ARTHRITIS & RHEUMATISM
Vol. 48, No. 11, November 2003, pp 3253–3265
DOI 10.1002/art.11299
© 2003, American College of Rheumatology
Generation and Characterization of LymphoStat-B, a Human
Monoclonal Antibody That Antagonizes the Bioactivities of
B Lymphocyte Stimulator
Kevin P. Baker,1 Bryan M. Edwards,2 Sarah H. Main,2 Gil H. Choi,1 Ruth E. Wager,1
Wendy G. Halpern,1 Patrick B. Lappin,3 Todd Riccobene,1 Donara Abramian,1
Les Sekut,1 Bonnie Sturm,1 Carol Poortman,1 Ralph R. Minter,2 Claire L. Dobson,2
Elizabeth Williams,2 Sara Carmen,2 Rodger Smith,1 Viktor Roschke,1 David M. Hilbert,1
Tristan J. Vaughan,2 and Vivian R. Albert1
Objective. To identify and characterize a fully
human antibody directed against B lymphocyte stimu-
lator (BLyS), a tumor necrosis factor–related cytokine
that plays a critical role in the regulation of B cell
maturation and development. Elevated levels of BLyS
have been implicated in the pathogenesis of auto-
immune diseases.
Methods. A human phage display library was
screened for antibodies against human BLyS. A human
monoclonal antibody, LymphoStat-B, specific for hu-
man BLyS was obtained from the library screening and
subsequent affinity optimization mutagenesis. The an-
tibody was tested for inhibition of human BLyS in vitro
and in an in vivo murine model. Additionally, the
consequences of BLyS inhibition were tested in vivo by
administration of LymphoStat-B to cynomolgus mon-
keys.
Results. LymphoStat-B bound with high affinity
to human BLyS and inhibited the binding of BLyS to its
1
Kevin P. Baker, PhD, Gil H. Choi, PhD, Ruth E. Wager,
PhD, Wendy G. Halpern, DVM, PhD, Todd Riccobene, PhD, Donara
Abramian, MS, Les Sekut, DVM, Bonnie Sturm, BS, Carol Poortman,
BS, Rodger Smith, PhD, Viktor Roschke, PhD, David M. Hilbert,
PhD, Vivian R. Albert, PhD: Human Genome Sciences, Inc., Rock-
ville, Maryland; 2Bryan M. Edwards, PhD, Sarah H. Main, MSc, Ralph
R. Minter, PhD, Claire L. Dobson, PhD, Elizabeth Williams, BSc, Sara
Carmen, BSc, Tristan J. Vaughan, PhD: Cambridge Antibody Tech-
nology, Granta Park, Cambridge, UK; 3Patrick B. Lappin, DVM, PhD:
Charles River Laboratories, Sierra Biomedical Division, Sparks, Ne-
vada.
Address correspondence and reprint requests to Kevin P.
Baker, PhD, Department of Antibody Discovery and Development,
Human Genome Sciences, Inc., 9410 Key West Avenue, Rockville,
MD 20850. E-mail: kevin_baker@hgsi.com.
Submitted for publication March 5, 2003; accepted in revised
form July 3, 2003.
3253
3 receptors, TACI, BCMA, and BLyS receptor
3/BAFF-R. LymphoStat-B potently inhibited BLyS-
induced proliferation of B cells in vitro, and ad-
ministration of LymphoStat-B to mice prevented
human BLyS-induced increases in splenic B cell num-
bers and IgA titers. In cynomolgus monkeys, adminis-
tration of LymphoStat-B resulted in decreased B cell
representation in both spleen and mesenteric lymph
nodes.
Conclusion. A fully human monoclonal antibody
has been isolated that binds to BLyS with high affinity
and neutralizes human BLyS bioactivity in vitro and in
vivo. Administration of this antibody to cynomolgus
monkeys resulted in B cell depletion in spleen and
lymph node. This antibody may prove therapeutically
useful in the treatment of autoimmune diseases in
humans.
B lymphocyte stimulator (BLyS; trademark of
Human Genome Sciences, Rockville, MD) protein (also
known as BAFF, THANK, TALL-1, TNFSF13B, and
zTNF4), a member of the tumor necrosis factor (TNF)
ligand superfamily, is synthesized as a 285–amino acid
type II membrane protein and exists in both membrane
and cleaved 152–amino acid soluble forms (1–6). BLyS
is expressed on monocytes, macrophages, and monocyte-
derived dendritic cells, and is up-regulated in response
to interferon-␥ and interleukin-10 (IL-10) (7). In vitro,
recombinant human BLyS enhances B cell proliferation
and Ig secretion through interaction with receptors
expressed predominantly on B cells (1). In vivo, recom-
binant human BLyS causes splenic hyperplasia in mice,
3254
primarily due to increases in the number of mature B
cells. BLyS administration to mice also causes increases
in the total serum Ig concentration and enhanced
humoral responses to both T cell–independent and T
cell–dependent antigens (1,2,8).
BLyS has been shown to bind with high affinity to
3 receptors, all of which are members of the TNF
receptor family (6,9,10). Two of the receptors, BCMA
and TACI, also bind APRIL, another TNF family mem-
ber that is the most homologous to BLyS (11–13). The
precise function of these two receptors is not well
understood, but studies in knockout mice suggest that
BCMA is functionally redundant, since BCMA-deficient
mice have a normal B cell phenotype (14). TACI has
been shown to have an inhibitory role in B cell develop-
ment, since TACI knockout animals exhibit increased
peripheral B cells, reduced responses to T cell–
independent antigens (15,16), and lymphoproliferative
disorders and autoimmune disease (17). In contrast,
BLyS-deficient mice show a phenotype of severe loss of
mature B cells in the spleen, peripheral blood, and
lymph nodes (18,19). A third receptor, BLyS receptor 3
(BR-3; BAFF-R), is specific for BLyS (9,10). The
A/WySnJ mouse strain, which harbors a naturally occur-
ring truncation of BR-3, exhibits a phenotype similar to
that of BLyS-deficient mice (9,10,20), suggesting that
BR-3 is the primary mediator of the effects of BLyS on
B cell survival and maturation.
Several lines of evidence suggest that elevated
levels of BLyS may be involved in the pathogenesis of B
cell–mediated autoimmune diseases. First, constitutive
overexpression of BLyS in transgenic animals results in
manifestations of autoimmune-like symptoms, including
anti-DNA antibodies, rheumatoid factor, circulating im-
mune complexes, and deposition of immune complexes
in the kidney leading to glomerulonephritis. These
symptoms resemble those of systemic lupus erythemato-
sus (SLE) and some aspects of rheumatoid arthritis
(RA) (21,22). Second, elevated levels of BLyS have been
found in other murine models of SLE, including MRL-
lpr/lpr and (NZB ⫻ NZW)F1 strains (6). Finally, ele-
vated levels of BLyS have been observed in the serum of
patients with SLE and RA (23,24) as well as Sjögren’s
syndrome (25,26). Significant correlations between
BLyS levels and autoantibody production were shown in
these studies.
The association of BLyS overproduction with
manifestations of several autoimmune diseases suggests
that modulation of BLyS levels could be a novel thera-
peutic approach to the treatment of such diseases.
Indeed, studies with soluble BLyS receptors as antago-
BAKER ET AL
nists have shown efficacy in reducing disease manifesta-
tions in murine models of both SLE and RA (6,27,28).
With the aim of developing a therapeutic agent for
autoimmune disease, LymphoStat-B (Human Genome
Sciences) antibody, a human, neutralizing monoclonal
antibody against human BLyS, was generated. We de-
scribe herein the generation of human antibodies against
BLyS using phage display, as well as the in vitro and in
vivo characterization of LymphoStat-B.
MATERIALS AND METHODS
Phage display libraries and selections. Nonimmunized
human single-chain variable fragment (scFv) phage display
libraries, recently expanded from 1010–1011 clones (29), were
used for lead-scFv isolation. BLyS, at 10 ␮g/ml in phosphate
buffered saline (PBS), was immobilized on immunotubes
(Nunc, Naperville, IL), and BLyS-binding phage was isolated
by 3 sequential rounds of panning (29). The optimized variants
were isolated by selection from randomized libraries in solu-
tion using biotinylated BLyS captured on streptavidin-coated
paramagnetic beads (Dynal, New Hyde Park, NY) (30). The
hBLySsc-1 and hBLySsc-2 V H third complementarity-
determining region (CDR3) randomized repertoires were con-
structed by polymerase chain reaction (PCR) using mutagenic
oligonucleotides to replace the last 6 VH CDR3 amino acids
with randomized codons. The mutated DNA for both lineages
was ligated into the phagemid vector pCANTAB6 (31) and
electroporated into Escherichia coli TG1 (32). Libraries of 2 ⫻
108 and 6 ⫻ 108 individual clones were generated for
hBLySsc-1 and hBLySsc-2, respectively. The randomized li-
braries were subjected to 1 round of panning on 10 ␮g/ml of
immobilized BLyS, followed by 12 rounds of soluble selection
using decreasing concentrations of biotinylated BLyS, from 50
nM down to 100 pM.
Preparation and screening of scFv in a receptor bind-
ing inhibition assay. Expression of scFv was induced in 2TY
medium (1.6% weight/volume tryptone, 1% w/v yeast extract,
0.5% w/v NaCl) containing 1 mM IPTG for 3 hours at 30°C.
Periplasmic extracts (33) were prepared, and His-tagged scFv
were purified on nickel agarose (Qiagen, Chatsworth, CA) and
buffer exchanged into PBS. BLyS was biotinylated using
N-hydroxysuccinimide (NHS)–biotin (Pierce, Rockford, IL) at
a molar ratio of 20:1 of biotin to BLyS. IM9 cells were
immobilized onto poly-L-lysine–coated 96-well plates at a
density of 1 ⫻ 105 cells/well, and scFv was added in the
presence of 25 ng/ml (0.5 nM) of biotinylated BLyS. Bound
biotinylated BLyS was detected via streptavidin–Delfia (Perkin
Elmer, Boston, MA). The 50% inhibition concentration (IC50)
values for the competition of each scFv for BLyS binding to its
receptor were then determined.
Cloning and expression of antibodies. Variable heavy
and light chain fragments from scFv clones were subcloned as
separate heavy and light chains into expression vectors ob-
tained from Cambridge Antibody Technology (34). Expression
constructs were used for transfection of mammalian cell lines
(HEK293T or NSO) to produce whole IgG. Conditioned
media were collected 72 hours posttransfection, and antibodies
were purified by protein A chromatography.
LYMPHOSTAT-B, HUMAN MONOCLONAL ANTIBODY AGAINST BLyS
Binding to BLyS in solid phase (direct enzyme-linked
immunosorbent assay [ELISA]). ELISA plates (Immulon-II;
Dynatech, Chantilly, VA) were coated with 1 ␮g/ml of strepta-
vidin (Sigma, St. Louis, MO) in PBS. After washing in PBS and
blocking with 3% bovine serum albumin (BSA), 100 ␮l of
biotinylated BLyS solution in PBST (1 ␮g/ml in PBS, 0.1%
Tween 20, 0.1% BSA) was added to each well, and the wells
were incubated for 1 hour at room temperature. After washing,
100 ␮l of serial dilutions of antibodies (66–6.6 ⫻ 10–6 nM) in
diluent buffer (PBS, 0.1% Tween 20, 0.1% BSA) was dis-
pensed into individual wells of BLyS-coated plates. The plates
were incubated for 2 hours at room temperature, washed 4
times with PBST, and then 100 ␮l of peroxidase-conjugated
goat anti-human IgG (1 ␮g/ml in diluent buffer; Vector,
Burlingame, CA) was added to each well. Plates were incu-
bated for 1 hour at room temperature, washed 4 times with
PBST, and developed with tetramethylbenzidine substrate
(Sigma). Absorption at 450 nm was measured with a Spectra-
Max 3000 instrument (Molecular Devices, Sunnyvale, CA).
Binding to BLyS in solution (competition ELISA).
Serial dilutions of BLyS (103–10–4 nM) were prepared in
diluent buffer, and 50-␮l aliquots of each dilution were distrib-
uted to individual wells of BLyS-coated plates (prepared as
described above). Antibodies were diluted to the 50% maxi-
mum response concentration (EC50), and 50 ␮l of the resultant
dilution was added to the wells already containing BLyS. The
plates were further processed as described above.
Murine splenocyte in vitro proliferation assay. Serial
3-fold antibody dilutions were prepared in a 96-well plate. The
final antibody concentrations ranged from 100 nM to 0.01 nM;
a preparation containing no antibody served as a medium
control. BLyS (3 ng/ml final concentration) that had been
diluted in complete medium (RPMI 1640 with 10% fetal
bovine serum containing 100 units/ml of penicillin, 100 ␮g/ml
of streptomycin, 4 mM L -glutamine, and 5 ⫻ 10 –5 M
␤-mercaptoethanol) was added to each well. Medium alone
(without BLyS) was used as the negative control for the assay
background. Human IgG1 isotype controls were used as
negative controls for the antibodies.
Plates were incubated for 30 minutes at 37°C in an
atmosphere containing 5% CO2. Splenocytes (final concentra-
tion 1 ⫻ 106/ml) containing Staphylococcus aureus Cowan
strain (Calbiochem, La Jolla, CA) were added to the wells. The
plates were further incubated for 72 hours at 37°C in an
atmosphere containing 5% CO2. Each well was supplemented
with complete medium containing 0.5 mCi of 3H-thymidine
(6.7 Ci/mmole; Amersham, Arlington Heights, IL), and the
cells were incubated for an additional 20–24 hours at 37°C. The
proliferation of each sample was measured by 3H-thymidine
incorporation and graphed as a function of antibody concen-
tration. A 4-parameter regression analysis was performed for
each data series, and IC50 values were derived.
Neutralizing activity of LymphoStat-B antibody in
mice. All animal experimentation was done in accordance with
the Guide for the Care and Use of Laboratory Animals and
under the supervision of the Institutional Animal Care and
Use Committee. On day 1, female BALB/c mice (ages 8–9
weeks) were injected intravenously with diluent (PBS) or with
LymphoStat-B or control human monoclonal IgG1 antibody at
doses of 0.05, 0.15, 0.5, 1.5, or 5 mg/kg. Recombinant human
BLyS at 0.3 mg/kg or BLyS vehicle (12.5 mM citrate, 125 mM
3255
NaCl, pH 6) was injected subcutaneously 1 hour after antibody
injection. On day 3, mice received a second dose of antibody (1
hour before BLyS injection). BLyS was injected subcutane-
ously once a day for 4 consecutive days. On day 5, all mice were
killed 24 hours after the last BLyS injection. Sera were
analyzed for IgA content by ELISA, wet spleen weights were
recorded, and splenocytes were analyzed by flow cytometry
using phycoerythrin (PE)–conjugated anti–B220 (anti-
CD45R) and fluorescein isothiocyanate (FITC)–conjugated
anti-ThB (anti-Ly6D) as markers, as previously described
(1,35).
Effects of LymphoStat-B in cynomolgus monkeys.
Studies with cynomolgus monkeys (Macaca fascicularis) were
performed at Charles River Laboratories, Sierra Biomedical
Division (Sparks, NV). Monkeys (n ⫽ 10 [5 males and 5
females] per dosage group) were given an intravenous injec-
tion of vehicle or LymphoStat-B at 5, 15, or 50 mg/kg once a
week for 4 weeks. On day 29 (4 weeks of treatment), 3 monkeys
of each sex from each dosage group were necropsied. The
remaining monkeys were necropsied on day 57, after an
additional 4-week treatment-free period. Periodically during
the study, the mononuclear cell populations in peripheral
blood were evaluated by flow cytometry using a Coulter Epics
XL-MCL instrument (Beckman-Coulter, Miami, FL). In addi-
tion, relative mononuclear cell populations in the spleen and
mesenteric lymph nodes were determined for each monkey at
the time of necropsy. For both peripheral blood and tissues,
the following markers were used to distinguish immunopheno-
type: CD2 for total lymphocytes, CD20 for B cells, CD20 and
CD21 for mature B cells, CD3 for total T cells, CD3 and CD4
for helper T cells, CD3 and CD8 for suppressor/cytotoxic T
cells, and CD14 (CD3–) for monocytes. The PE-labeled
anti-CD2 antibody was obtained from Beckman-Coulter.
FITC-labeled anti-CD3 and anti-CD20 antibodies, as well
as PE-labeled anti-CD4, anti-CD8, anti-CD14, and
anti-CD21 antibodies, were obtained from BD PharMingen
(San Diego, CA).
Inhibition of BLyS binding to BLyS receptor proteins.
BLyS was labeled with the electrochemiluminescent reporter
ORI-TAG–NHS (IGEN International, Gaithersburg, MD),
and binding to each of the 3 known BLyS receptors was
measured in the presence of LymphoStat-B, human IgG1
isotype control antibody, free receptor fusion protein, or
control fusion protein herpesvirus entry mediator ([HVEM]–
Fc) essentially as previously described (36). Briefly, fusion
proteins of the extracellular domain of each of the 3 BLyS
receptors (TACI, BCMA, and BR-3) and HVEM were biotin-
ylated with NHS–LC–biotin (Pierce) at molar ratios ranging
from 9:1 to 6.6:1. Twelve 2-fold serial dilutions of
LymphoStat-B, receptor, or controls at final concentrations
ranging from 53 nM to 0.006 nM were prepared in assay
diluent, and 50-␮l aliquots of each dilution were added to
individual wells of a 96-well plate. Fifty microliters of ORI-
TAG–labeled BLyS at a final concentration of 40 ng/ml in
assay diluent was added to each well, and the plate was
incubated for 60–90 minutes at room temperature, with gentle
shaking.
The biotinylated receptor proteins were each sepa-
rately diluted to 50 ng/ml in assay diluent, and Dynabeads
M280–streptavidin (Dynal) were used at a final concentration
of 250 ␮g/ml. One hundred microliters of this mixture was
3256
Figure 1. Identification of neutralizing antibodies to human B lym-
phocyte stimulator (BLyS). A, Purified BLyS-specific single-chain
variable fragments (scFv) were evaluated for the ability to inhibit
binding of biotinylated BLyS to its receptor on IM9 cells. The 50%
inhibition concentration (IC50) values are as follows: for hBLySsc-5,
14.12 nM; for hBLySsc-4, 18.23 nM; for hBLySsc-3, 20.63 nM; for
hBLySsc-2, 13.18 nM; and for hBLySsc-1, 2.69 nM; hBLySsc-6 did not
inhibit BLyS binding. B, Converted full IgG were evaluated for their
ability to neutralize BLyS-induced murine splenocyte proliferation as
measured by 3H-thymidine incorporation. The IC50 values are as
follows: for hBLySmAb-5, 0.09 nM; for hBLySmAb-4, 0.16 nM; for
hBLySmAb-3, 0.14 nM; for hBLySmAb-2, 0.08 nM; and for
hBLySmAb-1, 0.05 nM. A 4-parameter logistic model was used for
curve fitting and calculation of binding parameters. Values are the
mean ⫾ SEM of triplicate samples.
added to each well of the assay plate, and the plate was
incubated for 45 minutes at room temperature, with vigorous
shaking. The plate was read on an Origen M8 series electro-
chemiluminescence (ECL) analyzer (IGEN International),
and the resulting ECL signal was plotted against the log
nanomolar antibody or receptor concentration.
Inhibition of receptor binding in a cellular system
(flow cytometry). Flow cytometry was used to assess the
capacity of LymphoStat-B to inhibit binding of BLyS to
HEK293T cells transfected with complementary DNA en-
coding full-length BLyS receptors. Cells were transfected with
TACI-, BCMA-, or BR-3–encoding plasmids and used for
binding studies 24 hours posttransfection, as previously
described (12). Cells were stained with 1 ␮g/ml of biotinylated
BAKER ET AL
BLyS either alone or in the presence of a 5-fold molar
excess of LymphoStat-B or an isotype control antibody.
BLyS binding was assessed with streptavidin–PE conjugate and
analyzed on a FACScan instrument (Becton Dickinson,
Mountain View, CA).
Flow cytometry analysis of LymphoStat-B binding.
Peripheral blood mononuclear cells were isolated from whole
human blood using lymphocyte separation medium (ICN
Biochemicals, Irvine, CA). Cells (10 6 in 100 ␮ l) in
fluorescence-activated cell sorting (FACS) buffer (PBS with
0.1% sodium azide and 0.1% BSA) were dispensed to FACS
tubes. Human IgG1 (20 ␮g in 10 ␮l) was added to each tube,
and the tubes were incubated for 20 minutes at 4°C to block
nonspecific Fc receptor–mediated binding. Biotinylated anti-
body or isotype control (1–2 ␮g) was added to each tube along
with FITC-conjugated lineage-specific marker antibody. Anti-
Figure 2. Improved potency of optimized BLyS single-chain antibod-
ies in the receptor inhibition assay. Purified scFv were evaluated for
their ability to inhibit biotinylated BLyS binding to its receptors on
IM9 cells, as measured by europium-labeled streptavidin. A, Compar-
ison of hBLySsc-1 and hBLySsc-1.1 (LymphoStat-B), showing that
hBLySsc-1.1 results in a 10-fold improvement in potency compared
with the parental scFv hBLySsc-1. B, Comparison of hBLySsc-2 and
hBLySsc-2.1, showing that hBLySsc-2.1 results in a 20-fold improve-
ment in potency compared with the parental scFv hBLySsc-2. The IC50
values are as follows: for hBLySsc-1, 6.3 nM; for hBLySsc-1.1, 0.5 nM;
for hBLySsc-2, 5.86 nM; and for hBLySsc-2.1, 0.33 nM. A 4-parameter
logistic model was used for curve fitting and calculation of binding
parameters. Values are the mean ⫾ SEM of triplicate samples. See
Figure 1 for definitions.
LYMPHOSTAT-B, HUMAN MONOCLONAL ANTIBODY AGAINST BLyS 3257

demonstrated to specifically recognize BLyS and did not

bind to other TNF ligand family members, including
APRIL, LIGHT, TNF␣, Fas ligand, TRAIL, and TNF␤,
or to IL-4 or IL-18 (data not shown).
Optimization of lead scFv. Optimization of the
scFv corresponding to the lead candidates hBLySmAb-1
and hBLySmAb-2 was performed to identify antibodies
with enhanced inhibitory activity. Randomized libraries
were generated from the hBLySsc-1 and hBLySsc-2
sequences in which the terminal 6 residues of the VH
CDR3 were randomly replaced in order to increase
diversity within the VH domain. Optimized scFv were
again selected for binding to human BLyS.
The ability of the selected scFv to inhibit binding
of biotinylated BLyS to its receptors on the surface of
IM9 cells was assessed, and ⬎30 scFv were identified
with improved inhibitory profiles. Figure 2 illustrates the
Figure 3. Improved activity of optimized BLyS IgG in the murine
splenocyte proliferation assay. Converted full IgG molecules were
evaluated for their ability to inhibit BLyS-induced murine splenocyte
proliferation, as measured by 3H-thymidine incorporation. The IC50
values are as follows: for LymphoStat-B, 0.02 nM; for hBLySmAb-1,
0.05 nM, hBLySmAb-2.1, 0.05 nM; and for hBLySmAb-2, 0.08 nM. A
4-parameter logistic model was used for curve fitting and calculation of
binding parameters. Values are the mean ⫾ SEM of triplicate samples.
See Figure 1 for definitions.

body binding was assessed with streptavidin–PE (Dako,

Carpinteria, CA) measured in a FACScan instrument. The cell
line K562 was used as a positive control for the presence of
membrane-bound BLyS.

RESULTS
Isolation of BLyS binding antibodies. Large non-
immunized repertoires of human scFv fragments dis-
played on phage (29) were used for selection of single-
chain antibodies that bound to human BLyS protein.
More than 1,200 distinct BLyS-binding scFv were iden-
tified and were ranked by their ability to inhibit BLyS
binding to receptors on IM9 cells, a myeloma cell line
expressing significant levels of BLyS receptors. A panel
of 50 candidate scFv demonstrating the greatest inhibi-
tory activity was selected for further analysis. Inhibition
of binding activity for 6 of these scFv is shown in Figure
1A. These scFv were converted to full IgG molecules Figure 4. Binding of LymphoStat-B to B lymphocyte stimulator
and assessed for their ability to neutralize the activity of (BLyS), as determined by enzyme-linked immunosorbent assay. A,
human BLyS protein in a murine splenocyte in vitro Direct binding of LymphoStat-B to immobilized BLyS. B, Inhibition of
proliferation assay. Examples of antibodies demonstrat- LymphoStat-B binding to immobilized BLyS by BLyS in solution. The
calculated values for the 50% maximum response concentration and
ing the greatest neutralizing activity in this assay as full
the 50% inhibition concentration are 0.02 ⫾ 0.001 nM and 8.53 ⫾
IgG are shown in Figure 1B. Antibodies demonstrating 1.015 nM (mean ⫾ SEM of triplicate cultures), respectively. A
the best inhibitory profile as full IgG molecules were 4-parameter logistic model was used for curve fitting and calculation of
hBLySmAb-1 and hBLySmAb-2. Both antibodies were binding parameters. OD ⫽ optical density.
3258 BAKER ET AL

Table 1. Analysis of LymphoStat-B binding to membrane-bound Specificity of LymphoStat-B for soluble BLyS. In
BLyS* studies using a series of monoclonal antibodies against
Mean fluorescence intensity BLyS, differential recognition of membrane-bound
Human Mouse
BLyS has previously been reported (7), although the
IgG1 IgG1 biologic basis for this remains unclear. The ability of
Cell type control† LymphoStat-B control‡ 12D6 LymphoStat-B to recognize membrane-bound BLyS on
K562 cell line 7.19 5.26 3.48 6.34 the surface of cells was examined by flow cytometry
T lymphocytes (CD3⫹) 1.78 1.76 2.24 1.99 using primary human cells as well as the K562 myelog-
B lymphocytes (CD20⫹) 1.60 1.67 1.95 3.62 enous leukemia cell line that was previously character-
Natural killer cells 3.35 3.31 3.06 3.48
(CD56⫹) ized as expressing membrane-bound BLyS (Table 1).
Monocytes (CD14⫹) 4.06 3.29 2.93 9.20
Monocytes (CD14⫹) 8.46 5.13 3.87 21.55
plus interferon-␥

* Two-color fluorescence-activated cell sorter analysis was performed

on human peripheral blood mononuclear cells (PBMCs) or the cell
line K562 (positive control for the presence of membrane-bound B
lymphocyte) using biotinylated LymphoStat-B and fluorescein
isothiocyanate–labeled lineage-specific antibodies. PBMCs from 3
different donors were studied. A representative binding profile for 1
donor is presented. In all 3 donors, the 4 subpopulations of PBMCs
were negative for LymphoStat-B binding. In 2 of the 3 donors, the
murine monoclonal antibody detected BLyS on CD14⫹ monocytes
and possibly on a minor subpopulation of CD20⫹ cells.
† Negative control for LymphoStat-B binding.
‡ Negative control for 12D6 binding.

improvements that were observed for the 2 most potent

scFv from each lineage, hBLySsc-1.1 and hBLySsc-2.1,
compared with their respective parents. The hBLySsc-
1.1 isolate (IC50 0.7 nM) showed ⬃10-fold improvement
over hBLySsc-1 (Figure 2A), and hBLySsc-2.1 (IC50 0.2
nM) was improved ⬃20-fold over hBLySsc-2 (Figure
2B).
The top 30 scFv candidates were converted to full
IgG molecules and assayed for neutralization activity in
the murine splenocyte proliferation assay. Most full IgG
showed increased activity over the scFv form. The most
potent antibody in the neutralization assay, hBLySmAb-
1.1, was designated LymphoStat-B, and selected for
further characterization (Figure 3). The hBLySmAb-2.1
antibody, which showed the greatest inhibitory activity
as an scFv, did not demonstrate the greatest activity as a Figure 5. LymphoStat-B inhibition of B lymphocyte stimulator
full IgG. (BLyS) binding to A, TACI, B, BCMA, and C, BLyS receptor 3 (BR-3).
In vitro characterization of LymphoStat-B. Bind- LymphoStat-B was evaluated for its ability to inhibit the binding of
BLyS to extracellular domain fusion proteins of the 3 known BLyS
ing characteristics of LymphoStat-B were determined
receptors using an ORI-TAG–based electrochemiluminescence assay.
using a BLyS-specific ELISA in which biotinylated BLyS Binding is shown as electrochemiluminescence (ECL) counts. The
was immobilized in wells coated with streptavidin. As calculated 50% inhibition concentration (IC50) for LymphoStat-B
shown in Figure 4, LymphoStat-B bound to BLyS with inhibition of BLyS binding to TACI, BCMA, and BR-3 was 0.11, 0.10,
an EC50 value of 0.024 nM (Figure 4A). The binding of and 0.11 nM, respectively. The calculated IC50 value for each free
receptor fusion protein in the corresponding receptor-binding assay
LymphoStat-B was readily inhibited with soluble BLyS,
was 0.395 nM (TACI), 0.74 nM (BCMA), and 0.39 nM (BR-3). A
with an IC50 value of 8.5 nM (Figure 4B). Thus, 4-parameter logistic model was used for curve fitting and calculation of
LymphoStat-B binds BLyS in both a solid-phase capture binding parameters. Values are the mean ⫾ SEM of triplicate samples.
assay and in solution. Ab ⫽ antibody.
LYMPHOSTAT-B, HUMAN MONOCLONAL ANTIBODY AGAINST BLyS 3259

Figure 6. LymphoStat-B inhibition of B lymphocyte stimulator (BLyS) binding to full-length

TACI, BCMA, and BLyS receptor 3 (BR-3) expressed in HEK293T cells. Plasmids encoding the 3
full-length BLyS receptors or vector control were transfected into HEK293T cells. Cells were
subsequently stained with biotinylated BLyS, and binding was detected using a streptavidin–
phycoerythrin (PE) conjugate. Where indicated, a 5-fold molar excess (over BLyS) of
LymphoStat-B or human IgG1 control antibody was added to the staining reaction. Profiles marked
“No BLyS” represent staining of the streptavidin–PE conjugate alone (represented as the broken
line in the other plots).

The presence of membrane-bound BLyS was demon-
strated by staining with a murine monoclonal antibody,
12D6, which was previously characterized as recognizing
membrane-bound BLyS. LymphoStat-B failed to stain
cells that were positive for monoclonal antibody 12D6.
Furthermore, interferon-␥, which has previously been
demonstrated to enhance BLyS expression on mono-
cytes, led to increased specific binding of the 12D6
antibody but not LymphoStat-B. These results suggest
that LymphoStat-B does not recognize membrane-
bound BLyS.
LymphoStat-B inhibition of BLyS binding to
TACI, BCMA, and BR-3. Recent studies have deter-
mined that BLyS binds to 3 cellular receptors, TACI,
BCMA, and BR-3. Because the role of these receptors in
B cell development and function is not fully understood,
it was important to assess the ability of LymphoStat-B to
inhibit BLyS binding to each receptor. Accordingly, the
extracellular domains of TACI, BCMA, and BR-3 were
expressed as fusion proteins, and the ability of
LymphoStat-B to inhibit labeled BLyS binding to each
was evaluated in an electrochemiluminescence detection
assay. As shown in Figures 5A–C, LymphoStat-B inhib-
ited BLyS binding to all 3 receptors with equivalent
potency (IC50 0.10–0.11 nM).
The ability of LymphoStat-B to inhibit ligand–
receptor interactions was also evident in a cell-based
assay in which full-length TACI, BCMA, and BR-3 were
transiently expressed on the surface of HEK293T cells
(Figure 6). BLyS-specific binding was readily detected
on all receptor-expressing cells but not on cells trans-
fected with the corresponding vector control. Addition
3260 BAKER ET AL

Figure 7. Inhibitory effects of LymphoStat-B on the responses of BALB/c mice stimulated with human B
lymphocyte stimulator (BLyS). BLyS was administered to mice over a 5-day period, with or without
LymphoStat-B or control IgG1 on days 1 and 3. The effects of BLyS on spleen weight, splenic B cell
representation, and serum IgA levels were determined on day 5. A, Effect of LymphoStat-B on
BLyS-induced increases in spleen weight. B, Effect of LymphoStat-B on BLyS-induced increases in the
number of mature splenic B cells (ThB⫹/B220⫹). These markers are the murine equivalents of Ly6D and
CD45R, respectively. Data are reported as the mean of the B220⫹/ThB⫹ cell population, as determined
by flow cytometry analysis. C, Effect of LymphoStat-B on BLyS-induced elevations of serum IgA levels.
Values are the mean ⫾ SEM (n ⫽ 10 mice per treatment group). ### ⫽ P ⬍ 0.0005 for recombinant
human BLyS versus buffer; ⴱ ⫽ P ⬍ 0.05 for LymphoStat-B versus the corresponding dose of human IgG1;
ⴱⴱⴱ ⫽ P ⬍ 0.0005 for LymphoStat-B versus the corresponding dose of human IgG1; # ⫽ P ⬍ 0.05 for
recombinant human BLyS versus buffer. hu ⫽ human; Ab ⫽ antibody.

of a 5-fold molar excess of LymphoStat-B, but not LymphoStat-B inhibition of the pharmacologic
control IgG, blocked BLyS binding. Taken together, effects of human BLyS in mice. Although human and
these findings demonstrate that LymphoStat-B blocks murine BLyS share significant amino acid homology
BLyS binding to cells via TACI, BCMA, and BR-3. (63% identity), LymphoStat-B does not neutralize the
LYMPHOSTAT-B, HUMAN MONOCLONAL ANTIBODY AGAINST BLyS 3261

Table 2. Effect of LymphoStat-B on tissue immunophenotype in cynomolgus monkeys*

MNCs analyzed,
study day,
LymphoStat-B dose CD3⫹ CD3⫹/CD4⫹ CD3⫹/CD8⫹ CD3⫺/CD14⫹ CD20⫹ CD20⫹/CD21⫹
Spleen MNCs
Day 29 (n ⫽ 6/group)
None (control) 25.9 ⫾ 9.3 8.8 ⫾ 3.9 8.2 ⫾ 4.3 1.04 ⫾ 0.6 51.5 ⫾ 13.0 10.7 ⫾ 5.3
5 mg/kg/dose 45.6 ⫾ 9.0† 16.9 ⫾ 5.4† 16.2 ⫾ 3.6 0.45 ⫾ 0.3 24.9 ⫾ 16.7† 6.2 ⫾ 5.4
15 mg/kg/dose 48.4 ⫾ 11.7† 15.0 ⫾ 5.5 15.5 ⫾ 6.9 0.65 ⫾ 0.4 30.5 ⫾ 15.0 6.0 ⫾ 4.6
50 mg/kg/dose 44.1 ⫾ 19.8 11.9 ⫾ 4.4 14.3 ⫾ 7.7 1.31 ⫾ 0.7 33.9 ⫾ 12.9 6.4 ⫾ 4.5
Day 57 (n ⫽ 4/group)
None (control) 33.6 ⫾ 10.1 15.2 ⫾ 4.0 13.6 ⫾ 3.1 2.21 ⫾ 1.1 39.3 ⫾ 12.6 8.2 ⫾ 0.9
5 mg/kg/dose 44.8 ⫾ 4.3 21.6 ⫾ 2.6 17.5 ⫾ 3.1 1.92 ⫾ 0.8 18.0 ⫾ 5.7† 3.1 ⫾ 0.7†
15 mg/kg/dose 41.9 ⫾ 10.6 18.3 ⫾ 7.2 18.5 ⫾ 4.0 1.72 ⫾ 1.1 22.5 ⫾ 7.7† 4.9 ⫾ 2.1†
50 mg/kg/dose 55.3 ⫾ 5.5† 23.8 ⫾ 3.3 24.7 ⫾ 2.6† 1.02 ⫾ 0.5 20.8 ⫾ 7.5† 3.5 ⫾ 1.4†

Lymph node MNCs

Day 29 (n ⫽ 6/group)
None (control) 67.0 ⫾ 6.7 46.6 ⫾ 6.3 18.2 ⫾ 5.6 1.18 ⫾ 0.6 28.7 ⫾ 9.2 5.2 ⫾ 1.9
5 mg/kg/dose 77.9 ⫾ 4.5† 56.0 ⫾ 5.6† 20.9 ⫾ 5.9 0.66 ⫾ 0.6 19.7 ⫾ 3.9 3.2 ⫾ 1.6
15 mg/kg/dose 82.3 ⫾ 6.0† 60.6 ⫾ 2.0† 20.7 ⫾ 5.8 0.43 ⫾ 0.1 16.0 ⫾ 4.5 3.4 ⫾ 1.6
50 mg/kg/dose 78.1 ⫾ 7.4† 55.5 ⫾ 4.9† 21.7 ⫾ 4.6 1.19 ⫾ 1.7 18.2 ⫾ 7.1 2.4 ⫾ 1.8
Day 57 (n ⫽ 4/group)
None (control) 70.9 ⫾ 7.9 47.9 ⫾ 8.0 22.3 ⫾ 3.6 1.66 ⫾ 0.4 26.7 ⫾ 5.9 10.8 ⫾ 3.8
5 mg/kg/dose 79.5 ⫾ 8.3 59.4 ⫾ 6.7 21.4 ⫾ 3.1 0.78 ⫾ 0.4 15.9 ⫾ 6.3† 5.0 ⫾ 2.7†
15 mg/kg/dose 84.7 ⫾ 2.3† 61.2 ⫾ 3.4† 25.1 ⫾ 7.7 1.72 ⫾ 0.7 11.5 ⫾ 3.0† 4.0 ⫾ 0.8†
50 mg/kg/dose 87.5 ⫾ 1.4† 59.4 ⫾ 5.3 30.0 ⫾ 6.0 0.69 ⫾ 0.7 11.2 ⫾ 1.9† 2.9 ⫾ 1.2†

* LymphoStat-B was administered at the indicated doses once every 7 days for 4 treatment cycles. Lymph node and spleen mononuclear cells
(MNCs) were analyzed by flow cytometry using the indicated markers. Values are the mean ⫾ SD percentage of spleen or lymph node MNCs.
† P ⬍ 0.05 versus control group, by Dunnett’s test (analysis of variance).

effects of murine BLyS (Human Genome Sciences:
unpublished data) and is therefore unable to antagonize
endogenous BLyS activity in either normal mice or
murine models of autoimmune disease. In order to
assess the in vivo efficacy of human-specific BLyS an-
tagonists, a mouse model was developed in which the
ability of LymphoStat-B to inhibit the pharmacologic
effects of exogenously administered human BLyS was
tested. This model allows the biologic actions of BLyS to
be measured as increases in spleen weight, increases in
serum IgA levels, and increases in the population of
mature B cells in the spleen (B220⫹/ThB⫹ splenocytes;
the human equivalent of the markers are CD45R and
Ly6D, respectively) (1,35).
As shown in Figure 7, subcutaneous administra-
tion of 0.3 mg/kg of human BLyS for 4 consecutive days
resulted in increases in spleen weights, in the represen-
tation of CD45R⫹ (B220⫹)/ThB⫹ splenocytes, and in
the total serum IgA concentrations. Coadministration of
LymphoStat-B intravenously resulted in a dose-dependent
inhibition of these human BLyS-induced effects, with
complete inhibition observed between 1.5 and 5.0 mg/kg
of monoclonal antibody. No inhibition of BLyS-induced
effects was achieved by administration of the Ig isotype
control monoclonal antibody (human IgG1). These re-
sults demonstrate that LymphoStat-B is an effective in
vivo antagonist of human BLyS bioactivity.
Effects of administration of LymphoStat-B to
cynomolgus monkeys. The in vivo consequences of BLyS
inhibition was evaluated in cynomolgus monkeys. Cyno-
molgus monkeys were considered a suitable animal
model since cynomolgus BLyS is 96% identical to hu-
man BLyS, and LymphoStat-B inhibits the in vitro
activity of cynomolgus monkey BLyS with similar effi-
ciency as human BLyS (Human Genome Sciences: un-
published data). Animals were injected with 5, 15, or 50
mg/kg of LymphoStat-B or vehicle once every 7 days for
4 treatment cycles. Animals were necropsied on study
day 29 or, following a 28-day treatment-free recovery
period, on day 57.
Consistent with the phenotype of BLyS-deficient
mice (18,19), a decrease in B cell representation was
observed in spleen and mesenteric lymph nodes from
LymphoStat-B–treated monkeys on both day 29 and day
57, as monitored by flow cytometry (Table 2). As might
be expected, there was a reciprocal increase in the
percentage of tissue T cells, but there was no corre-
sponding increase in circulating T cells, nor was there a
consistent T cell immunophenotype that was increased
(i.e., CD8⫹ versus CD4⫹). Therefore, it is likely that
3262
the observed increase in the relative percentage of T
cells simply reflects the B cell depletion in these tissues.
Consistent with this, histologic analysis of these tissues
demonstrated a mild lymphoid depletion primarily in B
cell–dependent areas (data not shown). No significant
changes were observed in peripheral blood mononuclear
cells detected by flow cytometry or in serum Ig concen-
trations (data not shown). These results demonstrate
that LymphoStat-B is able to inhibit BLyS in vivo and
that this inhibition results in depletion of B cell popula-
tions after a relatively short course of treatment.
DISCUSSION
BLyS plays a critical role in the normal regulation
of B cell development and immune response. This was
demonstrated by the severely depleted B cell phenotype
observed in BLyS-deficient animals (18,19). As is the
case for other cytokines, BLyS levels must be carefully
regulated in order to maintain “immune homeostasis”
(37). Consistent with this notion are findings that over-
production of BLyS is associated with increased immu-
noglobulin production and autoimmunity. Transgenic
mice overexpressing either murine or human BLyS show
increased numbers of mature B cells in the spleen and
lymph nodes, elevated levels of total immunoglobulins,
rheumatoid factor, anti–double-stranded DNA antibod-
ies, and circulating immune complexes, all of which are
manifestations associated with autoimmune disease. The
mice eventually develop proteinuria and glomerulone-
phritis, symptoms that are specifically associated with
SLE (21,22). Elevated levels of BLyS have also been
detected in the (NZB ⫻ NZW)F1 and MRL-lpr/lpr
mouse models of lupus. BLyS levels in these animals
increase with age, in parallel with increased autoanti-
body production, proteinuria, and other disease manifes-
tations, again suggesting an important role for BLyS in the
development of autoimmune disease in animals (6).
Evidence that BLyS is associated with human
autoimmune disease comes from several studies demon-
strating elevated levels of circulating BLyS in serum
samples from patients with SLE and RA (23,24). In RA
patients, elevated levels of BLyS were also found in
synovial fluid and were consistently greater than the
serum levels in the same patient, possibly suggesting that
there is local production of BLyS at sites of inflamma-
tion (38). Increased BLyS levels have also been found in
sera from patients with Sjögren’s syndrome, another
autoimmune disease characterized by B cell hyperreac-
tivity and autoantibody production (25,26). Mice trans-
genic for BLyS also develop symptoms of Sjögren’s
BAKER ET AL
syndrome (25), consistent with clinical observations of
the association of Sjögren’s syndrome in subsets of
patients with SLE (39). Taken together, these observa-
tions suggest that elevated levels of BLyS may contribute
to B cell hyperactivity and autoantibody production in
multiple autoimmune diseases by enhancing the survival
of B cells. Specific targeting of BLyS may therefore
provide a novel treatment for these diseases. Toward
that end, we have generated a human antibody specific
for human BLyS that can neutralize the biologic func-
tions of BLyS in vitro and in vivo.
LymphoStat-B was generated from a naive hu-
man Ig library using phage display technology. More
than 1,200 antibodies with distinct amino acid sequences
were isolated that specifically bound soluble human
BLyS. Optimization of selected main candidates allowed
further improvements in activity, leading to the selection
of the clinical development candidate, LymphoStat-B.
Interactions between BLyS and its receptors are
complex. At least 3 distinct receptors exist for BLyS. The
major receptor for mediating BLyS-dependent B cell
development appears to be the BLyS-specific receptor
BR-3. This is based on data showing that the A/WySnJ
mouse strain, which has a defect in BR-3, mirrors the
severe deficits in peripheral mature B cell populations
seen in BLyS-deficient mice (9,10,18–20). Although
BLyS binds to 2 additional receptors, BCMA and TACI,
analyses of mice deficient in these receptors show no B
cell deficits in the BCMA knockout mice (14) and
increased levels of B cells in TACI-deficient mice (15),
suggesting a redundant role for BCMA and an inhibitory
role for TACI in the regulation of B cell maturation
(17). Importantly, we show by 2 independent methods
that LymphoStat-B is able to neutralize BLyS interac-
tions with all 3 BLyS receptors.
BLyS, like many TNF family members, exists in
both soluble and membrane-bound forms. Most avail-
able data support the action of BLyS as a soluble
cytokine (1), and elevated levels of soluble BLyS are
correlated with autoimmune disease (23,24). Any spe-
cific activity of membrane-bound BLyS remains to be
determined. In fact, the exact nature of membrane-
bound BLyS is not fully understood, but it has been
shown that a panel of antibodies can differentially
recognize membrane-bound BLyS (7). The BLyS anti-
bodies described in the present study were selected
specifically for their ability to recognize and inhibit the
effects of soluble human BLyS. LymphoStat-B does not
recognize membrane-bound BLyS and does not bind to
the surface of human cells that bear membrane-bound
BLyS. Thus, the specific activity of soluble BLyS is
LYMPHOSTAT-B, HUMAN MONOCLONAL ANTIBODY AGAINST BLyS
targeted, and any nonspecific effects that might be
associated with antibody binding to BLyS-producing
cells should be prevented.
BLyS has also been shown to exist in a complex
with APRIL to form heterotrimers. Although the bio-
logic significance of these heterotrimers is not clearly
defined, they have in vitro biologic activity and interact
with BLyS receptors (40). While LymphoStat-B was
raised against soluble “homotrimeric” BLyS, interac-
tions with heterotrimers have been shown in in vitro
assays (Smith R, Roschke V: unpublished observations),
although the affinity of these interactions was 7–8-fold
lower than that of the heterotrimer recognizing antibod-
ies reported previously. The significance of this inhibi-
tion awaits a clearer delineation of the in vivo role of
BLyS/APRIL heterotrimers in B cell regulation.
In an in vitro assay, LymphoStat-B was found to
inhibit human BLyS-induced stimulation of B cells from
both murine splenocytes and human tonsillar B cells
(data not shown). In vivo, LymphoStat-B neutralizes the
observable effects of exogenously administered human
BLyS in mice. These effects include stimulation of B cell
proliferation, as evidenced by increases in spleen weight,
representation of mature marginal zone B cells, as
demonstrated by increases in B220⫹/ThB⫹ B cells, and
immunoglobulin secretion, as evidenced by increases in
IgA levels. LymphoStat-B at dosages of 1–5 mg/kg
completely prevented these BLyS-induced activities. In
this model system, LymphoStat-B has similar efficiencies
when administered via the intravenous or the subcuta-
neous route (Hilbert D: unpublished observations). It
should also be noted that the dosing schedule of 4 daily
injections of BLyS used in the present study has only
modest effects on increasing IgM levels and little, if any,
effects on IgG levels (35). Nevertheless, LymphoStat-B
can reduce BLyS-induced changes in IgM (data not
shown).
Administration of LymphoStat-B to normal cyno-
molgus monkeys resulted in tissue B cell depletion that
was sustained for up to 4 weeks after the treatment
period. Although no significant depletion of circulating
B cells was observed at these time points, it has been
suggested that the spleen and lymphoid tissue act as
reservoirs for the supply of B cells into the bloodstream
(41), and effects on circulating B cells may not be
observed until reserves of B cells in lymphoid tissues are
adequately suppressed. No significant changes were
observed in serum immunoglobulin levels following
LymphoStat-B administration. However, since the B
cell–depleting drug rituximab results in severe (⬎90%)
and prolonged B cell depletion, with only minor con-
3263
comitant effects on serum immunoglobulin levels (42), it
is perhaps not surprising that changes in serum immu-
noglobulins were not observed in the study reported
here. Although the effects of LymphoStat-B administra-
tion over a short period of time might not be expected to
fully reflect the phenotypes of BLyS knockout mice, one
key similarity with the BLyS knockout mice, namely a
reduction of tissue B cell populations, was observed
following drug administration.
LymphoStat-B possesses many properties that
make it ideally suited for use as a therapeutic agent.
First, it binds with high affinity to soluble human BLyS
and inhibits its activities both in vitro and in vivo.
Second, it does not recognize other TNF family mem-
bers, including its closest homolog human APRIL, and is
therefore specific for the factor implicated in the patho-
genesis of several autoimmune diseases. As an antibody,
an additional advantage of LymphoStat-B is its long
terminal half-life. In mice and monkeys, the half-life of
LymphoStat-B is 2.5 days and 11–14 days, respectively
(Riccobene T: unpublished observations).
Overproduction of BLyS has been associated
with SLE, RA, and Sjögren’s syndrome. The etiologies
of these diseases are unknown, but they share the
common features of B cell hyperactivity and autoanti-
body production, which are strongly implicated in the
pathogenesis of the disease process. Support for the
ability of an antagonist of BLyS to affect these diseases
comes from several animal studies with soluble BLyS
receptors. Administration of TACI-Fc or BCMA-Fc has
been shown to inhibit antigen-specific antibody re-
sponses and decrease B cell numbers and germinal
centers (43,44). TACI-Fc and BR-3-Fc have also been
shown to inhibit the development of SLE disease man-
ifestations in the (NZB ⫻ NZW)F1 mouse (6,28).
TACI-Fc has also prevented disease onset in the
collagen-induced arthritis model in mice (27).
The critical role BLyS plays in the regulation of B
cell homeostasis suggests that deregulation of BLyS may
underlie these as well as additional B cell–mediated
diseases. LymphoStat-B is being developed as a novel
treatment for autoimmune diseases and is currently
being evaluated in a phase I clinical trial in SLE patients.
Studies are also under way to identify additional diseases
in which BLyS might play a role and LymphoStat-B may
have therapeutic potential.
ACKNOWLEDGMENTS
The authors wish to thank members of the DNA
sequencing facility at Human Genome Sciences for antibody
3264
sequencing, Steve Barash for bioinformatics analysis, Steve
Ullrich for antibody purification, and Jeff Carrell, Ernest
Boyd, and Dana Bresette for flow cytometry.
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