Summary
We recently developed a particle concentration fluoroimmunoassay for the measurement of serum antibodies to bovine serum albumin in patients with Type 1 (insulin-dependent) diabetes mellitus. We observed elevated IgG-anti-bovine serum albumin antibodies in 100% of newly-diagnosed diabetic children and in 2.5% of matched control children. Here we compare the fluoroimmunoassay and the more commonly available enzyme linked immunoassay technique, exchanging coded serum samples from 40 newly-diagnosed diabetic children and 179 control children between two laboratories. Particle concentration fluoroimmunoassay detected elevated IgG-anti-bovine serum albumin antibodies in all diabetic children, enzyme immunoassay in 25% (p <0.0001). Fluoroimmunoassay detected elevated levels in 2.2% and enzyme immunoassay in 10% of control children (p <0.002). Elevated IgA-antibovine serum albumin antibodies in patients were slightly more often detected by fluoroimmunoassay than by enzyme immunoassay, while in control children enzyme immunoassays detected elevated levels three times more often (p <0.01). Values measured in either assay showed overall no correlation in either patient (IgG: rs = 0.28; IgA: rs = 0.11) or control sera (IgG: rs = 0.02; IgA: rs = -0.05). Fluoroimmunoassay for IgG was 100% disease-sensitive (enzyme immu-noassay: 25%, p <0.0001) and more disease-specific (IgG; p <0.02). Our findings demonstrate that these assay techniques detected distinct subsets of anti-bovine serum albumin antibodies with little (IgG) or some (IgA) overlap. In fluoroimmunoassay procedures, antigen: antibody binding occurs within 1–2 min while hours are allowed in an enzyme immunoassay. Antibodies with high on-off binding rates typical for immune responses following hyperimmunization are therefore measured preferentially by particle concentration fluoroimmunoassay and it is these antibodies which appear to be associated with diabetes. These observations emphasize the need for epidemiological surveys to validate immunoassay procedures used for clinical purposes.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Dosch H-M, Karjalainen J, Morkowski J, Martin JM, Robinson BH (1992) Nutritional triggers of IDDM. In: Levy-Marchal C, Czernichow P (eds) Pediatric and adolescent endocrinology. Karger, Basel, pp 202–217
Glerum M, Robinson BH, Martin JM (1989) Could bovine serum albumin be the initiating antigen ultimately responsible for the development of insulin dependent diabetes mellitus? Diabetes Res 10: 103–107
Martin JM, Trink B, Daneman D, Dosch H-M, Robinson BH (1991) Milk proteins in the etiology of insulin-dependent diabetes mellitus (IDDM). Ann Med 23: 447–452
Elliott RB, Reddy SN, Bibby NJ, Kida K (1988) Dietary prevention of diabetes in the non-obese diabetic mouse. Diabetologia 36: 62–64
Scott FW, Daneman D, Martin JM (1988) Evidence for a critical role of diet in the development of insulin-dependent diabetes mellitus. Diabetes Res 7: 153–157
Karjalainen J, Martin JM, Knip M et al. (1992) Evidence for a bovine albumin peptide as a candidate trigger of Type 1 diabetes. N Engl J Med 327: 302–307
Dosch HM, Lam P, Guerin D, Leeder JS (1988) Characteristics of particle concentration fluorescence immunoassay (PCFIA): novel alternatives to ELISA and RIA, In: Gembala P-L (ed) Proceedings of the 1987 Pandex Symposium on particle concentration fluorescence immunoassay, Baxter Healthcare Corporation, Chicago, pp 5–22
Hui MF, Lam P, Dosch HM (1989) Properties and heterogeneity of human fetal pre-B cells transformed by EBV. J Immunol 143: 2470–2479
Dosch HM, Lam P, Hui MF, Hibi T (1989) Concerted generation of Ig isotype diversity in human fetal bone marrow. J Immunol 143: 2464–2469
Dosch H-M, Lam P, Hui MF, Hibi T, Cheung RK (1990) EBV utilizes a unique activation pathway for the transformation of human B cells. Int Immunol 2: 833–848
Walter MH, Dosch H-M, Cox DW (1991) A deletion map of the human immunoglobulin heavy chain region. J Exp Med 174: 335–349
Beppu H, Winter WE, Atkinson MA, Maclaren NK, Fujita K, Takahashi H (1987) Bovine albumin antibodies in NOD mice. Diabetes Res 6: 67–69
Scott FW, Cloutier HE, Souligny J, Riley WJ, Hoorfar J, Brogren CH (1989) Diet and antibody production in the diabetes-prone BB rat. In: Larkins R, Zimmet P, Chisholm D (eds) Diabetes 1988. Proceedings of the 13th International Diabetes Federation congress. Elsevier Science, Amsterdam, pp 763–767
Colman PG, Campbell IL, Kay TWH, Harrison LC (1987) 64 000-Mrauto-antigen in type 1 diabetes: evidence of its location on human islets. Diabetes 36: 1432–1440
Yokota A, Yamaguchi Y, Ueda Y et al. (1990) Comparison of islet cell antibodies, islet cell surface antibodies and anti-bovine serum albumin antibodies in type 1 diabetes. Diabetes Res Clin Prac 9: 211–217
Savilahti E, Åkerblom HK, Tainio V-M, Koskimies S (1988) Children with newly diagnosed insulin dependent diabetes mellitus have increased levels of cow's milk antibodies. Diabetes Res 7: 137–140
Pocecco M, Nicoloso F, Tonini G, Presani G, Marinoni S (1991) Increased levels of cow's milk antibodies in children with newly diagnosed insulin-dependent diabetes mellitus (IDDM). Horm Res 35: 67
Tainio VM, Savilahti E, Arjomaa P, Salmenperä L, Perheentupa J, Siimes M (1988) Plasma antibodies to cow's milk are increased by early weaning and consumption of unmodified milk, but production of plasma IgA and IgM cow's milk antibodies is stimulated even during exclusive breast-feeding. Acta Paediatr Scand 77: 807–811
Cheung RK, Dosch H-M (1991) The tyrosine kinase lck is critically involved in the growth transformation of human B lymphocytes. J Biol Chem 266: 8667–8670
Borch-Johnsen K, Joner G, Mandrup-Poulsen T et al. (1984) Relation between breast-feeding and incidence rates of insulin-dependent diabetes mellitus. Lancet II: 353–360
Scott FW (1990) Cow milk and insulin-dependent diabetes mellitus: is there a relationship? Am J Clin Nutr 51: 489–491
Virtanen SM, Räsänen L, Aro A et al. (1991) Infant feeding in Finnish children less than 7 yr of age with newly diagnosed IDDM. Childhood Diabetes in Finland Study Group. Diabetes Care 14: 415–417
Elliott RB, Martin JM (1984) Dietary protein: a trigger of insulin-dependent diabetes in the BB rat? Diabetologia 26: 297–299
Daneman D, Fishman L, Clarson C, Martin JM (1987) Dietary triggers of insulin-dependent diabetes in the BB rat. Diabetes Res 5: 93–97
Scott FW, Marliss EB (1991) Conference summary: diet as an environmental factor in development of insulin-dependent diabetes mellitus. Can J Physiol Pharmacol 69: 311–319
Pilcher CC, Dickens K, Elliott RB (1991) ICA only develop in early childhood. Diabetes Res Clin Pract 14 [Suppl 1]: S82 (Abstract)
Dosch H-M, Hui MF, Doherty PJ (1992) Development of normal and abnormal immune function in man. Dev Immunol 2 (in press)
Olerup O, Smith CIE, Hamarström L (1990) Different amino acids at position 57 of the HLA-DQ beta chain associated with susceptibility and resistance to IgA deficiency. Nature 347: 289–290
Caudwell J, Stojanovski C, Colma P (1991) Caution against the use of commercial ELISA kit assays for preclinical IDDM screening. Diabetes Res Clin Pract 14 [Suppl 1]: S89 (Abstract)
Kuglin B, Kolb H, Greenbaum C, Maclaren NK, Lernmark Å, Palmer JP (1990) The fourth international workshop on the standardisation of insulin autoantibody measurement. Diabetologia 33: 638–639
Levy-Marchal C, Bridel MP, Sodoyez-Goffaux F et al. (1991) Superiority of radiobinding assay over ELISA for detection of IAAs in newly diagnosed type 1 diabetic children. Diabetes Care 14: 61–63
Greenbaum GJ, Palmer JP, Kuglin B, Kolb H (1992) Insulin autoantibodies measured by RIA methodology are more related to IDDM than those measured by ELISA. J Clin Endocrinol Metab 74: 1040–1044
Sodoyez-Goffaux F, Koch M, Dozio N, Brandenburg D, Sodoyez J-C (1988) Advantages and pitfalls of radioimmune and enzyme linked immunosorbent assays of insulin antibodies. Diabetologia 31: 694–702
Sodoyez J-C, Sodoyez-Goffaux F, Koch M, Bouillenne C, Francois-Gerard C, Bosi E (1990) Clonally restricted insulin auto-antibodies in a cohort of 2200 healthy blood donors. Diabetologia 33: 719–725
Foote J, Milstein C (1991) Kinetic maturation of an immune response. Nature 352: 530–532
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Karjalainen, J., Saukkonen, T., Savilahti, E. et al. Disease-associated anti-bovine serum albumin antibodies in Type 1 (insulin-dependent) diabetes mellitus are detected by particle concentration fluoroimmunoassay, and not by enzyme linked immunoassay. Diabetologia 35, 985–990 (1992). https://doi.org/10.1007/BF00401430
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00401430