Skip to main content
SpringerLink
Log in

Ontogenetic analysis of some surface markers on pig lymphocytes using fluorescence-activated cell sorter

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Surface markers were demonstrated on pig lymphocytes using anti-T call-IgG and anti-Helix pomatia (HP) IgG during prenatal and postnatal development. A fluoreseenceactivated cell sorter analysis of T-cell surface markers was accompanied by an image analysis to prove the association of T antigenic determinants with the plasma membrane only. We found development-dependent changes in both anti-T cell and HP surface markers in both primary and secondary lymphatic organs.

  1. 1.

    The number of T-positive (T+) cells estimated by anti-T cell-IgG was very similar to the results obtained by spontaneous E-rosette forming tests. At all selected age intervals, changes in the number of T+ cells were not significant in the thymus, but a marked inerease in T+ cells was found in both spleen and lymph nodes. The image analysis confirmed the expression of T cell markers on the cell surface. The distribution of T cell markers was uneven,i.e. various degree of fluorescence intensity on whole ring-pattern projeetion of the cell surface image was estimated.

  2. 2.

    In second lymphatic organs especially, fluorescence intensity of cells,i.e. total number of T cell markers estimated by anti-T cell-IgG, increased with age. On fetal day 73, T cell markers were slightly expressed, but very high fluorescence intensity and heterogenous distribution of T cell markers on lymphocytes were found on fetal day 107 and postnatal day 56. The results indicate the possibility of functional maturation of various T cell markers on T cell subsets, furthermore a different degree of expression of T cell markers on various T cell subsets can be suggested.

  3. 3.

    The number of HP+ cells increased with age in both primary and secondary lymphatic organs.

  4. 4.

    In the prenatal period, the expression of HP receptors was very weak in both primary and secondary organs in contrast to the marked inerease in HP+ cells during the postnatal interval. Differences in fluorescence intensity of cells were found, representing the inerease by 22% in thymus cells comparing to cells of secondary lymphatic organs. Heterogeneity of HP+ cell populations in thymus was shown by the Scatchard plot, indicating at least two subpopulations of HP+ cells with different avidity to HP. Cells with low HP avidity could include a subset with cytolytic activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Binns R. M.: Cellular immunology in the pig.Proc. Roy. Soc. Med. 66, 1 (1973).

    Google Scholar 

  • Binns R. M., Symons D.B.A.: The ontogeny of lymphocyte immunoglobulin determinants and their relationship to antibody production in the fetal pig.Res. Vet. Sci. 16, 260 (1974).

    PubMed  CAS  Google Scholar 

  • Binns R.M.: Organisation of the lymphoreticular system and lymphocyte markers in the pig.Vet. Immunol. Immunopathol. 3, 95 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Fišar. Z., Kovářů F., Kovářů H.: Ontogenetical dependence of some properties of thymocyte membranes, p. 356 inProc. 16 th FEBS Meeting, Moscow 1984.

  • Hartman J., šrajer J., Fleischmannová V., Lodin Z.: Adhesion of dissociated mouse embryonic brain cells. A new method of quantitative evaluation.Physiol. Bohemoslov. 30, 295 (1981).

    PubMed  CAS  Google Scholar 

  • Herzenberg L.A., Herzenberg L.A.: Analysis and separation using the fluorescence activated cell sorter (FACS), p. 22 inHandbook of Experimental Immunology, 3rd edition (D. M. Weir, Ed.). Blackwell Sci. Publ., Oxford (England) 1978.

    Google Scholar 

  • Jarošková L., Trebichavský I., Říha I., Kovářů F., Holub M.: Immunoglobulin determinants in lymphocytes in germ-free piglets.Eur. J. Immunol. 3, 818 (1973).

    Article  PubMed  Google Scholar 

  • Jarošková L.: Membrane receptors on lymphocytes. Ontogenetic studies in the fetal pig.Acta Vet. Brno 46, Suppl. 4, 55 (1977).

    Google Scholar 

  • Jarošková L., Kovářů F.: Identification of T and B lymphocytes in pigs by combined E-rosette test and surface Ig labelling.J. Immunol. Meth. 22, 253 (1978).

    Article  Google Scholar 

  • Jarošková L., Trebichavský I., KovÁŘťr F., Prokešová L.: Precursors of B-lymphocytes with surface IgM and their possible migration from bone marrow to peripheral lymphatic tissues.Lymphology 11, 81 (1978).

    PubMed  Google Scholar 

  • Kim Y.B., Huh N.D.: Natural killer and killer (NK/K) cell system in gnotobiotic miniature swine, p. 585 inRec. Adv. Germfree Res. (S. Sasaki, A. Ozawa, K. Hashimoto, Eds.). Tokai Univ. Press 1981.

  • KovÁŘťr F., Stožický V., Kruml J., Dlabač V., Donát J., Novotná J.: Experimental surgery in the foetal period of mammals.Acta Vet. Brno, Suppl.3, 1 (1972).

    Google Scholar 

  • Kovářů F., Pospíšil M., Hopman J., Trebichavský I.: The morphological development and some functional features of the lymphatic tissue in pig fetuses, p. 252 inLymphology — Proe. éth Internat. Congr. (P. Málek, V. Bartoš, H. Weissleder, M. H. Witte, Eds.). G. Thieme Publ., Stuttgart and Avicenum, Prague 1978.

    Google Scholar 

  • Kovářů F., Štěpánková R., Kruml J., Mandel L., Kenig E.: Development of lymphatic and haemopoietic organs in germfree models.Folia Microbiol. 24, 32 (1979).

    Article  Google Scholar 

  • Kovářů F., Jarošková L.: Development of E-rosette formation in ontogeny of pigs.Folia Biol. (Prague) 24, 399 (1979).

    Google Scholar 

  • Kovářů H., Kovářů F., Plášek J., Fišar Z., Mandel L.: Lectin-induced surface changes in lymphocytes of gnotobiotic pigs, p. 315 inLedins — Biology, Biochemistry, Clinical Biochemistry, Vol. 2 (T. C. Bog-Hansen, Ed.). W. de Gryuter Co., Berlin- New York 1982.

    Google Scholar 

  • Kovářů H., Kovářů F., Mareš V., Kozáková H., Fišar Z., Hartman J.: Using various types of Percoll gradients for isolation of populations and subpopulations of cells from lymphoid and non-lymphoid tissues and their characterization. (In Czech)čs. fyziol., in press (1985).

  • Kruml J., Kovářů F., Pospíšil M., Trebichavský I.: The development of lymphatic tissue during ontogenesis, p. 35 inDevelopmental Aspects of Antibody Formation and Structure, Vol. 1 (J. Šterzl, I. Říha, Eds.). Academia, Prague 1970.

    Google Scholar 

  • Lu C.Y., Changelian P.S., Unanue E.R.: Alpha-fetoprotein inhibits macrophage expression of la antigens.J. Immunol. 132, 1722 (1984).

    PubMed  CAS  Google Scholar 

  • Mareš V., Kovářů F., Kovářů H.: Alphafetoprotein in the brain of developing rats and pigs. An immunofluorescent study of cell-and-tissue differentiation.Bas. Appl. Histochem. 26, 53 (1982).

    Google Scholar 

  • Mareš V., Kovářů F., Kovářů H., Müller L., Žižkovský V.: Alpha-fetoprotein in the brain of embryonic pigs.Biomed. Biochim. Acta, in press (1985).

  • Mattes M. J., Holden H.T.: The distribution ofHelix pomatia lectin receptors on mouse lymphoid cells and other tissues.Eur. J. Immunol. 11, 358 (1981).

    Article  PubMed  CAS  Google Scholar 

  • Poros A., Åhrlund-Richter L., Klein E., Hammarström S., Koide N.: Expression ofHelix pomatia haemagglutinin receptors on cytolytio receptors activated in mixed cultures.J. Immunol. Meth. 57, 9 (1983).

    Article  CAS  Google Scholar 

  • Pospíšil M.,Kubrycht J.,Bednář M.,Bezoušek K.,Táborský O.: Modulation of K and NK cell activities in pigs, p. 379 inProc. 6th Eur. Immunol. Meeting, Interlaken (Switzerland) 1984.

  • Prokešová L., Trebichavský I., Kovářů F., Kostka J., Rejnek J.: Ontogeny of immunoglobulin synthesis. Production of IgM, IgG and IgA in pig foetuses.Develop. Comp. Immunol. 5, 491 (1981).

    Google Scholar 

  • Rejnek J., Tučkova L., Trávníček J., Tlaskalová H., Kovářů F.: Isolation and properties of antigen-specific receptors from rabbit and pig lymphoid cells.Mol. Immunol. 17, 65 (1980).

    Article  PubMed  CAS  Google Scholar 

  • Říha I., Trávníček J., Šírová M.: Lymphocyte immunoglobulin receptors in newborn piglets.Develop. Comp. Immunol. 7, 759 (1983).

    Article  Google Scholar 

  • Scatchard G.: The attraction of protein for small molecules and ions.Ann. N. Y. Acad. Sci. 51, 660 (1949).

    Article  CAS  Google Scholar 

  • Slavík J.: Intracellular pH topography: Determination by a fluorescent probe.FEBS Lett. 156, 227 (1983).

    Article  PubMed  Google Scholar 

  • Šterzl J.:Inductive Phase of Antibody Formation. Avieenum, Prague 1960.

    Google Scholar 

  • Šterzl J.: Quantitative and qualitative aspect of the inductive phase of antibody formation.J. Hyg. Epid. Microbiol. Immunol. 7, 301 (1963).

    Google Scholar 

  • Šterzl J., Rejnek J., Trávníček J.: Impermeability of pig placenta for antibodies.Folia Microbiol. 11, 7 (1966).

    Article  Google Scholar 

  • Šterzl J., Silverstein A.M.: Developmental aspects of immunity.Adv. Immunol. 5, 337 (1967).

    Article  Google Scholar 

  • Šterzl J., Šíma P., Medliín J., Tlaskalová H., Mandel L., Nordin A.: Induction of the primary response, preparation of the secondary response and tolerance, p. 865 inDevelopmental Aspects of Antibody Formation and Structure (J. Šterzl, I. Říha, Eds.). Academia, Prague 1970.

    Google Scholar 

  • Šterzl J., Kovářů F.: Development of lymphatic tissue and immunocompetency in pig foetuses and germ-free piglets.Acta Vet. Brno 46, Suppl. 4, 13 (1977).

    Google Scholar 

  • Tlaskaxová H., Hájek P., Šterzl J., Pospíšil M., Říha I., Marvanova H., Medlín J., Mandel L., Kbuml J., KovÁŘú F.: The development and antibody formation during embryonal and postnatal period, p. 767 inDevelopmental Aspects of Antibody Formation and Structure (J. Šterzl, I. Říha, Eds.). Academia, Prague 1970.

    Google Scholar 

  • Williams C.A., Chase M.W.:Methods in Immunology and Immunochemistry, Vol. 1, p. 319. Academic Press, New York - London 1967.

    Google Scholar 

  • Zajicek G., Shohat M., Melnik Y., Yeger A.: Image analysis of nucleated red blood cells.Comp. Biomed. Res. 16, 347 (1983).

    Article  CAS  Google Scholar 

  • Zimmerman B., Swain S.L., Dutton R.W.: Immunosuppressive ATS. V. Analysis of the effect of anti-thymocyte serum on T-lymphocyte subsets.J. Immunol. 129, 515 (1982).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Gnotobiology, Institute of Microbiology, Czechoslovak Academy of Sciences, 142 20, Prague 4

    F. Kovářů, H. Kovářů & Z. Fišar

Authors
  1. F. Kovářů
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Kovářů
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Z. Fišar
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The authors are grateful to Prof. R. Hamers (Institute of Molecular Biology, University of Brussels), for the possibility to perform the FACS analysis in the course of a stay of one of us (F.K.) and for his valuable criticism. They thank Mrs. K. Rybínová for her skilled technical assistance.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kovářů, F., Kovářů, H. & Fišar, Z. Ontogenetic analysis of some surface markers on pig lymphocytes using fluorescence-activated cell sorter. Folia Microbiol 30, 277–290 (1985). https://doi.org/10.1007/BF02923521

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02923521

Keywords

Search

Navigation