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Ultrastructural morphometry of blastogenesis I: Transformation of small lymphocytes stimulated in vivo with dinitrochlorobenzene

Cell and Tissue Research volume 200, pages 495–509 (1979)Cite this article

Summary

Changes in the volumes and surfaces of subcellular compartments of unstimulated small lymphocytes and immunoblasts in mouse axillary lymph nodes have been established using stereological techniques. Blast transformation was induced in vivo with dinitrochlorobenzene (DNCB). Cell samples were obtained by random sampling regimes applied at light and electron microscopic levels.

From electron micrographs the volume densities of euchromatin, heterochromatin, nucleoli, mitochondria, Golgi apparatus and rough endoplasmic reticulum were determined. Cell surface/volume ratios were also computed. By estimating mean nuclear volumes using light microscopy, it was possible to calculate absolute compartmental volumes and to evaluate the plasma membrane surface areas of average cells.

Transformation in this model was characterized by a considerable cellular hypertrophy and a substantial increase in plasmalemma surface. Hypertrophy was the consequence of increases in the volumes of all measured intracellular compartments, notably euchromatin and “residual cytoplasm” (including ground cytoplasm and free ribosomes). These changes are discussed in the context of the altered metabolic status of cells.

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References

  • Al-Hamdani, M.M., Atkinson, M.E., Mayhew, T.M.: Changes in the plasma membrane surface of lymphocytes stimulated in vivo with DNCB. Experientia 35, 398–400 (1979)

    Google Scholar 

  • André-Schwartz, J.: The morphological responses of the lymphoid system to homografts III: Electron microscope study. Blood 24, 113–133 (1964)

    Google Scholar 

  • Bach, F.H., Bach, M.L., Sondel, P.M.: Differential function of major histocompatability complex antigens in T-lymphocyte functions. Nature 259, 273–281 (1976)

    Google Scholar 

  • Biberfeld, P.: Morphogenesis in blood lymphocytes stimulated with phytohaemagglutinin (PHA). A light and electron microscopic study. Acta Pathol. Microbiol. Scand. (A) Suppl. 223, 2–70 (1971)

    Google Scholar 

  • Biberfeld, P., Hellstedt, H.: Selective activation of human B-lymphocytes by suboptimal doses of pokeweed mitogen (PWM). Quantitation and ultrastructure of the stimulated cell. Exp. Cell Res. 89, 377–388 (1974)

    Google Scholar 

  • Binet, J.L., Mathé, G.: Optical and electron microscope studies of “immunologically competent cells” in graft reactions. Nature 193, 992–993 (1962)

    Google Scholar 

  • Birbeck, M.S.C., Hall, J.G.: Transformation, in vivo, of basophilic lymph cells into plasma cells. Nature 214, 183–185 (1967)

    Google Scholar 

  • Burri, P.H., Giger, H., Gnägi, H.R., Weibel, E.R.: Application of stereologic methods to cytophysiologic experiments on polarized cells. In: Electron microscopy, Vol. 1 (D.S. Bocciarelli, ed.), p. 953, IV. European Conference, Rome 1968

  • Chalkley, H.W., Cornfield, J., Park, H.: A method for estimating volume-surface ratios. Science 110, 295–297 (1949)

    Google Scholar 

  • Cline, M.J.: The White Cell. Cambridge, Massachusetts: Harvard University Press 1975

    Google Scholar 

  • Cope, G.H., Williams, M.A.: Exocrine secretion in the parotid gland: A stereological analysis at the electron microscopic level of the zymogen granule content before and after isoprenaline-induced degranulation. J. Anat. 116, 269–284 (1973)

    Google Scholar 

  • De Petris, S., Karlsbad, G., Perins, B., Turk, J.L.: Ultrastructure of cells present in lymph nodes during the development of contact sensitivity. Int. Arch. Allergy Appl. Immunol. 29, 112–130 (1966)

    Google Scholar 

  • Douglas, S.D.: Electron microscopic and functional aspects of human lymphocyte response to mitogens. Transplant. Rev. 11, 39–59 (1972)

    Google Scholar 

  • Douglas, S.D., Hoffman, P.F., Borjeson, J., Chessin, L.N.: Studies on human peripheral blood lymphocytes in vitro III. Fine structural features of lymphocyte transformation by pokeweed mitogen. J. Immunol. 98, 17–30 (1967)

    Google Scholar 

  • Dunlap, B., Bach, F.H., Bach, M.L.: Cell surface changes in alloantigen activated T-lymphocytes. Nature 271, 253–255 (1978)

    Google Scholar 

  • Elias, H., Hennig, A., Schwartz, D.E.: Stereology: applications to biomedical research. Physiol. Res. 51, 158–200 (1971)

    Google Scholar 

  • Gowans, J.L., McGregor, D.D., Cowen, D.M., Ford, C.E.: Initiation of immune responses by small lymphocytes. Nature 196, 651–655 (1962)

    Google Scholar 

  • Greaves, M.F., Janossy, G.: Elicitation of selective T and B lymphocyte responses by cell surface binding ligands. Transplant. Rev. 11, 87–130 (1972)

    Google Scholar 

  • Hally, A.D.: A counting method for measuring the volumes of tissue components in microscopical sections. Q. J. Microsc. Sci. 105, 503–517 (1964)

    Google Scholar 

  • Hamburger, J., Dimitriu, A., Banker, L., Debray-Sachs, M., Auvert, J.: Collection of lymph from kidneys homotransplanted in man: cell transformation in vivo. Nature 232, 633–634 (1971)

    Google Scholar 

  • Hayri, P., Defendi, V.: Mixed lymphocyte cultures produce effector cells: model in vitro for allograft rejection. Science 168, 133–135 (1970)

    Google Scholar 

  • Janossy, G., Shohat, M., Greaves, M.F., Dourmashkin, R.R.: Lymphocyte activation: IV. The ultrastructural pattern of response of mouse T and B cells to mitogenic stimulation in vitro. Immunology 24, 211–227 (1972)

    Google Scholar 

  • Konwinski, M., Kozlowski, T.: Morphometric study of normal and phytohaemagglutinin-stimulated lymphocytes. Z. Zellforsch. 129, 500–507 (1972)

    Google Scholar 

  • Mayhew, T.M.: Isolated peritoneal macrophages: component-biased sampling. In: Stereological Methods for Biomorphometry (E.R. Weibel, eds.). New York: Academic Press, 1979, in press

    Google Scholar 

  • Mayhew, T.M., Cruz, L.-M.: Stereological correction procedures for estimating true volume proportions from biased samples. J. Microsc. 99, 287–299 (1973)

    Google Scholar 

  • Mayhew, T.M., Williams, M.A.: A comparison of two sampling procedures for stereological analysis of cell pellets. J. Microsc. 94, 195–204 (1971)

    Google Scholar 

  • Mayhew, T.M., Williams, M A.: A quantitative morphological analysis of macrophage stimulation. I. A study of subcellular compartments and of the cell surface. Z. Zellforsch. 147, 567–588 (1974a)

    Google Scholar 

  • Mayhew, T.M., Williams, M.A.: A quantitative morphological analysis of macrophage stimulation. II. Changes in granule number, size and size distributions. Cell Tissue Res. 150, 529–543 (1974b)

    Google Scholar 

  • Oort, J., Turk, J.L.: A histochemical and autoradiographical study of lymph nodes during the development of contact sensitivity in the guinea-pig. Brit. J. Exp. Pathol. 46, 147–154 (1965)

    Google Scholar 

  • Parrott, D.M.V.: The response of draining lymph nodes to immunological stimulation in intact and thymectomised animals. Suppl. J. Clin. Pathol. 20, 456–465 (1967)

    Google Scholar 

  • Parrott, D.M.V., de Sousa, M.A.B., East, J.: Thymus dependent areas in the lymph organs of neonatally thymectomised mice. J. Exp. Med. 123, 191–203 (1966)

    Google Scholar 

  • Petrzilka, G.E., Graf-de Beer, M., Schroeder, H.E.: Stereological model system for free cells and baseline data for human peripheral blood-derived small T-lymphocytes. Cell Tissue Res. 192, 121–142 (1978)

    Google Scholar 

  • Schroeder, H.E., Graf-de Beer, M.: Stereological analysis of chronic lymphoid cell infiltrates in human gingiva. Arch. Oral Biol. 21, 527–537 (1976)

    Google Scholar 

  • Scothorne, R.J., McGregor, I.A.: Cellular changes in lymph nodes and spleen following skin homografts in the rabbit. J. Anat. 89, 283–292 (1955)

    Google Scholar 

  • Sörén, L., Biberfeld, P.: Quantitative studies on RNA accumulation in human PHA-stimulated lymphocytes during blast transformation. Exp. Cell Res. 79, 359–367 (1973)

    Google Scholar 

  • Turk, J.L.: Cytology of the induction of hypersensitivity. Br. Med. Bull. 23, 3–8 (1967)

    Google Scholar 

  • Underwood, E.E.: Quantitative Stereology. Massachusetts: Addison-Wesley 1970

    Google Scholar 

  • Weibel, E.R.: Stereologic principles for morphometry in electron microscopic cytology. Int. Rev. Cytol. 26, 235–302 (1969)

    Google Scholar 

  • Weibel, E.R.: Coherent test systems for stereological analysis by point counting. In: Newsletter '75 in Stereology (G. Ondracek, ed.), pp. 15–44, Kernforschungszentrum Karlsruhe (1975)

  • Wiener, J., Shapiro, D., Russell, P.S.: An electron microscope study of the homograft reaction. Am. J. Pathol. 44, 319–347 (1964)

    Google Scholar 

  • Wiener, J., Lattes, R.G., Shapiro, D.: An electron microscopic study of leucocyte emigration and vascular permeability in tuberculin sensitivity. Am. J. Path. 50, 485–521 (1967)

    Google Scholar 

  • Woodward, W.D.H.: A stereological ultrastructural study of peritoneal macrophages from germ-free and conventionally-reared mice. Cell Tissue Res. 192, 157–166 (1978)

    Google Scholar 

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Author notes

  1. M. M. Al-Hamdani

    Present address: Department of Anatomy, University of Basrah, Basrah, Iraq

Affiliations

  1. Department of Human Biology and Anatomy, University of Sheffield, S102TN, Sheffield

    M. M. Al-Hamdani,  M. E. Atkinson & T. M. Mayhew

Authors
  1. M. M. Al-Hamdani
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  2. M. E. Atkinson
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  3. T. M. Mayhew
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Additional information

M.M. Al-Hamdani was supported by a grant from the Ministry of Higher Education, Republic of Iraq

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Al-Hamdani, M.M., Atkinson, M.E. & Mayhew, T.M. Ultrastructural morphometry of blastogenesis I: Transformation of small lymphocytes stimulated in vivo with dinitrochlorobenzene. Cell Tissue Res. 200, 495–509 (1979). https://doi.org/10.1007/BF00234859

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  • DOI: https://doi.org/10.1007/BF00234859

Key words

  • T-lymphocytes
  • Blast cells
  • Lymph node
  • Electron microscopy
  • Stereology