Advertisement

Virchows Archiv B

, 19:269 | Cite as

Histological and autoradiographical findings in the immunologically stimulated spleen

  • B. Helpap
  • V. Grouls
  • K. Yamashita
Article
  • 10 Downloads

Summary

In order to estimate the role of a possible immunological coeffect on the splenic cellular proliferation during wound healing after mechanical or thermal lesions of internal organs, light microscopical and autoradiographical investigations with tritiated thymidine were performed on spleens of non-germfree rats, immunized by a single injection of sheep erythrocytes. Two days after an initial dissociation of preexisting germinal centers in the spleen a steep rise of the percentages of labeled cells in newly formed germinal centers occurs with a maximum on the fifth day. In the marginal zone an increased cell proliferation (mostly large lymphoid blasts) starts at 12 hours. In the lymphatic mantle zone a marked increase of labeled basophilic blasts (immunoblasts) can be observed with a maximum on the second day. The values are higher in the perifollicular B cell region than in the periarteriolar T cell region. In the red pulp of the spleen the highest percentages of labeled cells occur five days after the antigen injection.

These findings in the spleen after a strong antigenic stimulation are characteristic for an immunological reaction of anamnestic type. In comparison with investigations of the spleen after cryolesions in internal organs such studies may be particularly helpful in judging the proportion of an unknown immunological reaction in the spleen after such an operation, which by tissue necroses and cellular destruction may have caused denaturation of self components.

Key words

Injection of sheep erythrocytes Immunology 3H-thymidine autoradiography Spleen reaction 

Zusammenfassung

Um den Anteil einer möglichen immunologischen Reaktion in der Milz bei Wundheilungsvergängen nach mechanischen oder thermischen Läsionen an inneren Organen abschätzen zu können, wurden histologische und autoradiographische Untersuchungen mit H-3-Thymidin an der weißen und roten Pulpa von Ratten nach Gabe von Schaferythrozyten durchgeführt.

Histologisch kommt es zwischen 12–48 Stunden nach der Antigeninjektion zu einer Keimzentrumsatrophie bzw. einem vollständigen Verlust bestehender Keimzentren, die jedoch ab 3. Tag neugebildet und zunehmend hyperplastisch werden. Zum Zeitpunkt dieser sog. Keimzentrumsdissoziation steigt die Zahl großer pyroninophiler lymphoider Zellen (Immunoblasten) in der Marginalzone und im Lymphozytenwall an. Autoradiographisch läßt sich eine deutliche Zunahme der Prozentsätze radioaktiv markierter Zellen in der Marginalzone und in der perifollikulären Zone 12 Stunden nach der Antigeninjektion beobachten. In der periarteriolären Zone des Lymphozytenwalles setzt der Anstieg markierter Zellen etwas verspätet ein. Die höchsten Werte liegen in der B- und T-Zellregion nach 2 Tagen vor. In den Keimzentren erreicht die Proliferation ihren Höhepunkt zwischen dem 5. und 10. Versuchstag. In der roten Pulpa sind die höchsten Prozentsätze radioaktiv-markierter Zellen am 5. Tag nach antigener Stimulation zu beobachten.

Die Befunde in der Milz nach Injektion von Schaferythrozyten sprechen für das Vorliegen einer anamnestischen immunologischen Reaktion. Wegen dieses charakteristischen proliferativen Verhaltens der Zellen in der weißen und roten Pulpa nach antigener Stimulation können derartige morphologisch-zellkinetischen Untersuchungen hilfreich sein in der analogen Beurteilung von zellulären Milzreaktionen bei Wundheilungsvorgängen, bei denen ein immunologischer Prozeß vermutet wird, wie z.B. nach Kryotherapie.

References

  1. Bürki, H., Cottier, H., Hess, M. W., Laissue, J., Stoner, R. D.: Distinctive medullary and germinal center proliferative patterns in mouse lymph nodes after regional primary and secondary stimulation with tetanus toxoid. J. Immunol.112, 1961–1970 (1974)Google Scholar
  2. Bürki, H., Luscieti, P., Pedrinis, E., Schädeli, J., Hess, H. W., Cottier, H.: Milz und Antikörperbildung. Schweiz. med. Wschr.104, 1351–1360 (1974)PubMedGoogle Scholar
  3. Congdon, C. C.: Effect of injection of foreign bone marrow on the lymphatic tissues of normal mice. J. Natl. Cancer Inst.28, 305–329 (1962)PubMedGoogle Scholar
  4. Congdon, C. C.: The early histologic effects of antigenic stimulation. Arch. Path.78, 83–96 (1964)PubMedGoogle Scholar
  5. Congdon, C. C., Makinodan, T.: Splenic white pulp alteration after antigen injection: relation to time of serum antibody production. Amer. J. Path.39, 697–709 (1961)PubMedGoogle Scholar
  6. Grobler, P., Bürki, H., Cottier, H., Hess, M. W., Stoner, R. D.: Cellular bases for relative radioresistance of the antibody forming system at advanced stages of the secondary response to tetanus toxoid in mice. J. Immunol.112, 2154–2165 (1974)PubMedGoogle Scholar
  7. Hanna, M. G., Jr.: An autoradiographic study of the germinal center in spleen white pulp during early intervals of the immune response. Lab. Invest.13, 95–104 (1964)PubMedGoogle Scholar
  8. Hanna, M. G., Jr., Congdon, C. C., Wust, C. J.: Effect of antigen dose on lymphatic tissue germinal center changes. Proc. Soc. exp. Biol. Med.121, 286–290 (1966)PubMedGoogle Scholar
  9. Hanna, M. G., Jr., Swartzendruber, D. C., Congdon, C. C.: Morphologic and autoradiographic studies of spleen white pulp germinal centers after antigen injection. In: H. Cottier, N. Odartschenko, R. Schindler, C. C. Congdon (Eds.): Germinal centers in immunse response, pp. 189–198. Berlin-Heidelberg-New York: Springer 1967Google Scholar
  10. Hanna, M. G., Jr., Szakal, A. K., Walburg, H. E.: The relation of antigen and virus localization to the development and growth of lymphoid germinal centers. Adv. exp. Med. Biol.5, 149–164 (1969)Google Scholar
  11. Helpap, B., Grouls, V., Yamashita, K., Breining, H., Lymberopoulos, S.: The cellular reaction of the spleen after cryolesions in parenchymal organs. Histologic and autoradiographic studies with3H-thymidine. Minerva-med.65, 3655–3658 (1974a)PubMedGoogle Scholar
  12. Helpap, B., Yamashita, K., Breining, H., Lymberopoulos, S.: Autoradiographische Untersuchungen mit3H-Thymidin an der Rattenmilz nach Kryonekrose. Experientia 30, 273–275 (1974b)PubMedCrossRefGoogle Scholar
  13. Helpap, B., Yamashita, K., Grouls, V., Kasprzyk, R.: Zur zellulären Reaktion der Mäusemilz nach traumatischer Schädigung der Leber. Virchows Arch. B. Zellpath.15, 237–250 (1974c)Google Scholar
  14. Helpap, B., Grouls, V., Yamashita, K., Breining, H.: The proliferative response of the spleen in cryosurgery. Cryobiology (in press)Google Scholar
  15. Helpap, B., Grouls, V., Yamashita, K.: Zum zellulären Verhalten der Rattenmilz nach großem Parenchymverlust der Leber. Beitr. Path.156, 16–31 (1975)Google Scholar
  16. Jacobson, E. B., Thorbecke, G. J.: Relationship of germinal centres in lymphoid tissue to immunologic memory. IV. Formation of 19 S and 7 S antibody by splenic white and red pulp during the secondary response in vitro. Lab. Invest.19, 635–642 (1968)PubMedGoogle Scholar
  17. Keuning, F. J., Bos, W. H.: Regeneration patterns of lymphoid follicles in the rabbit spleen after sublethal X-irradiation. In: H. Cottier, N. Odartchenko, R. Schindler, C. C. Congdon (Eds.): Germinal centers in immune response, pp. 250–257, Berlin-Heidelberg-New York: Springer 1967Google Scholar
  18. Koburg, E.: Cell production and cell migration in the tonsil. In: H. Cottier, N. Odartschenko, R. Schindler, C. C. Congdon (Eds.): Germinal centers in immune response, pp. 176–182, Berlin-Heidelberg-New York: Springer 1967Google Scholar
  19. Langevoort, H. L.: The histophysiology of antibody response. I. Histogenesis of plasma cell reaction in rabbit spleen. Lab. Invest.12, 106–118 (1963)PubMedGoogle Scholar
  20. Lewerenz, M., Laissue, J., Bürki, H., Bürki, K.: Das Verhalten von Keimzentrenzellen der Mäusemilz nach Sekundärinjektion von Tetanustoxoid in die Fußsohlen. Autoradiographische Untersuchungen mit Hilfe von Thymidin-3H. Virchows Arch. B Zellpath.2, 203–219 (1969)Google Scholar
  21. Lukes, R. J.: The pathology of the white pulp of the spleen. In: K. Lennert, D. Harms (Eds.): The spleen, pp. 130–138, Berlin-Heidelberg-New York: Springer 1970Google Scholar
  22. Mellbye, O. J.: Spleen and immunity. In: K. Lennert, D. Harms (Eds.): The spleen, pp. 139–144, Berlin-Heidelberg-New York: Springer 1970)Google Scholar
  23. Miller, J. F. A. P.: Lymphocyte interactions in antibody responses. Intern. Rev. Cytol.33, 77–130 (1972)CrossRefGoogle Scholar
  24. Mitchell, J., Abbot, A.: Antigens in immunity. XVI. A light and electron microscope study of antigen localization in the rat spleen. Immunology21, 207–224 (1971)PubMedGoogle Scholar
  25. Nieuwenhuis, P.: Histophysiology of germinal centers and their role in antibody response: an autoradiographic study in the rabbit. Adv. exp. Med. Biol.5, 113–120 (1969)Google Scholar
  26. Nieuwenhuis, P., Keuning, F. J.: Germinal centres and the origin of the B-cell system. II. Germinal centers in the rabbit spleen and popliteal lymph nodes. Immunology26, 509–519 (1974a)PubMedGoogle Scholar
  27. Nieuwenhuis, P., van Nouhuijs, C. E., Eggens, J. H., Keuning, F. J.: Germinal centres and the origin of the B-cell system. I. Germinal centres in the rabbit appendix. Immunology26, 497–507 (1974b)PubMedGoogle Scholar
  28. Nossal, G. J. V., Austin, C. M., Pye, J., Mitchell, J.: Antigens in immunity. XII. Antigen trapping in the spleen. Int. Arch. Allergy29, 368–383 (1966)PubMedGoogle Scholar
  29. Nossal, G. J. V., Abbot, A., Mitchell, J., Lummus, Z.: Antigens in immunity XV. Ultrastructural features of antigen capture in primary and secondary lymphoid follicles. J. exp. Med.127, 277–290 (1968)PubMedCrossRefGoogle Scholar
  30. Pelc, S. R.: Incorporation of (3H)-thymidine in mouse spleen. J. Cell Sci.3, 263–272 (1968)PubMedGoogle Scholar
  31. Pettersen, J. C., Borgen, D. F., Graupner, K. C.: A morphological and histochemical study of the primary and secondary immune responses in the rat spleen. Amer. J. Anat.121, 305–318 (1967)PubMedCrossRefGoogle Scholar
  32. Richardson, K. C., Jartett, L., Finke, E. H.: Embedding in epoxy resins for ultrathin sectoning in electron microscopy. Stain Technol.35, 313–323 (1960)PubMedGoogle Scholar
  33. Rooijen, van N.: The vascular pathways in the white pulp of the rabbit spleen. Acta morph. neerl.-scand.10, 351–357 (1972)Google Scholar
  34. Rooijen, van N.: Antigens in the spleen: localization of labelled antigen in the spleen of normal rabbits and rabbits preimmunized with a non related antigen. Z. Immun. Forsch.144, 442–449 (1973a)Google Scholar
  35. Rooijen, van N.: Mechanism of follicular antigen trapping. Migration of antigen-antibody complexes from marginal zone towards follicle centres. Immunology25, 847–852 (1973b)PubMedGoogle Scholar
  36. Rooijen, van N.: Nonspecific nature of antigen trapping in lymphoid follicles. A comparative study on the localization of the antigens3H-DNP-BGG and125J-HGG in the spleen of rabbits, using autoradiographic technique for distinction between two isotopes in the same tissue section. Immunology25, 853–867 (1973c)PubMedGoogle Scholar
  37. Rooijen, van N.: Mechanism of follicular antigen trapping. Evidence for a two cell mechanism using double isotope autoradiography. Immunology27, 617–622 (1974)PubMedGoogle Scholar
  38. Sargent, A. U., Myers, J., Richter, M.: The immune response to circulating autologous hepatocellular antigens. J. Immunol.96, 268–272 (1966)PubMedGoogle Scholar
  39. Thunold, S.: Antigen binding cells in spleen sections from mice immunized with sheep erythrocytes. Int. Arch. Allergy44, 529–543 (1973)PubMedGoogle Scholar
  40. Veerman, A. J. P., van Ewijk, W.: White pulp compartments in the spleen of rats and mice. A light and electron microscopic study of lymphoid and non-lymphoid celltypes in T- and B-areas. Cell Tiss. Res.156, 417–441 (1975)CrossRefGoogle Scholar
  41. Wakefield, J., Thorbecke, G. J.: Relationship of germinal centers in lymphoid tissue to immunological memory. II. The detection of primed cells and their proliferation upon cell transfer to lethally irradiated syngeneic mice. J. exp. Med.128, 171–185 (1968)PubMedCrossRefGoogle Scholar
  42. Yamashita, K., Helpap, B.: A staining method of epoxy-embedded lymphatic and hemopoietic tissues in autoradiograms. Histochemistry40, 275–279 (1974)PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • B. Helpap
    • 1
  • V. Grouls
    • 1
  • K. Yamashita
    • 1
  1. 1.Institute of Pathology of the University of BonnGermany

Personalised recommendations