Immunological Characterization and Therapeutic Activity of an Altered-Peptide Ligand, NBI-6024, Based on the Immunodominant Type 1 Diabetes Autoantigen Insulin B-Chain (9–23) Peptide

• Published in: Diabetes (New York, N.Y.) Vol. 51; no. 7; pp. 2126 - 2134
• Main Authors: Alleva, David G., Gaur, Amitabh, Jin, Liping, Wegmann, Dale, Gottlieb, Peter A., Pahuja, Anil, Johnson, Eric B., Motheral, Theresa, Putnam, Amy, Crowe, Paul D., Ling, Nicholas, Boehme, Stefen A., Conlon, Paul J.
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Diabetes Association 01.07.2002
• Subjects: Amino Acid Substitution; Animals; Autoantigens - blood; Biological and medical sciences; Cells, Cultured; Clone Cells; Development and progression; Diabetes; Diabetes mellitus; Diabetes Mellitus, Type 1 - blood; Diabetes Mellitus, Type 1 - drug therapy; Diabetes Mellitus, Type 1 - immunology; Diabetes. Impaired glucose tolerance; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Etiopathogenesis. Screening. Investigations. Target tissue resistance; Female; Humans; Hypoglycemic Agents - pharmacology; Insulin - immunology; Insulin - pharmacology; Ligands; Lymphocyte Activation - drug effects; Medical sciences; Mice; Mice, Inbred NOD; Peptide Fragments - immunology; Peptide Fragments - pharmacology; Spleen - drug effects; Spleen - immunology; T-Lymphocytes - immunology; Type 1 diabetes; United States; Endocrinopathy; Characterization; Immunopathology; Pancreatic hormone; Autoantigen; Peptide fragment; Ligand; Insulin dependent diabetes; Autoimmune disease; Insulin
• ISSN: 0012-1797; 1939-327X; 1939-327X
• DOI: 10.2337/diabetes.51.7.2126

Immunological Characterization and Therapeutic Activity of an Altered-Peptide Ligand, NBI-6024, Based on the Immunodominant Type 1 Diabetes Autoantigen Insulin B-Chain (9–23) Peptide David G. Alleva 1 , Amitabh Gaur 1 , Liping Jin 1 , Dale Wegmann 2 , Peter A. Gottlieb 2 , Anil Pahuja 1 , Eric B. Johnson 1 , Theresa Motheral 2 , Amy Putnam 2 , Paul D. Crowe 1 , Nicholas Ling 1 , Stefen A. Boehme 1 and Paul J. Conlon 1 1 Neurocrine Biosciences, Inc., San Diego, California 2 University of Colorado Health Sciences Center, Barbara Davis Center for Childhood Diabetes, Denver, Colorado Abstract The nonobese diabetic (NOD) mouse is a good model for human type 1 diabetes, which is characterized by autoreactive T-cell-mediated destruction of insulin-producing islet β-cells of the pancreas. The 9–23 amino acid region of the insulin B-chain [B (9–23) ] is an immunodominant T-cell target antigen in the NOD mouse that plays a critical role in the disease process. By testing a series of B (9–23) peptide analogs with single or double alanine substitutions, we identified a set of altered peptide ligands (APLs) capable of inhibiting B (9–23) -induced proliferative responses of NOD pathogenic T-cell clones. These APLs were unable to induce proliferation of these clones. However, vaccinations with the APLs induced strong cellular responses, as measured by in vitro lymphocyte proliferation and Th2 cytokine production (i.e., interleukin [IL]-4 and IL-10, but not γ-interferon [IFN-γ]). These responses were cross-reactive with the native antigen, B (9–23) , suggesting that the APL-induced Th2 responses may provide protection by controlling endogenous B (9–23) -specific Th1 (i.e., IFN-γ-producing) pathogenic responses. One of these APLs that contained alanine substitutions at residues 16 and 19 (16Y→A, 19C→A; NBI-6024) was further characterized for its therapeutic activity because it consistently induced T-cell responses (e.g., T-cell lines and clones) that were of the Th2 type and that were cross-reactive with B (9–23) . Subcutaneous injections of NBI-6024 to NOD mice administered either before or after the onset of disease substantially delayed the onset and reduced the incidence of diabetes. This study is the first to report therapeutic activity of an APL derived from an islet β-cell-specific antigen in type 1 diabetes. Footnotes Address correspondence and reprint requests to Dr. David G. Alleva, Neurocrine Biosciences, Inc., 10555 Science Center Dr., San Diego, CA 92121-1102. E-mail: dalleva{at}neurocrine.com . Received for publication 4 December 2001 and accepted in revised form 3 April 2002. D.G.A., L.J., A.P., E.B.J., N.L., S.A.B., and P.J.C. are employed by and hold stock in Neurocrine Biosciences, Inc.; A.G. holds stock in Neurocrine Biosciences, Inc. APL, altered peptide ligand; B (9–23) , 9–23 amino acid region of the insulin B-chain; βCA, β-cell target antigen; CFA, complete Freund’s adjuvant; EAE, experimental autoimmune encephalomyelitis; ELISA, enzyme-linked immunosorbent assay; ELISPOT, enzyme-linked immunosorbent spot; IFA, incomplete Freund’s adjuvant; IFN-γ, γ-interferon; IL, interleukin; MBP, myelin basic protein; MHC, major histocompatibility complex; MS, multiple sclerosis; NT, neurotensin; PLP, proteolipid protein; SI, stimulation index; SWM (110–121) , sperm whale myoglobin (110–121); TCR, T-cell receptor. DIABETES