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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
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)
André-Schwartz, J.: The morphological responses of the lymphoid system to homografts III: Electron microscope study. Blood 24, 113–133 (1964)
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)
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)
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)
Binet, J.L., Mathé, G.: Optical and electron microscope studies of “immunologically competent cells” in graft reactions. Nature 193, 992–993 (1962)
Birbeck, M.S.C., Hall, J.G.: Transformation, in vivo, of basophilic lymph cells into plasma cells. Nature 214, 183–185 (1967)
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)
Cline, M.J.: The White Cell. Cambridge, Massachusetts: Harvard University Press 1975
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)
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)
Douglas, S.D.: Electron microscopic and functional aspects of human lymphocyte response to mitogens. Transplant. Rev. 11, 39–59 (1972)
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)
Dunlap, B., Bach, F.H., Bach, M.L.: Cell surface changes in alloantigen activated T-lymphocytes. Nature 271, 253–255 (1978)
Elias, H., Hennig, A., Schwartz, D.E.: Stereology: applications to biomedical research. Physiol. Res. 51, 158–200 (1971)
Gowans, J.L., McGregor, D.D., Cowen, D.M., Ford, C.E.: Initiation of immune responses by small lymphocytes. Nature 196, 651–655 (1962)
Greaves, M.F., Janossy, G.: Elicitation of selective T and B lymphocyte responses by cell surface binding ligands. Transplant. Rev. 11, 87–130 (1972)
Hally, A.D.: A counting method for measuring the volumes of tissue components in microscopical sections. Q. J. Microsc. Sci. 105, 503–517 (1964)
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)
Hayri, P., Defendi, V.: Mixed lymphocyte cultures produce effector cells: model in vitro for allograft rejection. Science 168, 133–135 (1970)
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)
Konwinski, M., Kozlowski, T.: Morphometric study of normal and phytohaemagglutinin-stimulated lymphocytes. Z. Zellforsch. 129, 500–507 (1972)
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
Mayhew, T.M., Cruz, L.-M.: Stereological correction procedures for estimating true volume proportions from biased samples. J. Microsc. 99, 287–299 (1973)
Mayhew, T.M., Williams, M.A.: A comparison of two sampling procedures for stereological analysis of cell pellets. J. Microsc. 94, 195–204 (1971)
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)
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)
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)
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)
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)
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)
Schroeder, H.E., Graf-de Beer, M.: Stereological analysis of chronic lymphoid cell infiltrates in human gingiva. Arch. Oral Biol. 21, 527–537 (1976)
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)
Sörén, L., Biberfeld, P.: Quantitative studies on RNA accumulation in human PHA-stimulated lymphocytes during blast transformation. Exp. Cell Res. 79, 359–367 (1973)
Turk, J.L.: Cytology of the induction of hypersensitivity. Br. Med. Bull. 23, 3–8 (1967)
Underwood, E.E.: Quantitative Stereology. Massachusetts: Addison-Wesley 1970
Weibel, E.R.: Stereologic principles for morphometry in electron microscopic cytology. Int. Rev. Cytol. 26, 235–302 (1969)
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)
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)
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)
Author information
Authors and Affiliations
Additional information
M.M. Al-Hamdani was supported by a grant from the Ministry of Higher Education, Republic of Iraq
Rights and permissions
About this article
Cite this article
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
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00234859