human MOG. MOG and fragments thereof are useful for diagnosing and treating autoimmune diseases. In addition, methods of screening and developing therapeutic agents useful for the treatment of autoimmune diseases are also disclosed.

Description

USES OF MYELIN OLGODENDROCYTE GLYCOPROTEIN AND PEPTIDE PORTIONS THEREOF IN PROTOCOLS RELATED TO AUTOIMMUNE DESEASE

Since this is an open matter, no full text was included.

Figure 1 shows the complete nucleotide sequence (SEQ ID NO: 1) and the deduced amino acid sequence of DNA encoding the human MOG protein (SEQ ID NO: 2).

3 is a table showing the effect of each of the natural amino acids at a specific position on a peptide that binds to the MHC class II product of DR3.

5a corresponds to SEQ ID NOS: 73-93 and is designated by the amino acid sequence useful in the present invention (20mer),

5B is a peptide of human MOG1-12 designated by an amino acid sequence containing at least one T cell epitope.

FIG. 6 shows that the X-axis represents four different cell lines, the seven tested peptides are represented by different legends, and the 3H-thymidine inserted by each vertical line in which the Y-acting data responds to each individual peptide. Bar graph of data from Example 3 showing the coefficients per minute expressed in CPM).

Claims (27)

An isolated nucleic acid molecule comprising a nucleotide sequence encoding human myelin oligodendrosheet glycoprotein (MOG) or at least one antigen fragment thereof. The nucleic acid molecule according to claim 1, wherein the nucleotide is composed of nucleotides encoding amino acids 1-218 (SEQ ID NO: 2) or allelic variants thereof. An oligomer comprising the nucleotide sequence represented by the nucleotide of FIG. An expression system, wherein the expression system is a DNA molecule comprising a nucleotide sequence encoding a human MOG or antigenic fragment thereof operably linked to modulate a sequence for expression in a host. The expression system of claim 4, wherein the nucleotide sequence encodes amino acids 1-218 (SEQ ID NO: 2) or antigen fragment portion thereof of FIG. 1. Recombinant host cell modified to contain the expression system of claim 4. Isolated and purified human MOG or antigen fragments thereof. a) culturing the host cell modified to contain an expression system directing the expression of the nucleotide sequence encoding the human MOG or fragment thereof in an appropriate medium to produce a culture containing the human MOG or fragment thereof. b) recovering said human MOG or fragment thereof from culture. 9, wherein the peptide is any of the amino acid sequences shown in FIG. 2 (SEQ ID NOs: 4-9, 11, 15, 16 and 42-72), and the amino acid sequences shown in FIGS. 5A and 5B. Peptides, including those. A peptide comprising at least two T cell epitopes of human MOG. Modified form of a human MOG or antigenic fragment thereof, wherein said modification comprises substituting one or more amino acids of said peptide or protein, a different amino acid that does not interfere with the biological serum of said peptide or protein. A nucleic acid molecule comprising a nucleotide sequence encoding the peptide of claim 9. An expression system that is a DNA molecule comprising a nucleotide sequence encoding a peptide of any one of claims 9 to 12 operably linked to modulate a sequence for expression in a persistent host. A recombinant host cell modified to contain the expression system of claim 14. A method for producing a peptide of any one of claims 9 to 12, wherein the host cell is modified to contain an expression system that directs the expression of a nucleotide sequence encoding the peptide. Culturing in a suitable medium, and recovering the peptide from the culture. A modified form of a human MOG or an antigen fragment thereof or the peptide of any one of claims 9 to 12, wherein the modification purifies residues, proteins, fragments or peptides that enhance the solubility of polyethylene glycol, proteins, fragments or peptides. A modified form comprising linking said protein, fragment or peptide to an additional residue selected from the group consisting of residues comprising residues and proteolytic cleavage sites that facilitate said residues. A therapeutic composition comprising isolated human MOG and / or at least one antigen fragment thereof and / or at least one peptide of any of claims 9-12 or 16 and a pharmaceutically acceptable carrier or diluent. The therapeutic composition of claim 11, wherein the human MOG comprises an allelic variant of the amino acid sequence of positions 1-128 shown in FIG. 1 (SEQ ID NO: 2) or the nucleotide sequence of FIG. 1 or an antigen fragment thereof. 20. A method for treating a demyelinized autoimmune disease comprising administering to a patient a therapeutically effective amount of claim 19. Human MOG or antigen fragments thereof or peptides of claims 9 to 12 and 16 or isolated from a blood sample obtained from the patient to be tested are bound under conditions suitable for binding the blood component and MOG or fragment thereof and the extent to which such binding occurs. A method of diagnosing demyelinized autoimmune disease comprising determining a. The method of claim 21, wherein the extent to which the binding takes place comprises the cellular function of the protein or fragment thereof, T cell proliferation, B cell function; The method is determined by evaluating binding to or combination of antibodies present in the blood. Monoclonal antibodies or immunoreactive fragments thereof that are specifically reactive with MOG or antigen fragments thereof or the peptides of claims 9-12 and 16. Identifying hydrophobic amino acids contained in human MOG and determining the properties of amino acids in N-C sequenced from human hydrophobic amino acids by said four amino acids, wherein said amino acids are glycine, alanine, serine, framework; A small amino acid selected from the group consisting of onin and cysteine or a different amino acid is determined, and the peptide containing the hydrophobic small amino acid and the small amino acid separated by 4 NCs from the hydrophobic amino acid is identified as a T cell epitope. A method for identifying T cell epitopes in human MOG, including. An isolated peptide of human MOG comprising at least one T-cell epitope, wherein the peptide is MOG 1-13 (SEQ ID NO. 42), MOG 103-115 (SEQ ID NO. 75), MOG 1-121 (SED DI First 121 amino acids of NO2), MOG 1-20 (SEQ ID NO. 73), MOG 11-30 (SEQ ID NO. 74), MOG 21-40 (SEQ ID NO. 75), MOG 31-50 (SEQ ID NO. 76), MOG 41-60 (SEQ ID NO. 77), MOG 51-70 (SEQ ID NO. 78), MOG 61-80 (SEQ ID NO. 79), MOG 71-90 (SEQ ID NO 80), an isolated peptide of human MOG comprising a sequence of amino acid residues selected from the group consisting of MOG 91-110 (SEQ ID NO. 82), and MOG 101-120 (SEQ ID NO. 83). A mixture of peptide human MOGs comprising a mixture of peptide human MOGs, wherein the mixture comprises at least one T cell epitope. Administering the human MOG to the mouse in the form of an immunogen causing induction in the mouse; Treating EAE-induced mice with at least one therapeutic composition comprising human MOG at least one antigenic fragment before or after initiation of EAE symptoms in a scam mouse; A method for identifying a therapeutic composition for the treatment of multiple sclerosis comprising determining whether the at least one therapeutic composition prevents the onset or development of symptoms of EAE in the mouse induced with EAE. ※ Note: The disclosure is based on the initial application.