Summary
Although CD5+B lymphocytes are mostly committed to the production of polyreactive natural autoantibodies, CD5+B lymphocytes committed to the production of somatically mutated and monoreactive high-affinity IgM autoantibodies have been also shown. Increased proportions of CD5+B lymphocytes in some autoimmune diseases, including insulin-dependent diabetes mellitus (IDDM), have been noticed. The present study was undertaken to analyse the differences between CD5+ and CD5-B lymphocyte subsets for production of IDDM-related autoantibodies, i.e. anti-human insulin antibodies (IA) and anti-human islet cell antibodies (ICA). For this purpose, Epstein-Barr Virus (EBV)-transformation of FACS cell-sorted CD5+ and CD5-B lymphocytes and unfractionated enriched B lymphocytes from nine IDDM patients treated exclusively with recombinant human insulin, and from four healthy control subjects was performed; a mean of 102–216 microcultures with a mean of 1,000–2,333 cells/microculture for each B-lymphocyte fraction and individual was established. Data show that both CD5+ and CD5-B-lymphocyte subsets from either normal subjects or from IDDM patients receiving recombinant human insulin, contain B lymphocytes committed to the production of IA-IgM as a common element of their repertoire. In contrast, cells committed to the production of IA-IgG were only detected among the CD5-B lymphocyte subset from some IDDM patients. Only one microculture, out of a total of 6,211 screened (from control subjects and patients), in the CD5-B-cell subset from a recently-diagnosed IDDM patient, was found to produce ICA-IgMλ. This might suggest that the frequency of circulating B lymphocytes committed to the production of ICA is very low even in IDDM patients bearing serum ICA. EBV-transformed B cells producing the ICA-IgMψ were stabilized and cloned by somatic hybridization technique. This ICA-IgMψ human monoclonal antibody, designated HY1-MB91, is not polyreactive, but shows a restricted reactivity with human pancreatic islets, failing to react with other human tissues including cerebellar cortex, and lacking rheumatoid factor and anti-DNA antibody activities. It also lacks reactivity with pancreatic islets from other mammalian species (rat, mouse and monkey) as well as with other rat tissues, including cerebellar cortex. The antigen recognized by HY1-MB91 antibody in human islet cells is a cytoplasmic component mostly found in beta cells. [Diabetologia (1995) 38:62–72]
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Abbreviations
- EBV:
-
Epstein-Barr virus
- IDDM:
-
insulin-dependent diabetes mellitus
- GAD:
-
glutamic acid decarboxylase
- IA:
-
anti-insulin antibodies
- ICA:
-
islet cell antibodies
- mAbs:
-
monoclonal antibodies
- NAA:
-
natural autoantibodies
- PBMC:
-
peripheral blood mononuclear cells
- RF:
-
Rheumatoid factor
References
Castaño L, Eisenbarth GS (1990) Type 1 diabetes: a chronic autoimmune disease of human, mouse and rat. Ann Rev Immunol 8: 647–679
Srikanta S, Ricker AT, McCulloch DK, Soeldner JS, Eisenbarth GS, Palmer JP (1986) Autoimmunity to insulin, β cell dysfunction and development of insulin-dependent diabetes mellitus. Diabetes 35: 139–142
Bottazzo GF, Dean BM, McNally JM, MacKay EH, Swift PGF, Gamble DR (1985) In situ characterization of autoimmune phenomena and expression of HLA molecules in the pancreas in diabetic insulitis. N Engl J Med 313: 353–360
Bottazzo GF, Florien-Christensen A, Doniach D (1974) Islet cell antibodies in diabetes mellitus with autoimmune polyendocrine deficiencies. Lancet II: 1279–1283
Palmer JP, Asplin CM, Clemons P, Lyon K, Iapati O, Raghu P, Paquette TL (1983) Insulin autoantibodies in insulin-dependent diabetes before insulin treatment. Science 222: 1337–1339
Thai A-Ch, Eisenbarth GS (1993) Natural history of IDDM. Diabetes Rev 1: 1–14
Avrameas S (R) Natural autoantibodies: from “horror autotoxicus” to “gnothi seauton”. Immunol Today 154: 154–159
Kantor AB, Herzenberg LA (1993) Origin of murine B cell lineages. Ann Rev Immunol 11: 501–538
Hayakawa K, Hardy RR, Parks DR, Herzenberg LA (1983) The “Ly-1 B” cell subpopulation in normal immunodefective, and autoimmune mice. J Exp Med 157: 202–218
Kantor AB (1991) The development and repertoire of B1 cells (CD5 B cells). Immunol Today 12: 389–391
Hayakawa K, Hardy RR (1988) Normal, autoimmune, and malignant CD5+B cells: the Ly-1 B lineage? Ann Rev Immunol 6: 197–218
Kipps TJ (1989) The CD5+B cell. Adv Immunol 47: 117–185
Casali P, Notkins AL (1989) Probing the human B-cell repertoire with EBV: polyreactive antibodies and CD5+B lymphocytes. Ann Rev Immunol 7: 513–535
Hardy RR (1992) Variable gene usage, physiology and development of Ly-1+(CD5+) B cells. Curr Opin Immunol 4: 181–185
Kipps T, Carson DA (1993) Autoantibodies in chronic lymphocytic leukemia and related systemic autoimmune diseases. Blood 81: 2475–2487
Plater-Zyberk C, Maini RN, Lam K, Kennedy TD, Janossy G (1985) A rheumatoid arthritis B cell subset expresses a phenotype similar to that in chronic lymphocytic leukemia. Arthritis Rheum 28: 971–976
Dauphinée M, Tovar Z, Talal N (1988) B cells expressing CD5 are increased in Sjögren's syndrome. Arthritis Rheum 31: 642–647
Iwatani Y, Amino N, Kaneda T et al. (1989) Marked increase of CD5+B cells in hyperthyroid Graves' disease. Clin Exp Immunol 78: 196–200
Nicoletti F, Meroni PL, Barcellini W et al. (1989) Enhanced percentage of CD5+B lymphocytes in newly diagnosed IDDM patients. Immunol Lett 23: 211–216
Muñoz A, Gallart T, Viñas O, Gomis R (1991) Increased CD5-positive B lymphocytes in type 1 diabetes. Clin Exp Immunol 83: 304–308
Schatz DA, Lang F, Cantor AB et al. (1991) CD5+B lymphocytes in high-risk islet cell-antibody-positive and newly diagnosed IDDM subjects. Diabetes 40: 1314–1318
Sidman CHL, Shultz LD, Hardy RR, Hayakawa K, Herzenberg LA (1986) Production of immunoglobulin isotypes by Ly-1+B cells in viable motheaten and normal mice. Science 232: 1423–1425
Painter C, Monestier M, Bonin B, Bona AC (1986) Functional and molecular studies of V genes expressed in autoantibodies. Immunol Rev 94: 75–98
Casali P, Burastero SE, Nakamura M, Inghirami G, Notkins AL (1987) Human lymphocytes making rheumatoid factor and antibody to ssDNA belong to Leu-1+ B-cell subset. Science 236: 77–81
Hardy RR, Hayakawa K, Shimizu M, Yamasaki K, Kishimoto T (1987) Rheumatoid factor secretion from human Leu-1+B cells. Science 236: 81–83
Casali P, Burastero S, Balow JE, Notkins AL (1989) High-affinity antibodies to ssDNA are produced by CD5-B cells in systemic lupus erythematosus patients. J Immunol 143: 3476–3483
Shutte MEM, Ebeling SB, Akkermans KE, Gmelig-Meyling FHJ, Logtenberg T (1991) Antibody specificity and immunoglobulin VH gene utilization of human monoclonal CD5+B cell lines. Eur J Immunol 21: 1115–1121
Burastero SE, Casali P, Wilder RL, Notkins AL (1988) Monoreactive high affinity and polyreactive low affinity rheumatoid factors are produced by CD5+B cells from patients with rheumatoid arthritis. J Exp Med 168: 1979–1992
Mantovani L, Wilder RL, Casali P (1993) Human rheumatoid B-1a (CD5+B) cells make somatically hypermutated high affinity IgM rheumatoid factors. J Immunol 151: 473–488
Van der Heijden RWJ, Bunschoten H, Hoek A et al. (1991) A human CD5+B cell clone that secretes an idiotype-specific high-affinity IgM monoclonal antibody. J Immunol 146: 1503–1508
Kaushik A, Mayer R, Fidanza V, Zaghouani H, Lim A, Bona C, Dighiero G (1990) Ly1 and V-gene expression among hybridomas secreting natural autoantibody. J Autoimmunity 3: 687–700
Kasaian MT, Ikemnatsu H, Casali P (1992) Identification and analysis of a novel CD5-B lymphocyte subset producting natural antibodies. J Immunol 148: 2690–2695
National Diabetes Data Group (1979) Classification and diagnosis of diabetes mellitus and other categories of glucose intolerance. Diabetes 29: 1039–1057
Bergua M, Solé J, Marion G et al. (1987) Prevalence of islet cell antibodies, insulin antibodies and hyperglycaemia in 2291 school-children. Diabetologia 30: 724–726
Peig M, Gomis R, Ercilla G, Casamitjana R, Bottazzo GF, Pujol-Borrell R (1989) Correlation between residual β-cell function and islet cell antibodies in newly diagnosed type 1 diabetes. Diabetes 38: 1396–1401
Anegón I, Vilella R, Gallart T et al. (1986) B-C1, B-C2, B-C3: monoclonal antibodies against B cell differentiation antigens. In: Reinherz EL, Haynes BF, Nadler LM, Berstein ID (eds) Human B lymphocytes. Vol 2. Springer-Verlag, New York, Berlin, pp 121–139
Ling NR, MacLennan ICM, Mason DY (1987) B cell-antigens: new and previously defined clusters. In: McMichael AJ, Beverley PCL, Cobbold S et al. (eds) Oxford University Press. Oxford, New York, pp 302–335
Casali P, Nakamura M, Ginsberg-Fellner F, Notkins AL (1990) Frequency of B cells committed to the production of antibodies to insulin in newly diagnosed patients with insulin-dependent diabetes mellitus and generation of high affinity human monoclonal IgG to insulin. J Immunol 144: 3741–3747
De La Fuente MA, Egile C, Pereira A et al. (1994) Molecular characterization of a monoclonal IgMK(GAS) anti-Gd cold agglutinin (CA). Its co-existence with a monoclonal IgG3K(GAS) without CA activity that might be clonally related to IgMGAS. Blood 83: 1310–1322
Gallart T, Bladé J, Martínez Quesada J, Sierra J, Rozman C, Vives J (1985) Multiple myeloma with monoclonal IgG and IgD of lambda type exhibiting, under treatment, a shift from mainly IgG to mainly IgD. Immunology 55: 45–57
Lozano F, Parés A, Borche L, Plana M, Gallart T, Rodés J, Vives J (1988) Autoantibodies against nuclear envelope-associated proteins in primary biliary cirrhosis. Hepatology 8: 930–938
Vives-Pi M, Somoza N, Vargas F, Armengol P, Sarri Y, Wu JY, Pujol-Borrell R (1993) Expression of glutamic acid decarboxylase (GAD) in the α, β and δ cells of normal and diabetic pancreas: implications for the pathogenesis of type I diabetes. Clin Exp Immunol 92: 391–396
Randen I, Pasqual V, Victor K, Thompson KM, Forre O, Capra JD, Natvig JB (1993) Synovial IgG rheumatoid factors show evidence of an antigen-driven immune repertoire compared to IgM rheumatoid factors. Eur J Immunol 23: 1220–1225
Casali P, Prabhakar BS, Notkins AL (1988) Characterization of multireactive autoantibodies and identification of Leu-1+B lymphocytes as cells making antibodies binding multiple self and exogenous molecules. Intern Rev Immunol 3: 17–45
Satoh J, Prabhakar BS, Haspel MV, Ginsberg-Fellner F, Notkins AL (1983) Human monoclonal autoantibodies that react with multiple endocrine organs. New Engl J Med 309: 217–220
Punnonen J, Aversa GG, Vandekerckhove B, Roncarolo M-G, de Vries JE (1992) Induction of isotype switching and Ig production by CD5+and CD10+human fetal B cells. J Immunol 148: 3398–3404
Braun J, Krall WJ, Krall WJ, Clark ME, Gowan III HL, Chen U (1988) Inducible Ig heavy chain switching in an IgM+Ly-1 B cell line. Evidence for a state of switch commitment. J Mol Cell Immunol. 4: 105–119
Sarfati M, Luo H, Delespesse G (1989) IgE synthesis by chronic lymphocytic leukemia. J Exp Med 170: 1775–1780
Houdayer M, Bouvet JP, Wolff A et al. (1993) Simultaneous presence, in one serum, of four monoclonal antibodies that might correspond to different steps in a clonal evolution from polyreactive to monoreactive antibodies. J Immunol 150: 311–319
Van Es JH, Gmelig Myeling FHJ, Van der Akker WRM, Aaanstoot H, Derksen RHWM, Logtenberg T (1991) Somatic mutations in the variable regions of a human IgG anti-double-stranded DNA autoantibody suggest a role for antigen in the induction of systemic lupus erythematosus. J Exp Med 173: 461–470
Winker TH, Fehr H, Kalden JR (1992) Analysis of immunoglobulin variable region genes from human IgG anti-DNA hybridomas. Eur J Immunol 22: 1719–1728
Caligaris-Cappio F, Riva M, Tesio L, Schena M, Gaidano G, Bergui L (1989) Human normal CD5+B lymphocytes can be induced to differentiate to CD5-B lymphocytes with germinal center cell features. Blood 73: 1259–1263
Banchereau J, Rousset F (1992) Human B lymphocytes: phenotype, proliferation and differentiation. Adv Immunol 52: 125–262
Stewart AK, Huang C, Long AA, Stollar BD, Schwartz RS (1992) VH-gene representation in autoantibodies reflects the normal human B-cell repertoire. Immunol Rev 128: 101–122
Thomas JW (1993) V region diversity in human anti-insulin antibodies. Preferential use of a VHIII gene subset. J Immunol 150: 1375–1382
Davis SN, Thompson CJ, Peak M, Brown MD, Alberti KGMM (1992) Effects of human insulin on insulin binding antibody production in nondiabetic subjects. Diabetes Care 15: 124–126
Richter W, Endl J, Eiermann TH et al. (1992) Human monoclonal islet cell antibodies from a patient with insulin-dependent diabetes mellitus reveal glutamate decarboxylase as the target antigen. Proc Natl Acad Sci USA 89: 8467–8471
Einsenbarth GS, Linnenbach A, Jackson R, Scearce R, Croce CM (1982) Human hybridomas secreting anti-islet autoantibodies. Nature 300: 264–267
Nayak RC, Omar OAK, Rabizadeh A, Srikanta S, Eisenbarth GS (1985) “Cytoplasmic” islet cell antibodies: evidence that the target antigen is a sialoglycoconjugate. Diabetes 34: 617–619
Golman PG, Nayak RC, Campbell IL, Eisenbarth GS (1988) Binding of cytoplasmic islet cell antibodies is blocked by human pancreatic glycolipid extracts. Diabetes 37: 645–652
Richter W, Shi Y, Baekkeskov S (1993) Autoreactive epitopes defined by diabetes-associated human monoclonal antibodies are localized in the middle and C-terminal domains of the smaller form of glutamate decarboxylase. Proc Natl Acad Sci USA 90: 2832–2836
Richter W, Eiermann TH, Endl J et al. (1993) Human monoclonal islet specific autoantibodies share features of islet cell and 64 kDA antibodies. Diabetologia 36: 785–790
Inman LR, McAllister CT, Chen L et al. (1993) Autoantibodies to the GLUT-2 glucose transporter of B cells in insulin-dependent diabetes mellitus of recent onset. Proc Natl Acad Sci USA 90: 1281–1284
Solimena M, De Camilli P (1993) Spotlight on a neuronal enzyme. Nature 366: 15–17
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Muñoz, A., Gallart, T., Usac, E.F. et al. Anti-islet cell and anti-insulin antibody production by CD5+ and CD5- B lymphocytes in IDDM. Diabetologia 38, 62–72 (1995). https://doi.org/10.1007/BF02369354
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DOI: https://doi.org/10.1007/BF02369354