Production of a recombinant hybrid molecule of cholera toxin-B-subunit and proteolipid-protein-peptide for the treatment of experimental encephalomyelitis.
Mucosal administration of experimental autoimmune encephalomyelitis (EAE)-specific autoantigens can reduce the onset of disease. To examine whether cholera toxin-B-subunit (CTB)-conjugated EAE-specific T-cell epitope can reduce development of the autoimmune disease in mice, we produced a recombinant hybrid molecule of CTB fusion protein linked with proteolipid-protein (PLP)-peptide139-151(C140S) at levels up to 0.1 gram per liter culture media in Bacillus brevis as a secretion-expression system. Amino acid sequencing and GM1-receptor binding assay showed that this expression system produced a uniformed recombinant hybrid protein. EAE was induced in SJL/J mice by systemic administration with the PLP-peptide. When nasally immunized 5 times with 70 microg rCTB PLP-peptide hybrid protein, mice showed a significantly suppressed development of ongoing EAE and an inhibition of both the PLP-peptide-specific delayed-type hypersensitivity (DTH) responses and leukocyte infiltration into the spinal cord. In contrast, all mice given the PLP-peptide alone or the PLP-peptide with the free form of CTB did not suppress the development of EAE and DTH responses. These results suggest that nasal treatment with the recombinant B. brevis-derived hybrid protein of CTB and autoantigen peptide could prove useful in the control of multiple sclerosis.
Yuki
Y
Y
JCR Pharmaceuticals Co., 2-2-10 Murotani, Nishi-Ku, Kobe 651-2241 Japan. yukiez@jcrpharm.co.jp
Byun
Y
Y
Fujita
M
M
Izutani
W
W
Suzuki
T
T
Udaka
S
S
Fujihashi
K
K
McGhee
J R
JR
Kiyono
H
H
D000281:Administration, Intranasal; D000595:Amino Acid Sequence; D001407:Bacillus; D002772:Cholera Toxin / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use*; D016503:Drug Delivery Systems; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000188:drug therapy*; D005822:Genetic Vectors; D006968:Hypersensitivity, Delayed; D008969:Molecular Sequence Data; D018991:Myelin Proteolipid Protein / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use*; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use; D013116:Spinal Cord / Q000276:immunology / Q000378:metabolism / Q000473:pathology
D018991:Myelin Proteolipid Protein; D010446:Peptide Fragments; D011994:Recombinant Proteins; C435023:myelin proteolipid protein (139-151); D002772:Cholera Toxin