(EBV) Type-l Variant Strains in Both Malignant and

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Common Epstein-Barr Virus (EBV) Type-l Variant Strains in Both
Malignant and Benign EBV-Associated Disorders
By Volker Schuster, German Ott, Silvia Seidenspinner, and Hans Wolfgang Kreth
In the present study, Epstein-Barr virus (EBV) isolates from
l 8 malignant tumors (angioimmunoblastic lymphadenopathy [AILD], n = 4; Hodgkin‘s disease [HDI,n = 3; pleomorphic
T-cell non-Hodgkin‘s lymphoma [l-NHLI, n = 1; &cell nonHodgkin‘s lymphoma [B-NHL], n = 8; gastric carcinoma, n =
2) as well as from 10 tonsils of EBV-seropositive children
and from peripheral blood mononuclear cells of 12 children
with uncomplicated infectious mononucleosis (IM) and of a
boy with severe chronic active EBV infection ware genotyped in the EBV nuclear antigen-2 (EBNA-2) gene. A total
of 40 of 41 isolates harbored EBV type 1; in 1 specimen
(tonsil), only EBV type 2 was found. Further molecular chartp
wild-type isolates in the EBNA-2
acterization of EBV y
gene and in the 40-kb distant EBV-encoded small RNAs
(EBER) region showed that different groups of stable EBV
strains exist in vivo both in benign and malignant lymphatic tissue. Group 1 is composed of EBV type-l
isolates (B-NHL, n = 3;T-NHL, n = 1; HD, n = 1; IM, n = 4)
that showed a 695-8-Iike DNAsequence pattern in both
viral genes. Group 2 isolates (HD, n = 1; AILD, n = 1; B-NHL,
n = 1; tonsils of EBV-seropositive children, n= 9; IM, n = 2)
showed a nucleotide change at position 49095 in the EBNA2 gene, leading t o an amino acid substitution (Pro -,Ser),
and EBV type9 sequences in the EBER region. EBV type-l
isolates that fall into
group 3 (AILD, n = 3; HD, n = 1; B-NHL,
n = 4; gastric carcinoma, n = 2; IM, n = 6; severe chronic
active EBV infection, n = 1) were characterized by typical
nucleotidechanges and a 3-bp
insertion (CTC;extra Leu residue) in the EBNA-2 gene and anEBV type-2-specific sequence pattern in the EBER region. These EBV type-l variant
strains may represent the most prevalent circulating EBV
type-l strains in the exposed population and seem not t o
be restricted t o a certain EBV-associated disease or tumor
type. However, analysis of more EBV isolates from benign
and malignant lesions must show whether more EBV type1 substrains exist in vivo.
0 1996 b y The American Society of Hematology.
a common phenomenon of EBV type-l wild-type isolates
derived from patients with different malignant lymphomas
and from children with benign clinical conditions. We extended this analysis to another locus, the 40-kb distant EBER
region, using single-strand conformation polymorphism
PSTEIN-BARR VIRUS (EBV), the causative agent of
infectious mononucleosis (M),
has been associated
with endemic Burkitt’s lymphoma (BL), with nasopharyngeal carcinoma, with certain B- and T-cell lymphomas, with
approximately 50% of Hodgkin’s disease (m),
with different types of gastric carcinomas, and with B-cell lymphoproliferative disorders in immunosuppressed individuals.”’
According to their DNA sequence (and protein) divergence within the EBV nuclear antigens (EBNA-2, -3a, -3b,
-3c) and the 40-kb distant EBV-encoded small RNAs
(EBER) region, two types of EBV (EBV type 1, EBV type
2) have been identified.”” EBNA-2 is of particular interest
because this protein plays an essential role in the transformation process of B lymphocytes by EBV”*” and seems to be
a critical determinant for EBV-induced lymphoma tumor
growth in immunodeficient severe combined immunodeficiency disease mice.I3 The biological differences between
EBV type 1 and EBV type 2 appear to be mainly caused by
the divergence within the EBNA-2 protein, because EBV
type 2, when compared with EBV type 1, has a lower transforming efficiency, a poorer initial outgrowth, and a higher
cell-density dependence for cell viability in ~ i t r 0 . lBoth
EBV types occur worldwide, with different geographical distributions. In general, EBV type 1 is the predominant type
in lymphatic cells of EBV-infected individuals. Immunodeficient hosts, especially those with acquired immunodeficiency syndrome, show increased infection rates with EBV
type 2.15However, little is known of whether both strains
really differ in their biological activities in vivo. Recently,
Aitken et a l l 6 found EBNA-2 sequence heterogeneity in EBV
type-l isolates from four African and New Guinea endemic
BL cases, suggesting that variant EBV type-l substrains may
be associated with these tumors. Different point mutations
in the EBNA-2 gene have been described in some EBV
type-l isolates from human immunodeficiency virus-infected
patients but not from human immunodeficiency virus-negative subjects.”
The present study was undertaken to evaluate if stable
and constant nucleotide changes in the EBNA-2 gene are
Blood, Vol 87, No 4 (February 15). 1996: pp 1579-1585
Clinical samples. Biopsy specimens of 16 malignant tumors
(centroblastic B-cell non-Hodgkin’s lymphoma [B-NHL], n = 7;
HD, n = 2; angioimmunoblastic lymphadenopathy [AILD], n = 4;
pleomorphic T-cell NHL [T-NHL], n = 1; and gastric carcinoma, n
= 2) were selected from the files of the lymph node registry at the
Department of Pathology in Wiirzburg, Germany. In addition, 2
biopsy samples (Hodgkin’s disease [HD-21 and small-cell lymphocytic B-NHL with plasmacytoid features [B-NHL-4], respectively),
were derived from 2 brothers with X-linked lymphoproliferative
disease, an inherited immunodeficiency with a high vulnerability to
EBV infection^.'^,'^ Furthermore, peripheral blood mononuclear cells
(PBMCs) from 12 children with uncomplicated acute IM and from
a 6-year-old boy with severe chronic active EBV infection
(SCAEBV) as well as tonsils from 10 tonsillectomized EBV-seropositive children were examined for the presence of EBV type-l
variant strains.
Cell lines. The B95-8 line was used as a source of EBV type1 prototype.” This EBV strain was originally established from an
From the Children’s Hospital and Institute of Pathology, University of Wiirzburg, Wiirzburg, Germany.
Submitted June 5, 1995; accepted October 12, 1995.
Supported by the Deutsche Forschungsgemeinschft (grant no.
Schu NOD-3) and in part the Sondegorschungsbereich 172, C8.
Address reprint requests to Volker Schuster, MD, Children’s Hospital, University of Wiirzburg, Josef-Schneider-Strasse 2, D-97080
Wiirzburg, Germany.
The publication costs of this article were defrayed in part by page
charge payment. This article must therefore be hereby marked
“advertisement” in accordance with 18 V.S.C. section 1734 solely to
indicate this fact.
0 1996 by The American Society of Hematology.
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1 2 3 45 6 7 8 9l011 1213 141516 17 1819
IM patient.” The BL cell line Jijoye was used as a source of EBV
type-:! prototype.”
Polymerase chain reaction (PCR) of o portion of theEBNA-2
gene. Total DNAwas extracted from biopsy material, PBMCs, or
cell lines and was then amplified in duplicates either with EBNA-2
primers specificfor EBV type 1 (S’-TCTTGATAGGGATCCGCTAGGATA-3’. nucleotide positions 48839-48862: and S’-ACCGTGGTTCTGGACTATCTGGATC-3’. nucleotide positions 49335-4931
or with EBNA-2 primers specific for
EBV type 2 (S’-ACTGGATATGAATCCCCTGGGCAG-3’. nucleotide positions 48766-48789; and
S‘-GAGTCCTGTACTATCAGAACTACAATG-3‘, nucleotidepositions 49231-4920S).’”.’2 The amplified products had a size of 497
bp (EBV type-l sequences) or 466 bp (EBVtype-2 sequences),
respectively. PCR conditions have been reported eisewhere.”
Southern blot h,vbridi:atior~ ona/ysi.r. To controlthespecificity
ofEBNA-2 genotyping, PCR products were electrophoresed in a
2% agarosegelandwere
Southem-blotted onto a nylonsupport
(Gene Screen Plus; DuPont, Braunschweig, Germany). Membranes
an internal”P-labeledEBNA-2
probe that was either specfic for EBV type I or specific for EBV
type 2. The internalprobespecificfor EBVtype 1 (155 bp) was
1 2 3 4 5 6 7 8 9 1011 1213 141518171819
generated by PCR using the primers S’-CAACCACTCATGATGCCACCAAGGC-3’ (nucleotide positions 49077-49101) and S’-GATGGTGTGGGTTGAAGTTCGGTAG-3’ (nucleotide positions 4923 149207) and as template EBV type-lprototypestrainB95-8.
internal probe specific for EBV type 2 (IS5 bp) was generated by
CATA-3’(nucleotide positions 49004-49028) and S‘-AGACTTAGTTGATGCCCTAGTGTGA-3’ (nucleotide positions 49158-49134)
and as template EBV type-2 prototype strain Jijoye. Hybridization
probes were radiolabeled by inclusion of 2 pCi [a-“P]-deoxycytidine triphosphate (3.000 Cilmmol) in the PCR. PCR conditions were
the same as indicated above.” After washing under stringentconditions blots were dried and subjected
to autoradiography at -70°C
with intensifying screens.
Direct DNAseqrcencirtg
uf EBNA-2 PCR products. Doublestranded PCR products were purified with PrimeErase Quick Push
Columns (Stratagene, Heidelberg, Germany) and were directly cycle-sequenced by the dideoxy-termination method using ”S-deoxyadenosine triphosphate and theExo(-)Pfu Cyclist DNA Sequencing
Fig 1. (a and b) Genotyping of clinical EBV isolates in the EBNAKit (Stratagene) according to the manufacturer’s protocol. Samples
2 gene. DNA was amplified in duplicates either by primers specific
wereelectrophoresed on6% polyacrylamide/8.3 mol/L urea sefor EBV type 1 (A) or by primers specific for EBV type 2 (B)?’ Lane
quencing gels (Roth, Karlsruhe, Germany), dried, andexposed to
1, 100 bp ladder; lane 2, EBV type-l prototype strain 895-8; lane 3,
Kodak XAR-S films (Eastman-Kodak, Rochester. NY).
EBV type-2 prototype strain Jijoye; lane 4, HP; lane 5, EBV-negative
SSCP analysis .f theEBER region. Forgenotypingof
control DNA (BJAB); 14 different clinical isolates (tonsils, lanes 6-9
isolates in the EBER gene, which reveals 6 type-specific point mutaand 15-19; PBMC from children with IM, lanes 11-14; and HD-3, lane
tions: we used SSCP analysis similar to that describede l s e ~ h e r e . ~ ” . ~ 10)
~ that all contain EBV type 1 are shown. (a) Ethidium bromide
staining. (b) Southern blot hybridization with an internal (EBV typeThe genotype-specificpointmutationscaneasilybedetected
l-specific) probe (A); Southern blot hybridization with an internal
characteristic shifts in mobility because of conformational changes
fEBV type-2-specific) probe (B).
of the (single-stranded) DNA sequences.
PCR fragments with a size of190 bp were amplified from 200
ngof genomic DNA in a S0 pL mixture containing I pmol/L of
both EBER-specific primers S’-GTGGTCCGCATGTTTTGATC-3‘ cyanol) and was heat-denatured for 10 minutes at 80°C. A total of
(nucleotide positions 6780 - 6799) and S’-GCAACGGCTGTCCTG4 pL of themixture was electrophoresed in a nondenaturing 5%
TTTGA-3’ (nucleotide positions 6969 - 6950),’1200 prnol/L of each
polyacrylamidegel (Roth) containing S% glycerol in 0.SX Trisdeoxynucleotidetriphosphate, 0. I S pL “P-labeled deoxycytidine triborate-EDTA (TBE) buffer at room temperature for 7 hours at 10
phosphate (3.000 Cilmmol; Amersham Buchler, Braunschweig, GerW. The gel was dried and exposed to Kodak XAR-S films.
many), 10 mmol/L Tris-HCI (pH 8.3). S0 mmol/L KCI, I .S mmol/L
MgCI?, and 2.5 U of Taq DNA polymerase (Boehringer, Mannheim,
Germany). Initial denaturation was for S minutes at 94°C. SubseDNA was isolated from biopsy specimens of 18 malignant
quently, 35 cycles with denaturation for 30 seconds at 94°C. annealtumors (AILD, B- and T-NHL, HD, and gastric carcinoma),
ing for 30 seconds at 57°C. and extension for I minute at 72°C were
from 10 tonsils (of EBV-seropositive childrenwho were
performed in a PerkinElmer-Cetusthermocycler(PerkinElmertonsillectomized for recurrent chronic tonsillitis), from
Cetus, Nonvalk. CT). A total of S pL of the final product was mixed
PBMCs of 12 children with uncomplicated acute IM, and
and diluted with 45 pL formamide-loading buffer (95% formamide,
from PBMCs of a boy with SCAEBV and were
10 mmol/LEDTA, 0.05% bromophenolblue,and 0.05% xylene
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Table 1. EBNA-2 Sequences and EBVGenotype in the EBER Region Present in 41 EBV Wild-Type lsolatw
EBNA-2 Nucleotide‘ Sequences
Group 1
895-8 (EBV type-l strain)
B-NHL-1 (CB, 3844B9)
B-NHL-2 (CB, 1005/90)
B-NHL-3 (CB composite, 1826/91)
T-NHL-1 (9594/89)
HD-3 (379)
IM (n = 4)
Group 2
HD-2, B-NHL-4t
AILD-2 (2769190). tonsils (n = 9)
IM (n = 2)
Group 3a
(H209/89) AILD-1
(1966/91) AILD-3
B-NHL-5 (CB, H4088/91)
B-NHL-6 (14760/91)
IM (n = 4)
Group 3b
HD-1 (376)
AILD-4 (24618/91)
B-NHL-7 (H2141/91)
B-NHL-8 (9945/92)
GC-l (13811/92)
GC-2 (17965/92)$
IM (n = 2)
W91 (African BL)”,”§
Group 3c
FWA (African BL)Ӥ
Group 3d
ODH (African BL)16§
(EBV AG876
type-2 strain)
Jijoye (EBV type-2 strain)
Tonsils (n = 1)
EBER Genotype
EBV type l
EBV type 2
EBV type 2
EBV type 2
(W91 NT)
A la
EBV type 2
Abbreviations: CB, centroblastic; GC, gastric carcinoma; SCAEBV, severe chronic active EBV infection.
Numbering of nucleotides is rendered according to Baer et al?’
t Tumor biopsy specimens from two brothers withX-linked lymphoproliferative disease.”
This EBV isolate shows an additional base exchange at position 49159 (G + A) leading to an amino acid substitution (Arg His).
5 Data are taken from the literature.”~’2~’E
EBV isolates RNA and ODH show an additional base exchange at position 49110 (C T) leading to
an amino acid substitution (Pro -,Sed.
for the presence of EBV. All but 1 of the biopsy samples
contained EBV type- 1 genomes as determined by FCR using
genotype-specific EBNA-2 primers (Figs l a and b). Only 1
isolate from benign tonsil tissue contained EBVtype 2. Double infections with both strains, EBV type 1 and EBV type
2, were not detected in any of the pathological specimens.
To detect DNA sequence variations within the EBNA-2
gene that may possibly allow the definition of EBV type-l
variant strains, EBNA-2 PCR products of all isolates were
directly sequenced and compared with known EBNA-2 sequences of EBV type-l prototype strain B95-8andEBV
type-2 prototype strains AG876 and Jijoye (Table 1).In
addition, all isolates were simultaneously genotyped in a
gene 40-kb distant, the EBER region. As shown in Table 1,
the isolates could be classified into three different groups
according to their individual EBNA-2 sequence pattern and
their EBER genotype.
Isolates showing a B95-&like pattern at both gene loci
fall into group 1 (Fig 2%right [B-NHL-l]; Fig 2b, middle
[T-NHL-l]; and Fig 3, lanes 12 and 9, respectively; see
Table l]).
EBV type-l substrains of group 2 were characterized by
a nucleotide mismatch inthe EBNA-2 gene at position 49095
(C -+ T) leading toanamino acid exchange (Pro + Ser)
and EBV type-2 sequences in the EBER region. Figure 2a
(middle) shows the EBNA-2 sequence pattern of a group-2
isolate (AILD-2). Inthe EBER region, this isolate was found
to contain EBV type-2 sequences (Fig 3, lane 10).
Interestingly, in 2 brothers with X-linkedlymphoproliferative disease who developed histologically different malig-
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(C+A) 49091 *
b lT+C
(G-A 1
491 98
1C+A 49091
Fig 2. (a) Nucleotide sequence pattern of EBNA2 PCR products of three different
EBV type-l isolates
(the noncoding, complementary strand is shown).
Numbering of nucleotides is consistent
with that
used by Baer et 81.2" (Left panel) This EBV type-l
isolate derived fromAILD-1 (H209/89) is a representative for the group3 EBV type-l substrain characterized by nucleotidechanges I") at positions 49091
(C A),49113 (T A), and 49170 (G A) and a
3-bp insertion (GAG) between position 49136 and
49137. In addition, a nucleotide mismatch was found
C) in this isolate. (Middle
at position 49119(G
panel) This EBV type-l isolate derived from AILD-2
(2769/90) shows a nucleotide mismatch at position
49095 (G A), which ischaracteristic for thegroup2 EBV type-l substrain. (Right panel) The EBNA-2
nucleotide sequence of this EBV type-l isolate derived from centroblasticB-NHL-1 (3844/89) is identical with thecorresponding sequence of 895-8 prototype lie, group-l EBV type-l substrain). (b)
Nucleotide sequence pattern of EBNA-2 PCR products of three different EBV type-l isolates (the noncoding, complementary strand is shown). Numbering of nucleotides is consistent with that used by
Baer et 81.2' (Left and rightpanels) These EBV type1 isolates derived fromHD-1 (376)and AILD-3 (1966/
91) are representatives for the group-3 EBV type-l
substrains characterized by nucleotide changes (*I
at positions 49091 IC A), 49113 (T -Al. and 49170
(G -A) and a 3-bp insertion(GAG) between position
49136 and 49137. The isolate from AILD-3 showed
an additional nucleotide mismatch
at position 49119
(G C). (Middle panel) The EBNA-2 nucleotide sequence of this EBV type-l isolate derived from TNHL-1 (9594/89) is identical with the corresponding
sequence of 895-8 prototype lie, group-l EBV type1 substrain).
L9136 I
AlLD- 3
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-ds DNA
12 3 4 5 6 7 8 9
12131415 16 17 18
Fig 3. PCR-SSCP analysis of EBER region. PCR products of 190 bp were amplified usingEBER-specific primers, were denatured, and were
subjected t o SSCP analysis in a nondenaturing 5% polyacrylamide gel, as described in the Materials and Methods. Genotype-specific point
mutations were visualized as shifts in mobility. The single-stranded DNA resolved into the threeupper bands, which is caused by thefaster
mobility of the
sense strand when compared
with thatof the antisense strand." The reason for the third
(fainter) upper band remains unclear.
Probably, it contains a partially denaturedcomplex of senselantisense strands. The thick band with the fastest mobility
represents the doublestranded DNA (dsDNA). Lane 1, 895-8 (EBV type-l prototype strain); lane 2, Jijoye (EBV t y p e 9 prototype strain); lane 3, BJAB (EBV-negative
BL cell line); lane 4, Jijoye; lane 5, HD-1 (376); lane 6, HD-3 (379); lane 7, AILD-1 (H209/89); lane 8, T-NHL-2 (H101/90); lane 9, T-NHL-1 (95941
89); lane 10, AILD-2 (2769/90); lane 11, AILD-3 (1966/911; lane 12, B-NHL-1 (centroblastic, 3844/89); lane 13, B-NHL-3 (centroblastic composite,
1826/91); lane 14, B-NHL-6 (14760/91); lane 15, B-NHL-5 (polymorphic centroblastic, H4088/91); lane 16, B-NHL-2 (centroblastic, 1005/901; lane
17, double-stranded DNA control 1895-8); and lane 18, 895-8 (EBV type-l prototype).
nant tumors, namely HD (HD-2) and small-cell lymphocytic
B-NHLwith plasmacytoid features (B-NHL-4), the same
EBV type-l substrain (group 2) was found in tumor biopsy
specimens. This might suggest that the strain was transmitted
from one family member to another.
Somewhat surprisingly, all 9 EBV type-l isolates from
tonsils of children with chronic tonsillitis examined so far
could be classified as group 2 EBV type-l substrains.
Group 3 is composed ofEBV
isolates with EBNA-2
nucleotide mismatches at positions 49057 (A + G ) , 49091
(G + T), 491 13 (A T) and 49170 (C T), leading to
three amino acid substitutions (Gln
Thr + Ser) and an insertion of three nucleotides (CTC) between position 49 l36 and 49 137 resulting in an extra amino
acid (Leu) (Figs 2a and b and Table 1). All these isolates
contained EBV type-2 sequences in the EBER region (Fig
3). In 9 isolates (AILD-I; AILD-3; B-NHLJ; B-NHL-6;
SCAEBV; IM, n = 4) an additional nucleotide mismatch
was found at position 491 19 (C + G), leading to an amino
exchange (Leu
Val; see Fig 2a, left, andFig 2b, right,
Table l).
Recently, a region of the EBNA-2 gene of the African
endemic BLs W91 ," FWA and ODH'" has been sequenced
and compared with the corresponding region of EBV type1 strain B95-8. According to the characteristic nucleotide
mismatches in EBNA-2, these three strains most probably
belong to the category of group 3 substrains (Table l ) . However, genotyping in the EBER region has been not performed
in any of these isolates.
Our results show that distinct stable EBV type-l variant
strains are common in vivo. According to their characteristic
sequence pattern in the EBNA-2 and the EBER genes, we
can define three groups of EBV type-l substrains. Group 1
is composed of EBV isolates with a B95-8-like pattern at
the EBNA-2 and the EBER gene locus. Group 2 isolates are
characterized by one nucleotide mismatch (C T) in the
EBNA-2 gene at position 49095 andbyEBV type-2-like
sequences in the EBER region. Group 3 isolates show a
W91-like pattern in the EBNA-2 gene and EBV type-2-like
sequences in the EBER region.
These EBV type-l variant strains seem not to be restricted
to a certain clinical disorder or tumor type. We could also
show that the same EBV type-l substrain (of group 2) occurred in histologically different malignant tumors of 2
brothers whoboth carried the defective gene of X-linked
lymphoproliferative disease,"' suggesting intrafamilial transmission of this EBV substrain.
During IM, the infection is not only restricted to B cells,
which morphologically include Reed-Stemberg-like cells,
immunoblasts, medium-sized lymphoid cells, andplasma
cells. In addition, varying numbers of T lymphocytes are
also infected by EBV.'"' Therefore, it is tempting to assume
that these EBV-infected B- and T-cell populations in IM
might represent thebenign counterparts of EBV-infected
tumor cells in BL, HD, and different B- and T-cell lymphomas. Our findings that the EBV type-l variant strains found
in benign clinical conditions (such as 1M) matched the EBV
type-l variant strains isolated from different malignant tumors might support this suggestion. In addition, EBV variant
strains with certain deletions/mutations within the gene of
EBVlatent membrane protein-l have beenfound in both
malignant andbenign
EBV-associated diseases." These
findings and our findings might suggest that these EBV variant strains may represent the most prevalent circulating EBV
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strains to which the patient population is exposed. However,
analysis of more EBV isolates from benign and malignant
lesions must showwhether more EBV type-l substrains exist
in vivo.
The biological consequences of EBV variant strains in
vivo are still not clear. Because the W91 strain has a “normal” transforming capacity in vitro”.” and in vivo,” we
suggest that other EBV isolates from group 3 also do not
significantly differ in their biological behavior when compared with B95-8-like group-l EBV type-l strains.
Intactand deleted W91 strain-like EBNA-2 sequences
have been recently identified in two patients with oral hairy
leukoplakia,**suggesting that group-3 (and most probably
also group-2) EBV type-l variant strains may also persist
within the epithelial compartment.
It has been suggested that the sequences of the EBNA-2
and EBER genes normally belong to the same genotype.’
However, all isolates investigated in this study and classified
as belonging to group 2 and 3 of EBV type-l variant strains
carried EBV type-l sequences inthe EBNA-2 gene while
maintaining EBV type-2 sequences in the EBER region.
Similar “discrepant” results of genotyping atboth loci,
EBNA-2 and EBER, have been recently described in some
BL and nasopharyngeal carcinoma cell lines and in a higher
percentage ofbiopsy
posttransplantation lymphoproliferative lesions.29It is conceivable that these
“mixed” isolates, when sequenced in the EBNA-2 gene,
may also represent EBV type-1 variant strains of either group
2 or group 3 . It isvery likely thatall these mixedEBV
variant strains with type-l sequences in the EBNA-2 gene
show the biological behavior of the EBV type-l prototype
strain B95-8.
The pathogenesis of EBV-associated diseases is complex,
involving viral, immunologic, and genetic factors. To what
extent certain EBV type-l variant strains may influence the
course of EBV infection is not known at present. Functional
assays are needed to test and compare the biological consequences of EBV type-l variant strains.
We would like to thank Prof Helms (Department of Otorhinolaryngology, University of Wurzburg, Germany) for supplying tonsil
tissue from tonsillectomized children. EBNA-2 primers were kindly
provided from Prof Jilg (Department of Medical Microbiology, University of Regensburg, Germany). We thank Dr Johnston (Department of Virology, University of Wurzburg, Germany) for critically
reading the manuscript.
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1996 87: 1579-1585
Common Epstein-Barr virus (EBV) type-1 variant strains in both
malignant and benign EBV-associated disorders
V Schuster, G Ott, S Seidenspinner and HW Kreth
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