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Elektronenmikroskopische und immunhistochemische Untersuchungen an menschlichen Lymphozyten

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Zusammenfassung

Lymphozyten von Gesunden und von Patienten mit CLL wurden elektronenmikroskopisch-immunhistochemisch untersucht. Mit Hilfe Peroxidase-konjugierter spezifischer heterologer Antiseren wurden die Immunglobulin-Determinanten des B-Lymphozyten sowie spezifische Membranantigene des T-Lymphozyten dargestellt. T-Lymphozyten wurden zusätzlich durch ihre Fähigkeit zur spontanen Rosettenbildung mit Schaferythrozyten markiert. Der Anteil der mit den verschiedenen Methoden dargestellten B- und T-Lymphozyten bei Gesunden und bei CLL-Patienten wird mitgeteilt. Entsprechend ihrem zunehmenden Differenzierungsgrad lassen sich 3 Arten von B-Lymphozyten und 2 Arten von T-Lymphozyten unterscheiden: Nur weitgehend differenzierte B-Lymphozyten sind aufgrund ihres Reichtums an Ergastoplasma allein durch morphologische Merkmale zu identifizieren. Durch die immunhistochemische Methodik hingegen läßt sich jede einzelne Zelle morphologisch beurteilen und gleichzeitig als B- oder T-Lymphozyt identifizieren.

Summary

Lymphocytes from the blood of healthy individuals and of patients suffering from CLL were investigated by electron microscopy and peroxidase-immunohistochemistry. B-lymphocytes were labelled by heterologous, peroxidase-conjugated antisera directed against the Ig-determinants of their membranes. T-lymphocytes were labelled by an indirect method: specific incubation with a specific anti-T-cell-globulin from the rabbit; labelling-incubation with a peroxidase-conjugated anti-rabbit-IgG-globulin from the sheep. In addition, T-lymphocytes were identified by their ability to form rosettes with sheep erythrocytes spontaneously. The quantitative results were: about 80% T-lymphocytes and about 24% B-lymphocytes in normal persons, the opposite results in CLL. T- and B-lymphocytes were photographed electron microscopically; the number of organelles in the single cells was evaluated: lysosomes in the average are more numerous in T-lymphocytes, ergastoplasm in B-lymphocytes, mitochondria are equally distributed in both groups of cells. There is so much overlapping, however, that the single cell only with the aid of immunochemistry or rosette formation can be identified as a B- or T-cell. In both, the T- and the B-cell-series, different forms of lymphocytes can be distinguished according to the degree of cell differentiation. Some further problems, as specificity of the antisera and labelling of the cells by means of their Fc-receptor are discussed.

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Abbreviations

ATCG:

Anti-T-Zell-Globulin

AKanlg(-Pox):

(Peroxidase-konjugiertes) Anti-Kaninchen-IgG-Globulin

AHumlg(-Pox):

(Peroxidase-konjugiertes) gegen menschliches IgG gerichtetes Globulin

Kan-N(-Pox):

(Peroxidase-konjugiertes) Kaninchen-Normal-IgG

CLL:

chronische lymphatische Leukämie

PBS:

phosphatgepufferte physiologische NaCl-Lösung

MEM:

Eagle's Minimal Essential Medium

Pox:

Peroxidase

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Author notes

  1. Dieter Huhn

    Present address: Lehrstuhl für Innere Medizin (speziell Hämatologie) der Universität München, Deutschland

Authors and Affiliations

  1. Institut für Hämatologie der GSF, München

    Dieter Huhn, Hans Rodt, Eckhard Thiel & Walter Ruppelt

  2. I. Med. Klinik re. d. Isar d. Techn. Univ. München, Deutschland

    Ulrich Fink

Authors
  1. Dieter Huhn
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  2. Hans Rodt
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  3. Eckhard Thiel
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  4. Ulrich Fink
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  5. Walter Ruppelt
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Additional information

Wir danken Frl. G.Wochinger und Frl. M.Darsow für hervorragende technische Hilfe. Studie im Rahmen der Assoziation Hämatologie-EURATOM-GSF Nr. 031-64/1 BIAD.

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Huhn, D., Rodt, H., Thiel, E. et al. Elektronenmikroskopische und immunhistochemische Untersuchungen an menschlichen Lymphozyten. Blut 32, 87–102 (1976). https://doi.org/10.1007/BF00995936

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