Diabetologia

, Volume 32, Issue 5, pp 282–289 | Cite as

The natural history of lymphocyte subsets infiltrating the pancreas of NOD mice

  • A. Signore
  • P. Pozzilli
  • E. A. M. Gale
  • D. Andreani
  • P. C. L. Beverley
Originals

Summary

A longitudinal study of lymphocytic infiltration in the endocrine pancreas of non-obese diabetic mice was performed to investigate the role of different lymphocyte subsets in the pathogenesis of diabetes. The incidence of insulitis and the percentage of mononuclear cell subsets in the pancreas were evaluated in non-obese diabetic mice of various ages (5, 9, 13, 17, 22, 29 and 36 weeks). Cryostat sections of pancreas were stained with heamatoxilin-eosin or with different monoclonal antibodies against total T lymphocytes, helper T lymphocytes, cytotoxic/suppressor T lymphocytes, activated interleukin 2 receptor positive lymphocytes and B lymphocytes. A monoclonal antibody against Class-II antigens was also used. Positive cells were revealed by the immunoperoxidase technique. Insulitis was found in 5 weeks old mice but to a lesser extent than in adult animals. No significant variation between infiltrating cell subsets was found in different age groups. T lymphocytes ranged between 20.4% and 28.1%, B lymphocytes between 28.8% and 30.8% and Class-II positive cells between 22.8% and 32.2%. Interleukin 2 receptor positive cells ranged between 5.5% and 8.5% as detected with AMT-13 monoclonal antibody which recognise the interleukin 2 binding site. A higher percentage of activated cells was observed using another monoclonal antibody (7D4) directed against a different epitope of the interleukin 2 receptor, suggesting the presence of activated lymphocytes with interleukin 2 receptors saturated by interleukin 2. No insulin-containing cells were found to express Class-II molecules as demonstrated by a double immunofluorescence technique. Most infiltrating mononuclear cells were found to be positive for Class-II and L3T4 antigens or to be Class-II positive and express surface immunoglobulins. We conclude that pancreatic infiltration is an early expression of autoimmune phenomena occurring in these mice and that monocytes and B-lymphocytes predominate in the infiltrate. The role of interleukin 2 and interleukin 2 receptor positive lymphocytes in inducing and maintaining the immune response towards B cells is discussed.

Key words

Non-obese diabetic mouse insulitis lymphocyte subsets Class-II antigens interleukin 2 receptor insulin-dependent diabetes 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Fujita F, Yui R, Kusumoto Y, Serizawa Y, Makino S, Tochino Y (1982) Lymphocytic insulitis in ‘Non-Obese Diabetic (NOD)’ strain of mice: an immunohistochemical and electron microscope investigation. Biomed Res 3: 429–443Google Scholar
  2. 2.
    Miyazaki A, Hanafusa T, Yamada K, Miyagawa J, Fujino-Kurihara H, Nakajima H, Nonaka K, Tarai S (1985) Predominance of T lymphocytes in the pancreatic islets and spleen of prediabetic non obese diabetic (NOD) mice: a longitudinal study. Clin Exp Immunol 60: 622–630Google Scholar
  3. 3.
    Hattori M, Buse JB, Jackson RA, Glimcher L, Dorf ME, Minami M, Makino S, Moriwaki K, Kuzuya H, Imura H, Strauss WM, Seidman JG, Eisenbarth GS (1986) The NOD mouse: recessive diabetogenic gene in the major histocompatibility complex. Science 231: 733–735Google Scholar
  4. 4.
    Prochazka M, Leiter EH, Serreze DV, Coleman DL (1987) Three recessive loci required for insulin-dependent diabetes in nonobese diabetic mice. Science 237: 286–289Google Scholar
  5. 5.
    Kanazawa Y, Komeda K, Sato S, Mori S, Akanuma K, Takaku F (1984) Non-Obese-Diabetic mice: immune mechanisms of pancreatic B-cell destruction. Diabetologia 27: 113–115Google Scholar
  6. 6.
    Pontesilli O, Carotenuto P, Gazda LS, Pratt PF, Prowse SJ (1987) Circulating lymphocyte populations and autoantibodies in nonobese diabetic (NOD) mice: a longitudinal study. Clin Exp Immunol 70: 84–93Google Scholar
  7. 7.
    Makino S, Harada M, Kishimoto Y, Hayashi Y (1986) Absence of insulitis and overt diabetes in athymic nude mice with NOD genetic background. Exp Anim 35: 495–498Google Scholar
  8. 8.
    Harada M, Makino S (1986) Suppression of overt diabetes in NOD mice by anti-thymocyte serum or anti-Thy 1,2 antibody. Exp Anim 35: 501–504Google Scholar
  9. 9.
    Bendelac A, Carnaud C, Boitard C, Bach JF (1987) Syngeneic transfer of autoimmune diabetes from diabetic NOD mice to healthy neonates. J Exp Med 166: 823–832Google Scholar
  10. 10.
    Wang Y, Hao L, Gill RG, Lafferty KJ (1987) Autoimmune diabetes in NOD mouse is L3T4 T-Iymphocyte dependent. Diabetes 36: 535–538Google Scholar
  11. 11.
    Shizuru JA, Taylor-Edwardd C, Banks BA, Gregory AK, Fathman CG (1988) Immunotherapy of the Nonobese Diabetic mouse: treatment with an antibody to T-helper lymphocytes. Science 240: 659–662Google Scholar
  12. 12.
    Momburg F, Koch N, Moller P, Moldenhauer G, Butcher G, Hammerling GJ (1986) Differential expression of Ia and Ia-associated invariant chain in mouse tissues after in vivo treatment with IFN-γ. J Immunol 136: 940–948Google Scholar
  13. 13.
    Osawa H, Diamenstein T (1984) A rat monoclonal antibody that binds specifically to mouse T lymphoblasts and inhibits IL2 receptor function: a putative anti-IL2r Ab. J Immunol 132: 2445–2450Google Scholar
  14. 14.
    Malek TR, Robb RJ, Shevach EM (1983) Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex. Proc Natl Acad Sci USA 80: 5694–5698Google Scholar
  15. 15.
    Rowe DJ, Isemberg DA, Beverley PCL (1983) Monoclonal antibodies to human leukocyte antigens in polymyositis and muscolar dystrophy. Clin Exp Immunol 54: 327–336Google Scholar
  16. 16.
    Fujino-Kurihara H, Fujita H, Hakura A, Nonaka K, Tarui S (1985) Morphological aspects on pancreatic islets of non-obese diabetic (NOD) mice. Virchows Arch [Cell Pathol] 49: 107–120Google Scholar
  17. 17.
    Yale JF, Marliss EB (1984) Altered immunity and diabetes in the BB rat. Clin Exp Immunol 57: 1–11Google Scholar
  18. 18.
    Dean BM, Walker R, Bone AJ, Baird JD, Cooke A (1985) Prediabetes in the spontaneously diabetic BB/E rat: lymphocyte subpopulations in the pancreatic infiltrate and expression of rat MHC class II molecules in endocrine cells. Diabetologia 28: 464–466Google Scholar
  19. 19.
    Kataoka S, Satoh J, Fujiya H, Toyota T, Suzuki R, Itoh K, Kumagai K (1983) Immunologic aspects of the Nonobese Diabetic (NOD) mouse. Abnormalities of cellular immunity. Diabetes 32: 247–253Google Scholar
  20. 20.
    Pozzilli P, Sensi M, Al-Sakkaf L, Tarn A, Zuccarini O, Bottazzo GF (1984) Prospective study of lymphocyte subsets in genetically susceptible subjects to Type 1 diabetes. Diabetologia 27: 132–135Google Scholar
  21. 21.
    Tarn A, Dean BM, Thomas JM, Ingram D, Schewarz G, Bottazzo GF, Gale EAM (1988) Predicting insulin dependent diabetes. Lancet I: 845–850Google Scholar
  22. 22.
    Bottazzo GF, Dean BM, McNally JM, MacHay 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: 353–360Google Scholar
  23. 23.
    Foulis AK, Liddle CN, Farquharson MA, Richmond JA, Weir RS (1986) The histopathology of the pancreas in Type 1 (insulin-dependent) diabetes mellitus: a 25 year review of deaths in patients under 20 years of age in United Kingdom. Diabetologia 29: 267–274Google Scholar
  24. 24.
    Signore A, Cooke A, Pozzilli P, Butcher G, Simpson E, Beverley PCL (1987) Class-II and IL2 receptor positive cells in the pancreas of NOD mice. Diabetologia 30: 902–905Google Scholar
  25. 25.
    Talle MA, Rao PE, Westberg E, Allegar N, Makowski M, Mittler RS, Goldstein G (1983) Patterns of antigenic expression on human monocytes as defined by monoclonal antibodies. Cell Immunol 78: 83–99Google Scholar
  26. 26.
    Flavell RA, Alien H, Burkly LC, Scherman DH, Waneck GL, Widera G (1986) Molecular biology of the H-2 histocompatibility complex. Science 233: 437–443Google Scholar
  27. 27.
    Araneo BA, Yowell RL (1985) MHC-linked immune responce suppression mediated by T cells bearing I-A-encoded determinants. J Immunol 135: 73–79Google Scholar
  28. 28.
    Lee K, Amano K, Yoon J (1988) Evidence for initial involvement of macrophage in the development of insulitis in NOD mice. Diabetes 37: 989–991Google Scholar
  29. 29.
    Kiesel U, Oschilewski M, Kantwerk G, Maruta M, Hanenberg H, Treichel U, Kolb-Bachofen V, Hartung HP, Kolb H (1986) Essential role of macrophages in the development of Type 1 diabetes in BB rats. Transplant Proc 18: 1525–1527Google Scholar
  30. 30.
    Kolb H, Zielasek J, Treichel U, Freitag G, Wrann M, Kiesel U (1986) Recombinant interleukin 2 enhances spontaneous insulin-dependent diabetes in BB rats. Eur J Immunol 16: 209–212Google Scholar
  31. 31.
    Kelley VE, Gaulton GN, Hattori M, Ikegami H, Eisenbarth G, Strom TB (1988) Anti-interleukin 2 receptor antibody suppresses murine diabetic insulitis and lupus nephritis. J Immunol 140: 59–61Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • A. Signore
    • 1
    • 3
  • P. Pozzilli
    • 2
    • 3
  • E. A. M. Gale
    • 2
  • D. Andreani
    • 3
  • P. C. L. Beverley
    • 1
  1. 1.ICRF Human Tumour Immunology GroupUniversity College and Middlesex School of MedicineUK
  2. 2.Department of Diabetes and ImmunogeneticsSt. Bartholomew's HospitalLondonUK
  3. 3.Cattedra Endocrinologia (I), onClinica Medica (II)University of Rome “La Sapienza”RomaItaly

Personalised recommendations