U.S. patent application number 16/118909 was filed with the patent office on 2018-12-27 for compositions and methods for beta-glucan immunotherapy.
The applicant listed for this patent is Biothera, Inc.. Invention is credited to Mary A. Antonysamy, Nandita Bose, Michael E. Danielson, William J. Grossman, Kyle S. Michel, Mariana I. Nelson, Richard M. Walsh.
Application Number | 20180369376 16/118909 |
Document ID | / |
Family ID | 49514726 |
Filed Date | 2018-12-27 |
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United States Patent
Application |
20180369376 |
Kind Code |
A1 |
Grossman; William J. ; et
al. |
December 27, 2018 |
COMPOSITIONS AND METHODS FOR BETA-GLUCAN IMMUNOTHERAPY
Abstract
This disclosure describes, in one aspect, a composition that
includes a .beta.-glucan component and an antibody component that
specifically binds to the .beta.-glucan. In another aspect, this
disclosure describes a method of increasing a subject's response to
.beta.-glucan immunotherapy. Generally, the method includes
identifying the subject as a low binder of .beta.-glucan and
administering to the subject a composition that comprises a
.beta.-glucan moiety conjugated to the therapeutic antibody. In
some cases, the therapeutic antibody can be an anti-tumor
antibody.
Inventors: |
Grossman; William J.; (Third
Lake, IL) ; Antonysamy; Mary A.; (Woodbury, MN)
; Walsh; Richard M.; (Lino Lakes, MN) ; Nelson;
Mariana I.; (Rosemount, MN) ; Bose; Nandita;
(Plymouth, MN) ; Danielson; Michael E.; (St. Paul,
MN) ; Michel; Kyle S.; (Eagan, MN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Biothera, Inc. |
Eagan |
MN |
US |
|
|
Family ID: |
49514726 |
Appl. No.: |
16/118909 |
Filed: |
August 31, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14398020 |
Oct 30, 2014 |
10092646 |
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PCT/US2013/031625 |
Mar 14, 2013 |
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16118909 |
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61640397 |
Apr 30, 2012 |
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61640834 |
May 1, 2012 |
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61640842 |
May 1, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 37/02 20180101;
A61P 43/00 20180101; C07K 16/12 20130101; G01N 33/56966 20130101;
A61P 37/04 20180101; A61K 39/39 20130101; C07K 16/14 20130101; A61K
2039/55583 20130101; G01N 33/554 20130101; A61K 39/39558 20130101;
A61K 39/39583 20130101; A61K 45/06 20130101; A61P 35/02 20180101;
G01N 2400/24 20130101; A61K 39/39575 20130101; A61K 47/6835
20170801; A61K 31/716 20130101; A61P 35/00 20180101; A61K 31/716
20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 31/716 20060101 A61K031/716; G01N 33/569 20060101
G01N033/569; G01N 33/554 20060101 G01N033/554; C07K 16/14 20060101
C07K016/14; A61K 39/39 20060101 A61K039/39; A61K 47/68 20170101
A61K047/68; A61K 45/06 20060101 A61K045/06; C07K 16/12 20060101
C07K016/12 |
Claims
1. A composition comprising: a soluble .beta.-glucan component; and
an antibody component that specifically binds to the soluble
.beta.-glucan.
2. The composition of claim 1 wherein the soluble .beta.-glucan is
derived from yeast.
3. The composition of claim 1 wherein the soluble .beta.-glucan
comprises a .beta.-1,3/1,6 glucan.
4. The composition of claim 1 wherein the soluble .beta.-glucan
comprises
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
5. The composition of claim 1 wherein the antibody component
comprises a monoclonal antibody that specifically binds to the
soluble .beta.-glucan.
6. The composition of claim 5 wherein the monoclonal antibody
comprises BfD I, BfD II, BfD III, or BfD IV.
7. The composition of claim 1 wherein the soluble .beta.-glucan
component and the antibody component are provided in a single
formulation.
8. The composition of claim 1 wherein the soluble .beta.-glucan
component and the antibody component are provided in separate
formulations.
9. The composition of claim 1 further comprising an anti-tumor
antibody.
10-31. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority to U.S. Provisional Patent
Application Ser. No. 61/640,834, filed May 1, 2012, and U.S.
Provisional Patent Application Ser. No. 61/640,397, filed Apr. 30,
2012, each of which is incorporated herein by reference.
SUMMARY
[0002] This disclosure describes, in one aspect, a composition that
includes a .beta.-glucan component and an antibody component that
specifically binds to the .beta.-glucan. In some embodiments, the
.beta.-glucan may be derived from yeast. In some embodiments, the
.beta.-glucan can include a .beta.-1,3/1,6 glucan such as
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
[0003] In some embodiments, the antibody component can include a
monoclonal antibody that specifically binds to the .beta.-glucan.
In some embodiments, the monoclonal antibody can include BfD I, BfD
II, BfD III, or BfD IV.
[0004] In some embodiments, the .beta.-glucan component and the
antibody component can be provided in a single formulation. In
other embodiments, the .beta.-glucan component and the antibody
component can be provided in separate formulations.
[0005] In another aspect, this disclosure describes a method that
generally includes co-administering to a subject a .beta.-glucan
and an antibody preparation that specifically binds to the
.beta.-glucan. In some embodiments, the method can further include
administering to the subject an anti-tumor antibody.
[0006] In another aspect, this disclosure describes a method of
increasing a subject's response to .beta.-glucan immunotherapy.
Generally, the method includes co-administering to the subject a
composition that comprises a .beta.-glucan and an antibody
preparation that specifically binds to the .beta.-glucan. In some
embodiments, the method can further include identifying the subject
as a low binder and administering a composition that comprises a
.beta.-glucan and an antibody preparation that specifically binds
to the .beta.-glucan.
[0007] In some embodiments of any of these methods, the
.beta.-glucan and the antibody preparation may be co-administered
simultaneously. In other embodiments of any of the methods, the
antibody preparation may be co-administered at different times. In
some embodiments of any of the methods, the .beta.-glucan and the
antibody preparation may be co-administered at different sites.
[0008] In some embodiments of these methods, the .beta.-glucan may
be derived from yeast. In some embodiments of these methods, the
.beta.-glucan can include a .beta.-1,3/1,6 glucan such as
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
[0009] In some embodiments of these methods, the antibody component
can include a monoclonal antibody that specifically binds to the
.beta.-glucan such as BfD I, BfD II, BfD III, or BfD IV.
[0010] In another aspect, this disclosure describes a method of
increasing a subject's response to .beta.-glucan immunotherapy
involving an antibody. Generally, the method includes administering
to the subject a composition that includes a .beta.-glucan moiety
conjugated to the antibody. In some embodiments, the .beta.-glucan
moiety may conjugated to a therapeutic antibody such as, for
example, an anti-tumor antibody. In some embodiments, the method
further includes identifying the subject as a low binder of
.beta.-glucan.
[0011] In some embodiments, the .beta.-glucan moiety may be derived
from yeast. In some embodiments, the .beta.-glucan moiety can be,
or is derived from, a .beta.-1,3/1,6 glucan such as
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
[0012] In some embodiments, the .beta.-glucan therapy can include
administering to a subject a .beta.-glucan, antibody that
specifically binds .beta.-glucan, and a therapeutic antibody. In
some cases, the therapeutic antibody can include an anti-tumor
antibody.
[0013] The above summary of the present invention is not intended
to describe each disclosed embodiment or every implementation of
the present invention. The description that follows more
particularly exemplifies illustrative embodiments. In several
places throughout the application, guidance is provided through
lists of examples, which examples can be used in various
combinations. In each instance, the recited list serves only as a
representative group and should not be interpreted as an exclusive
list.
BRIEF DESCRIPTION OF THE FIGURES
[0014] FIG. 1. Flow cytometry data showing differential
.beta.-glucan (PGG) binding to polymorphonuclear leukocytes in
healthy human whole blood.
[0015] FIG. 2. Data showing differential .beta.-glucan binding to
neutrophils in healthy human whole blood.
[0016] FIG. 3. Data showing differential .beta.-glucan binding to
monocytes in healthy human whole blood.
[0017] FIG. 4. Data comparing anti-.beta.-glucan antibody titers of
low binders and high binders.
[0018] FIG. 5. Data showing that high binder serum can increase
.beta.-glucan binding to PMNs obtained from a low binder.
[0019] FIG. 6. Data showing the anti-.beta.-glucan antibodies can
increase .beta.-glucan binding to PMNs from a low binder.
[0020] FIG. 7. Data showing intravenous immunoglobulin can increase
.beta.-glucan binding to PMNs from a low binder.
[0021] FIG. 8. Data showing conversion of a low binder to a high
binder by treatment with intravenous immunoglobulin that includes a
combination of .beta.-glucan and anti-.beta.-glucan antibodies.
[0022] FIG. 9. Comparison of the average number of days on therapy
for patients in control and investigational arms of two-armed,
open-label, randomized, multi-center study.
[0023] FIG. 10. Data showing binding of PGG-antibody conjugates to
PMNs.
[0024] FIG. 11. Data showing binding of PGG-IVIG conjugates to
PMNs.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0025] This disclosure describes methods related to the use of
.beta.-glucan as a component of immunotherapy. The compositions and
methods described herein exploit the observation of differential
binding of .beta.-glucan by immune cells in different populations
of healthy humans. Surprisingly, "high binders" of .beta.-glucan
exhibit higher titers of anti-.beta.-glucan antibodies than "low
binders." Thus, this disclosure describes compositions that include
a .beta.-glucan component and an antibody component that
specifically binds to the .beta.-glucan. This disclosure also
describes methods that generally include co-administering a
.beta.-glucan and an antibody or antibody component that
specifically binds the .beta.-glucan, or a .beta.-glucan moiety
conjugated to an antibody or antibody fragment. Such methods can
convert a "low binder" to a "high binder" and, thus, increase the
population for whom .beta.-glucan-based immunotherapy can be
effective.
[0026] .beta.-glucans are polymers of glucose derived from a
variety of microbiological and plant sources including, for
example, yeast, bacteria, algae, seaweed, mushroom, oats, and
barley. Of these, yeast .beta.-glucans have been extensively
evaluated for their immunomodulatory properties. Yeast
.beta.-glucans can be present as various forms such as, for
example, intact yeast, zymosan, purified whole glucan particles,
solubilized zymosan polysaccharide, or highly-purified soluble
.beta.-glucans of different molecular weights. Structurally, yeast
.beta.-glucans are composed of glucose monomers organized as a
.beta.-(1,3)-linked glucopyranose backbone with periodic
.beta.-(1,3) glucopyranose branches linked to the backbone via
.beta.-(1,6) glycosidic linkages. The different forms of yeast
.beta.-glucans can function differently from one another. The
mechanism through which yeast .beta.-glucans exert their
immunomodulatory effects can be influenced by the structural
differences between different forms of the .beta.-glucans such as,
for example, its particulate or soluble nature, tertiary
conformation, length of the main chain, length of the side chain,
and frequency of the side chains. The immune stimulating functions
of yeast .beta.-glucans are also dependent upon the receptors
engaged in different cell types in different species, which again,
is dependent on the structural properties of the
.beta.-glucans.
[0027] In one aspect, this disclosure describes a composition that
includes, generally, a .beta.-glucan component and an antibody
component that specifically binds to the .beta.-glucan.
[0028] The .beta.-glucan component may include any suitable form of
.beta.-glucan or any combination of two or more forms of
.beta.-glucan. Suitable .beta.-glucans and the preparation of
suitable .beta.-glucans from their natural sources are described
in, for example, U.S. Patent Application Publication No.
US2008/0103112 A1. In some embodiments, the .beta.-glucan may be
derived from a yeast such as, for example, Saccharomyces
cerevisiae. In certain specific embodiments, the .beta.-glucan may
be or be derived from
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e, also referred to herein as PGG (IMPRIME PGG, Biothera, Inc.,
Eagan, Minn.), a highly purified and well characterized form of
yeast-derived .beta.-glucan. Thus, the .beta.-glucan component can
include, for example, a modified and/or derivatized .beta.-glucan
such as those described in International Patent Application No.
PCT/US12/36795. In other embodiments, the .beta.-glucan component
can include, for example, a particulate-soluble .beta.-glucan or a
particulate-soluble .beta.-glucan preparation, each of which is
described in, for example, U.S. Pat. No. 7,981,447.
[0029] The antibody component of the composition can include any
antibody preparation that specifically binds to the .beta.-glucan
component of the composition. As used herein, "specific" and
variations thereof refer to having a differential or a non-general
(i.e., non-specific) affinity, to any degree, for a particular
target. Thus, the antibody component can include a polyclonal
antibody preparation (e.g., derived from serum), a monoclonal
antibody preparation, or any antibody fragment such as an Fc
portion. Exemplary monoclonal antibodies that specifically bind
.beta.-glucan include, for example, monoclonal antibodies
identified as BfD I, BfD II, BfD III, and/or BfD IV (Biothera,
Inc., Eagan, Minn.), each of which is described in U.S. Pat. No.
6,294,321.
[0030] The antibody or antibody fragment (e.g., the Fc portion)
conjugated to the .beta.-glucan component can be any suitable
antibody or antibody fragment that can be linked to the
.beta.-glucan component.
[0031] The .beta.-glucan component, the antibody component, and/or
the combination of both components may be formulated in a
composition along with a "carrier." As used herein, "carrier"
includes any solvent, dispersion medium, vehicle, coating, diluent,
antibacterial agent and/or antifungal agent, isotonic agent,
absorption delaying agent, buffer, carrier solution, suspension,
colloid, and the like. The use of such media and/or agents for
pharmaceutical active substances is well known in the art. Except
insofar as any conventional media or agent is incompatible with the
.beta.-glucan or the antibody, its use in the therapeutic
compositions is contemplated. Supplementary active ingredients also
can be incorporated into the compositions.
[0032] By "pharmaceutically acceptable" is meant a material that is
not biologically or otherwise undesirable, i.e., the material may
be administered to an individual along with the .beta.-glucan
and/or the antibody without causing any undesirable biological
effects or interacting in a deleterious manner with any of the
other components of the pharmaceutical composition in which it is
contained.
[0033] The .beta.-glucan component, the antibody component, and/or
the combination of both components may be formulated into a
pharmaceutical composition. In some embodiments, the .beta.-glucan
component of the composition and the antibody component of the
composition may be provided in a single formulation. In other
embodiments, the .beta.-glucan component and the antibody component
may be provided in separate formulations. The composition may be
formulated in a variety of and/or a plurality of forms adapted to
one or more preferred routes of administration. Thus, a composition
can be administered via one or more known routes including, for
example, oral, parenteral (e.g., intradermal, transcutaneous,
subcutaneous, intramuscular, intravenous, intraperitoneal, etc.),
or topical (e.g., intranasal, intrapulmonary, intramammary,
intravaginal, intrauterine, intradermal, transcutaneous, rectally,
etc.). A composition, or a portion thereof, can be administered to
a mucosal surface, such as by administration to, for example, the
nasal or respiratory mucosa (e.g., by spray or aerosol). A
composition, or a portion thereof, also can be administered via a
sustained or delayed release.
[0034] A formulation may be conveniently presented in unit dosage
form and may be prepared by methods well known in the art of
pharmacy. Methods of preparing a composition with a
pharmaceutically acceptable carrier include the step of bringing
the .beta.-glucan and/or the antibody into association with a
carrier that constitutes one or more accessory ingredients. In
general, a formulation may be prepared by uniformly and/or
intimately bringing the active compound into association with a
liquid carrier, a finely divided solid carrier, or both, and then,
if necessary, shaping the product into the desired
formulations.
[0035] The .beta.-glucan component, the antibody component, and/or
the combination of both components may be provided in any suitable
form including but not limited to a solution, a suspension, an
emulsion, a spray, an aerosol, or any form of mixture. The
composition may be delivered in formulation with any
pharmaceutically acceptable excipient, carrier, or vehicle. For
example, the formulation may be delivered in a conventional topical
dosage form such as, for example, a cream, an ointment, an aerosol
formulation, a non-aerosol spray, a gel, a lotion, and the like.
The formulation may further include one or more additives including
such as, for example, an adjuvant, a skin penetration enhancer, a
colorant, a fragrance, a flavoring, a moisturizer, a thickener, and
the like.
[0036] In another aspect, the invention provides a method that
generally includes co-administering to a subject, in effective
amounts with one another, a .beta.-glucan and an antibody
preparation that specifically binds the .beta.-glucan. As used
herein, "co-administered" refers to two or more components of a
combination administered so that the therapeutic or prophylactic
effects of the combination can be greater than the therapeutic or
prophylactic effects of either component administered alone. Two
components may be co-administered simultaneously or sequentially.
Simultaneously co-administered components may be provided in one or
more pharmaceutical compositions. Sequential co-administration of
two or more components includes cases in which the components are
administered so that both components are simultaneously
bioavailable after both are administered. Regardless of whether the
components are co-administered simultaneously or sequentially, the
components may be co-administered at a single site or at different
sites. Also as used herein, "an effective amount" refers to the
amount of .beta.-glucan and antibody that specifically binds to the
.beta.-glucan effective to increase binding of the .beta.-glucan to
immune cells--e.g., polymorphonuclear leukocytes (PMNs), monocytes,
or neutrophils--or to increase production of cytokines and/or
chemokines associated with binding of .beta.-glucans--e.g., IL-8
production.
[0037] .beta.-glucans suitable for use in the methods include one
or more of those described as suitable for use as the .beta.-glucan
component of the compositions described above. Also, the antibody
preparation can include one or more antibodies described as
suitable for use as the antibody component of the compositions
described above.
[0038] The amount of .beta.-glucan and antibody effective to induce
one or more of the desired effects can vary depending on various
factors including, but not limited to, the weight, physical
condition, and/or age of the subject, and/or the route of
administration. Thus, the absolute amount of .beta.-glucan and
antibody that specifically binds the .beta.-glucan that are
included in a given unit dosage form can vary widely, and depends
upon factors such as the species, age, weight and physical
condition of the subject, as well as the method of administration.
Accordingly, it is not practical to set forth generally the amount
that constitutes an amount of .beta.-glucan and antibody effective
for all possible applications. Those of ordinary skill in the art,
however, can readily determine the appropriate amount with due
consideration of such factors.
[0039] In some embodiments, the method can include administering
sufficient .beta.-glucan to provide a dose of, for example, from
about 100 ng/kg to about 50 mg/kg to the subject, although in some
embodiments the methods may be performed by administering the
.beta.-glucan in a dose outside this range. In some embodiments,
the method includes administering sufficient .beta.-glucan to
provide a dose of from about 10 .mu.g/kg to about 10 mg/kg to the
subject such as, for example, a dose of about 1 mg/kg, about 2
mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg, about 6 mg/kg,
about 7 mg/kg, about 8 mg/kg, about 9 mg/kg, or about 10 mg/kg. In
one particular embodiment, the method includes administering
sufficient .beta.-glucan to provide a dose of 4 mg/kg.
[0040] Alternatively, the dose may be calculated using actual body
weight obtained just prior to the beginning of a treatment course.
For the dosages calculated in this way, body surface area (m.sup.2)
is calculated prior to the beginning of the treatment course using
the Dubois method: m.sup.2=(wt kg.sup.0.425.times.height
cm.sup.0.725).times.0.007184. In some embodiments, therefore, the
method can include administering sufficient .beta.-glucan to
provide a dose of, for example, from about 0.01 mg/m.sup.2 to about
10 mg/m.sup.2.
[0041] In some embodiments, the method can include administering
sufficient antibody that specifically binds the .beta.-glucan to
provide a dose of, for example, from about 100 ng/kg to about 50
mg/kg to the subject, although in some embodiments the methods may
be performed by administering the antibody in a dose outside this
range. In some embodiments, the method includes administering
sufficient antibody to provide a dose of from about 10 .mu.g/kg to
about 5 mg/kg to the subject, for example, a dose of from about 100
.mu.g/kg to about 1 mg/kg. In some embodiments, antibody that
specifically binds the .beta.-glucan can be administered in the
form of intravenous immunoglobulin (IVIG), a blood product that
contains pooled polyvalent IgG from many donors (typically many
hundreds, even thousands, of donors and, thus, naturally containing
anti-.beta.-glucan antibodies). In such embodiments, IVIG may be
administered in a dose of from about 0.1 g/kg to about 2.0 g/kg
such as, for example, 0.1 g/kg, 0.2 g/kg, 0.3 g/kg, 0.4 g/kg, 0.5
g/kg, 0.6 g/kg, 0.7 g/kg, 0.8 g/kg, 0.9 g/kg, 1.0 g/kg, 1.1 g/kg,
1.2 g/kg, 1.3 g/kg, 1.4 g/kg, 1.5 g/kg, 1.6 g/kg, 1.7 g/kg, 1.8
g/kg, 1.9 g/kg, or 2.0 g/kg. In certain embodiments, IVIG may be
administered to provide a dose of about 0.4 g/kg to about 1.0
g/kg.
[0042] Alternatively, the dose may be calculated using actual body
weight obtained just prior to the beginning of a treatment course.
For the dosages calculated in this way, body surface area (m.sup.2)
is calculated prior to the beginning of the treatment course using
the Dubois method: m.sup.2=(wt kg.sup.0.425.times.height
cm.sup.0.725).times.0.007184. In some embodiments, therefore, the
method can include administering sufficient antibody to provide a
dose of, for example, from about 0.01 mg/m.sup.2 to about 10
mg/m.sup.2.
[0043] In some embodiments, the .beta.-glucan and antibody may be
co-administered, for example, from a single dose to multiple doses
per week, although in some embodiments the method may be performed
by co-administering the .beta.-glucan and antibody at a frequency
outside this range. In certain embodiments, the .beta.-glucan and
antibody may be administered from about once per year to once per
week.
[0044] As noted above, yeast .beta.-glucans have been extensively
evaluated for their immunomodulatory properties. We discovered,
however, that distinct populations of individuals exist: one
population exhibits relatively high capacity of .beta.-glucan
binding to innate immune cells in whole blood; another population
exhibits relatively low capacity of .beta.-glucan binding to innate
immune cells in whole blood. This observation was wholly unexpected
based on data from mouse models of immunity and studies involving
isolated human immune cells. Many individuals exhibit some level of
.beta.-glucan binding to immune cells from native, low level
exposure to .beta.-glucans. (e.g., FIG. 1, "De novo"). When
exogenous .beta.-glucan is administered, "low binders" exhibit a
modest increase in the percentage of innate immune cells that bind
.beta.-glucan, while "high binders" exhibit a marked increase in
the percentage of innate immune cells that bind .beta.-glucan.
(FIG. 1, "+ Exogenous PGG"). FIG. 1 and FIG. 2 show data reflecting
.beta.-glucan binding to polymorphonuclear leukocytes (PMNs), and
FIG. 3 (monocytes) shows that the differential binding applies to
other immune cell populations as well. In addition, "high binders"
also tend to produce more cytokines and/or chemokines such as, for
example, IL-8, MCP, MIP-1, etc.
[0045] As used herein, status as a "high binder" refers to an
individuals who exhibit a predetermined percentage of a particular
immune cell population that binds exogenously provided
.beta.-glucan. The immune cell population used to determine whether
an individual is a "high binder" or a "low binder" can be, for
example, polymorphonuclear lymphocytes (PMNs) or monocytes. An
individual can be considered a "high binder" if at least 10% of the
PMNs or monocytes in a blood sample from the individual bind
exogenously provided .beta.-glucan. Thus, an individual may be a
"high binder" if at least 10%, at least 12%, at least 15%, at least
20%, at least 15%, or at least 40% of PMNs or monocytes in a blood
sample from the individual bind exogenously provided .beta.-glucan.
(See, e.g., FIG. 2 and FIG. 3). In some cases, the exogenously
provided .beta.-glucan can include PGG provided to final
concentration of 10 .mu.g/mL to 100 .mu.g/mL. Status as a "low
binder" refers to an individual who fails to exhibit "high binder"
status.
[0046] Moreover, "high binders" can exhibit higher titers of
anti-.beta.-glucan antibodies than "low binders." (FIG. 4). A
typical anti-.beta.-glucan antibody titer for a "high binder" can
be a titer of at least 25,000 such as, for example, at least
30,000, at least 35,000, at least 40,000, at least 45,000, at least
50,000, at least 55,000, or at least 60,000. (See, e.g., FIG. 4).
Anti-.beta.-glucan antibody titers typically refers to IgG. In some
cases, however, the presence of IgM can compensate for a lower IgG
titer to help establish "high binder" status.
[0047] .beta.-glucans are known to bind a lectin-like domain within
the COOH-terminal region of the CD11b subunit of leukocyte
complement receptor 3 (CR3; CD11b/CD18, aMh2 integrin, Mac-1; refs.
Thorton et al., J Immunol 156:1235-46, Xia et al., J Immunol
162:2281-90). .beta.-glucans can prime CR3 of neutrophils,
macrophages, and natural killer cells for cytotoxicity against
tumors opsonized with iC3b. Dual occupancy of leukocyte CR3 by the
I-domain ligand iC3b and the lectin-like domain ligand
.beta.-glucan can lead to degranulation and cytotoxic responses (Li
et al., J Immunol 177:1661-9; Tsikitis et al., J Immunol
173:1284-91). Thus, one might suspect that "low binder" individuals
might possess higher natural titers of anti-.beta.-glucan
antibodies that can disrupt binding between .beta.-glucan and
CR3.
[0048] We found, however, exactly the opposite. "High binders"
exhibited higher titers of anti-.beta.-glucan antibodies than "low
binders." (FIG. 4). Thus, higher titers of anti-.beta.-glucan
antibodies are associated with enhanced .beta.-glucan binding to
CR3 on immune cells.
[0049] Moreover, the effect is transferable. "High binder" serum
can increase .beta.-glucan binding to immune cells (e.g., PMNs) of
a "low binder." (FIG. 5). Increasing amounts of anti-.beta.-glucan
monoclonal antibody also can increase .beta.-glucan binding to
immune cells (e.g., PMNs) in serum from a "low binder." (FIG. 6).
Also, intravenous immunoglobulin, a blood product that contains
pooled, polyvalent IgG from many donors (typically many hundreds,
even thousands, of donors) and high natural anti-.beta.-glucan
titers, also can increase .beta.-glucan binding to immune cells
(e.g., PMNs) in serum from a "low binder." (FIG. 7).
[0050] The effect also is demonstrable in vivo. A subject with
recurrent metastatic colorectal cancer exhibited as a "low binder"
over five cycles of therapy that included administration of
.beta.-glucan. The subject exhibited .beta.-glucan binding to
<5% of PMNs and monocytes and an anti-.beta.-glucan antibody
titer in the bottom 10% of the distribution curve for healthy
individuals (1:1,600 to 1:3,200). The subject was treated multiple
times with intravenous immunoglobulin (IVIG) (0.4 g/kg-1 g/kg).
Pre- and post-treatment samples were obtained before and after the
second treatment. FIG. 8 shows that the subject exhibited a low
capacity to bind .beta.-glucan in PMNs and monocytes in the
pre-treatment samples (FIG. 8, Pre-infusion Cycle 7), but had a
significant increase in the capacity to bind .beta.-glucan in
post-IVIG treatment samples (FIG. 8, Post-infusion Cycle 7). In the
post-treatment sample, the subject's titer of anti-.beta.-glucan
antibodies also increased to 1:25,600, demonstrating the transfer
of anti-.beta.-glucan antibodies with the IVIG treatment.
[0051] In addition, in a two-armed, open-label, randomized,
multi-center study, 795 subjects with recurrent/progressive
colorectal cancer after at least two previous chemotherapeutic
treatments were divided into a control arm and an investigational
arm. Subjects in the control arm received treatment with cetuximab.
Subjects in the investigational arm received treatment with
cetuximab+4 mg/kg PGG .beta.-glucan. FIG. 9 shows that while
subjects receiving .beta.-glucan as part of their immunotherapy
remained on therapy for a longer average period than subjects
receiving only cetuximab, the effect was greatest in those subjects
that were "high binders." In this context, length of therapy is an
indication of therapy success so that a longer therapy time
indicates a positive therapeutic outcome while a shorter length of
therapy indicates poorer outcomes. Thus, there is a clinical
consequence to "high binder" status versus "low binder" status.
[0052] Thus, in another aspect, this disclosure describes
immunotherapy that includes administering to a subject
.beta.-glucan co-administered with antibody that specifically binds
.beta.-glucan and, in addition, an anti-tumor antibody. As used
herein, "anti-tumor" antibody refers to an antibody that
specifically binds neoplastic cells, regardless of whether the
neoplastic cells form a solid tumor or include leukemic or
lymphomic cells. The .beta.-glucan and antibody that specifically
binds the .beta.-glucan may be administered as described in detail
above. The anti-tumor antibody may be any suitable anti-tumor
antibody administered as directed by the manufacturer or health
professional. In this context, co-administering the .beta.-glucan
and the antibody preparation can increase the efficacy of the
immunotherapy. For example, PGG .beta.-glucan has demonstrated
preclinical activity against a variety of cancer types when
administered in combination with anti-tumor monoclonal antibodies
(mAbs). Exemplary types of cancer and their associated anti-tumor
mAbs include, for example, T-cell lymphoma (anti-MUC1, anti-GD2),
non-Hodgkin's lymphoma (rituximab), chroninc lymphocytic leukemia
(rituximab), lung carcinoma (anti-MUC1), breast adenocarcinoma
(anti-MMTV), ovarian carcinoma (bevacizumab), non-small-cell lung
carcinoma (bevacizumab, cetuximab), colorectal cancer (cetuximab),
and pancreatic carcinoma (cetuximab, anti-MUC1). For some subjects,
the immunostimulatory effect of PGG .beta.-glucan may be enhanced
by co-administering antibody that specifically binds the
.beta.-glucan.
[0053] A similar conversion of status from "low binder" to "high
binder" can occur by administering to the subject a composition
that includes a .beta.-glucan moiety conjugated to any antibody or
a portion of an antibody. FIG. 10 shows data illustrating
relatively low PGG binding by PMNs in whole blood (Imp Ref, second
panel) changing to high binding status by conjugating the PGG to
either BTH1704 (anti-MUC1, U.S. Pat. No. 6,204,366, Biothera, Inc.,
Eagan, Minn., third panel) or ERBITUX (Eli Lilly and Co.,
Indianapolis, Ind., fourth panel) anti-tumor antibodies. FIG. 11
also illustrates relatively low PGG binding by PMNs in whole blood
(Imp Ref, second panel) changing to high binding status by
conjugating the PGG to intravenous immunoglobulin (IVIG, Biolegend,
San Diego, Calif.).
[0054] Thus, in another aspect, this disclosure describes
immunotherapy that includes administering to a subject a
composition that includes a .beta.-glucan moiety conjugated to an
antibody, a therapeutic antibody, an anti-tumor antibody, or an
antibody fragment such as the Fc portion of an antibody. Modified
and/or derivatized PGG, including PGG conjugates of a PGG moiety
and an antibody are described in International Patent Application
No. PCT/US12/36795, which may also be applied to conjugates of
antibody fragments. The PGG moiety may be, or be derived from a
.beta.-1,3/1,6 glucan. In this context, "derived from" acknowledges
that a conjugate may necessarily be prepared by creating a covalent
linkage that replaces one or more atoms of the PGG .beta.-glucan.
As used herein, "derived from a .beta.-1,3/1,6 glucan" refers to a
portion of the PGG .beta.-glucan that remains as part of a
conjugate after replacing one or more atoms of the PGG to form the
covalent linkage of the conjugate.
[0055] The therapeutic antibody may be any therapeutic antibody
capable of being combined with .beta.-glucan for immunotherapy.
Thus, the therapeutic antibody also can include any of the
anti-tumor antibodies-described above in connection with other
aspects of this disclosure--in order to provide immunotherapy
against various forms of cancer.
[0056] As used herein, the term "and/or" means one or all of the
listed elements or a combination of any two or more of the listed
elements; the terms "comprises" and variations thereof do not have
a limiting meaning where these terms appear in the description and
claims; unless otherwise specified, "a," "an," "the," and "at least
one" are used interchangeably and mean one or more than one; and
the recitations of numerical ranges by endpoints include all
numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5,
2, 2.75, 3, 3.80, 4, 5, etc.).
[0057] In the preceding description, particular embodiments may be
described in isolation for clarity. Unless otherwise expressly
specified that the features of a particular embodiment are
incompatible with the features of another embodiment, certain
embodiments can include a combination of compatible features
described herein in connection with one or more embodiments.
[0058] For any method disclosed herein that includes discrete
steps, the steps may be conducted in any feasible order. And, as
appropriate, any combination of two or more steps may be conducted
simultaneously.
[0059] The present invention is illustrated by the following
examples. It is to be understood that the particular examples,
materials, amounts, and procedures are to be interpreted broadly in
accordance with the scope and spirit of the invention as set forth
herein.
EXAMPLES
Example 1
Materials
[0060] Imprime PGG (Biothera, Inc., Eagan, Minn.) was provided as a
preservative-free, soluble .beta.-glucan formulation prepared at a
concentration of 1 mg/mL in 0.8% sodium chloride and 0.2% sodium
citrate monobasic, at a pH of 6.4. The compound was stored at
4-8.degree. C. until use.
Preparation of Samples
[0061] Whole Blood. Fresh whole blood (WB) was obtained from
healthy volunteers that had provided informed consent prior to
donation (New England Institutional Review Board, May 2007). The
blood was collected in a Vacutainer.RTM. containing 158 USP Units
Freeze-Dried Sodium Heparin (BD Biosciences; San Jose, Calif.).
[0062] Serum and Plasma.
[0063] Whole blood was processed into serum or plasma by
Vacutainer.RTM. tubes (BD Biosciences; San Jose, Calif.) collection
with either serum separator (red top) or sodium heparin (green top)
tubes. Tubes were mixed well, incubated at room temperature for 30
minutes, and then centrifuged at 2000 rpm (.about.1150.times.g) for
10 minutes. The supernatant (either serum or plasma) was then
transferred to a fresh polycarbonate storage conical tube.
Anti-BG ELISA Method
[0064] A preliminary ELISA method modified from the monkey
anti-.beta.-glucan method (Noss et al., 2012 Int. Arch. Allergy
Immunol., 157:98-108) was used to test the human sera samples.
Costar universal binding plates were coated with 50 .mu.L of
.beta.-glucan at 1 .mu.g/mL purified .beta.-glucan diluted in
purified water and incubated at 37.degree. C. for 30 minutes. The
coated plate was then exposed to high intensity ultraviolet light
at >1500 .mu.W/cm.sup.2 for five minutes at room temperature and
placed in a 50.degree. C. forced air oven until dry before a second
exposure to ultraviolet light at >1500 .mu.W/cm.sup.2 for five
minutes at room temperature. The plate was then blocked with a 0.5%
solution of Bovine Serum Albumin for >30 minutes before washing
with wash buffer (phosphate buffered saline [PBS] with 0.05%
Tween-20). Human serum samples were diluted into wash buffer added
to the plate and subsequently serially diluted in wash buffer on
the plate. Test samples diluted 1:400 were pipetted onto the test
plate with seven additional serial 1:2 dilutions (serum dilutions
between 1:400 and 1:12,800). Samples were incubated at room
temperature for 30 minutes to permit human IgG to bind to the
plate-bound .beta.-glucan antigen. Following incubation the wells
were washed with wash buffer and an enzyme labeled secondary
antibody (horseradish peroxidase conjugated affinity purified goat
anti human IgG, Fc gamma specific) was incubated in the wells to
bind with the human IgG bound to the .beta.-glucan antigen. The
secondary antibody was allowed to incubate for 30 minutes before
washing with wash buffer. After the entire wash buffer was removed
from the wells a peroxidase substrate was incubated in the wells
and color development was quenched with .about.1 M phosphoric acid
at five minutes color development. The optical density (OD) at 450
nm was measured using a microtiter plate reader.
Determining Anti-.beta.-Glucan Ab Titer
[0065] Resulting OD from replicate wells were averaged and the mean
assay background subtracted. The greatest dilution giving a
background adjusted OD greater than or equal to 0.100 was
considered the samples titer and was expressed as the inverse of
that dilution. For definition of assay performance a value was
assigned the standard reference serum and a reference curve was
constructed on each assay plate. For example a test sample giving a
background adjusted OD of 0.100 at a dilution of 1:12,800 was
considered to have a titer of 12,800. Where samples were tested
multiple times and the average of their titers fell between the
serial 1:2 titer levels from 1:400 the next lowest titer level was
reported as its titer. For example, one donor's serum from four
donations was tested in five separate assays resulting in a mean
titer of 28,160; its titer was reported to be 25,600.
[0066] Assay Standard Curve.
[0067] A value of 160 Arbitrary Units per mL (AU/mL) was assigned
to the standard human anti-.beta.-glucan antibody. Thus a 1:400
dilution in the assay method results in a value of 400 mAU/mL as
the highest point of a standard dilution curve additional serial
1:2 dilutions were prepared on the assay plate. Assay controls were
diluted 1:100 in ELISA wash buffer for testing. Furthermore two
dilutions of each control level were independently prepared for
testing on each plate in parallel.
[0068] Statistical Analysis.
[0069] Plotting standard concentration in mAU/mL versus mean
background corrected optical density resulted in a standard
reference curve. Using the ELISA software a 4-parameter fit was
computed from the standard dose response curve to determine unknown
values for samples, controls and test serum. Assay response values
falling between the upper and lower inflection points of the
standard curve (linear portion) were used to determine a samples
test value. To compute the coefficient of variation (% CV); the
standard deviation of a set of values was divided by the mean of
the same set of values and the result multiplied by 100.
Binding of PGG to Cells of Whole Blood (WB)
[0070] One hundred microliters of WB from healthy donors was
aliquoted into 5 mL polystyrene fluorescence activated cell sorter
(FACS) tubes. These WB samples were stimulated with either Imprime
PGG (10 .mu.g/mL or 100 .mu.g/mL) or citrate buffer, the vehicle
control. The FACS tubes containing the samples were loosely covered
with the corresponding caps and incubated for 30 minutes or two
hours, at 37.degree. C. in a humidified incubator (5%
CO.sub.2).
TABLE-US-00001 TABLE 1 Antibody Cocktail Used To Stain Whole Blood
Samples Dilution or Final For identifi- Antibody Company; Clone #
Concentration cation of: Anti-CD15 Biolegend; W6D3 0.2 .mu.g/mL
neutrophils Anti-CD19 Biolegend; HIB19 0.63 .mu.g/mL B cells
Anti-CD14 Biolegend; HCD14 5 .mu.g/mL monocytes Anti-CD14
Invitrogen; TuK4 1:50 monocytes Anti-CD3 Biolegend; HIT3a 0.25
.mu.g/mL T cells Anti-CD45 Biolegend; HI30 0.25 .mu.g/mL hematopoi-
etic cells excluding erythrocytes and platelets Goat F(ab')2 anti-
Jackson Immunolab 5 .mu.g/mL mouse anti-.beta. mouse IgM glucan
antibody Proceeding incubation with the anti-.beta.-glucan antibody
BfD IV, the cells were incubated with the antibody cocktail which
contains a secondary antibody for the recognition of BfD IV as well
as antibodies for the recognition of various cell surface
markers
[0071] After incubation, all samples were washed by adding 2 mL of
1.times. Dulbecco's phosphate buffered saline (DPBS) and
centrifuged at 1500-1700 rpm at 4.degree. C. for five minutes.
After two rounds of washes and aspirations, 5 .mu.L of the
anti-.beta.-glucan antibody BfD IV (.about.100 .mu.g/mL), was mixed
into each tube and incubated at room temperature for 30 minutes.
This primary antibody was washed off twice with 1.times.DPBS as
described above and a cocktail of antibodies containing the
secondary antibody as well as the specific cell surface markers
(Table 1) was added and incubated for 30 minutes at room
temperature in the dark. To lyse the red blood cells, 2 mL of
1.times.BD lysing solution (BD Biosciences; San Jose, Calif.) was
added to each sample and gently vortexed. After an incubation
period of one hour at room temperature, the samples were
centrifuged at 1500-1700 rpm at 4.degree. C. for five minutes. The
BD lysing solution was aspirated and the cells were washed once
with 1.times.DPBS and aspirated as described above. For fixation,
300-400 .mu.L of 1% paraformaldehyde was added to each sample. The
samples were acquired on the LSR II (BD Biosciences; San Jose,
Calif.) within 20 hours of fixation. Data was analyzed using FlowJo
software (Tree Star, Ashland, Oreg.).
Example 2
Materials
[0072] Imprime PGG (Biothera, Inc., Eagan, Minn.) was provided in a
preservative-free, soluble .beta.-glucan formulation prepared at a
concentration of 1 mg/mL in 0.8% sodium chloride and 0.2% sodium
citrate monobasic, at a pH of 6.4. The compound was stored at
4-8.degree. C. until use.
Whole Blood (WB) Binding Assay
[0073] Fresh WB was obtained from healthy volunteers that had
provided informed consent prior to donation (New England
Institutional Review Board. Blood Donation Protocol No. 07-124).
The blood was collected in a Vacutainer.RTM. containing 158 USP
Units Freeze-Dried Sodium Heparin (BD Biosciences; San Jose,
Calif.). Serum was collected in a Vacutainer.RTM. containing a
thrombin-based clot activator (BD Biosciences; San Jose, Calif.).
Approximately 20 minutes after collection, the vial was centrifuged
at 2000 rpm for 10 minutes at room temperature. Serum was harvested
from this vial and stored at 4.degree. C. for use within 8 hours or
at -80.degree. C. for use after 8 hours.
[0074] The whole blood binding assay was performed by incubating
whole blood samples with Imprime PGG for 30 minutes or two hours at
37.degree. C. in a humidified incubator. After washing with
1.times. Dulbecco's phosphate buffered saline (DPBS), BfDIV, a
mouse anti-.beta.-glucan antibody was added and incubated with the
WB for 30 minutes at room temperature. After more rounds of
washing, an antibody cocktail including a goat anti-mouse detection
antibody and antibodies to surface molecules were added and
incubated at room temperature in the dark for 30 minutes.
Erythrocytes were lysed with BD Lyse and samples were resuspended
in 1% paraformaldehyde. Samples were acquired on a flow cytometer
and analyzed using FlowJo software (Ashland, Oreg.).
WB and Serum Crossover Studies
[0075] For serum crossover studies, whole blood was spun down at
1200 rpm for 10 minutes and plasma removed. Blood cells were washed
1-2 times with 1.times.DPBS to remove remaining plasma. 50 .mu.L of
serum was added and mixed before addition of Imprime.
[0076] For incubation with anti-.beta.-glucan IgG (BioSupplies,
Australia), the lyophilized antibody was resuspended to 1 mg/mL
with 1.times.DPBS and stored at -80.degree. C. or 4.degree. C. as a
stock solution. Before being added to blood samples, the stock was
diluted 1:10 to 100 .mu.g/mL and 10 .mu.L of this solution was
added to 100 .mu.L of blood. For incubation with IVIG, 10% IVIG
(100 mg/mL) (PRIVIGEN, CSL Behrling, King of Prussia, Pa.) was
added to the whole blood sample at the indicated final
concentrations.
Example 3
[0077] Fresh whole blood was obtained from healthy volunteers that
had provided informed consent prior to donation. The blood was
collected in a Vacutainer.RTM. containing 158 USP Units
Freeze-Dried Sodium Heparin (BD Biosciences; San Jose, Calif.). 100
.mu.L of whole blood from the healthy donors was aliquoted into 5
mL polystyrene FACS tubes. The samples were stimulated with either
the vehicle control, or PGG reference standard, or PGG-Muc1
conjugate (Imp-BTH1704), PGG-Erbitux conjugate (10 .mu.g/mL), or
PGG-IVIG conjugate. PGG-anti-tumor antibody conjugates were
prepared as described in International Patent Application No.
PCT/US12/36795.
[0078] Tubes containing samples were loosely covered with parafilm
and incubated for 30 minutes at 37.degree. C. in a humidified
incubator (5% CO.sub.2). After incubation, all samples were washed
two times with 2 mL of 1.times.DPBS and centrifuged at 1500-1700
rpm at 4.degree. C. for five minutes. After aspiration, 5 .mu.L of
the anti-.beta.-glucan antibody BfD IV (Biothera, Inc., Eagan,
Minn.; U.S. Pat. No. 6,294,321), was mixed into each tube and
incubated at room temperature for 30 minutes. This antibody was
washed twice and a cocktail of antibodies containing the secondary
Ab FITC-conjugated goat anti-mouse IgM (Southern Biotech;
Birmingham, Ala.) as well as the specific cell surface markers,
CD15, CD14, CD19, CD3 and CD45 (Biolegend, San Diego, Calif.) were
added and incubated for 30 minutes at room temperature in the dark.
To lyse the red blood cells, 2 mL of 1.times.BD Lysing solution (BD
Bioscience; San Jose, Calif.) was added to each sample and
vortexed. After incubating at room temperature for 30 minutes the
samples were centrifuged as described above and the pellet was
washed with 2 mL 1.times.DPBS. Cells were fixed with 300 .mu.L of
1% paraformaldehyde and acquired on the LSR II (BD Biosciences, San
Jose, Calif.). Data was analyzed with FlowJo software (Tree Star,
Ashland, Oreg.). The cells were assessed for their capacity to bind
PGG by comparing the median fluorescence intensity (MFI) of the
cells stained with BfD IV and the percentage of cells positive for
BfD IV relative to that of the vehicle treated control group.
Exemplary Embodiments
Embodiment 1
[0079] A composition comprising:
[0080] a soluble .beta.-glucan component; and
[0081] an antibody component that specifically binds to the soluble
.beta.-glucan.
Embodiment 2
[0082] The composition of Embodiment 1 wherein the soluble
.beta.-glucan is derived from yeast.
Embodiment 3
[0083] The composition of Embodiment 1 or Embodiment 2 wherein the
soluble .beta.-glucan comprises a .beta.-1,3/1,6 glucan.
Embodiment 4
[0084] The composition of any preceding Embodiment wherein the
soluble .beta.-glucan comprises
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
Embodiment 5
[0085] The composition of any preceding Embodiment wherein the
antibody component comprises a monoclonal antibody that
specifically binds to the soluble .beta.-glucan.
Embodiment 6
[0086] The composition of Embodiment 5 wherein the monoclonal
antibody comprises BfD I, BfD II, BfD III, or BfD IV.
Embodiment 7
[0087] The composition of any preceding Embodiment wherein the
soluble .beta.-glucan component and the antibody component are
provided in a single formulation.
Embodiment 8
[0088] The composition of any preceding Embodiment wherein the
soluble .beta.-glucan component and the antibody component are
provided in separate formulations.
Embodiment 9
[0089] The composition of any preceding Embodiment and further
comprising an anti-tumor antibody.
Embodiment 10
[0090] A method comprising co-administering to a subject a soluble
.beta.-glucan and an antibody preparation or antibody component
that specifically binds to the soluble .beta.-glucan.
Embodiment 11
[0091] A method of increasing a subject's response to soluble
.beta.-glucan immunotherapy, the method comprising co-administering
to the subject a composition that comprises a soluble .beta.-glucan
and an antibody preparation that specifically binds to the soluble
.beta.-glucan.
Embodiment 12
[0092] The method of Embodiment 11 further comprising identifying
the subject as a low binder of .beta.-glucan.
Embodiment 13
[0093] The method of any one of Embodiments 10-12 wherein the
soluble .beta.-glucan and the antibody preparation are
co-administered simultaneously.
Embodiment 14
[0094] The method of any one of Embodiments 10-12 wherein the
soluble .beta.-glucan and the antibody preparation are
co-administered at different times.
Embodiment 15
[0095] The method of any one of Embodiments 10-12 wherein the
soluble .beta.-glucan and the antibody preparation are
co-administered at different sites.
Embodiment 16
[0096] The method of any one of Embodiments 10-15 wherein the
soluble .beta.-glucan is derived from yeast.
Embodiment 17
[0097] The method of any one of Embodiments 10-16 wherein the
soluble .beta.-glucan comprises a .beta.-1,3/1,6 glucan.
Embodiment 18
[0098] The method of any one of Embodiments 10-17 wherein the
soluble .beta.-glucan comprises
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
Embodiment 19
[0099] The method of any one of Embodiments 10-18 wherein the
antibody component comprises a monoclonal antibody that
specifically binds to the .beta.-glucan.
Embodiment 20
[0100] The method of Embodiment 19 wherein the monoclonal antibody
comprises BfD I, BfD II, BfD III, or BfD IV.
Embodiment 21
[0101] A method of increasing a subject's response to soluble
.beta.-glucan immunotherapy, the method comprising administering to
the subject a composition that comprises a soluble .beta.-glucan
moiety conjugated to an antibody or antibody fragment.
Embodiment 22
[0102] The method of Embodiment 21 and further comprising
identifying the subject as a low binder of soluble
.beta.-glucan.
Embodiment 23
[0103] The method of Embodiment 21 or Embodiment 22 wherein the
soluble .beta.-glucan moiety is derived from yeast.
Embodiment 24
[0104] The method of Embodiments 21-23 wherein the soluble
.beta.-glucan moiety comprises, or is derived from, a
.beta.-1,3/1,6 glucan.
Embodiment 25
[0105] The method of any one of Embodiments 21-24 wherein the
.beta.-glucan moiety comprises, or is derived from,
.beta.(1,6)-[poly-(1,3)-D-glucopyranosyl]-poly-.beta.(1,3)-D-glucopyranos-
e.
Embodiment 26
[0106] The method of any one of Embodiments 21-25 wherein the
antibody comprises a therapeutic antibody.
Embodiment 27
[0107] The method of any one of Embodiments 21-26 and further
comprising administering an anti-tumor antibody.
Embodiment 28
[0108] The method of Embodiment 27 wherein the anti-tumor antibody
specifically binds to leukemic or lymphomic cells.
Embodiment 29
[0109] The method of Embodiment 27 wherein the anti-tumor antibody
binds to cells of solid tumor.
Embodiment 30
[0110] A method of treating a tumor comprising co-administering to
a subject having a tumor a composition that comprises:
[0111] a soluble .beta.-glucan;
[0112] an antibody preparation that specifically binds to the
soluble .beta.-glucan; and an anti-tumor antibody preparation.
Embodiment 31
[0113] The method of Embodiment 30 and further comprising
identifying the subject as a low binder of soluble
.beta.-glucan.
[0114] The complete disclosure of all patents, patent applications,
and publications, and electronically available material (including,
for instance, nucleotide sequence submissions in, e.g., GenBank and
RefSeq, and amino acid sequence submissions in, e.g., SwissProt,
PIR, PRF, PDB, and translations from annotated coding regions in
GenBank and RefSeq) cited herein are incorporated by reference in
their entirety. In the event that any inconsistency exists between
the disclosure of the present application and the disclosure(s) of
any document incorporated herein by reference, the disclosure of
the present application shall govern. The foregoing detailed
description and examples have been given for clarity of
understanding only. No unnecessary limitations are to be understood
therefrom. The invention is not limited to the exact details shown
and described, for variations obvious to one skilled in the art
will be included within the invention defined by the claims.
[0115] Unless otherwise indicated, all numbers expressing
quantities of components, molecular weights, and so forth used in
the specification and claims are to be understood as being modified
in all instances by the term "about." Accordingly, unless otherwise
indicated to the contrary, the numerical parameters set forth in
the specification and claims are approximations that may vary
depending upon the desired properties sought to be obtained by the
present invention. At the very least, and not as an attempt to
limit the doctrine of equivalents to the scope of the claims, each
numerical parameter should at least be construed in light of the
number of reported significant digits and by applying ordinary
rounding techniques.
[0116] Notwithstanding that the numerical ranges and parameters
setting forth the broad scope of the invention are approximations,
the numerical values set forth in the specific examples are
reported as precisely as possible. All numerical values, however,
inherently contain a range necessarily resulting from the standard
deviation found in their respective testing measurements.
[0117] All headings are for the convenience of the reader and
should not be used to limit the meaning of the text that follows
the heading, unless so specified.
* * * * *