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NEOPLASIA
From the Institute of Pathology, Consultation and
Reference Centre for Lymph Node Pathology and Haematopathology,
University Hospital Benjamin Franklin, Free University, Berlin,
Germany; Department of Physiological Chemistry, Universität Ulm,
Germany; and Istituto di Ematologia, Universita` di Perugia, Perugia,
Italy.
In contrast to the tumor cells (L&H cells) of lymphocyte
predominant Hodgkin disease (LPHD), Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin disease (cHD) are unable to transcribe immunoglobulin, despite the presence of rearranged immunoglobulin genes. Although initial studies have suggested crippling immunoglobulin gene mutations to be the cause of absent immunoglobulin expression in
cHD, recent work of our group has demonstrated an impaired activation
of the immunoglobulin promoter as a superior mechanism. As
immunoglobulin transcription is mainly regulated by the B-cell transcription factors Oct2 and BOB.1/OBF.1, we analyzed 35 cases of
LPHD, 32 cases of cHD, and 2 Hodgkin disease cell lines for the
expression of these transcription factors and also in parallel for
immunoglobulin expression. Our results demonstrate an absence of Oct2
and/or BOB.1/OBF.1 in cHD and a striking overexpression of Oct2 in
LPHD. Immunoglobulin expression was lacking in cHD but present in LPHD.
Furthermore, the reintroduction of BOB.1/OBF.1 and Oct2 into cultured
HRS cells restored the activity of cotransduced immunoglobulin promoter
constructs. Our findings dismiss the concept that the different
immunoglobulin expression in cHD and LPHD is due to disrupting
mutations of immunoglobulin V genes in cHD but is most likely due to a
down-regulation of Oct2 and/or BOB.1/OBF.1. This study further revealed
Oct2 as a new and valuable marker for the identification of L&H cells
and their distinction from HRS cells. The impairment of immunoglobulin
transcription with a down-regulated synthesis of Oct2 and
BOB.1/OBF.1 is the first established general recurrent defect
found in HRS cells.
(Blood. 2001;97:496-501) Hodgkin disease (HD) is one of the most common
categories of malignant lymphoma in Europe and North America. Two
distinct types, the rare lymphocyte predominant (LPHD) form and the
common classical form (cHD), have been distinguished. LPHD and cHD have both common features and marked differences. The features that they
have in common are (1) a few large tumor cells in an abundant admixture
of non-neoplastic inflammatory cells, (2) T-cell rosettes around the
tumor cells, (3) a high proliferation rate, and (4) the presence of
clonal immunoglobulin gene rearrangements in the tumor cells (5) with a
high load of somatic mutations. The features that differentiate them
mainly concern the gene expression of the tumor cells. In LPHD the
tumor cells (designated L&H cells) constantly express
B-cell-associated molecules (eg, CD20, CD79a, J-chain, and
immunoglobulin transcripts in the absence of CD30 and CD15), whereas
the tumor cells of cHD (termed Hodgkin and Reed-Sternberg [HRS]
cells) carry CD30 and CD15 but lack CD20, CD79a, J-chain, and
immunoglobulin transcripts in most or all instances (reviewed by Stein
et al1 and Anagnostopoulos et al2). The
different immunoglobulin expression has raised particular interest
because the tumor cells of both LPHD and cHD contain rearranged
immunoglobulin genes.3-7 This difference was explained by
Rajewsky and colleagues7,8 by the presence of intact
immunoglobulin gene coding sequences in L&H cells and crippled
immunoglobulin genes in HRS cells. In contrast to this, our studies
have shown that the immunoglobulin gene coding capacity is intact not
only in LPHD but also in the majority of cHD cases,4
prompting the conclusion that the absent immunoglobulin expression in
HRS cells is due to a defect in the transcription machinery rather than to a disruption of the immunoglobulin genes themselves. This was confirmed by our recent transfection studies which showed that immunoglobulin gene promoter luciferase reporter constructs exhibited no activity in cultured HRS cells.4
The transcription factors Oct1 and Oct2 and their
coactivator BOB.1/OBF.1 regulate immunoglobulin gene
transcription. Oct1 and Oct2 bind to the immunoglobulin gene promoter
octamer motif and activate immunoglobulin gene transcription when the
coactivator BOB.1/OBF.1 is recruited into the transcription
complex.9,10 BOB.1/OBF.1 was also shown to act as a
molecular clamp that holds together the POU subdomains of Oct1
or Oct2.11 Finally, according to knockout experiments
performed, Oct1 and Oct2 can replace each other, whereas BOB.1/OBF.1 is
unconditionally required for the activation of the immunoglobulin gene
promoter.12 This implies that Oct1 and Oct2 or their
coactivator BOB.1/OBF.1 are regularly expressed in L&H cells but are
absent from, or are defective in, HRS cells. This paper reports on
studies designed to check this claim. The results obtained confirm the
claim concerning the absence of Oct2 and/or BOB.1/OBF.1 in HRS cells
with the additional interesting observation that Oct2 is enormously
overexpressed by the L&H cells. These findings, in association with our
demonstration of the restoration of the immunoglobulin promoter
activity by cotransfection of BOB.1/OBF.1 and Oct2 into cultured HRS
cells, strengthen our hypothesis that the non-expression of
immunoglobulin by HRS cells is caused by a down-regulated synthesis of
immunoglobulin transcription factors required for immunoglobulin
transcription rather than by crippling mutations of immunoglobulin
genes. Our findings also add more knowledge to the molecular
differences found between LPHD and cHD. Finally, the overexpression of
Oct2 in L&H cells is a new and valuable feature for their distinction
from classical HRS cells.
Tissues and cell lines
Immunohistochemistry
In situ hybridization The detection of messenger RNA (mRNA) for Igµ, Ig , Ig ,
Ig , Ig , and Oct2 was carried out, using a highly sensitive
radioactive in situ hybridization method as previously
described.4,15 The Oct2 probe was prepared from
complementary DNA (cDNA) that had been amplified by reverse
transcriptase-polymerase chain reaction (RT-PCR) from a B-cell line and
cloned into the appropriate plasmids. After linearization of the
plasmids with restriction enzymes, anti-sense and sense (control)
transcripts were generated, using T7 or SP6 RNA-polymerase with the
incorporation of [35S]-dUTP. Cells with a 4-fold signal
intensity, as compared to background signal, were scored positive.
Hybridization with sense-transcripts showed only very few homogeneously
distributed background signals.
Northern and Western blot analyses Total cytoplasmic RNA was prepared and analyzed as described elsewhere.16,17 Western blot analysis was carried out according to previously published protocols.16,18Cell culture and transfection For electroporation, 5 × 106 cultured cells were pelleted and resuspended in a final volume of 300 µL RPMI medium supplemented with 10% FCS. Fifteen micrograms of the respective luciferase reporter plasmid (details about the wild-type immunoglobulin octamer enhancer and the synthetic multimerized octamer reporter constructs can be found in Laumen et al12 and Pfisterer et al19,20) were added to the cells. The mixture was transferred to a cuvette with a gap of 0.4 cm and electroporated at 230 V and 950 µF with a Gene Pulser (Biorad, Munich, Germany). The cells were immediately transferred to a Petri dish containing 10 mL RPMI/10% FCS and incubated at 37°C in 5% CO2 for 20 hours. The cells were harvested, and the lysates were analyzed with a dual-luciferase reporter assay system (Promega, Mannheim, Germany). All luciferase values were calculated against the renilla-luciferase values to correct for different transfection efficiencies.
Expression of immunoglobulin in cHD and LPHD The expression of immunoglobulin in classical HRS cells and L&H cells was investigated by radioactive in situ hybridization and immunohistochemistry. As in a previous study,4 there were no immunoglobulin-specific transcripts demonstrable in the HRS cells of 25 cases of cHD (Table 1). This result held true even when the exposure time was prolonged considerably, and the internal control cells (plasma cells and B cells) became very intensely positive. Immunostaining with immunoglobulin-specific antibodies often produced a cytoplasmic labeling of many HRS cells. The staining, however, spared the nuclear space of most HRS cells although this space was strongly positive in the bystander B lymphocytes and plasma cells in all instances. As revealed by Northern blotting and immunohistochemistry, the cell lines L428 and KM-H2 derived from HRS cells of cHD cases were devoid of immunoglobulin-specific transcripts and protein (Table 2).
In contrast, L&H cells of all but 2 cases were labeled with
immunoglobulin-specific probes with an intensity between that generally
seen in plasma cells and B lymphocytes (Table 1, Figure 1A). Immunostaining with antibodies
directed at immunoglobulin isotypes revealed a very strong labeling of
the nuclear space and the cytoplasm of the L&H cells in 26 of 32 cases
(Figure 1B-D,F; Table 1). The tumor cells of 8 cases of follicular
lymphoma studied for control purposes showed positive labeling for
immunoglobulin mRNA and immunoglobulin protein in all instances (Table
1). The positive immunoglobulin staining included the nuclear space in all 8 cases. The frequency with which the different IgH classes and the
IgL types were found to be expressed by the L&H cells is shown in Table
3.
Expression of BOB.1/OBF.1 and Oct2 in cHD and LPHD The expression of BOB.1/OBF.1 and Oct2 was studied by immunohistochemistry. BOB.1/OBF.1 and Oct2 were completely absent from the HRS cells of 24 (75%) and 28 (87.5%) of the 32 cHD cases, respectively (Table 1; Figure 2B,D). In the remaining cases only a small proportion of the HRS cells showed a weak labeling for these proteins (Figure 2C) with the exception of one case in which all HRS cells showed a strong labeling for Oct2 protein. To check the validity of the negative labeling for Oct2 in HRS cells, 11 cases were examined by in situ hybridization with a radioactive Oct2 probe. In 9 cases there were no signals over the HRS cells, although internal control cells were labeled (Table 1, Figure 2A). In difference to cHD, expression of BOB.1/OBF.1 and Oct2 was demonstrable in all L&H cells of all LPHD cases studied (Table 1, Figure 3A). The immunostaining of L&H cells for BOB.1/OBF.1 was as strong as in normal and neoplastic germinal center cells. However, the labeling for Oct2 proved to be much more intense in the L&H cells when compared with normal cells and the tumor cells of follicular lymphoma (Figure 3A,B).
Co-transfection of cultured HRS cells with immunoglobulin promoter-reporter constructs and Oct2 and/or BOB.1/OBF.1 The above findings prompted the question as to whether direct evidence can be provided showing that the lack of the Oct2 transcription factor and the BOB.1/OBF.1 coactivator is decisive for deficient immunoglobulin gene transcription in HRS cells. For this purpose we cotransfected the HD-derived cell line L428 with wild-type and mutated (point mutation in the promoter octamer motif) immunoglobulin reporter constructs either with BOB.1/OBF.1 or Oct2 expression vectors individually or with a combination of both. As shown in Figure 4A, the wild-type immunoglobulin reporter was slightly stimulated by BOB.1/OBF.1 and to a lower extent induced by Oct2. The combination of both factors, however, resulted in a significantly higher stimulation of immunoglobulin reporter gene activity. This activity was virtually identical to that obtained in a normal B-cell line expressing endogenous levels of these factors (data not shown).
These results were confirmed by transfection of L428 with immunoglobulin reporter constructs that contain multimerized octamer motifs upstream of a minimal promoter consisting of only a TATA box (Figure 4B). Transfection of the multimerized immunoglobulin reporter construct with BOB.1/OBF.1 resulted in a strong stimulation of wild-type octamer promoter activity, whereas transfection with Oct2 displayed only a marginal effect on immunoglobulin reporter activity. The BOB.1/OBF.1-induced immunoglobulin promoter activity could be increased further by cotransfection of BOB.1/OBF.1 together with Oct2 (Figure 4B). Mutated multimerized immunoglobulin reporter constructs showed no significant activity under all conditions tested (Figure 4B).
In this study we have analyzed how consistent cHD and LPHD differ
from each other in their expression of immunoglobulin and which
molecular mechanisms might account for this difference. We have
confirmed our previous findings that HRS cells of all (or nearly all)
cases of cHD lack transcripts specific for IgL and IgH
chains4 and have found that the L&H cells of LPHD
overexpress immunoglobulin at the protein and/or transcriptional level
in most instances when compared with normal follicular mantle cells and
follicular lymphoma cells. We have in addition found that the octamer
transcription factor Oct2 is highly overexpressed in L&H cells but
totally absent from 87.5% of the cases or only partially (9.4%)
present in classical HRS cells. There was only one exception to this in
which all HRS cells expressed Oct2 at a relatively high level.
BOB.1/OBF.1, the coactivator of Oct2, proved to be consistently
expressed in the L&H cells at a level equivalent to that seen in
reactive and neoplastic germinal center B cells, whereas there was no
(75% of the cases) or only very little expression (25%) of
BOB.1/OBF.1 in classical HRS cells. Co-transfection experiments
performed revealed that the reintroduction of BOB.1/OBF.1 and Oct2 into
cultured HRS cells can restore the transcription activity of the
immunoglobulin promoter. On the basis of these findings, we conclude
that the difference in the immunoglobulin expression between LPHD and
cHD does not lie in the absence of crippling immunoglobulin gene
mutations in L&H cells and their presence in HRS cells In the past, many studies21-25 have been performed to
analyze the immunoglobulin production by L&H cells and HRS cells.
However, in none of these previous studies has a larger series of cases of LPHD and cHD been investigated in parallel with a highly efficient antigen retrieval procedure and a highly sensitive radioactive in situ
hybridization method. Furthermore, all previous in situ hybridization
studies that were applied to LPHD cases were restricted to IgL
chains.3,26-28 To clarify how frequently and at which level the immunoglobulin synthesis in the tumor cells of cHD is disturbed, it is necessary to obtain representative results to investigate the same collection of cases not only for the
immunoglobulin light chain types In contrast to other reports7,8,29 we were recently able
to demonstrate that in most instances the rearranged immunoglobulin genes are not only functional in L&H cells but also in classical HRS
cells, prompting the conclusion that the absent expression of
immunoglobulin molecules in HRS cells is not caused by crippling mutations of the rearranged immunoglobulin genes but by silencing the
immunoglobulin promoter.4 We confirmed the lack of
immunoglobulin promoter activity in HRS cells by analyzing the HD cell
lines L428 and KM-H2 in transient transfection experiments. An
immunoglobulin promoter/enhancer reporter construct showed virtually no
activity in these cells in contrast to control B-cell
lines.4 We hypothesized that the loss of immunoglobulin
promoter activity in the HRS cells might be due to an absence of Oct2
and/or BOB.1/OBF.1 and, if true, that HRS cells should not only differ
from L&H cells in immunoglobulin expression but also in the expression
of Oct2 and/or BOB.1/OBF.1. To verify this hypothesis we investigated a
larger series of cHD and LPHD cases by immunohistochemistry for Oct2 and BOB.1/OBF.1 protein expression and correlated the results with the
presence of immunoglobulin protein and/or transcripts. For comparison
we included 8 cases of follicular lymphoma with monotypic IgH and IgL
expression and 3 non-neoplastic tonsils (data not shown). Our
investigations revealed that L&H cells The absence of the Oct2 protein from classical HRS cells came as a surprise because in one study30 its constant presence in primary HRS cells and HD cell lines, including the L428 and the KM-H2 lines, was reported. This discrepancy prompted us to examine the validity of our immunohistological results by investigating 11 cases of cHD, 4 cases of follicular lymphoma, and 3 hyperplastic tonsils for Oct2 mRNA with our very sensitive radioactive in situ hybridization procedure. In contrast to L&H cells, follicular lymphoma cells, and normal germinal center B cells, the HRS cells consistently failed to give rise to any Oct2-specific transcript signals. We also re-evaluated Oct2 expression in the HD-derived cell lines L428 and KM-H2, using various RNA and protein detection methods. We were unable to detect any Oct2-specific signals with any of the methods employed. In light of the highly positive correlation between immunoglobulin production and expression of Oct2 and BOB.1/OBF.1, it is tempting to assume that the absent transcription of immunoglobulin genes in HRS cells is due to the lack of Oct2 and/or BOB.1/OBF.1 in these cells. To strengthen or dismiss this conclusion, we performed cotransfection experiments in which we reintroduced BOB.1/OBF.1 and/or Oct2 into cultured HRS cells (L428) in association with immunoglobulin promoter-reporter constructs. The results obtained demonstrate that the lack of these transcription factors is indeed the likely cause for the defective immunoglobulin gene transcription in HRS cells. The strongest effect was seen when both the BOB.1/OBF.1 coactivator and the Oct2 transcription factor were cotransduced. However, the transfection of BOB.1/OBF.1 by itself also showed a significant effect. This observation is consistent with previous studies demonstrating that BOB.1/OBF.1 can also function in the absence of Oct2 as a coactivator because the function of Oct2 can be replaced by the ubiquitous Oct1 transcription factor and vice versa.12,31 In line with this assumption, Oct1 is expressed in the cultured HRS cells used for our transfection experiments (data not shown). The finding that the effect of BOB.1/OBF.1 is augmented by Oct2 in the
cultured HRS cells suggests that in more advanced differentiated B
cells Because the strong overexpression of Oct2 correlates to the high level of immunoglobulin in LPHD in the majority of cases, it is suggestive that the enormous amount of Oct2 in L&H cells might be responsible for, or contribute to, the frequent overexpression of immunoglobulin by these cells. However, what is more difficult to understand are the 2 cases of LPHD in which Oct2 is overexpressed and BOB.1/OBF.1 is normally expressed, but no immunoglobulin production is detectable. This points to the possibility that a further as yet unidentified factor (or factors) is involved in the regulation of immunoglobulin expression.
We thank H. Protz, A. Foerster, E. Berg, E. Seibt, and L. Oehring for their excellent technical assistance as well as L. Udvarhelyi for his help with the preparation of the manuscript.
Submitted May 31, 2000; accepted September 15, 2000.
Supported by grants of the Deutsche Forschungsgemeinschaft and Deutsche Krebshilfe, AIRC (Associazione Italiana per la Ricerca sul Cancro), and MURST.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Harald Stein, Institute of Pathology, Benjamin Franklin University Hospital, Free University Berlin, Hindenburgdamm 30, 12200 Berlin, Germany; e-mail: stein{at}ukbf.fu-berlin.de.
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 S. E. Coupland, C. Loddenkemper, J. R. Smith, R. M. Braziel, F. Charlotte, I. Anagnostopoulos, and H. Stein Expression of Immunoglobulin Transcription Factors in Primary Intraocular Lymphoma and Primary Central Nervous System Lymphoma Invest. Ophthalmol. Vis. Sci., November 1, 2005; 46(11): 3957 - 3964. [Abstract] [Full Text] [PDF]
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 D. Re, R. Kuppers, and V. Diehl Molecular Pathogenesis of Hodgkin's Lymphoma J. Clin. Oncol., September 10, 2005; 23(26): 6379 - 6386. [Abstract] [Full Text] [PDF]
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D. Re, R. K. Thomas, K. Behringer, and V. Diehl From Hodgkin disease to Hodgkin lymphoma: biologic insights and therapeutic potential Blood, June 15, 2005; 105(12): 4553 - 4560. [Abstract] [Full Text] [PDF]
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Y. Natkunam, I. S. Lossos, B. Taidi, S. Zhao, X. Lu, F. Ding, A. S. Hammer, T. Marafioti, G. E. Byrne Jr, S. Levy, et al. Expression of the human germinal center-associated lymphoma (HGAL) protein, a new marker of germinal center B-cell derivation Blood, May 15, 2005; 105(10): 3979 - 3986. [Abstract] [Full Text] [PDF]
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 C. Atayar, S. Poppema, T. Blokzijl, G. Harms, M. Boot, and A. van den Berg Expression of the T-Cell Transcription Factors, GATA-3 and T-bet, in the Neoplastic Cells of Hodgkin Lymphomas Am. J. Pathol., January 1, 2005; 166(1): 127 - 134. [Abstract] [Full Text] [PDF]
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A. Ushmorov, O. Ritz, M. Hummel, F. Leithauser, P. Moller, H. Stein, and T. Wirth Epigenetic silencing of the immunoglobulin heavy-chain gene in classical Hodgkin lymphoma-derived cell lines contributes to the loss of immunoglobulin expression Blood, November 15, 2004; 104(10): 3326 - 3334. [Abstract] [Full Text] [PDF]
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I. Bonzheim, E. Geissinger, S. Roth, A. Zettl, A. Marx, A. Rosenwald, H. K. Muller-Hermelink, and T. Rudiger Anaplastic large cell lymphomas lack the expression of T-cell receptor molecules or molecules of proximal T-cell receptor signaling Blood, November 15, 2004; 104(10): 3358 - 3360. [Abstract] [Full Text] [PDF]
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 A. Dutton, J. D. O'Neil, A. E. Milner, G. M. Reynolds, J. Starczynski, J. Crocker, L. S. Young, and P. G. Murray Expression of the cellular FLICE-inhibitory protein (c-FLIP) protects Hodgkin's lymphoma cells from autonomous Fas-mediated death PNAS, April 27, 2004; 101(17): 6611 - 6616. [Abstract] [Full Text] [PDF]
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T. Marafioti, M. Pozzobon, M.-L. Hansmann, G. Delsol, S. A. Pileri, and D. Y. Mason Expression of intracellular signaling molecules in classical and lymphocyte predominance Hodgkin disease Blood, January 1, 2004; 103(1): 188 - 193. [Abstract] [Full Text] [PDF]
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 M. Herling, G. Z. Rassidakis, L. J. Medeiros, T. P. Vassilakopoulos, K.-O. Kliche, G. Nadali, S. Viviani, V. Bonfante, R. Giardini, M. Chilosi, et al. Expression of Epstein-Barr Virus Latent Membrane Protein-1 in Hodgkin and Reed-Sternberg Cells of Classical Hodgkin's Lymphoma: Associations with Presenting Features, Serum Interleukin 10 Levels, and Clinical Outcome Clin. Cancer Res., June 1, 2003; 9(6): 2114 - 2120. [Abstract] [Full Text] [PDF]
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A. Brauninger, H.-H. Wacker, K. Rajewsky, R. Kuppers, and M.-L. Hansmann Typing the Histogenetic Origin of the Tumor Cells of Lymphocyte-rich Classical Hodgkin's Lymphoma in Relation to Tumor Cells of Classical and Lymphocyte-predominance Hodgkin's Lymphoma Cancer Res., April 1, 2003; 63(7): 1644 - 1651. [Abstract] [Full Text] [PDF]
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T. Marafioti, S. Ascani, K. Pulford, E. Sabattini, M. Piccioli, M. Jones, P. L. Zinzani, G. Delsol, D. Y. Mason, and S. A. Pileri Expression of B-Lymphocyte-Associated Transcription Factors in Human T-Cell Neoplasms Am. J. Pathol., March 1, 2003; 162(3): 861 - 871. [Abstract] [Full Text] [PDF]
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I. Schwering, A. Brauninger, U. Klein, B. Jungnickel, M. Tinguely, V. Diehl, M.-L. Hansmann, R. Dalla-Favera, K. Rajewsky, and R. Kuppers Loss of the B-lineage-specific gene expression program in Hodgkin and Reed-Sternberg cells of Hodgkin lymphoma Blood, February 15, 2003; 101(4): 1505 - 1512. [Abstract] [Full Text] [PDF]
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S. A. Pileri, G. Gaidano, P. L. Zinzani, B. Falini, P. Gaulard, E. Zucca, F. Pieri, E. Berra, E. Sabattini, S. Ascani, et al. Primary Mediastinal B-Cell Lymphoma: High Frequency of BCL-6 Mutations and Consistent Expression of the Transcription Factors OCT-2, BOB.1, and PU.1 in the Absence of Immunoglobulins Am. J. Pathol., January 1, 2003; 162(1): 243 - 253. [Abstract] [Full Text] [PDF]
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 V. Diehl, H. Stein, M. Hummel, R. Zollinger, and J. M. Connors Hodgkin's Lymphoma: Biology and Treatment Strategies for Primary, Refractory, and Relapsed Disease Hematology, January 1, 2003; 2003(1): 225 - 247. [Abstract] [Full Text] [PDF]
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G. Z. Rassidakis, L. J. Medeiros, T. P. Vassilakopoulos, S. Viviani, V. Bonfante, G. Nadali, M. Herling, M. K. Angelopoulou, R. Giardini, M. Chilosi, et al. BCL-2 expression in Hodgkin and Reed-Sternberg cells of classical Hodgkin disease predicts a poorer prognosis in patients treated with ABVD or equivalent regimens Blood, December 1, 2002; 100(12): 3935 - 3941. [Abstract] [Full Text] [PDF]
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 G. Muller, U. E. Hopken, H. Stein, and M. Lipp Systemic immunoregulatory and pathogenic functions of homeostatic chemokine receptors J. Leukoc. Biol., July 1, 2002; 72(1): 1 - 8. [Abstract] [Full Text] [PDF]
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D. F. Dukers, C. J. L. M. Meijer, R. L. ten Berge, W. Vos, G. J. Ossenkoppele, and J. J. Oudejans High numbers of active caspase 3-positive Reed-Sternberg cells in pretreatment biopsy specimens of patients with Hodgkin disease predict favorable clinical outcome Blood, June 17, 2002; 100(1): 36 - 42. [Abstract] [Full Text] [PDF]
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F. Jundt, I. Anagnostopoulos, R. Forster, S. Mathas, H. Stein, and B. Dorken Activated Notch1 signaling promotes tumor cell proliferation and survival in Hodgkin and anaplastic large cell lymphoma Blood, May 1, 2002; 99(9): 3398 - 3403. [Abstract] [Full Text] [PDF]
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F. Jundt, K. Kley, I. Anagnostopoulos, K. Schulze Probsting, A. Greiner, S. Mathas, C. Scheidereit, T. Wirth, H. Stein, and B. Dorken Loss of PU.1 expression is associated with defective immunoglobulin transcription in Hodgkin and Reed-Sternberg cells of classical Hodgkin disease Blood, April 15, 2002; 99(8): 3060 - 3062. [Abstract] [Full Text] [PDF]
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R. K. Thomas, A. Kallenborn, C. Wickenhauser, J. L. Schultze, A. Draube, M. Vockerodt, D. Re, V. Diehl, and J. Wolf Constitutive Expression of c-FLIP in Hodgkin and Reed-Sternberg Cells Am. J. Pathol., April 1, 2002; 160(4): 1521 - 1528. [Abstract] [Full Text] [PDF]
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 S A Pileri, S Ascani, L Leoncini, E Sabattini, P L Zinzani, P P Piccaluga, A Pileri Jr, M Giunti, B Falini, G B Bolis, et al. Hodgkin's lymphoma: the pathologist's viewpoint J. Clin. Pathol., March 1, 2002; 55(3): 162 - 176. [Abstract] [Full Text] [PDF]
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U. E. Hopken, H.-D. Foss, D. Meyer, M. Hinz, K. Leder, H. Stein, and M. Lipp Up-regulation of the chemokine receptor CCR7 in classical but not in lymphocyte-predominant Hodgkin disease correlates with distinct dissemination of neoplastic cells in lymphoid organs Blood, February 15, 2002; 99(4): 1109 - 1116. [Abstract] [Full Text] [PDF]
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E. Torlakovic, A. Tierens, H. D. Dang, and J. Delabie The Transcription Factor PU.1, Necessary for B-Cell Development Is Expressed in Lymphocyte Predominance, But Not Classical Hodgkin's Disease Am. J. Pathol., November 1, 2001; 159(5): 1807 - 1814. [Abstract] [Full Text] [PDF]
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F. Leithauser, M. Bauerle, M. Q. Huynh, and P. Moller Isotype-switched immunoglobulin genes with a high load of somatic hypermutation and lack of ongoing mutational activity are prevalent in mediastinal B-cell lymphoma Blood, November 1, 2001; 98(9): 2762 - 2770. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
-mercaptoethanol at 37°C and 5%
CO2.
131,
µEtm.luc contains the same promoter element with point mutations in
the octamer motif (see C). Relative activity is shown and the
cotransfections with empty expression vectors were arbitrarily set to
1. (B) L428 cells were cotransfected with wild type (4 × wt) or
mutant (4 × mut) octamer-dependent luciferase reporters together
with expression vectors for BOB.1/OBF.1 and/or Oct2 as indicated.
Relative activity is shown and the cotransfections with empty
expression vectors were arbitrarily set to 1. (C) Schematic
representation of the reporter constructs used in A and B. Wild-type
octamer motifs in the immunoglobulin promoter or synthetic promoter are
indicated as filled circles; mutant octamer motifs are shown as open
circles. Details about the reporter constructs can be found in Laumen
et al
as recently
assumed