Abstract Rosetting of CD4+ T cells around the neoplastic
Hodgkin and Reed-Sternberg (H&RS) cells is a character-
istic feature of Hodgkin’s disease (HD). To answer the
question whether this phenomenon is solely due to
chemokine-mediated attraction of T cells or whether the
rosetting T cells in addition recognize antigens presented
by the H&RS cells, we examined the T cells adherent to
H&RS cells. Cells from five cases of HD [four classic HD
and one lymphocyte-predominant (LP) HD] were examined
by single-cell analysis for the T-cell receptor (TCR) γ
gene. Between 5 and 17 rosettes containing one to ten
rosetting lymphocytes and the corresponding H&RS cells
were amplified in separate plastic tubes. Of the resulting
119 TCRγ polymerase chain reaction (PCR) products, 87
were sequenced. While no evidence of a clonal expansion
was obtained in the lymph nodes from four of five patients
with classic HD, clonal TCRγ sequences were found in
the lymph node from the patient within LPHD in two
independent experiments analyzing seven and ten different
rosetting complexes, respectively. Of 13 products, 11
showed identical Vγ9 sequences. Unrelated products were
found in all other TCRγ family subgroups in this case.
Single H&RS cells picked as controls were negative for
TCRγ rearrangements. Our results demonstrate that clonal
proliferations on a polyclonal background can occur
among the T cells forming rosettes with Hodgkin cells and
lend support to the view that Hodgkin cells may also
function as cells presenting antigens to the adhering T cells.
Keywords Hodgkin’s disease ·
T-cell receptor rearrangement · Reed-Sternberg cell ·
Antigen presenting cell · Clonality
Introduction
A characteristic feature that distinguishes Hodgkin’s
disease (HD) from most other neoplasms is the fact that
the neoplastic Hodgkin and Reed-Sternberg (H&RS)
cells represent a minority of the cellular elements in
involved tissues [13]. The majority of the reactive cells
in lymph nodes involved in classic HD of the nodular
sclerosing or mixed cellularity subtype are CD4
+
T cells
that express the memory phenotype CD45R0 [10, 18,
19]. These cells surround the neoplastic H&RS cells in
vivo and display the “rosetting phenomenon,” i.e., they
form aggregates with autologous H&RS cells in vitro
[16, 17, 28]. In nodular lymphocyte predominance HD,
the characteristic neoplastic Reed-Sternberg-like cells
(L&H cells, lymphocytic and histiocytic) are morphologi-
cally similar but immunologically different from classic
H&RS cells but are also surrounded by a characteristic
rim of T cells [14]. Two hypotheses that are not mutually
exclusive have been put forward to explain the intimate
contact between the neoplastic H&RS and the CD4
+
T
cells. According to the first hypothesis, this interaction is
antigen-independent and nonspecific, reflecting the
strong cytokine-secreting activity of H&RS cells [7],
among others the thymus and activation-regulated
chemokine TARC [19], and the complementarity of
adhesion ligands and receptors on H&RS cells and the
surrounding CD4
+
T cells, e.g., CD2/LFA-3. Alternatively,
T cells rosetting around H&RS cells may reflect a
cellular antitumor response. This hypothesis is supported
by the fact that H&RS cells express MHC class II mole-
cules on their surface by means of which H&RS may
present antigen-derived peptides which are specifically
recognized by the adherent CD4
+
T cells. Moreover,
evidence for a clonal expansion of T cells in tissues
L. Trümper (
✉
) · W. Jung · F. von Bonin
Department of Hematology/Oncology, Georg-August University,
Robert-Koch-Str. 40, 37075 Göttingen, Germany
e-mail: lorenz.truemper@med.uni-goettingen.de
Tel.: +49-551-398535, Fax: +49-551-398587
G. Mechtersheimer
Department of Pathology, University of Heidelberg,
69115 Heidelberg, Germany
H. Daus · M. Pfreundschuh
Department of Internal Medicine I, Saarland University,
66421 Homburg/Saar, Germany
Ann Hematol (2001) 80:653–661
DOI 10.1007/s002770100370
ORIGINAL ARTICLE
L. Trümper · W. Jung · H. Daus · G. Mechtersheimer
F. von Bonin · M. Pfreundschuh
Assessment of clonality of rosetting T lymphocytes in Hodgkin’s
disease by single-cell polymerase chain reaction: detection of clonality
in a polyclonal background in a case of lymphocyte predominance
Hodgkin’s disease
Received: 27 September 2000 / Accepted: 19 July 2001 / Published online: 29 September 2001
© Springer-Verlag 2001