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Demonstration of C3 receptors on frozen sections of normal lymphoid tissue and malignant lymphomas using various EAC complexes

Demonstration von C3-Rezeptoren an Gefrierschnitten von normalem lymphatischen Gewebe und von malignen Lymphomen mit verschiedenen EAC-Komplexen

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Zusammenfassung

Das Vorkommen und die Verteilung von Komplement-Rezeptoren (C3R) wurde an Gefrierschnitten von normalem lymphatischem Gewebe und von 185 malignen Lymphomen mit verschiedenen EAC-Komplexen [vitale EA und glutaraldehydfixierte EA, beschichtet mit Komplement von der Maus (EAC-Maus, Glu-EAC-Maus), vom Menschen (EAC-Human, Glu-EAC-Human) und zum Teil von C6-defekten Kaninchen (EAC-KC6d, Glu-EAC-KC6 d)] untersucht. Die verschiedenen EAC-Komplexe zeigten eine unterschiedliche Affinität zu den C3-Rezeptoren: Glu-EAC-Human > EAC-Maus = EAC-KC6d > Glu-EAC-Maus = Glu-EAC-KC6d > EAC-Human. Die EAC-Komplexe differierten nicht nur in ihrer Rezeptoraffinität, sondern auch im Haftungsmuster. Die EAC-Maus, EAC-KC6d und EAC-Human hafteten an Gefrierschnitten von Tonsillen ausschließlich im Bereich der Keimzentren und des Follikelwalls, während die Glu-EAC-Human, die Glu-EAC-Maus und die Glu-EAC-KC6d sowohl im Bereich der Keimzentren und des Follikelwalls als auch parafollikulär und bisweilen auch zwischen den Follikeln hafteten. Die Austestung der Reaktivität der verschiedenen EAC-Komplexe gegenüber C3bR von humanen Erythrozyten und C3R humaner Tonsillenzellen ergab, daß sich EAC-Maus, EAC-Human und EAC-KC6d konstant negativ gegenüber humanen Erythrozyten (C3bR+ und C3dR−), aber positiv gegenüber Tonsillenzellen (C3bR+ und C3dR+) verhielten, während die Glu-EAC-Human, Glu-EAC-Maus und Glu-EAC-KC6d eine starke Affinität zu den C3bR der Humanerythrozyten und zu den C3R der Tonsillenzellen zeigten. In Verbindung mit kürzlich mitgeteilten Daten geht aus diesen Befunden hervor, daß a) EAC-Maus, EAC-Human und EAC-KC6d nur mit Rezeptoren für C3d reagieren, während Glu-EAC-Human, Glu-EAC-Maus und Glu-EAC-KC6d eine Bindung mit CSbR- und wahrscheinlich auch mit C3dR eingehen und b) Keimzentrumszellen und Follikelwall-Lymphozyten C3bR und C3dR exprimieren und daß die Zellen der parafollikulären und zum Teil auch der interfollikulären Zone nur den C3bR besitzen. Am Gefrierschnitt eines fetalen Thymus ließen sich mit Glu-EAC-Human eindeutig C3R nachweisen. Der Nachweis von C3R an Gefrierschnitten maligner Lymphome ergab, daß mit dem Komplex Glu-EAC-Human bei allen Lymphomtypen, mit Ausnahme der Mycosis fungoides, C3R nachweisbar waren, allerdings in unterschiedlicher Häufigkeit und Dichte. Am häufigsten und in großer Dichte waren C3R bei den Keimzentrumszelltumoren, dem centroblastisch/centrocytischen Lymphom und dem centrocytischen Lymphom, zu demonstrieren. Die Analyse der C3R bei den lymphoblastischen Lymphomen vom T-Typ führte in Verbindung mit der Untersuchung der sauren Phosphatase und des Schafserythrozyten-Rosettentests zur Abgrenzung von drei Subtypen, nämlich einem Präthymozyten-, einem Prothymozyten- und einem reifen Thymozyten-Subtyp. An Hodgkin- und Sternberg-Reed-Zellen ließen sich in keinem der untersuchten Fälle eindeutig C3R nachweisen.

Summary

The occurrence and distribution of complement receptors (C3R) were investigated on frozen sections from normal lymphoid tissue and from 185 cases of malignant lymphoma. Various EAC complexes were used: vital EA or glutaraldehyde-fixed EA coated with mouse complement (EAC mouse and Glu-EAC mouse, respectively), human complement (EAC human and Glu-EAC human), or complement from C6-deficient rabbits (EACra-C6-def, Glu-EACra-C6-def). The different EAC complexes showed varying affinity for C3R: Glu-EAC human > EAC mouse = EACra-C6-def > Glu-EAC mouse = Glu-EACra-C6-def > EAC human. The EAC complexes differed not only in their receptor affinity but also in their pattern of binding. On frozen sections of tonsils, EAC mouse, EAC human and EACra-C6-def adhered exclusively to germinal centers and the follicular mantle, whereas Glu-EAC mouse, Glu-EAC human and Glu-EACra-C6-def adhered not only to germinal centers and the follicular mantle but also to parafollicular areas and sometimes to interfollicular regions. The various EAC complexes were also assayed for reactivity to C3b receptors of human erythrocytes and C3R of human tonsil cells. Human erythrocytes (C3b receptor+ and C3d receptor− did not react with EAC mouse, EAC human or EACra-C6-def whereas tonsil cells (C3b and C3d receptor+) showed positive reactions with these complexes. In contrast, Glu-EAC mouse, Glu-EAC human and Glu-EACra-C6-def displayed marked affinity for both the C3b receptors of human erythrocytes and the C3Rof tonsil cells. In connection with previously reported data, these findings indicate that (a) EAC mouse, EAC human and EACra-C6-def react only with receptors for C3d, whereas Glu-EAC human, Glu-EAC mouse and Glu-EACra-C6-def are bound by C3b receptors and probably by C3d receptors as well, and (b) germinal center cells and follicular mantle lymphocytes express C3b and C3d receptors whereas cells in parafollicular areas and those in the interfollicular zone bear only C3b receptors.

On frozen sections from a fetal thymus C3R could be clearly demonstrated with Glu-EAC human. The demonstration of C3R on frozen sections from malignant lymphomas with the Glu-EAC human complex revealed C3R on cells from all types of lymphoma except mycosis fungoides, although there were variations in the number of positive cells and in the density of the reaction. C3R were most common and were expressed most densely in two types of germinal center cell tumors, viz. centroblastic/centrocytic lymphoma and centrocytic lymphoma. Combined with the investigation of acid phosphatase activity and the sheep erythrocyte rosette test, analysis of C3R in lymphoblastic lymphomas of the T type led to the distinction of three subtypes, viz. a prethymocytic, a prothymocytic, and a mature thymocytic subtype. In none of the cases of Hodgkin's disease tested could C3R be demonstrated unequivocally on Hodgkin or Sternberg-Reed cells.

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  1. Institut für Pathologie, Klinikum der Universität, Hospitalstr. 42, D-2300, Kiel 1, Federal Republic of Germany

    H. Stein, E. Grosskopf & K. Lennert

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  1. H. Stein
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  2. E. Grosskopf
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  3. K. Lennert
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This study was supported by the Deutsche Forschungsgemeinschaft SFB 111, Project CL1

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Stein, H., Grosskopf, E. & Lennert, K. Demonstration of C3 receptors on frozen sections of normal lymphoid tissue and malignant lymphomas using various EAC complexes. Blut 41, 101–118 (1980). https://doi.org/10.1007/BF01039654

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