Proc. Natl. Acad. Sci.
USA
Vol. 94, pp. 10827–10832, September 1997
Immunology
Telomere lengthening and telomerase activation during human B
cell differentiation
NAN-PING WENG*†‡, L AWRENCE GRANGER*, AND RICHARD J. HODES*†
*Experimental Immunology Branch, National Cancer Institute, and †National Institute on Aging, National Institutes of Health, Bethesda, MD 20892
Edited by Irving L. Weissman, Stanford University School of Medicine, Stanford, CA, and approved August 4, 1997 (received for review May 6, 1997)
ABSTRACT The function of the immune system is highly
dependent on cellular differentiation and clonal expansion of
antigen-specific lymphocytes. However, little is known about
mechanisms that may have evolved to protect replicative poten-
tial in actively dividing lymphocytes during immune differenti-
ation and response. Here we report an analysis of telomere length
and telomerase expression, factors implicated in the regulation
of cellular replicative lifespan, in human B cell subsets. In
contrast to previous observations, in which telomere shortening
and concomitant loss of replicative potential occur in the process
of somatic cell differentiation and cell division, it was found that
germinal center (GC) B cells, a compartment characterized by
extensive clonal expansion and selection, had significantly longer
telomeric restriction fragments than those of precursor naive B
cells. Furthermore, it was found that telomerase, a telomere-
synthesizing enzyme, is expressed at high levels in GC B cells (at
least 128-fold higher than those of naive and memory B cells),
correlating with the long telomeres in this subset of B cells.
Finally, both naive and memory B cells were capable of up-
regulating telomerase activity in vitro in response to activation
signals through the B cell antigen receptor in the presence of
CD40 engagement andyor interleukin 4. These observations
suggest that a novel process of telomere lengthening, possibly
mediated by telomerase, functions in actively dividing GC B
lymphocytes and may play a critical role in humoral immune
response by maintaining the replicative potential of GC and
descendant memory B cells.
A hallmark of the immune system is its ability to respond
effectively to the constantly changing antigenic and pathogenic
challenges in its environment. In the case of antibody-mediated
humoral responses, this ability is achieved by the existence of a
diverse repertoire of naive B cells and by an adaptive process that
allows extensive receptor diversification by somatic hypermuta-
tion, clonal expansion, and selection to generate optimized bind-
ing affinity of antibody to antigen (1). B lymphocytes derive from
bone marrow progenitor cells and undergo an ordered sequence
of differentiation with phenotypically distinct stages during lin-
eage development and during activation of an immune response
(2). During T cell-dependent immune responses, mature antigen-
naive B lymphocytes differentiate in a unique germinal center
(GC) environment into GC B cells, then to memory B cells or
plasma cells (3). In the GC, substantial cell division occurs during
a differentiation process that comprises several important pro-
cesses, including somatic hypermutation of variable domains of
immunoglobulin (Ig) genes (4–6), clonal selection of mutated
antibody-producing B cells with high antigen-binding affinity (7),
and Ig isotype switching (8). In recent years, substantial progress
has been made in understanding the process of Ig variable gene
rearrangement, the structure and function of GCs, and the
phenotypic changes that occur in the transition of immature to
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10827
mature B cells, and of naive B cells to GC to memory or plasma
cells. However, less is known about the mechanisms underlying
the replicative lifespan of lymphocytes and immunologic mem-
ory.
Telomerase and the regulation of telomere length have re-
cently drawn considerable attention for their potential roles in
critical biological functions, including the control of cellular
replication (9). Telomeres are specialized terminal chromosomal
structures that consist of tandem hexanucleotide repeats
(TTAGGG)n in vertebrates (10, 11) and telomere binding pro-
teins (12, 13). The primary function of telomeres appear to be in
maintaining the integrity of chromosomes and completing chro-
mosomal replication. Telomeres have also been implicated in
determining cellular replicative capacity. Due to the inability of
DNA polymerase to replicate the ends of eukaryotic chromo-
somes completely (14, 15), each cell division results in a loss of
50–200 bp of telomere repeats in normal human somatic cells
(16–21). If a minimal length of telomeres is essential for chro-
mosomal integrity and replication, telomere length could limit the
replicative lifespan of cells. Telomerase is a ribonucleoprotein
enzyme that is capable of synthesizing telomeric repeats (22, 23).
It has been proposed that telomerase is expressed in germ-line
and malignant cells but not in most normal human somatic cells
(24–26), and this selective expression of telomerase has been
hypothesized as a basis for the immortality of the germ line and
of malignant cells (27, 28). However, recent studies indicate that
telomerase activity is also detected in normal human somatic cells
that have self-renewal capability, such as hematopoietic stem cells
(29–31), lymphocytes (29, 32–34), and skin epithelial cells (35–
37). Moreover, the expression of telomerase in T lymphocytes is
highly regulated during development and activation (33, 38, 39).
We report here the analysis of telomere length and telomerase
expression in tonsil B cell subsets. Telomere length and telom-
erase activity were found to be closely regulated in the course of
B cell differentiation. In contrast to previously characterized
models of telomere shortening during somatic cell division,
telomeres are significantly longer in GC B cells than in either
precursor naive or descendant memory B cells. Telomerase
activity was markedly increased in GC B cells relative to these
other B cell populations, correlating with increased telomere
length. Furthermore, high levels of telomerase activity are in-
duced in naive and memory B cells by in vitro activation. These
results suggest that telomerase may play a critical role in immune
responses by elongating telomeres and thus preserving the rep-
licative lifespan of GC and progeny memory B cells.
MATERIALS AND METHODS
Antibodies. Monoclonal antibodies used for immunomag-
netic cell separations: anti-CD3 (OKT-3), CD4 (0516), CD8
(B9.8), CD14 (63D3), CD16 (3G8), CD44 (NIH44–1), platelet
This paper was submitted directly (Track II) to the Proceedings office.
Abbreviations: GC, germinal center; TRF, telomeric restriction frag-
ment; SAC, Staphylococcus aureus Cowan strain.
‡To whom reprint requests should be addressed at: Experimental
Immunology Branch, National Cancer Institute, National Institutes
of Health, Building 10, Room 4B17, 10 Center Drive, MSC 1360,
Bethesda, MD 20892-1360. e-mail: wengn@exchange.nih.gov.
10828 Immunology: Weng et al.
(37F9), and erythrocyte (10F7) antibodies were gifts from
Steve Shaw (National Institutes of Health). Purified anti-IgD,
IgG, and IgA antibodies, fluorescein isothiocyanate (FITC)-
conjugated anti-CD20, and phycoerythrin (PE)-conjugated
anti-IgG were purchased from PharMingen. Purified anti-
CD38 and CD40, FITC-conjugated anti-CD23, CD38, and
IgD; PE-conjugated anti-CD38, CD40, CD44, and IgD; and
Tri-color (TC)-conjugated anti-CD19, CD38, and CD2 anti-
bodies were purchased from Caltag (South San Francisco,
CA). Purified anti-CD77 was purchased from Immunotech
(Westbrook, ME). FITC-conjugated mouse anti-rat IgM was
purchased from Jackson ImmunoResearch.
Isolation of B Cell Subsets from Tonsils. Human tonsils
were obtained from tonsillectomies for chronic tonsillitis on
children and adults aged 3–31 years and mononuclear cells
were isolated by Ficoll gradient centrifugation (Organon
Teknika–Cappel). B cells were isolated by binding to anti-
CD19 beads (Dynal, Great Neck, NY) and subsets of B cells
were further isolated by negative immunomagnetic cell sepa-
ration as described (40). Monoclonal antibodies against CD38,
IgG, and IgA were used for isolation of naive B cells
(CD191IgD1IgG2IgA2CD382); antibodies against CD44 and
IgD, for isolation of GC B cells (CD191IgD2CD381CD442),
and antibodies against CD38 and IgD for isolation of memory
B cells (CD191IgD2CD382). For telomerase activity analysis,
subsets of B cells were isolated by fluorescent cell sorting
(FACStar; Becton Dickinson). Bm1 (CD191IgD1CD382-
CD232) and Bm2 (CD191IgD1CD382CD231) subsets were
isolated from enriched naive B cells based on CD23 expres-
sion; Bm29 (CD191IgD1CD381) and Bm5 (CD191IgD2-
CD382) were directly sorted from CD191 B cells; Bm3
(centroblast, CD191IgD2CD381CD771) and Bm4 (centro-
cyte, CD191IgD2CD381CD772) were isolated from enriched
GC B cells based on CD77 expression. Plasma cells
(CD32CD142CD38hiCD20lo) were isolated directly either from
mononuclear cells after depleting T cells (CD31, CD41 and
CD81) and monocytes (CD141) by immunomagnetic beads, or
from CD191 B cells (41). The purity of isolated B cells was
analyzed by flow cytometry as described previously (33), and it
was generally 90–95% for cells isolated from cell sorting, and
80–90% for cells isolated by immunomagnetic cell separation.
Measurement of Telomere Length. Mean telomeric restric-
tion fragment (TRF) length was analyzed by hybridization in
dried gels with a telomeric repeat oligonucleotide probe as
previously described (42). ImageQuant software was used to
measure mean TRF length with compensation for the telo-
mere length effect on the intensity of the signals by using DNA
standards as previously described (17). The statistical analysis
of TRF lengths difference was done by t test using software
(Statistica, StatSoft, Tulsa, OK) for analysis of difference
between dependent samples.
PCR-Based Telomerase Assay. The telomerase assay used here
was modified from the telomeric repeats amplification protocol
(TRAP) (24) described previously (33). Cell extracts were pre-
pared from 2 3 105 to 2 3 106 FACS-sorted cells of each of seven
B cell subsets at a concentration of 100 ml of 3-[(3-cholamido-
propyl)dimethylammonio]-1-propanesulfonate (CHAPS; Cal-
biochem) lysis buffer per 106 cells. Telomere synthesis was carried
out starting with 5 3 104 cell equivalents, and PCR amplification
was carried out starting with 1 3 104 cell equivalents in a separate
tube. The amplified products from 3.3 3 103 cell equivalents were
loaded as the original concentration on a 12% acrylamide gel
(NOVEX, San Diego), and the results were analyzed on a
PhosphorImager (Molecular Dynamics). Telomerase activity was
quantitated by serial dilution of cell extracts, and an internal
competitive control employed to monitor the reproducibility of
the assay and to serve as a reference for comparing results
between different experiments (43). Controls for the TRAP assay
included cell lysates from 293 cells as a positive control, DNase-
free RNase (Boehringer Mannheim) treatment of cell extracts,
Proc. Natl. Acad. Sci. USA 94 (1997)
single primer, Ts or Cx alone, (Genosys, Woodlands, TX), and
lysate buffer as template for PCR.
Activation of Naive and Memory Tonsil B Cells in Vitro. Freshly
isolated naive and memory tonsil B cells were stimulated as
follows: Cells were suspended at 2–5 3 106 cells per ml in RPMI
1640 (BioWhittaker) with 10% fetal bovine serum (Biofluids,
Rockville, MD) in the presence of one of the following: 0.2 mgyml
(1:10,000 dilution) of Staphylococcus aureus Cowan strain (SAC;
Pansorbin cell, Calbiochem), or 12 mgyml purified goat F(ab9)2
fragment against human IgM Fc5m (Jackson ImmunoResearch)
(44), or 0.5 mgyml anti-CD40 (45), or 10 unitsyml recombinant
interleukin 4 (rIL-4; Genzyme), or combinations of anti-IgM and
rIL-4, or anti-CD40. Cells were collected at day 2 after stimula-
tion for the analysis of telomerase activity and hTR expression.
Cell proliferation was assayed by [3H]thymidine incorporation as
previously described (46). In brief, 1–3 3 105 naive and memory
B cells were cultured in triplicate samples in flat-bottomed
96-well microtiter plates in RPMI 1640 medium containing 10%
fetal bovine serum. Cells were incubated for 24 hr, 1 mCi (1 mCi 5
37 kBq) of [3H]thymidine was added, and incubation was con-
tinued for another 24 hr before harvest. [3H]Thymidine (New
England Nuclear) incorporation was measured by liquid scintil-
lation counting.
Northern Blot Analysis of Telomerase RNA Template (hTR)
Expression. Total RNA was isolated and probed with hTR and
7SK as described previously (47). In brief, 10 mg of total RNA was
used for each sample. The membranes were hybridized first with
hTR probe in Quickhyb solution (Stratagene) and then with 7SK
after removal of hTR probe. The washed membranes were then
exposed to PhosphorImager screens and the results were ana-
lyzed using ImageQuant (Molecular Dynamics).
RESULTS
Telomere Length in Naive, GC, and Memory B Cells. The
process of somatic hypermutation and subsequent clonal selec-
tion in GC occurs in the context of substantial cell division and
clonal expansion (5, 7, 48–50). In the absence of compensating
mechanisms, clonal expansion would be expected to result in
significant loss of telomere repeats; based on the telomere
hypothesis, such reduction of telomere length during B cell
differentiation would in turn decrease the replicative potential of
memory or effector B cells. To examine the effect of differenti-
ation and cell division on telomere length in B cell subsets,
genomic DNA was isolated from naive, GC, and memory B cells
of five tonsils, and TRF length was measured (Fig. 1A). It was
found that mean TRF was longest in GC B cells, followed by
memory and naive B cells (Fig. 1B). The mean difference in TRF
between GC and naive B cells was statistically significant (GC
longer than naive cell TRF by 0.8 6 0.2 kb, P 5 0.001), with a less
significant difference between GC and memory B cells (GC
longer by 0.6 6 0.4 kb, P 5 0.05), and no significant difference
between memory and naive B cells (0.2 6 0.56 kb, P 5 0.4). Thus,
in a model in which naive B cells differentiate into GC cells and
GC cells into memory cells, a significant increase in telomere
length occurred in the transition from naive to GC B cells, and
significant shortening in the transition from GC to memory B
cells. This represents a unique model in which telomere length-
ening occurs in the course of B cell differentiation concurrent
with substantial cell division.
Telomerase Activity in Tonsil B Cell Subsets. The increased
telomere length found in GC B cells relative to other B cell
populations could reflect the existence of a mechanism capa-
ble of elongating telomeres in these B cells. A known candidate
for such activity is telomerase, which is capable of extending
telomeric repeats (22, 23). To determine whether telomerase
expression is regulated during B cell differentiation and is
related to telomere length in B cell subsets, we analyzed
telomerase activity in tonsil B cell subsets. Analysis revealed
that telomerase activity was tightly regulated during B cell
differentiation: the highest levels of telomerase activity were
 Immunology: Weng et al.
FIG. 1. Telomere length in tonsil B cell subsets. (A) TRF blot of naive
(N), GC (G), and memory (M) B cells from three tonsils. One microgram
of digested genomic DNA from each sample was loaded in a 0.6% agarose
gel and separated by electrophoresis. Hybridization was carried out in the
dried gel with a 32P-labeled (CCCTAA)3 oligonucleotide at 43°C over-
night. (B) Comparison of mean TRF length of naive, GC, and memory
B cells from five patients. The mean TRF was calculated by converting the
intensity of signals to molecular size based on DNA molecular weight
markers, using ImageQuant (software of PhosphorImager). Telomere
size effect on the signal intensity was taken into consideration in calcu-
lation as previously described (17).
found in centrocytes (Bm4) and centroblasts (Bm3), followed
by the naive to GC transition population (Bm29); and low to
undetectable levels of telomerase were found in memory B
cells (Bm5), activated naive B cells (Bm2), and virgin naive B
cells (Bm1) (Fig. 2). Based on serial dilution analysis and
competition of an internal PCR standard (42), and compared
with the level of telomerase activity in Bm1 cells, telomerase
activity was at least 128-fold higher in Bm3 and Bm4, 16-fold
higher in Bm29, and 2-fold higher in Bm2 and Bm5 (Fig. 2B).
Plasma cells, which represent terminally differentiated B cells,
were also isolated and were found to express telomerase
Proc. Natl. Acad. Sci. USA 94 (1997) 10829
Table 1. DNA synthesis by naive and memory B cells in vitro
[3H]Thymidine incorporation,
cpm
Conditions* Naive Memory
Medium 443 6 48 495 6 19
Anti-IgM 4,694 6 285 1,667 6 184
rIL-4 847 6 52 884 6 88
Anti-CD40 782 6 49 996 6 142
Anti-IgM 1 rIL-4 11,072 6 2,208 6,157 6 120
Anti-IgM 1 anti-CD40 14,977 6 1,376 9,061 6 816
rIL24 1 anti-CD40 3,197 6 70 2,986 6 442
Anti-IgM 1 rIL24 1 anti-CD40 15,355 6 722 11,055 6 2,728
SAC 15,002 6 345 11,330 6 339
The results are expressed as mean 6 SD.
*Cells were incubated at 37°C for 24 hr under various stimulation
conditions. After addition of 1 mCi of [3H]thymidine, cells were
incubated for another 24 hr and harvested for DNA synthesis analysis.
activity 16-fold higher than that in resting naive and memory
B cells, and 16-fold lower than GC B cells (Fig. 2).
Induction of Telomerase Activity in Vitro in Naive and Memory
B Cells. The GC B cells, populations in which the highest levels
of telomerase activity are expressed, consist of activated and
dividing cells as well as cells undergoing apoptosis (51, 52). To
ascertain whether telomerase activity is induced by signals that
induce activation and proliferation, andyor by the signals that
induce apoptosis, freshly isolated naive and memory tonsil B cells,
which expressed low levels of telomerase, were cultured under
various conditions for 2 days and collected for telomerase anal-
ysis. In both naive and memory B cells, SAC treatment resulted
in strong induction of telomerase (32-fold increase), as did
treatment with (anti-IgM 1 anti-CD40 1 rIL-4) (16-fold), (anti-
IgM 1 anti-CD40) (12-fold), and (anti-IgM 1 rIL-4) (8-fold). In
contrast, treatment with anti-CD40, or rIL-4 alone, or (anti-CD40
1 rIL-4) did not induce telomerase activity (Fig. 3). Treatment
with anti-IgM alone induced low levels of telomerase in naive
(4-fold) but not memory B cells (Fig. 3). The induction of
telomerase in naive and memory B cells generally correlated well
with induction of cell proliferation as measured by [3H]thymidine
incorporation under these stimulation conditions (Table 1).
Because apoptotic death is a common phenomenon occurring
in GC B cells, where telomerase activity is highest, it was
determined whether telomerase is induced by apoptosis-inducing
signals in vitro. Although dexamethasone and g-irradiation in-
duced substantial apoptosis in naive and memory B cells, neither
treatment induced telomerase (data not shown).
hTR Expression in B Cell Subsets. To assess the regulation
of telomerase activity in B cell subsets at a molecular level, we
examined the expression of hTR (53) in naive, GC, and
memory B cells. It was found that the level of hTR is higher
in GC B cells than in naive and memory B cells (Fig. 4A). When
the level of hTR expression was normalized to a constitutively
expressed 7SK gene (54), hTR in GC B cells was twice as
abundant as in naive and memory B cells (Fig. 4B). This result
indicates that regulation of hTR correlates with the levels of
telomerase activity in these B cell subsets. Up-regulation of
hTR expression was also observed in both naive and memory
B cells after 2-day in vitro activation with either (anti-IgM 1
anti-CD40 1 rIL-4) or SAC (data not shown).
DISCUSSION
The T cell-dependent response of mature B cells to antigenic
stimulation is marked by a complex set of differentiation events
coupled to proliferation, somatic hypermutation, Ig class switch-
ing, clonal selection, and clonal expansion. It has been estimated
that antigen-naive B cells undergo at least 4 cell divisions in the
transition to GC B cell phenotype (51) and that GC B cells
undergo at least 20 cell divisions to generate effector or memory
10830 Immunology: Weng et al. Proc. Natl. Acad. Sci. USA 94 (1997)

FIG. 2. Telomerase activity in tonsil B cell subsets. (A) Representative results of telomerase activity in B cell subsets. Seven subsets of B cells
[Bm1–5 and plasma cell (PC)] were isolated from two donors by a combination of immunomagnetic cell separation and fluorescent cell sorting,
and three subsets (naive, GC, and memory) were isolated from five donors by immunomagnetic cell separation alone. The relative activity of
telomerase was consistent and reproducible among subsets of B cells from these different donors and cell separations. A transformed cell line, 293,
was used as a positive control and standard for quantitation of relative levels of telomerase activity. (B) Comparison of the levels of telomerase
activity in tonsil B cell subsets. The relative levels of telomerase activity were determined by serial dilution and are minimal estimates, compared
with the levels of telomerase activity of Bm1 B cell subsets. The internal standard was used to monitor the reproducibility of the assay and served
as a reference for comparing results among experiments.

B cells (55). Memory B cells are believed to be generated
predominantly from a GC-dependent pathway, whereas plasma
cells are probably derived from both GC-dependent and -inde-
pendent paths (3). The persistence of memory B cells ensures the
capacity to generate an accelerated response to a subsequent
encounter with the same antigen, with production of antibody
previously selected for high affinity and isotype (56). However,
the mechanisms underlying many of these striking events are
poorly understood, including the mechanisms involved in regu-
lating the capacity for clonal expansion. In this report, we have
assessed telomere length and telomerase activity in B cell subsets
in human tonsils to evaluate the potential role of telomere
regulation in B cell differentiation and activation.
The loss of telomeric DNA in vivo as a function of age and in
vitro with cell division has been well documented in many types
of normal human somatic cells over the past several years,
including fibroblasts (17, 19), endothelial cells (21), lymphocytes
(41, 57, 58), and hematopoietic stem cells (20). If telomere length
serves as a mitotic clock as proposed (19), the capacity for cellular
replication will be restricted by the telomere length. Evidence
supporting this model has included the observation that the
replication potential of human fibroblasts is better correlated with
the initial length of telomeres than with other parameters such as
the age of the donors (19). An additional observation consistent
with telomere shortening during in vivo cell division is the finding
that human CD41 memory T cells have telomeres that are
consistently 1.4 kb shorter than the CD41 naive T cells from
which they appear to differentiate and that memory T cells have
correspondingly less replicative capacity than naive T cells (42).
Here we demonstrate that telomere length is also regulated
 Immunology: Weng et al. Proc. Natl. Acad. Sci. USA 94 (1997) 10831

provide unique evidence that telomere length can be increased

during the course of B cell differentiation, and they suggest that
telomerase may play a role in regulating telomere length in these
normal human somatic cells in vivo.
Substantial evidence now indicates that telomerase can be
expressed in normal human somatic cells (29–37). The expression
of telomerase in these somatic cells appears to be generally
down-regulated during in vivo development and differentiation,
as demonstrated in hematopoietic progenitor cells (30) and T
cells (33) and during the induction of differentiation in the
promyelocytic cell line HL60 in vitro (61–63). A low to undetect-
able level of telomerase activity has been reported in peripheral
blood B cells (29, 32), and a high level in tonsil GC cells (31, 34).
Our results provide a detailed picture of regulation of telomerase
during B cell differentiation from which several interesting points
emerge. Telomerase activity is low in naive B cells but present at
high levels in the transition from naive B cells to GC centroblasts
and centrocytes. Telomerase then decreases as GC cells transition
to memory cells. Plasma cells, which are terminally differentiated
cells, expressed moderate levels of telomerase, thus differing
from previous observations of other terminally differentiated
cells (61–63). The telomerase RNA component was regulated in
parallel with telomerase activity in tonsil B cell subsets both in
vivo and in vitro.

FIG. 3. In vitro activation induces telomerase in naive and memory

tonsil B cells. (A) The results shown are representative of telomerase
induction in naive and memory tonsil B cells with in vitro activation.
Similar results were obtained in three independent experiments. (B)
Relative levels of telomerase activity in naive and memory tonsil B cells
induced by different stimulation conditions. The relative levels of telom-
erase activity were determined as described in the text and in the legend
of Fig. 2.

during B cell differentiation. However, the longest telomeres are

found not in precursor naive B cells but rather in GC B cells that
are the descendants of naive B cells. In principal, the long
telomeres found in GC B cells either could reflect their selective
derivation from precursor naive B cells, which have long telo-
meres, or could reflect an actual elongation of telomeres by
mechanisms operating in GC B cells. Although it is difficult to
rule out the first possibility, it is not apparent that there would be
an initial selective advantage to those naive B cells that have
relatively longer telomeres. It has previously been shown in
dividing yeast that telomerase not only is essential for preventing
telomere shortening (59) but also can function to dramatically
lengthen telomeres when specific mutations are introduced into
telomerase RNA template (60). Thus, the expression of strikingly FIG. 4. Telomerase RNA template expression in B cell subsets. (A)
Representative Northern blot of hTR expression in naive (N), GC (G),
high levels of telomerase activity in GC B cells is consistent with and memory (M) B cells. (B) Abundance of hTR expression in tonsil
the possibility that telomeres have been elongated in GC B cells B cell subsets. The abundance of hTR was analyzed by normalization
by telomerase, serving to preserve the replicative lifespan of GC to 7SK level, presented as in arbitrary units (a.u.), and the results were
and their memory B cell descendants (Fig. 5). These results summarized from B cells of three tonsils.
10832 Immunology: Weng et al. Proc. Natl. Acad. Sci. USA 94 (1997)

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