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. 1983 Feb 1;157(2):559–571. doi: 10.1084/jem.157.2.559

Experimental autoimmune thyroiditis. In vitro cytotoxic effects of T lymphocytes on thyroid monolayers

PMCID: PMC2186939  PMID: 6296264

Abstract

Effector mechanisms in experimental autoimmune thyroiditis (EAT) were studied in vitro by establishing a cytotoxicity system with thyroid target cells. Lymph node cells (LNC) from popliteal and inguinal lymph nodes were obtained from CBA/J mice (8-10 wk old) 12-18 d after immunization with 120 micrograms mouse thyroglobulin (MTg) in complete Freund's adjuvant (0.2 ml to both hind footpads and thighs) and were cultured with MTg (10-50 micrograms/ml). On day 5 of culture, viable LNC were added to labeled thyroid monolayers and their cytoxicity was assayed after 16 h. Functional thyroid target cells, as reflected by MTg production for up to 9 d, were prepared by adding 1 mM dibutyryl adenosine 3',5'-cyclic monophosphate and 60 microU thyroid-stimulating hormone/ml to the culture medium. On days 5-7, confluent monolayers were labeled with 111In and used as targets. Specific 111In-release ranged from 56 to 85%. The cytotoxic response is MTg specific and H-2 restricted. Pretreatment of thyroid target cells with rabbit antiserum to MTg completely inhibited cytotoxicity. Pretreatment with mouse antiserum to either Kk or Dk products resulted in approximately 50% inhibition, whereas the combined use of both antisera led to total inhibition. No cytotoxicity was observed when control BALB/c thyroid cultures were the target cells. The kinetics of the expansion of Thy-1+ cytotoxic cells by in vitro exposure to MTg were then studied. The cytotoxic response required 5 d to develop and was abolished by treating LNC on day 4 with monoclonal antibody to Lyt-1.1, but not to Lyt-2.1, plus complement. In contrast, by day 5, cytotoxicity was abrogated by similar treatment with antiserum to Lyt-2.1, but not to Lyt-1.1. We conclude that cytotoxic cells derived from MTg-immunized mice are Lyt-2-bearing cells but require the presence of Lyt-1-bearing cells for their generation and/or differentiation.

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Selected References

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  1. Beisel K. W., David C. S., Giraldo A. A., Kong Y. M., Rose N. R. Regulation of experimental autoimmune thyroiditis: mapping of susceptibility to the I-A subregion of the mouse H-2. Immunogenetics. 1982;15(4):427–430. doi: 10.1007/BF00364266. [DOI] [PubMed] [Google Scholar]
  2. Ben-Nun A., Maron R., Ron Y., Cohen I. R. H-2 gene products influence susceptibility of target thyroid gland to damage in experimental autoimmune thyroiditis. Eur J Immunol. 1980 Feb;10(2):156–159. doi: 10.1002/eji.1830100216. [DOI] [PubMed] [Google Scholar]
  3. Bidey S. P., Marsden P., Anderson J., McKerron C. G., Berry H. In-vitro studies of normal human thyroid cells: responses to thyrotrophin and dibutyryl cyclic AMP. J Endocrinol. 1977 Jan;72(1):87–96. doi: 10.1677/joe.0.0720087. [DOI] [PubMed] [Google Scholar]
  4. Bourke J. R., Carseldine K. L., Ferris S. H., Huxham G. J., Manley S. W. Changes in membrane potential of cultured porcine and human thyroid cells in response to thyrotrophin and other agents. J Endocrinol. 1981 Feb;88(2):187–196. doi: 10.1677/joe.0.0880187. [DOI] [PubMed] [Google Scholar]
  5. Braley-Mullen H., Tompson J. G., Sharp G. C., Kyriakos M. Adoptive transfer of experimental autoimmune thyroiditis (EAT) with in vitro activated lymph node cells from thyroglobulin-sensitized guinea pigs: characterization of the cell that transfers EAT. J Immunol. 1981 Nov;127(5):1767–1771. [PubMed] [Google Scholar]
  6. Calder E. A., Irvine W. J., Davidson N. M., Wu F. T, B and K cells in autoimmune thyroid disease. Clin Exp Immunol. 1976 Jul;25(1):17–22. [PMC free article] [PubMed] [Google Scholar]
  7. Charreire J., Michel-Bechet M. Syngeneic sensitization of mouse lymphocytes on monolayers of thyroid epithelial cells. III. Induction of thyroiditis by thyroid-sensitized T lymphoblasts. Eur J Immunol. 1982 May;12(5):421–425. doi: 10.1002/eji.1830120512. [DOI] [PubMed] [Google Scholar]
  8. Charreire J. Syngeneic sensitization of mouse lymphocytes on monolayers of thyroid epithelial cells. II. T and B cell involvement in primary responses. Eur J Immunol. 1982 May;12(5):416–421. doi: 10.1002/eji.1830120511. [DOI] [PubMed] [Google Scholar]
  9. ElRehewy M., Kong Y. M., Giraldo A. A., Rose N. R. Syngeneic thyroglobulin is immunogenic in good responder mice. Eur J Immunol. 1981 Feb;11(2):146–151. doi: 10.1002/eji.1830110216. [DOI] [PubMed] [Google Scholar]
  10. Esquivel P. S., Kong Y. C., Rose N. R. Evidence for thyroglobulin-reactive T cells in good responder mice. Cell Immunol. 1978 Apr;37(1):14–19. doi: 10.1016/0008-8749(78)90169-7. [DOI] [PubMed] [Google Scholar]
  11. Esquivel P. S., Rose N. R., Kong Y. C. Induction of autoimmunity in good and poor responder mice with mouse thyroglobulin and lipopolysaccharide. J Exp Med. 1977 May 1;145(5):1250–1263. doi: 10.1084/jem.145.5.1250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frost P., Smith J., Frost H. The radiolabeling of lymphocytes and tumor cells with 111indium. Proc Soc Exp Biol Med. 1978 Jan;157(1):61–65. doi: 10.3181/00379727-157-39991. [DOI] [PubMed] [Google Scholar]
  13. Kong Y. C., Rose N. R., Elrehewy M., Michaels R., Giraldo A. A., Accavitti M. A., Leon M. A. Thyroid alloantigens in autoimmunity. Transplant Proc. 1980 Sep;12(3 Suppl 1):129–134. [PubMed] [Google Scholar]
  14. Kong Y. M., Okayasu I., Giraldo A. A., Beisel K. W., Sundick R. S., Rose N. R., David C. S., Audibert F., Chedid L. Tolerance to thyroglobulin by activating suppressor mechanisms. Ann N Y Acad Sci. 1982;392:191–209. doi: 10.1111/j.1749-6632.1982.tb36108.x. [DOI] [PubMed] [Google Scholar]
  15. Kong Y., David C. S., Giraldo A. A., Elrehewy M., Rose N. R. Regulation of autoimmune response to mouse thyroglobulin: influence of H-2D-end genes. J Immunol. 1979 Jul;123(1):15–18. [PubMed] [Google Scholar]
  16. Laryea E., Row V. V., Volpé R. The effect of blood leucocytes from patients with Hashimoto's disease on human thyroid cells in monolayer culture. Clin Endocrinol (Oxf) 1973 Jan;2(1):23–35. doi: 10.1111/j.1365-2265.1973.tb03481.x. [DOI] [PubMed] [Google Scholar]
  17. Lin M. S., Salvin S. B. In vitro and in vivo studies on the mechanism of experimental autoimmune thyroiditis in guinea pigs. Cell Immunol. 1976 Dec;27(2):177–187. doi: 10.1016/0008-8749(76)90227-6. [DOI] [PubMed] [Google Scholar]
  18. Livezey M. D., Sundick R. S., Rose N. R. Spontaneous autoimmune thyroiditis in chickens. II. Evidence for autoresponsive thymocytes. J Immunol. 1981 Oct;127(4):1469–1472. [PubMed] [Google Scholar]
  19. Okayasu I., Kong Y. M., David C. S., Rose N. R. In vitro T-lymphocyte proliferative response to mouse thyroglobulin in experimental autoimmune thyroiditis. Cell Immunol. 1981 Jun;61(1):32–39. doi: 10.1016/0008-8749(81)90351-8. [DOI] [PubMed] [Google Scholar]
  20. Podleski W. K. Cytodestructive mechanisms provoked by lymphocytes. Am J Med. 1976 Jul;61(1):1–8. doi: 10.1016/0002-9343(76)90023-1. [DOI] [PubMed] [Google Scholar]
  21. Rose N. R., Accavitti M., Pydyn E. F., Leon M. A., Brown R. K. The use of hybridoma antibodies to probe the antigenic determinants of thyroglobulin. Adv Exp Med Biol. 1982;150:23–35. doi: 10.1007/978-1-4684-4331-8_2. [DOI] [PubMed] [Google Scholar]
  22. Rose N. R., Kong Y. C., Okayasu I., Giraldo A. A., Beisel K., Sundick R. S. T-cell regulation in autoimmune thyroiditis. Immunol Rev. 1981;55:299–314. doi: 10.1111/j.1600-065x.1981.tb00346.x. [DOI] [PubMed] [Google Scholar]
  23. Rose N. R., Twarog F. J., Crowle A. J. Murine thyroiditis: importance of adjuvant and mouse strain for the induction of thyroid lesions. J Immunol. 1971 Mar;106(3):698–704. [PubMed] [Google Scholar]
  24. Tötterman T. H. Distribution of T-, B-, and thyroglobulin-binding lymphocytes infiltrating the gland in Graves' disease, Hashimoto's thyroiditis, and de Quervain's thyroiditis. Clin Immunol Immunopathol. 1978 Jul;10(3):270–277. doi: 10.1016/0090-1229(78)90181-2. [DOI] [PubMed] [Google Scholar]
  25. Vladutiu A. O., Rose N. R. Autoimmune murine thyroiditis relation to histocompatibility (H-2) type. Science. 1971 Dec 10;174(4014):1137–1139. doi: 10.1126/science.174.4014.1137. [DOI] [PubMed] [Google Scholar]
  26. Vladutiu A. O., Rose N. R. Cellular basis of the genetic control of immune responsiveness to murine thyroglobulin in mice. Cell Immunol. 1975 May;17(1):106–113. doi: 10.1016/s0008-8749(75)80010-4. [DOI] [PubMed] [Google Scholar]
  27. Yeni P., Charreire J. Syngeneic sensitization of mouse lymphocytes on monolayers of thyroid epithelial cells. I. Study of proliferative response. Cell Immunol. 1981 Aug;62(2):313–323. doi: 10.1016/0008-8749(81)90332-4. [DOI] [PubMed] [Google Scholar]

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