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An Islet-Cell Protein Tyrosine Phosphatase Is a Likely Precursor to the 37-kDa Autoantigen in Type 1 Diabetes: Human and Macaque Sequences, Tissue Distribution, Unique and Shared Epitopes, and Predictive Autoantibodies

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Abstract

Background

We sought to identify novel islet-cell autoantigens to better understand the pathogenesis, prediction, and immunotherapy of type 1 diabetes.

Materials and Methods

Macaque and human islet cDNA libraries expressed in mammalian cells were screened with human diabetes sera. A positive clone was sequenced directly and after 5′ rapid amplification of cDNA ends (RACE). Northern blotting and in situ hybridization revealed the tissue distribution of the corresponding protein. Antigen, expressed by in vitro translation, and tryptic peptides were analyzed by SDS-PAGE. For the immunoprecipitations, 183 diabetic, 60 prediabetic, and 91 control sera were used. Truncated antigens were used in immunoprecipitations for epitope mapping. Recombinant antigen expressed in transfected fibroblasts was used in competition assays.

Results

Sequencing yielded an 111-kDa, 1,013 amino acid, transmembrane protein (M1851) containing consensus protein tyrosine phosphatase (PTPase) sequence. M1851 was 77% identical in the intracellular domain, but only 31% identical extracellularly, to the islet-cell autoantigen ICA512. mRNA localized to brain, prostate, pancreatic islets, and adrenal medulla. After limited trypsinization, the in vitro translated antigen was 37 kDa. M1851 was recognized by 47% of prediabetes sera, 31% of new diabetes sera, but only 1% of healthy controls. Only 1/73 sera binding M1851 failed to bind ICA512, whereas 42/114 binding ICA512 did not bind M1851. M1851 reactivity was not fully displaced by ICA512 in 24/49 sera. Removing the C-terminal 27, 80, or 160 amino acids of M1851 decreased reactivity by 70%, 90%, and 100%, respectively.

Conclusions

This new islet-cell PTPase is likely to be the precursor to the 37-kDa tryptic fragment antigen. It is structurally related to ICA512 but has distinct diabetes autoantibody epitopes located at the C terminus.

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References

  1. Atkinson MA, Maclaren NK. (1994) The pathogenesis of insulin-dependent diabetes mellitus. N. Engl. J. Med. 331: 1428–1436.

    Article  CAS  PubMed  Google Scholar 

  2. Bottazzo GF, Florin-Christensen A, Doniach D. (1974) Islet cell antibodies in diabetes mellitus with autoimmune polyendocrine deficiency. Lancet ii: 1279–1283.

    Article  Google Scholar 

  3. Marshall MO, Hxyer PE, Petersen JS, et al. (1994) Contribution of glutamate decarboxylase antibodies to the reactivity of islet cell cytoplasmic antibodies. J. Autoimmun. 7: 497–508.

    Article  CAS  PubMed  Google Scholar 

  4. Bonifacio E, Lampasona V, Genovese S, Ferrari M, Bosi E. (1995) Identification of protein tyrosine phosphatase-like IA2 (islet cell antigen 512) as the IDDM-related 37/40K autoantigen and a target of ICA. J. Immunol. 155: 5419–5426.

    PubMed  CAS  Google Scholar 

  5. Hagopian W, Sanjeevi C, Kockum I, et al. (1995) Glutamate decarboxylase-, insulin- and islet cell-antibodies and HLA typing to detect diabetes in a general population-based study of Swedish children. J. Clin. Invest. 95: 1505–1511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kolb H. (1994) Immune intervention in type I diabetes mellitus—current clinical and experimental approaches. Exp. Clin. Endocrinol. 102: 269–272.

    Article  CAS  PubMed  Google Scholar 

  7. Christie MR, Vohra G, Champagne P, Daneman D, Delovitch TL. (1990) Distinct antibody specificities to a 64-kD islet cell antigen in type 1 diabetes as revealed by trypsin treatment. J. Exp. Med. 172: 789–794.

    Article  CAS  PubMed  Google Scholar 

  8. Christie M, Hollands J, Brown T, Michelsen B, Delovitch T. (1993) Detection of pancreatic islet 64,000 Mr autoantigens in insulin-dependent diabetes distinct from glutamate decarboxylase. J. Clin. Invest. 92: 240–248.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Christie MR, Genovese S, Cassidy D, et al. (1994) Antibodies to islet 37K antigen, but not glutamate decarboxylase, discriminate rapid progression to IDDM in endocrine autoimmunity. Diabetes 43: 1254–1259.

    Article  CAS  PubMed  Google Scholar 

  10. Ongagna J-C, Levy-Marchal C. (1995) Anti-37K antibodies are associated with the development of IDDM in individuals with ICA. Diabetologia 38: 370–375.

    Article  CAS  PubMed  Google Scholar 

  11. Bingley P, Christie M, Bonifacio E, et al. (1994) Combined analysis of autoantibodies improves prediction of insulin-dependent diabetes in islet cell antibody-positive relatives. Diabetes 43: 1304–1310.

    Article  CAS  PubMed  Google Scholar 

  12. Bonifacio E, Genovese S, Braghi S, et al. (1995) Islet autoantibody markers in IDDM: Risk assessment strategies yielding high sensitivity. Diabetologia 38: 816–822.

    Article  CAS  PubMed  Google Scholar 

  13. Hagopian W, Karlsen A, Petersen J, et al. (1993) Regulation of glutamic acid decarboxylase (GAD) diabetes autoantigen expression in highly purified isolated islets from Macaca nemestrina. Endocrinology 132: 2674–2681.

    Article  CAS  PubMed  Google Scholar 

  14. Komori M, Kikuchi O, Sakuma T, et al. (1992) Molecular cloning of monkey liver cytochrome P-450 cDNAs: Similarity of primary sequences to human cytochromes P-450. Biochim. Biophys. Acta 1171: 141–146.

    Article  CAS  PubMed  Google Scholar 

  15. Jelinek L, Lok S, Rosenberg G, et al. (1993) Expression cloning and signaling properties of the rat glucagon receptor. Science 259: 1614–1616.

    Article  CAS  PubMed  Google Scholar 

  16. Rabin D, Pleasic S, Palmer-Crocker R, Shapiro J. (1992) Cloning and expression of IDDM-specific human autoantigens. Diabetes 41: 183–186.

    Article  CAS  PubMed  Google Scholar 

  17. Rabin D, Pleasic S, Shapiro J, et al. (1994) Islet cell antigen 512 is a IDDM-specific islet autoantigen related to protein tyrosine phosphatases. J. Immunol. 152: 3183–3188.

    PubMed  CAS  Google Scholar 

  18. Lan M, Lu J, Goto Y, Notkins A. (1994) Molecular cloning and identification of a receptor-type protein tyrosine phosphatase, IA-2, from human insulinoma. DNA Cell Biol. 13: 505–514.

    Article  CAS  PubMed  Google Scholar 

  19. Verge C, Gianani R, Kawasaki E, et al. (1996) Prediction of Type 1 diabetes in first-degree relatives using a combination of insulin, GAD, and ICA512bdc/IA2 autoantibodies. Diabetes 45: 926–933.

    Article  CAS  PubMed  Google Scholar 

  20. Karlsen A, Hagopian W, Grubin C, et al. (1991) Cloning and primary structure of a human islet isoform of glutamic acid decarboxylase from chromosome 10. Proc. Natl. Acad. Sci. U.S.A. 88: 8337–8341.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Frohman M, Dush M, Martin G. (1988) Rapid production of full-length cDNAs from rare transcripts: Amplification using a single gene-specific oligonucleotide primer. Proc. Natl. Acad. Sci. U.S.A. 85: 8998–9002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hagopian W, Karlsen A, Gottsäter A, et al. (1993) Quantitative assay using recombinant human islet glutamic acid decarboxylase (GAD-64) shows 64K autoantibody positively at onset predicts diabetes type. J. Clin. Invest. 91: 368–374.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Dixon J. (1995) Structure and catalytic properties of protein tyrosine phosphatases. Ann. N.Y. Acad. Sci. 766: 18–22.

    Article  CAS  PubMed  Google Scholar 

  24. Chiang M-K, Flanagan J. (1996) PTP-NP, a new member of the receptor protein tyrosine phosphatase family, implicated in development of nervous system and pancreatic endocrine cells. Development 122: 2239–2250.

    PubMed  CAS  Google Scholar 

  25. Brady-Kalnay S, Tonks N. (1995) Protein tyrosine phosphatases as adhesion receptors. Curr. Opin. Cell Biol. 7: 650–657.

    Article  CAS  PubMed  Google Scholar 

  26. Wasmeier C, Hutton J. (1996) Molecular cloning of phogrin, a protein-tyrosine phosphatase homologue localized to insulin secretory granule membranes. J. Biol. Chem. 271: 8161–8170.

    Article  Google Scholar 

  27. Lu J, Notkins A, Lan M. (1994) Isolation, sequence and expression of a novel mouse brain cDNA, mIA-2, and its relatedness to members of the protein tyrosine phosphatase family. Biochem. Biophys. Res. Commun. 204: 930–936.

    Article  CAS  PubMed  Google Scholar 

  28. Christgau S, Aanstoot H-J, Schierbeck H, et al. (1992) Membrane anchoring of the autoantigen GAD65 to microvesicles in pancreatic β-cells by palmitoylation in the NH2-terminal domain. J. Cell Biol. 118: 309–320.

    Article  CAS  PubMed  Google Scholar 

  29. Aguilar-Diosdado M, Parkinson D, Corbett J, et al. (1994) Potential autoantigens in IDDM. Expression of carboxypeptidase-H and insulin but not glutamate decarboxylase on the beta-cell surface. Diabetes 43: 418–425.

    Article  CAS  PubMed  Google Scholar 

  30. Hartley S, Crosbie J, Brink R, Kantor A, Basten A, Goodnow C. (1991) Elimination from peripheral lymphoid tissues of self-reactive B lymphocytes recognizing membrane-bound antigens. Nature 353: 765–769.

    Article  CAS  PubMed  Google Scholar 

  31. Payton M, Hawkes C, Christie M. (1995) Relationship of the 37,000- and 40,000-Mr tryptic fragments of islet antigens in IDDM to the protein tyrosine phosphatase-like molecule IA-2 (ICA512). J. Clin. Invest. 96: 1506–1511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Lu J, Li Q, Xie H, et al. (1996) Identification of a second transmembrane protein tyrosine phosphatase, IA2β, as an autoantigen in insulin-dependent diabetes mellitus: Precursor of the 37kDA tryptic fragment. Proc. Natl. Acad. Sci. U.S.A. 93: 2307–2311.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lampasona V, Bearzatto M, Genovese S, Bosi E, Ferrari M, Bonifacio E. (1996) Autoantibodies in IDDM recognize distinct cytoplasmic domains in the protein tyrosine phosphatase-like IA2 antigen. J. Immunol. 157: 2707–2711.

    PubMed  CAS  Google Scholar 

  34. Palmer JP, Asplin CM, Clemons P, et al. (1983) Insulin antibodies in insulin-dependent diabetics before insulin treatment. Science 222: 1337–1339.

    Article  CAS  PubMed  Google Scholar 

  35. Hummel M, Durinovic-Bello I, Zeigler A-G. (1996) Relation between cellular and humoral immunity to islet cell antigens in type 1 diabetes. J. Autoimmun. 9: 427–430.

    Article  CAS  PubMed  Google Scholar 

  36. Roep B. (1996) T-cell responses to autoantigens in IDDM. The search for the Holy Grail. Diabetes 45: 1147–1156.

    Article  CAS  PubMed  Google Scholar 

  37. Baekkeskov S, Nielsen JH, Marner B, Bilde T, Ludvigsson J, Lernmark Å. (1982) Autoantibodies in newly diagnosed diabetic children immunoprecipitate human pancreatic islet cell proteins. Nature 298: 167–169.

    Article  CAS  PubMed  Google Scholar 

  38. Atkinson M, Kaufman D, Campbell L, et al. (1992) Response of peripheral blood mononuclear cells to GAD in IDDM. Lancet 339: 458–459.

    Article  CAS  PubMed  Google Scholar 

  39. Arden S, Roep B, Neophytou P, et al. (1996) Imogen 38: A novel 38 kD islet mitochondrial autoantigen recognized by T-cells from a newly diagnosed type 1 diabetic patient. J. Clin. Invest. 97: 551–561.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Aanstoot H-J, Kang S-M, Kim J, et al. (1996) Identification and characterization of Glima 38, a glycosylated islet cell membrane antigen, which together with GAD65ab and IA2 marks the early phases of the autoimmune response in type I diabetes. J. Clin. Invest. 97: 2772–2800.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Atkinson M, Bowman M, Kao K, et al. (1993) Lack of immune responsiveness to bovine serum albumin in insulin-dependent diabetes. N. Engl. J. Med. 329: 1853–1858.

    Article  CAS  PubMed  Google Scholar 

  42. Atkinson M, Holmes L, Scharp D, Lacy P, Maclaren N. (1991) No evidence for serological autoimmunity to islet cell heat shock proteins in insulin-dependent diabetes. J. Clin. Invest. 87: 721–724.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Thomas N, Ginsberg-Fellner F, McEvoy R. (1990) Strong association between diabetes and displacement of mouse anti-rat insulinoma cell monoclonal antibody by human serum in vitro. Diabetes 39: 1203–1211.

    Article  CAS  PubMed  Google Scholar 

  44. Karounas D, Odebunmi T, Hickman S, Stepiek-Biek P. (1992) Sequence determination of the diabetes-associated 52Kd autoantigen [abstract]. Diabetes 41: 97A.

    Article  Google Scholar 

  45. Buschard K, Josefsen K, Rygaard J, Spitalnik S. (1991) Pancreatic islet-cell epitope recognized by an anti-sulphatide monoclonal antibody. APMIS 99: 1151–1156.

    Article  CAS  PubMed  Google Scholar 

  46. Dotta F, Dionisi S, Gianani R, et al. (1993) Expression of autoantibodies to the GM-2 islet ganglioside precedes the onset of type 1 diabetes in high risk subjects [abstract]. Diabetologia 36: A24.

    Google Scholar 

  47. Shimada A, Charlton B, Taylor-Edwards C, Fathman CG. (1996) β-cell destruction may be a late consequence of the autoimmune process in nonobese diabetic mice. Diabetes 45: 1063–1067.

    Article  CAS  PubMed  Google Scholar 

  48. Harris M, Robbins D. (1994) Prevalence of adult-onset IDDM in the U.S. population. Diabetes Care 17: 1337–1340.

    Article  CAS  PubMed  Google Scholar 

  49. Kobayashi T, Nakanishi K, Murase T, Kosaka K. (1996) Small doses of subcutaneous insulin as a strategy for preventing slowly progressive β-cell failure in ICA-positive patients with clinical features of NIDDM. Diabetes 45: 622–626.

    Article  CAS  PubMed  Google Scholar 

  50. Björk E, Berne C, Kämpe O, Wibell L, Oskarsson P, Karlsson FA. (1996) Diazoxide treatment at onset preserves residual insulin secretion in adults with autoimmune diabetes. Diabetes 45: 1427–1430.

    Article  PubMed  Google Scholar 

  51. Fathman C. (1993) Peptides as therapy of autoimmune disease. Diabetes Metab. Rev. 9: 239–244.

    Article  CAS  PubMed  Google Scholar 

  52. Nag B, Mukku P, Arimilli S, Phan D, Deshpande S, Winkelhake J. (1994) Antigenic peptide binding to MHC class II molecules at increased peptide concentrations. Mol. Immunol. 31: 1161–1168.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Barbara Snowden, Regina Park, and Rashmi Patel for excellent technical assistance. We also thank Dr. J. P. Palmer for help in procurement of and ICA/IAA measurements on sera used for library screening. Dr. D. Rabin for generous gifts of recombinant intracellular ICA512 and rabbit antiserum to ICA512.1, Dr. D. Baskin for advice on in situ hybridizations, and Dr. G. T. Nepom for assistance in procurement of IDDM sera. This paper was presented in part as a poster at the General Clinical Research Centers Annual Conference, Washington DC, March 1996. WAH is supported by the American Diabetes Assn. by NIH grants to the CRC (RR00037), the DERC (DK17047) and the NW Regional Primate Center (RR00166).

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Authors and Affiliations

  1. Department of Medicine, University of Washington, Box 357710, Seattle, Washington, 98195, USA

    James LaGasse, John Breininger & William A. Hagopian

  2. ZymoGenetics, Inc., Seattle, Washington, USA

    Laura Jelinek, Shannon Sexson, Cathy Lofton-Day, Paul Sheppard & Wayne Kindsvogel

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  1. James LaGasse
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  2. Laura Jelinek
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  8. William A. Hagopian
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Correspondence to William A. Hagopian.

Additional information

The macaque sequence reported in this paper has been deposited in the GenBank data base (accession no. U91574).

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LaGasse, J., Jelinek, L., Sexson, S. et al. An Islet-Cell Protein Tyrosine Phosphatase Is a Likely Precursor to the 37-kDa Autoantigen in Type 1 Diabetes: Human and Macaque Sequences, Tissue Distribution, Unique and Shared Epitopes, and Predictive Autoantibodies. Mol Med 3, 163–173 (1997). https://doi.org/10.1007/BF03401670

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