Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation
Background: Infection of human B cells with Epstein—Barr virus (EBV) induces metabolic activation, morphological transformation, cell proliferation and eventual immortalization. Aim: To identify the nuclear receptors, which are the cellular interaction partners of EBNAs, that will help to elucidat...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2009
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| Цитувати: | Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation / S.P. Yenamandra, A. Lundin1, V. Arulampalam, M. Yurchenko, S. Pettersson, G. Klein, E. Kashuba // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 92–96. — Бібліогр.: 23 назв. — англ. |
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Yenamandra, S.P. Lundin, A. Arulampalam, V. Yurchenko, M. Pettersson, S. Klein, G. Kashuba, E. 2018-06-15T14:02:50Z 2018-06-15T14:02:50Z 2009 Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation / S.P. Yenamandra, A. Lundin1, V. Arulampalam, M. Yurchenko, S. Pettersson, G. Klein, E. Kashuba // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 92–96. — Бібліогр.: 23 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/135700 Background: Infection of human B cells with Epstein—Barr virus (EBV) induces metabolic activation, morphological transformation, cell proliferation and eventual immortalization. Aim: To identify the nuclear receptors, which are the cellular interaction partners of EBNAs, that will help to elucidate the mechanism of B cell transformation. Methods: We have compared the nuclear receptor profile in the naïve and EBV-transformed B-lymphocytes, using TaqMan LDA microfluidic card technology. Results: Out of 48 nuclear receptor, 17 showed differential expression at the mRNA level. The expression of 5 genes was elevated in EBV-transformed cells, whereas 12 genes were downregulated in lymphoblastoid cells (LCLs). 7 genes were studied at the protein level; 2 genes were up regulated (Nr2F2 and RARA) and 4 genes were down regulated (ERB, NUR77, PPARG, and VDR) in LCLs. Conclusion: The nuclear receptor profiling on EBV infected B cells showed alterations of nuclear receptors expression at both mRNA and protein levels compared with non infected peripheral blood cells. Further analysis on a possible role of each nuclear receptor in EBV induced cell transformation should be performed. Swedish Cancer Society, a matching grant from the Concern Foundation (Los Angeles), the Cancer Research Institute (New York), Swedish Institute, and Swedish Foundation for Strategic Research supported this work. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation Article published earlier |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
| title |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| spellingShingle |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation Yenamandra, S.P. Lundin, A. Arulampalam, V. Yurchenko, M. Pettersson, S. Klein, G. Kashuba, E. Original contributions |
| title_short |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| title_full |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| title_fullStr |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| title_full_unstemmed |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| title_sort |
expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation |
| author |
Yenamandra, S.P. Lundin, A. Arulampalam, V. Yurchenko, M. Pettersson, S. Klein, G. Kashuba, E. |
| author_facet |
Yenamandra, S.P. Lundin, A. Arulampalam, V. Yurchenko, M. Pettersson, S. Klein, G. Kashuba, E. |
| topic |
Original contributions |
| topic_facet |
Original contributions |
| publishDate |
2009 |
| language |
English |
| container_title |
Experimental Oncology |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| format |
Article |
| description |
Background: Infection of human B cells with Epstein—Barr virus (EBV) induces metabolic activation, morphological transformation, cell
proliferation and eventual immortalization. Aim: To identify the nuclear receptors, which are the cellular interaction partners of EBNAs,
that will help to elucidate the mechanism of B cell transformation. Methods: We have compared the nuclear receptor profile in the naïve
and EBV-transformed B-lymphocytes, using TaqMan LDA microfluidic card technology. Results: Out of 48 nuclear receptor, 17 showed
differential expression at the mRNA level. The expression of 5 genes was elevated in EBV-transformed cells, whereas 12 genes were downregulated
in lymphoblastoid cells (LCLs). 7 genes were studied at the protein level; 2 genes were up regulated (Nr2F2 and RARA) and
4 genes were down regulated (ERB, NUR77, PPARG, and VDR) in LCLs. Conclusion: The nuclear receptor profiling on EBV infected
B cells showed alterations of nuclear receptors expression at both mRNA and protein levels compared with non infected peripheral blood
cells. Further analysis on a possible role of each nuclear receptor in EBV induced cell transformation should be performed.
|
| issn |
1812-9269 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/135700 |
| citation_txt |
Expression profile of nuclear receptors upon epstein — barr virus induced b cell transformation / S.P. Yenamandra, A. Lundin1, V. Arulampalam, M. Yurchenko, S. Pettersson, G. Klein, E. Kashuba // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 92–96. — Бібліогр.: 23 назв. — англ. |
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| first_indexed |
2025-11-24T04:31:31Z |
| last_indexed |
2025-11-24T04:31:31Z |
| _version_ |
1850842156260917248 |
| fulltext |
92 Experimental Oncology 31, 92–96, 2009 (June)
Infection of human B cells with Epstein — Barr virus
(EBV) induces metabolic activation, morphological trans-
formation, cell proliferation and eventual immorta lization
[1, 2]. Six of the 9 EBV-encoded proteins (EBNA -1, -2,
-3, -5, -6, and LMP-1) are necessary for the efficient
transformation [3]. During transformation no genetic
aberrations were detected. However, signal transduction
pathways were ultimately changed — either blocked, ei-
ther activated. For example, LMP1 activates TNFα/CD40
downstream signaling pathways that can stimulate cell
growth and survival through activation of NFκB, jun and
p38/map kinase. LMP2A activates constitutively B-cell
receptor (BCR) (reviewed in [1]). Latency III genes ex-
pression leads to the change of gene expression profile.
EBNA-2 activates and regulates the transcription of Notch
and PU.1 responsive promoters of the cellular genes due
to a binding to RBP-Jκ. A more detailed description of
some cellular pathways and, moreover, nuclear recep-
tors that may be implicated in the EBV-induced B-cell
transformation, are reviewed in [4].
Anyway, many questions about the mechanism
of B cell transformation into lymphoblasts are unan-
swered yet. The identification of the cellular partners
of EBNAs and determination of the intervening cellular
pathways will help to elucidate the mechanism of B cell
transformation. The aim of the present paper was to
compare the nuclear receptor profile in the naïve and
EBV-transformed B-lymphocytes (freshly infected B
cells and long-term cultured LCLs).
MATERIALS AND METHODS
Western blotting. We prepared whole cell lysates
using NP40 lysis buffer (1% NP40, 150 mM NaCl,
50mM Tris, pH = 8) with a protease inhibitor cocktail
(Sigma-Aldrich, St Louis, MO, USA). Lysates were
cleared by centrifugation. Proteins were separated us-
ing the sodium dodecyl sulphate (SDS) polyacrylamide
gel electrophoresis (SDS-PAGE). After transfer the
membrane was probed with the specific antibodies:
mouse monoclonal anti-actin (Sigma-Aldrich), anti-
Nr2F2 (Abnova Corp., Taipei, Taiwan), anti-RARA (Ab-
nova Corp.), anti-VDR (Santa Cruz Biotechnology Inc.,
Santa Cruz, CA, USA), and polyclonal mouse serum
against PPARG (Abnova Corp.); rabbit polyclonal se-
rum against NUR77 (Santa Cruz Biotechnology Inc.).
Secondary antibodies (anti-rabbit and anti-mouse IgG
Horseradish-conjugated) were purchased from GE
Healthcare Bio-Sciences AB, Uppsala, Sweden.
Low density array. TaqMan LDA microfluidic card
technology from Applied Biosystems (Foster City, CA,
USA) allows the simultaneous assay of mRNA gene
expression of up to 384 targets on a single card. The
LDA used in this study was custom designed to consist
of 48 TaqMan Gene Expression Assays (Applied Bio-
systems) per loading port (48 genes × 5 samples run
three times for statistical significance). Each reaction
well contained all reagents specific for a given assay.
Each target assay consisted of a forward primer and
a reverse primer.
Cell culture, immunostaining and imaging.
All cells were cultured at 37 °C, in Iscove’s medium
containing 10% fetal bovine serum and appropriate
EXPRESSION PROFILE OF NUCLEAR RECEPTORS UPON
EPSTEIN — BARR VIRUS INDUCED B CELL TRANSFORMATION
S.P. Yenamandra1, 2, §, A. Lundin1, 2, §, V. Arulampalam1, 2,
M. Yurchenko3, S. Pettersson1, 2, G. Klein1, E. Kashuba1, 2, 3, *
1Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institute, Stockholm S17177, Sweden
2Center for Integrative Recognition in the Immune System (IRIS), Karolinska Institute, Stockholm
S17177, Sweden
3R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology NAS of Ukraine, Kiev
03022, Ukraine
Background: Infection of human B cells with Epstein — Barr virus (EBV) induces metabolic activation, morphological transformation, cell
proliferation and eventual immortalization. Aim: To identify the nuclear receptors, which are the cellular interaction partners of EBNAs,
that will help to elucidate the mechanism of B cell transformation. Methods: We have compared the nuclear receptor profile in the naïve
and EBV-transformed B-lymphocytes, using TaqMan LDA microfluidic card technology. Results: Out of 48 nuclear receptor, 17 showed
differential expression at the mRNA level. The expression of 5 genes was elevated in EBV-transformed cells, whereas 12 genes were down-
regulated in lymphoblastoid cells (LCLs). 7 genes were studied at the protein level; 2 genes were up regulated (Nr2F2 and RARA) and
4 genes were down regulated (ERB, NUR77, PPARG, and VDR) in LCLs. Conclusion: The nuclear receptor profiling on EBV infected
B cells showed alterations of nuclear receptors expression at both mRNA and protein levels compared with non infected peripheral blood
cells. Further analysis on a possible role of each nuclear receptor in EBV induced cell transformation should be performed.
Key Words: EBV, cell transformation, nuclear receptors, expression pattern, microarrays.
Received: April 3, 2009.
§Equal contribution.
*Correspondence: Fax: 468330498
E-mail: Elena.Kashuba@ki.se
Abbreviations used: EBV — Epstein — Barr Virus; EBNA — EBV-enco-
ded nuclear antigen; LCL — lymphoblastoid cell line; LMP — latent
membrane protein ; PBC — peripheral blood cells; TBC — tonsil B cells.
Exp Oncol 2009
31, 2, 92–96
Experimental Oncology 31, 92–96, 2009 (June) 93
antibiotics. Tonsil B cells (TBC) were isolated from
human tonsils obtained from routine tonsillectomy
(Karolinska Hospital, Stockholm). The tonsils were
cut into the fragments and passed through a metal
mesh. Peripheral blood B cells were isolated from
buffy coat blood (Karolinska Hospital) on Lymphoprep
gradients. An ethical permission was received for both
procedures of B cell isolation. Two subsequent rounds
of E-rosetting removed the T-cells. The B95.8 EBV
strain was used for B cell infection. Control B cells were
activated by anti-CD40 mouse monoclonal antibody
(Nordic Biosite AB, Täby, Sweden, 1 μg/ml) and IL4
(ImmunoTools GmbH, Friesoythe, Germany) 25 ng/ml
for 48 h. Prior to immunostaining experiments, the
cells were spun on glass slides, using Cytospin cen-
trifuge. Immunostaining and digital image capturing
was performed as described earlier [5]. Briefly, cells
on slides after cytospin were fixed in a 1 : 1 mixture of
cold methanol and acetone (–20 °C). After rehydra-
tion in phosphate buffer saline, cells were stained
with antibodies. Hoechst 33258 (Sigma-Aldrich) was
added at a concentration of 0.4 μg/ml to the seconda ry
antibody for DNA staining when necessary. The ima-
ges were captured using DAS microscope Leitz DM
RB with a dual mode cooled charged coupled device
(CCD) camera C4880 (Hamamatsu, Japan) or Zeiss
LSM 510 laser scanning confocal microscope with
ORCA-ER CCD camera (Hamamatsu).
RESULTS
Nuclear receptor expression profile on the
microfluidic cards. We have run Nuclear Receptor
profiling in freshly EBV-infected B cells (48 h) and
lymphoblastoid cell lines (LCLs) versus primary B
cells. We have used two different RNA samples of B
cells: peripheral blood B cells (PBC) and TBC, and
three different RNA samples of EBV-infected cells:
48 h post infection (EBV 48 h), and LCL that were cul-
tured for 2 month and 1.5 year. The set of 48 nuclear
receptors and GAPD were on the cards. Among them
12 receptors were not expressed, and 33 nuclear re-
ceptors were expressed differentially in normal PBC
and EBV-infected cells (level of expression differed
more than 2-fold). Standard deviation did not exceed
5.0%.
Only 17 genes showed consistent differences
in expression at the mRNA level — decreased in B
cells, upregulated in EBV-infected cells and LCLs, or
vice versa. The expression of 5 genes was elevated
in EBV-transformed cells, whereas 12 genes were
downregulated in LCLs (Table).
We have studied expression of 7 genes at the pro-
tein levels; 2 genes were upregulated and 4 genes were
downregulated, ER-α protein was not detected.
Genes upregulated upon EBV-infection. The
five genes: Nr2F2 (COUP2, NP_066285), Nr4A3
(MINOR, NP_775290), Nr6A1 (GCNF, NP_201591),
RARA (NP_000955), and RXRA (NP_002948) were
induced in LCLs at the mRNA level (Fig. 1, а). Slight
elevation of mRNA was observed in TBC, probably,
due to the activation by infection. Strong correlation
was observed at mRNA and protein level for Nr2F2
(trans criptional factor COUP2) and RARA (retinoic acid
receptor α), when primary PBC and TBC were com-
pared with freshly EBV infected cells and LCLs. Nr2F2
and RARA proteins were elevated in LCLs, compared
with primary B cells (Fig. 1, b, c, respectively).
Table. Differently expressing genes in EBV-infected compared to primary
B-cells
№ Name of the gene Accession number,
OMIM link
Upregulated receptors
1 Nr2F2, Nuclear receptor subfamily 2, group F,
member 2; TF COUP2; COUPTFII
NP_066285
*107773
2 Nr4A3, Nuclear receptor subfamily 4, group A,
member 3; Neuron-derived orphan receptor 1,
NOR1; Mitogen-induced orphan receptor, MINOR
NP_775290
+600542
3 Nr6A1, Nuclear receptor subfamily 6, group A,
member 1; Germ cell nuclear factor, GCNF
NP_201591
*602778
4 RARA, Retinoid acid receptor alpha NP_000955
*180240
5 RXRA, Retinoid X receptor alpha NP_002948
*180245
Downregulated receptors
1 PPAR-gamma, Peroxisome proliferator-acivated
receptor gamma; PPARG
NP_619725
*601487
2 ER-alpha, Estrogen receptor alpha; ESR; ESR1;
ER1
NP_000116
*133430
3 ER-beta, Estrogen receptor beta; ESR2; ER2 Q92731
*601663
4 Nr1H3, Nuclear receptor subfamily 1, group H,
member 3; Liver X receptor alpha; LXRA
NP_005684
*602423
5 Nr2F1, Nuclear receptor subfamily 2, group F,
member 1; Transcription factor COUP1; TFCOUP1
NP_005645
*132890
6 Nr3C1, Nuclear receptor subfamily 3, group C,
member 1; Glucoccorticoid receptor; GCCR
NP_001018087
+138040
7 Nr4A1, Nuclear receptor subfamily 4, group A,
member 1; NAK1; Nuclear hormone receptor
TR3; TR3; NUR 77 (homolog of mouse NUR77)
NP_775180
*139139
8 RARB, Retinoic acid receptor beta NP_000956
*180220
9 RORC, RAR-related orphan receptor gamma NP_005051
*602943
10 RXRB, Retinoid X receptor beta NP_068811
*180246
11 THRB, Thyroid hormone receptor beta NP_000452
+190160
12 VDR, Vitamin D3 receptor NP_000367
*601769
Genes downregulated in EBV-infected B cells.
Eleven genes were downregulated in LCLs compared
with primary B cells (Fig. 2, a, b). We have run Western
blotting for the 5 of them: ER-α and -β (Estrogen recep-
tor α (NP_000116) and -β (Q92731)), Nr4A1 (Nur77,
NP_775180), PPARG (peroxisome proliferator-activated
receptor γ, NP_619725), and VDR (vitamin D receptor,
NP_000367). ER-α protein was not detected by Wes-
tern blotting. ER-β protein level was very low in primary
B cells and LCLs. Nur77 and PPARG protein levels in
LCLs does not differ much from the protein level in the
primary B cells (Fig. 2, c). Different trends in mRNA
and protein levels (compare Fig. 2, a, c) could be due
to the protein stability. However, VDR protein expres-
sion (Fig. 2, d) followed a pattern of mRNA expression
(compare Fig. 2, a, d). Moreover, after a brief increase,
all 11 receptors were expressed at lower level in LCLs,
compared with primary B cells.
We have to mention, that not only the level of
expression, but also a cellular distribution of the nu-
clear receptors were changed after EBV infection. For
94 Experimental Oncology 31, 92–96, 2009 (June)
examp le, in primary TBC the PPARG was expressed as
small cytoplasmic patches (Fig. 3, a). Upon activation,
pattern of expression of PPARG was changed. Upon
LPS (lipopolysaccharide) stimulation of B cells PPARG
showed perinuclear localization, which could also be
found in the patches (Fig. 3, b). In B cells stimulated
by anti-CD40 and IL-4, PPARG expressed not only in
patches, but also at the membrane of the joined cells
(Fig. 3, c). Upon EBV-infection PPARG was observed
in the cellular membrane, and great portion of protein
was distributed in both, a nucleus and the cytoplasm
(Fig. 3, d). We may hypothesize that PPARG would
function differently depending on distribution pattern
in cells. However, this should be further elucidated.
DISCUSSION
EBV-encoded proteins expressed in latently infec-
ted B cells are known to interact with a cellular signaling
pathways to establish latency and ensure the growth of
Р 1
Р 4
-1.5
-1
-0.5
0
0.5
1
1.5
2
2.5
PB
C
TB
C
EB
V
(4
8
h)
LC
L
(2
m
on
th
)
LC
L
(1
.5
y
ea
rs
)
0
0.5
1
1.5
2
2.5
Nr2F2/Actin
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
RARA/Actin
Nr2F2
Actin
RARA
Actin
50 kDa –
98 kDa –
B-
ce
lls
CD
40
+
IL
4
EB
V
LC
L
48 h
B-
ce
lls
CD
40
+
IL
4
EB
V
LC
L
48 h
a
b
c
RARA
RXRA
Nr6A1(GCNF)
Nr2F2(COUP2)
Nr4A3(MINOR)
Fig. 1. a, mRNA expression values (log) of upregulated nuclear
receptors. Names of receptors are indicated in corresponding
colors. Notice increased expression in LCLs. b, Western blotting
of the Nr2F2 in primary, CD40+Il4 activated, freshly EBV infected
B cells, and LCLs (cultured for 1.5 years). Lower panel — relative
ratio of protein signal to actin. c, Similar to b, for RARA protein
a
b
c
d
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
PPARG/Actin Nur77/Actin
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
VDR/Actin
VDR
Actin
PPARG
Nur77
Actin
50 kDa –
64 kDa –
50 kDa –
98 kDa –
64 kDa –
B-
ce
lls
CD
40
+
IL
4
EB
V
LC
L
B-
ce
lls
CD
40
+
IL
4
EB
V
CD
40
+
IL
4
EB
V
LC
L
48 h
PB
C
TB
C
EB
V
(4
8
h)
LC
L
(2
m
on
th
)
LC
L
(1
.5
y
ea
rs
)
Р 1
Р 3
Р 5
-2
-1.5
-1
-0.5
0
0.5
1 ER-beta
ER-alpha
Nur77(Nr4A1)
LXRA(Nr1H3)
VDR
PPARG
Р 1
Р 3
Р 5
-3
-2.5
-2
-1.5
-1
-0.5
0
PB
C
TB
C
EB
V
(4
8
h)
LC
L
(2
m
on
th
)
LC
L
(1
.5
y
ea
rs
)
THRB
RARB
RORC
Nr3C1(GCCR)
Nr2F1(COUP1)
RXRB
Fig. 2. a, b, mRNA expression values (log) of downregulated
nuclear receptors. Names of receptors are indicated in cor-
responding colors. Notice decreased expression in LCLs.
c, Western blotting of the Nur77 and PPARG in primary, CD40+Il4
activated, freshly EBV infected B cells, and LCLs (cultured for 1,5
years). Lower panels — relative ratio of protein signal to actin.
d, Similar to c, for VDR protein
Experimental Oncology 31, 92–96, 2009 (June) 95
the transformed cells (reviewed in [6]). Beside latent
membrane proteins (LMPs), EBV-encoded nuclear
antigens (EBNAs) are implicated in this process as
well. For example, the EBNA-2 can bind to the nuclear
receptor Nur77 and block NUR77 mediated apoptosis
[7, 8]. EBNA-2 can also inhibit the pro-apoptotic and
anti-proliferative functions of the transforming growth
factor β1 (TGFβ1, NP_000651) cytokine, as shown in
the EBNA-2 inducible EREB system [9]. We have pre-
viously shown that EBNA-3 can bind to and regulate
the transactivation function of the cellular nuclear
receptor AhR [10]. Importantly, EBNA-3 influenced
the transcription of AhR dependent genes at the basal
receptor level and after ligands (xenobiotics) activa-
tion, including 2,3,7,8-tetrachloro-dibenzo-p-dioxin
(TCDD). The physiological role of the EBNA-3 — AhR
interaction was illuminated by treating B cell lines
with TCDD. EBNA-3 was found to protect cells from
TCDD-induced growth arrest and/or apoptosis.
However, the question about regulation of the
nuclear receptors upon EBV-infection of B cells is
not completely elucidated yet. In current study we
aimed to get an overview about relationship between
EBV-induced transformation of B cells and the expres-
sion profile of 48 nuclear receptors.
Summarizing, we have found that 17 nuclear
receptors were expressed differently in primary and
EBV-infected B cells; 12 of them were downregulated
and 5 were upregulated in LCls (see Table).
Some of these receptors previously were shown to be
implicated in the regulation of the EBV-infected cell fate.
For example, it was demonstrated that EBV lytic cycle
activation was inhibited upon retinoic acid treatment due
to direct binding between RARA and the EBV-encoded
lytic BZLF1 protein [11, 12]. It was also found that reti-
noids inhibited naïve B cell proliferation, but promoted
cell survival [13]. Noteworthy, here we show that RARA
is upregulated in LCLs (see Fig. 1, a, c).
Earlier, it was reported that in B cells (PBCs and
LCLs) the VDR-dependent gene regulation was
blocked [14]. Moreover, the active VDR pathway
could inhibit proliferation and enhance differentiation
of leukemic cells [15]. Interestingly, the level of VDR
expression (at the mRNA and protein levels) was found
to be lower in the EBV transformed cells compared
with primary B cells (see Fig. 2, a, d). Recently, an
anti-tumor effect was proposed for vitamin D and VDR
[16] (for review see [17, 18]).
Expression of functional PPARG was shown in
lymphocytes, and its activation led to apoptosis, or
growth arrest [19, 20], or differentiation [21]. PPARG
can have both, transactivating and transrepressing
activity (reviewed in [22]). PPARG can repress some
interferon-gamma and LPS-inducible genes, such,
as IL-12 and IP10, for example. In their turn, cytokines
can repress PPARG by inhibiting of DNA binding [23].
Here we showed that PPRG was downregulated in LCLs
compared with primary B cells. Moreover, cellular distri-
bution of this nuclear receptor was changed in LCLs.
Levels of nuclear receptors mRNA and protein ex-
pression vary from maximum to minimum before they
get stabilized, as it is seen from the Fig. 1 and 2. Note-
worthy, it was shown earlier that EBV infection often
results in temporary (0–72 h) up- or downregulation of
many cellular and viral genes (reviewed in [1]).
The nuclear receptor profiling on EBV infected B
cells showed alterations of nuclear receptors expression
at both mRNA and protein levels compared with non
infected peripheral blood cells. In most of the cases,
the mRNA levels observed via LDA are strongly corrobo-
rated by expression at protein level. Further analysis on
a possible role of each nuclear receptor in EBV induced
cell transformation should be carried out.
ACKNOWLEDGMENTS
Swedish Cancer Society, a matching grant from the
Concern Foundation (Los Angeles), the Cancer Research
Institute (New York), Swedish Institute, and Swedish Foun-
dation for Strategic Research supported this work.
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