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Ultrastructural studies of peripheral blood lymphocytes in T cell-depleted rabbits

A scanning- and transmission electron microscopic analysis

Cell and Tissue Research volume 213, pages 221–235 (1980)Cite this article

Summary

Density separation of purified peripheral blood leucocytes from T-cell depleted rabbits on a linear Ficoll-metrizoate gradient has been applied to obtain different leucocyte fractions. Two lymphocyte fractions separated on density seem to have different characteristics, both morphologically and immunologically. In this study these two fractions have been characterized ultrastructurally by using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and a relationship has been established between the surface architecture (SEM), the cell size (SEM/TEM) and surface-Ig/C3-receptors (LM, light microscopy). Finally three types of lymphocytes have been described in the two lymphocyte fractions separated on density. Morphometric information such as cell size, cell shape, eu-/heterochromatin ratio in the nucleus and the nucleus-/cell ratio have been correlated to the stage of activation of the B lymphocyte in a representative density separation.

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References

  • Alexander E, Wetzel B (1975) Human lymphocytes: Similarity of B and T cell surface morphology. Science 188:732–734

    Google Scholar 

  • Anderson TF (1951) Techniques for preservation of three dimensional structure in preparing specimens for the electron microscope. Trans N Y Acad Sci 13:130–134

    Google Scholar 

  • Barber TA, Burkholder PM (1975) Relation of surface and internal ultrastructure of thymus and bone marrow derived lymphocytes to specimens preparatory technique. Proceedings of the eighth annual scanning electron microscope symposium IIT research institute, Chicago 369–378

  • Bast BJEG, Catty D, Manten-Slingerlandt R, Jansen JTG, Veldhuis DH, Roholl PJM, Ballieux RE Surface Ig on rabbit lymphocytes. (Accepted for publication, Eur J Immunol)

  • Bell C, Wigzell H (1977) Rabbits possess conventional T lymphocytes. Eur J Immunol 7:726–731

    Google Scholar 

  • Bentwich Z, Kunkel HG (1973) Specific properties of human B and T lymphocytes and alterations in disease. Transplant Rev 16:29–50

    Google Scholar 

  • Bentwich Z, Douglas SD, Siegal FP, Kunkel HG (1973) Human lymphocytes-sheep erythrocyte rosette formation. Some characteristics of the interaction. Clin Immunol Immunopathol 1:511–517

    Google Scholar 

  • Cavaillon JM, Bona C, Cazenave PA, Cinader B (1977) Purification of rabbit T-derived (Ig−) lymphocytes. J Immunol Methods 14:355–359

    Google Scholar 

  • Darźynkiewicz Z, Andreeff M, Traganos F, Sharpless T, Melamed MR (1978) Discrimination of cycling and non-cycling lymphocytes by BUdR-suppressed acridine orange fluorescence in a flow cytometric system. Exp Cell Res 115:31–35

    Google Scholar 

  • Ewijk W van, Brons NHC, Rozing J (1975) Scanning electron microscopy of homing and recirculating lymphocyte populations. Cell Immunol 19:245–261

    Google Scholar 

  • Fujiwara S, Armstrong RM, Cinader B (1974) Age and organ dependent variations in the number of thymus derived RTLA bearing cells. Immunol Commun 3:275–284

    Google Scholar 

  • Grossi CE, Webb SR, Zicca A, Lydyard PM, Moretta L, Mingari MC, Cooper MD (1978) Morphological and histochemical analysis of two human T-cell subpopulations bearing receptors for IgM or IgG. J Exp Med 147:1405–1417

    Google Scholar 

  • Huhn D, Rodt H, Thiel E, Fink U, Ruppelt W (1976) Elektronenmikroskopische und immunhistochemische Untersuchungen an menschlichen Lymphozyten. Blut 32:87–102

    Google Scholar 

  • Kay MMB (1975) Multiple labelling technique used for kinetic studies of activated human B lymphocytes. Nature 254:424–426

    Google Scholar 

  • Keuning FJ, Van den Broek AA (1968) Role of marginal zone cells of lymphoid follicles in the immune response of rabbits (abstr.). Exp Haematol 17:4

    Google Scholar 

  • Lamelin JP, Lisowska-Bernstein B, Matter A, Ryser JE, Vassali P (1972) Mouse thymus-independent and thymus-derived lymphoid cells. I. Immunofluorescent and functional studies. J Exp Med 136:984–1007

    Google Scholar 

  • Leene W, Duyzings MJM, van Steeg C (1973) Lymphoid stem cell identification in the developing thymus and bursa of Fabricius of the chick. Z Zellforsch 136:521–533

    Google Scholar 

  • Leene W, Roholl PJM, de Groot C (1976) Lymphocyte differentiation in the rabbit thymus. Ann Immunol (Inst Pasteur) 127c, 911–921

    Google Scholar 

  • Leene W, Roholl PJM, Hoeben KA, Hogenes TM (1979) Two subpopulations of cortisone-resistant lymphoid cells in the rabbit thymus. Adv Exp Med Biol 114, Plenum Press, New York

  • Linthicum DS, Sell S (1974a) Surface immunoglobulin on rabbit lymphoid cells. I. Ultrastructural distribution and endocytosis of b4 allotypic determinants on peripheral blood lymphocytes. Cell Immunol 12:443–458

    Google Scholar 

  • Linthicum DS, Sell S (1974b) Surface immunoglobulin on rabbit lymphoid cells. II. Ultrastructural distribution of b4 allotypic determinants and morphology of spleen lymphocytes. Cell Immunol 12:459–71

    Google Scholar 

  • Linthicum DS, Sell S (1975) Topography of lymphocyte surface immunoglobulin using scanning immunoelectron microscopy. J Ultrastruct Res 51:55–68

    Google Scholar 

  • Linthicum DS, Sell S (1977) Surface immunoglobulin on rabbit lymphoid cells. V. Ultrastructural distribution of b4 allotypic determinants on thymus, lymph node and bone marrow lymphocytes. Int Arch Allergy Appl Immunol 54:385–399

    Google Scholar 

  • Matter A, Lisowska-Bernstein B, Ryser JE, Lamelin JP, Vassalli P (1972) Mouse thymus-independent and thymus-derived lymphoid cells. II. Ultrastructural studies. J Exp Med 136:1008–1030

    Google Scholar 

  • Newell DG, Roath S, Smith JL (1976) The scanning electron microscopy of normal human peripheral blood lymphocytes. Br J Haematol 32:309–316

    Google Scholar 

  • Nieuwenhuis P (1971) On the origin and fate of immunologically competent cells. Thesis, Groningen

  • Perkins WD, Karnovsky MJ, Unanue ER (1972) An ultrastructural study of lymphocytes with surfacebound immunoglobulin. J Exp Med 135:267–276

    Google Scholar 

  • Pernis B, Forni L, Amante L (1970) Immunoglobulin spots on the surface of rabbit lymphocytes. J Exp Med 132:1001–1018

    Google Scholar 

  • Polliack A, De Harven E (1975) An interpretative review. Surface features of normal and leukemic lymphocytes as seen by scanning electron microscopy. Clin Immunol Immunopathol 3:412–430

    Google Scholar 

  • Polliack A, Lampen N, Clarkson BD, De Harven E (1973) Bentwich Z, Siegal FP, Kunkel HG (1973) Identification of human B and T lymphocytes by scanning electron microscopy. J Exp Med 138:607–624

    Google Scholar 

  • Polliack A, Lampen N, De Harven E (1974) Scanning electron microscopy of lymphocytes of known B and T derivation. Proceedings of the seventh annual scanning electron microscope symposium IIT Research Institute, Chicago 673:682

  • Renau-Piqueras J, Cerdan F, Barbera E, Cervera J (1978) Electron microscopic morphometric analysis of human T and B peripheral blood lymphocytes. Virchows Archiv B Cell Pathol 28:47–57

    Google Scholar 

  • Reyes F, Lejonc JL, Gourdin MF, Mannoni P, Dreyfus B (1975) The surface morphology of human B lymphocytes as revealed by immunoelectron microscopy. J Exp Med 141:392–410

    Google Scholar 

  • Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–212

    Article  CAS  PubMed  Google Scholar 

  • Schuyt HC, Leene W (1978) An improved method in rabbit intubation and thymectomy. Eur Surg Res 10:362–372

    Google Scholar 

  • Siegal S (1956) Non parametric statistics for the behavioral sciences. International student edition, McGraw-Hill Kogakusha, Tokyo

    Google Scholar 

  • Stempak JG, Ward RT (1964) An improved staining method for electron microscopy. J Cell Biol 22:697–701

    Google Scholar 

  • Wilson AB, Gurner BW, Coombs RRA (1975) Observations on rabbit thymocytes and peripheral T cells. II. Rosette formation with rabbit erythrocytes. Int Arch Allergy Appl Immunol 48:383–394

    Google Scholar 

  • Zucker-Franklin D (1974) The percentage of monocytes among “mononuclear” cell fractions obtained from normal human blood. J Immunol 112:234–240

    Google Scholar 

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Affiliations

  1. Laboratory of Histology and Cell Biology, University of Amsterdam, 1e Constantijn Huygensstraat 20, 1054, Amsterdam, The Netherlands

    J. G. Vos, P. J. M. Roholl &  W. Leene

Authors
  1. J. G. Vos
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  2. P. J. M. Roholl
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  3. W. Leene
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Vos, J.G., Roholl, P.J.M. & Leene, W. Ultrastructural studies of peripheral blood lymphocytes in T cell-depleted rabbits. Cell Tissue Res. 213, 221–235 (1980). https://doi.org/10.1007/BF00234783

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  • DOI: https://doi.org/10.1007/BF00234783

Key words

  • Peripheral blood lymphocyte
  • Density separation
  • T cell depleted rabbits
  • Ultrastructure