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. 1998 Oct 1;102(7):1454–1462. doi: 10.1172/JCI2793

Serum from patients with type 2 diabetes with neuropathy induces complement-independent, calcium-dependent apoptosis in cultured neuronal cells.

S Srinivasan 1, M J Stevens 1, H Sheng 1, K E Hall 1, J W Wiley 1
PMCID: PMC508993  PMID: 9769338

Abstract

We hypothesized that sera from type 2 diabetic patients with neuropathy contains an autoimmune immunoglobulin that promotes complement-independent, calcium-dependent apoptosis in neuronal cell lines. Neuronal cells were cultured in the presence of complement-inactivated sera obtained from patients with type 2 diabetes with and without neuropathy and healthy adult control patients. Serum from diabetic patients with neuropathy was associated with a significantly greater induction of apoptosis, compared to serum from diabetic patients without neuropathy and controls. In the presence of calcium channel antagonists, induction of apoptosis was reduced by approximately 50%. Pretreatment of neuronal cells with serum from diabetic patients with neuropathy was associated with a significant increase in elevated K+-evoked cytosolic calcium concentration. Serum-induced enhancement in cytosolic calcium and calcium current density was blocked by treatment with trypsin and filtration of the serum using a 100,000-kd molecular weight filter. Treatment with an anti-human IgG antibody was associated with intense fluorescence on the surface of neuronal cells exposed to sera from patients with type 2 diabetes mellitus with neuropathy. We conclude that sera from type 2 diabetic patients with neuropathy contains an autoimmune immunoglobulin that induces complement-independent, calcium-dependent apoptosis in neuronal cells.

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Selected References

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  1. Behse F., Buchthal F., Carlsen F. Nerve biopsy and conduction studies in diabetic neuropathy. J Neurol Neurosurg Psychiatry. 1977 Nov;40(11):1072–1082. doi: 10.1136/jnnp.40.11.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brismar T. Neuropathy-functional abnormalities in the BB rat. Metabolism. 1983 Jul;32(7 Suppl 1):112–117. doi: 10.1016/s0026-0495(83)80023-7. [DOI] [PubMed] [Google Scholar]
  3. Bär P. R. Apoptosis--the cell's silent exit. Life Sci. 1996;59(5-6):369–378. doi: 10.1016/0024-3205(96)00315-3. [DOI] [PubMed] [Google Scholar]
  4. Cachia M. J., Peakman M., Zanone M., Watkins P. J., Vergani D. Reproducibility and persistence of neural and adrenal autoantibodies in diabetic autonomic neuropathy. Diabet Med. 1997 Jun;14(6):461–465. doi: 10.1002/(SICI)1096-9136(199706)14:6<461::AID-DIA380>3.0.CO;2-H. [DOI] [PubMed] [Google Scholar]
  5. Cece R., Barajon I., Tredici G. Cisplatin induces apoptosis in SH-SY5Y human neuroblastoma cell line. Anticancer Res. 1995 May-Jun;15(3):777–782. [PubMed] [Google Scholar]
  6. Chan J., Greenberg D. A. Effects of glucose on calcium channels in neural cells. Neurosci Lett. 1991 Jan 2;121(1-2):34–36. doi: 10.1016/0304-3940(91)90642-7. [DOI] [PubMed] [Google Scholar]
  7. Dowd D. R. Calcium regulation of apoptosis. Adv Second Messenger Phosphoprotein Res. 1995;30:255–280. doi: 10.1016/s1040-7952(05)80010-2. [DOI] [PubMed] [Google Scholar]
  8. Ellis T. M., Atkinson M. A. The clinical significance of an autoimmune response against glutamic acid decarboxylase. Nat Med. 1996 Feb;2(2):148–153. doi: 10.1038/nm0296-148. [DOI] [PubMed] [Google Scholar]
  9. Ewing D. J., Clarke B. F. Diagnosis and management of diabetic autonomic neuropathy. Br Med J (Clin Res Ed) 1982 Oct 2;285(6346):916–918. doi: 10.1136/bmj.285.6346.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fruttiger M., Montag D., Schachner M., Martini R. Crucial role for the myelin-associated glycoprotein in the maintenance of axon-myelin integrity. Eur J Neurosci. 1995 Mar 1;7(3):511–515. doi: 10.1111/j.1460-9568.1995.tb00347.x. [DOI] [PubMed] [Google Scholar]
  11. Gajewski T. F., Thompson C. B. Apoptosis meets signal transduction: elimination of a BAD influence. Cell. 1996 Nov 15;87(4):589–592. doi: 10.1016/s0092-8674(00)81377-x. [DOI] [PubMed] [Google Scholar]
  12. Gelperin D., Mann D., del Valle J., Wiley J. W. Bradykinin (Bk) increases cytosolic calcium in cultured rat myenteric neurons via Bk-2 type receptors coupled to mobilization of extracellular and intracellular sources of calcium: evidence that calcium influx is prostaglandin dependent. J Pharmacol Exp Ther. 1994 Oct;271(1):507–514. [PubMed] [Google Scholar]
  13. Giordano C., Stassi G., Todaro M., De Maria R., Richiusa P., Scorsone A., Giordano M., Galluzzo A. Low bcl-2 expression and increased spontaneous apoptosis in T-lymphocytes from newly-diagnosed IDDM patients. Diabetologia. 1995 Aug;38(8):953–958. doi: 10.1007/BF00400585. [DOI] [PubMed] [Google Scholar]
  14. Gold R., Schmied M., Giegerich G., Breitschopf H., Hartung H. P., Toyka K. V., Lassmann H. Differentiation between cellular apoptosis and necrosis by the combined use of in situ tailing and nick translation techniques. Lab Invest. 1994 Aug;71(2):219–225. [PubMed] [Google Scholar]
  15. Greene D. A., Lattimer S. A., Sima A. A. Are disturbances of sorbitol, phosphoinositide, and Na+-K+-ATPase regulation involved in pathogenesis of diabetic neuropathy? Diabetes. 1988 Jun;37(6):688–693. doi: 10.2337/diab.37.6.688. [DOI] [PubMed] [Google Scholar]
  16. Greene D. A., Sima A. A., Stevens M. J., Feldman E. L., Lattimer S. A. Complications: neuropathy, pathogenetic considerations. Diabetes Care. 1992 Dec;15(12):1902–1925. doi: 10.2337/diacare.15.12.1902. [DOI] [PubMed] [Google Scholar]
  17. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  18. Hall K. E., Sima A. A., Wiley J. W. Opiate-mediated inhibition of calcium signaling is decreased in dorsal root ganglion neurons from the diabetic BB/W rat. J Clin Invest. 1996 Mar 1;97(5):1165–1172. doi: 10.1172/JCI118530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hall K. E., Sima A. A., Wiley J. W. Voltage-dependent calcium currents are enhanced in dorsal root ganglion neurones from the Bio Bred/Worchester diabetic rat. J Physiol. 1995 Jul 15;486(Pt 2):313–322. doi: 10.1113/jphysiol.1995.sp020814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  21. Jensen L. M. Phenotypic differentiation of aphidicolin-selected human neuroblastoma cultures after long-term exposure to nerve growth factor. Dev Biol. 1987 Mar;120(1):56–64. doi: 10.1016/0012-1606(87)90103-5. [DOI] [PubMed] [Google Scholar]
  22. Joseph R., Li W., Han E. Neuronal death, cytoplasmic calcium and internucleosomal DNA fragmentation: evidence for DNA fragments being released from cells. Brain Res Mol Brain Res. 1993 Jan;17(1-2):70–76. doi: 10.1016/0169-328x(93)90074-y. [DOI] [PubMed] [Google Scholar]
  23. Juntti-Berggren L., Larsson O., Rorsman P., Ammälä C., Bokvist K., Wåhlander K., Nicotera P., Dypbukt J., Orrenius S., Hallberg A. Increased activity of L-type Ca2+ channels exposed to serum from patients with type I diabetes. Science. 1993 Jul 2;261(5117):86–90. doi: 10.1126/science.7686306. [DOI] [PubMed] [Google Scholar]
  24. Kappelle A. C., Biessels G., Bravenboer B., van Buren T., Traber J., de Wildt D. J., Gispen W. H. Beneficial effect of the Ca2+ antagonist, nimodipine, on existing diabetic neuropathy in the BB/Wor rat. Br J Pharmacol. 1994 Mar;111(3):887–893. doi: 10.1111/j.1476-5381.1994.tb14821.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Levy J., Gavin J. R., 3rd, Sowers J. R. Diabetes mellitus: a disease of abnormal cellular calcium metabolism? Am J Med. 1994 Mar;96(3):260–273. doi: 10.1016/0002-9343(94)90152-x. [DOI] [PubMed] [Google Scholar]
  26. Linnik M. D., Hatfield M. D., Swope M. D., Ahmed N. K. Induction of programmed cell death in a dorsal root ganglia X neuroblastoma cell line. J Neurobiol. 1993 Apr;24(4):433–446. doi: 10.1002/neu.480240403. [DOI] [PubMed] [Google Scholar]
  27. McNeil H. P., Chesterman C. N., Krilis S. A. Immunology and clinical importance of antiphospholipid antibodies. Adv Immunol. 1991;49:193–280. doi: 10.1016/s0065-2776(08)60777-4. [DOI] [PubMed] [Google Scholar]
  28. Morton A. J., Hammond C., Mason W. T., Henderson G. Characterisation of the L- and N-type calcium channels in differentiated SH-SY5Y neuroblastoma cells: calcium imaging and single channel recording. Brain Res Mol Brain Res. 1992 Mar;13(1-2):53–61. doi: 10.1016/0169-328x(92)90044-c. [DOI] [PubMed] [Google Scholar]
  29. Nagata S. Apoptosis by death factor. Cell. 1997 Feb 7;88(3):355–365. doi: 10.1016/s0092-8674(00)81874-7. [DOI] [PubMed] [Google Scholar]
  30. Nayak R. C., Omar M. A., Rabizadeh A., Srikanta S., Eisenbarth G. S. "Cytoplasmic" islet cell antibodies. Evidence that the target antigen is a sialoglycoconjugate. Diabetes. 1985 Jun;34(6):617–619. doi: 10.2337/diab.34.6.617. [DOI] [PubMed] [Google Scholar]
  31. Nobe S., Aomine M., Arita M., Ito S., Takaki R. Chronic diabetes mellitus prolongs action potential duration of rat ventricular muscles: circumstantial evidence for impaired Ca2+ channel. Cardiovasc Res. 1990 May;24(5):381–389. doi: 10.1093/cvr/24.5.381. [DOI] [PubMed] [Google Scholar]
  32. Patterson M. K., Jr Measurement of growth and viability of cells in culture. Methods Enzymol. 1979;58:141–152. doi: 10.1016/s0076-6879(79)58132-4. [DOI] [PubMed] [Google Scholar]
  33. Pittenger G. L., Liu D., Vinik A. I. The apoptotic death of neuroblastoma cells caused by serum from patients with insulin-dependent diabetes and neuropathy may be Fas-mediated. J Neuroimmunol. 1997 Jun;76(1-2):153–160. doi: 10.1016/s0165-5728(97)00042-8. [DOI] [PubMed] [Google Scholar]
  34. Pittenger G. L., Liu D., Vinik A. I. The neuronal toxic factor in serum of type 1 diabetic patients is a complement-fixing autoantibody. Diabet Med. 1995 May;12(5):380–386. doi: 10.1111/j.1464-5491.1995.tb00499.x. [DOI] [PubMed] [Google Scholar]
  35. Pittenger G. L., Liu D., Vinik A. I. The toxic effects of serum from patients with type 1 diabetes mellitus on mouse neuroblastoma cells: a new mechanism for development of diabetic autonomic neuropathy. Diabet Med. 1993 Dec;10(10):925–932. doi: 10.1111/j.1464-5491.1993.tb00008.x. [DOI] [PubMed] [Google Scholar]
  36. Ristic H., Wiley J. W., Hall K. E., Sima A. A. Failure of nimodipine to prevent or correct the long-term nerve conduction defect and increased neuronal Ca(2+)-currents in the diabetic BB/W-rat. Diabetes Res Clin Pract. 1996 May;32(3):135–140. doi: 10.1016/0168-8227(96)01250-8. [DOI] [PubMed] [Google Scholar]
  37. Seilheimer B., Persohn E., Schachner M. Antibodies to the L1 adhesion molecule inhibit Schwann cell ensheathment of neurons in vitro. J Cell Biol. 1989 Dec;109(6 Pt 1):3095–3103. doi: 10.1083/jcb.109.6.3095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shibasaki F., McKeon F. Calcineurin functions in Ca(2+)-activated cell death in mammalian cells. J Cell Biol. 1995 Nov;131(3):735–743. doi: 10.1083/jcb.131.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sima A. A., Brismar T. Reversible diabetic nerve dysfunction: structural correlates to electrophysiological abnormalities. Ann Neurol. 1985 Jul;18(1):21–29. doi: 10.1002/ana.410180105. [DOI] [PubMed] [Google Scholar]
  40. Sundkvist G., Lind P., Bergström B., Lilja B., Rabinowe S. L. Autonomic nerve antibodies and autonomic nerve function in type 1 and type 2 diabetic patients. J Intern Med. 1991 Jun;229(6):505–510. doi: 10.1111/j.1365-2796.1991.tb00386.x. [DOI] [PubMed] [Google Scholar]
  41. Trump B. F., Berezesky I. K. Calcium-mediated cell injury and cell death. FASEB J. 1995 Feb;9(2):219–228. doi: 10.1096/fasebj.9.2.7781924. [DOI] [PubMed] [Google Scholar]
  42. White R. E., Carrier G. O. Vascular contraction induced by activation of membrane calcium ion channels is enhanced in streptozotocin-diabetes. J Pharmacol Exp Ther. 1990 Jun;253(3):1057–1062. [PubMed] [Google Scholar]
  43. Williams A. F., Barclay A. N. The immunoglobulin superfamily--domains for cell surface recognition. Annu Rev Immunol. 1988;6:381–405. doi: 10.1146/annurev.iy.06.040188.002121. [DOI] [PubMed] [Google Scholar]
  44. Zhivotovsky B., Wade D., Nicotera P., Orrenius S. Role of nucleases in apoptosis. Int Arch Allergy Immunol. 1994 Dec;105(4):333–338. doi: 10.1159/000236778. [DOI] [PubMed] [Google Scholar]

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