Methods of treating inflammation in neuronal tissue
United States Patent: 8,105,590
Issued: January 31, 2012
Inventors: Yao; Yue
(Issaquah, WA), Bilsborough; Janine (Seattle, WA)
Inc. (Seattle, WA)
Appl. No.: 13/075,412
Filed: March 30, 2011
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Use of antagonists to IL-31 are used to
treat inflammation and pain by inhibiting, preventing, reducing,
minimizing, limiting or minimizing stimulation in neuronal tissues. Such
antagonists include antibodies and fragments, derivative, or variants
thereof. Symptoms such as pain, tingle, sensitization, tickle associated
with neuropathies are ameliorated.
Description of the
BACKGROUND OF THE INVENTION
The inflammatory process activates the nervous system causing inflammatory
pain and a disruption in motor function. Stimulation of sensory nerves
produces vasodialtion and plasma extravasation, leading to neurogenic
inflammation and stimulation causing sensory irritation, hypersensitivity
Neurogenic inflammation is caused by activation of nociceptive and
thermal-sensitive endings in tissues and can be caused by innate
conditions, such as autoimmune diseases, including allergy, by viral
infection, as well as by injury. The neurogenic inflammation from these
conditions can affect the somatosensory system, which consists of various
sensory receptors responsible for sensations such as pressure, touch,
temperature, pain, itch, tickle, tingle, and numbness. Activated nerves
can perpetuate chronic inflammation by inducing secretion of cytokines,
activating monocytes and chemotaxis.
Proteins active in neurogenic inflammation can serve as targets for
therapeutic approaches to diagnosis and treatment of diseases.
An example of a drug used to treat pain is Neurontin (gabapentin), which
is used to treat diabetic peripheral neuropathy as post-herpatic
neuralgia. Thus, there is a need for additional medication to treat
DESCRIPTION OF THE INVENTION
The present invention is based in part upon the discovery that the
subunits of the heterodimeric receptor which binds IL-31, e.g. IL-31Ra and
OSMRb, are expressed on neural cells such as dorsal root ganglion cells.
Thus the present invention encompasses the use of antagonists to IL-31 in
inhibiting pain and inflammation and the symptoms of inflammatory bowel
disease, Crohn's disease, pruritis, and neurogenic pain and sensitization.
The present invention also encompasses the use of IL-31 agonists in
improving sensitization through stimulation of the dorsal root gangion
IL-31 is the HUGO name for a cytokine that has been previously described
as Zcyto17rlig in a published U.S. patent application (See published U.S.
patent application number 20030224487, U.S. patent application Ser. No.
10/352,554, filed Jan. 21, 2003, now issued U.S. Pat. No. 7,064,186;
Sprecher, Cindy et al., 2003, incorporated herein by reference). The
heterodimeric receptor for IL-31, comprises a heterodimer formed between
IL-31Ra and OncostatinM receptor beta (OSMRb). IL-31Ra is the HUGO name
for a protein called zcytor17 in commonly-owned U.S. published patent
application number 20030215838, U.S. patent application Ser. No.
10/351,157, filed Jan. 21, 2003, herein incorporated by reference. The
polynucleotide and polypeptide sequences for human IL-31 are shown in SEQ
ID NOs: 1 and 2, respectively. The polynucleotide and polypeptide
sequences for murine IL-31 are shown in SEQ ID NOs: 3 and 4, respectively.
As used herein the term, IL-31 shall mean zcytor17lig as used in U.S.
patent publication number 20030224487, as shown above. IL-31Ra has been
previously described in commonly-owned U.S. patent application Ser. No.
09/892,949 filed Jun. 26, 2001, which is herein incorporated by reference.
The amino acid sequence for the OSMR, and IL-31RA receptors indicated that
the encoded receptors belonged to the Class 1 cytokine receptor subfamily
that includes, but is not limited to, the receptors for IL-2, IL-4, IL-7,
Lif, IL-12, IL-15, EPO, TPO, GM-CSF and G-CSF (for a review see, Cosman,
"The Hematopoietin Receptor Superfamily" in Cytokine 5(2): 95-106, 1993).
The zcytor17 receptor is fully described in commonly-owned PCT Patent
Application No. US01/20484 (WIPO publication No. WO 02/00721; herein
incorporated by reference).
The present invention includes the use of anti-IL-31, including
antagonists, antibodies, binding proteins, variants and fragments, having
anti-IL-31 activity. The invention includes administering to a subject the
anti-IL-31 molecule and contemplates both human and veterinary therapeutic
uses. Illustrative veterinary subjects include mammalian subjects, such as
farm animals and domestic animals.
The native polynucleotide and polypeptide sequences for the "long" form of
IL-31RA are shown in SEQ ID NOs:5 and 6, respectively. The native
polynucleotide and polypeptide sequences for the "short" form of IL-31RA
are shown in SEQ ID NOs:7 and 8, respectively. Additional truncated forms
of IL-31RA polypeptide appear to be naturally expressed. Both forms encode
soluble IL-31RA receptors. The "long" soluble IL-31RA polynucleotide and
polypeptide sequences are shown in SEQ ID NOs:9 and 10, respectively. The
"short" soluble IL-31RA polynucleotide and polypeptide sequences are shown
in SEQ ID NOs:11 and 12, respectively. The native polynucleotide and
polypeptide sequences for mouse IL-31RA are shown in SEQ ID NOs:13 and 14,
respectively. The native polynucleotide and polypeptide sequences for
human OSMRbeta are shown in SEQ ID NOs:15 and 16, respectively. See PCT
applications WO 02/00721 and WO 04/003140, both of which are incorporated
IL-31 antagonists include anti-IL31 molecules such as antibodies that bind
IL-31, including, variants, fragments or derivatives thereof and that
inhibit, limit, reduce, minimize, prevent, or neutralize the effect of
IL-31 has on binding its cognate receptor.
In situ expression analysis revealed that IL-31RA and OSMRbeta are
expressed in the spinal cord and dorsal root ganglion cells in humans. See
Example 1. Therefore, IL-31 molecules, their agonists, or antagonists play
a role in the maintenance of neurons and neurogenic inflammation and
stimulation. This indicates that IL-31 agonists, antagonists can be used
to treat a variety of neurodegenerative diseases such as amyotrophic
lateral sclerosis (ALS), Alzheimer's disease, Huntington's disease,
Parkinson's disease, peripheral neuropathies, and demyelinating diseases
including multiple sclerosis. The tissue specificity of IL-31RA and OSMRb
suggests that IL-31 may be a growth and/or maintenance factor in the
spinal cord and brain which can be used to treat spinal cord, brain or
peripheral nervous system injuries.
Methods of measuring the ability of IL-31 to stimulate pain are known to
one of skill in the art. For example, dorsal root gangion cells can be
isolated and cultured. See Voilley, N. et al., J. Neurosci.,
27(20):8026-8033, 2001. For example, dorsal root ganglion cells are
prepared from Wistar adult male (5-7 weeks) and newborn rats by 0.1%
collagenase dissociation and plating on collagen coated P35 dishes in DMEM
plus 5% fetal calf serum. Similarly methods of isolating dorsal root
ganglion cells are described by Steinhoff, M. et al. (See Steinhoff, M. et
al., Nature Medicine, 6(2):151-157, 2000). Briefly, dorsal root ganglion
cells are minced in cold Dulbeccos' modified Eagle's Medium (DMEM) and
incubated in DMEM containing 0.05 mg/ml trypsin, 1 mg/ml collagenase, and
0.01 mg/ml DNAse I for 45-60 minutes at 37 degrees C. SBTI is added to
neutralize trypsin and the suspension is centrifuged at about 1,000 g for
1 min. Neurons in the pellet are suspended in DMEM containing 10% fetal
bovine serum, 5 ng/ml nerve growth factor, 2 mM glutamine, 1 mg/ml
penicillin/streptomycin and DNAse I, and plated on glass coverslips coated
with Matrigel. Neurons are cultured for 3-5 days before use. Expression of
IL-31Ra at the plasma membranes is verified by immunofluorescence using an
To measure the effect of IL-31 on dorsal root ganglion stimulation,
intracellular calcium ion concentration is measured in the cultured
neurons as described by Steinhoff et al., supra. The neurons are incubated
in Hank's balanced salt solution, 20 mM HEPES, pH 7.4 containing 5 uM
Fura-2/AM (Molecular Probes, Eugene, Oreg.) for 1 h at 37 degrees C.
Coverslips are washed, mounted in a chamber (1 ml volume) on a Zeiss 100
TV inverted microscope and observed using a Zeiss x40 Fluar objective.
Fluorescence is measured at 340 nm and 380 nm to allow determination of
calcium. Cells are exposed to IL-31 with and without other sensitization
agents, and inhibition in the presence of IL-31 antagonists is measured.
To measure the ability of an IL-31 antagonist on effect of IL-31 binding
to its cognate heterodimeric receptor on dorsal root gangion, or neural
cells in general, on pain several mediators of pain can be measured, such
as for example, but not limited to, prostaglandins, substance P, CGRP,
galanin, Neuropeptide Y, histamine, bradykinin, cannabinoids, and
mediators of the arachinoid acid pathway.
In addition to the above in vitro methods to measure the ability of
antagonists to IL-31 pain-inducing effect of IL-31 on neural cells,
several in vivo models are also useful. See, for example, Honore, P. et
al., Neuroscience, 98(3):585-598, 2000. This article describes several
models for inflammatory pain, neuropathic pain and cancer pain. For
example, one model measures the effect of an antagonist to IL-31, such as
a subcutaneous injection of IL-31, with and with out the antagonist
molecule, into the plantar surface of the hindpaw of a mouse. The mouse is
euthanized 3 days after injection peripheral edema is measured. The effect
of the IL-31 antagonist molecule to inhibit, limit, minimize, reduce,
prevent, or neutralize the edema is measured. Additional in vivo models
are spinal nerve ligation, sciatic nerve transaction, sarcoma-induced bone
cancer, behavioral analysis, and effects of morphine.
Another mouse model of pain is mechanical allodynia. See for example,
Sweitzer, S. M. et al., J. Neuroimm., 125:82-93, 2002. Briefly, rats or
mice are tested for mechanical allodynia with 2- and/or 12-g von Frey
filaments. First the animals are acclimated to the procedure and baseline
measurement are taken. The IL-31 is administered in varying amounts.
Allodynia is characterized as an intense withdrawal of the paw to a
normally non-noxious stimuli in response to IL-31 administration.
Comparison is made with and without administration of the IL-31
A proinflammatory neuropeptide, Substance P (SP), is made the dorsal
ganglia and then transported to the periphery by nociceptive nerves A and
C (15). SP can induce itch by releasing histamine from the mast cell
granules. In the skin, SP can also cause erythema, edema and neurogenic
inflammation releasing histamine, IL-1, prostaglandins and lysosomal
enzymes but is quickly degraded in the dermis (16). The prior oral
administration of antihistamines inhibits the pruritus caused by SP.
Capsaicin obtained from hot pepper applied locally depletes SP from
cutaneous nerves, and so diminishes pruritus. As the receptor subunits for
IL-31 are expressed in the dorsal root ganglion cells, administration of
the IL-31 antagonist molecules can decrease the stimulation of these cells
and may decrease Substance P that may be induced by IL-31 administration.
The binding of IL-31 to its receptor, i.e., IL-31RA and OSMR beta, on
dorsal root ganglion cells can stimulate the somatosensory system, which
consists of various sensory receptors responsible for sensations such as
pressure, touch, temperature, pain, itch, tickle, tingle, and numbness.
The binding of IL-31 to its cognate receptor can result in neurogenic
inflammation and stimulation, which may lead to release of additional
factors that induce neurogenic stimulus. One group of factors that mediate
pain is the prostaglandins, which also contribute to local inflammation.
Thus, an IL-31 antagonist may have benefit in acute inflammatory pain
commonly treated with NSAIDs, such as myalgia, headache, joint pains from
acute injuries or chronic pain such as that caused by osteoarthritis. Such
neurogenic stimulus can be the result of inflammation caused by, for
example, autoimmune reactions, such as allergy, viral infection, such as
varicella, and injury, such as burn or trauma. Thus, antagonists that
interfere with signal transduction induced by the binding of the IL-31
ligand to its cognate receptor can be useful in reducing, limiting,
preventing, or minimizing neurogenic inflammation and the stimulation of
the somatosensory system. As such, antagonists of IL-31-induced signal
transduction in dorsal root ganglion cells can be used to treat pain,
itch, tingling, associated with autoimmune diseases, viral infection, and
trauma. Moreover, since neurogenic inflammation can result in a
hypersensitivity of the nerve after the initial insult, antagonists of
IL-31 can be effective treatment of symptoms. For example, some shingles
patients experience the sensory symptoms of pain and/or itch long after
the viral infection has been cleared or minimized. The neuralgia that
accompanies acute herpes zoster, and postherpetic neuralgia are likely due
to inflammation of the dorsal root ganglia and trigeminal ganglia, where
viral antigens attract T cells and other inflammatory cells. Long lasting
pain may result from persistent inflammation of the dermatome following a
robust antiviral response. Consequently, the level or stage of viral
infection may not be representative of the sensory perception of the
subject. Thus, the beneficial effect of antagonizing IL-31-induced signal
transduction may extend beyond the immediate state of viral infection or
Neuropathy and sensory deficiency involve pain and loss of sensitivity,
and can be related to such diseases as, atopy, diabetes, multiple
sclerosis, and hypertension, for example. As IL-31RA and OSBRbeta are
proteins that are expressed in the spinal cord and dorsal root ganglion
cells, antagonists of IL-31 may be useful to treat pain and sensory
deficiencies. For example, IL-31 antagonists can be delivered topically,
subcutaneously, centrally, or systemically, to treat diabetic
peripherineuropathy, postherpatic peripheral neuropathy, as well as pain,
in general, including pain as a symptom in burn patients.
Burn injuries cause intense and prolonged pain that is intensified when
the wound dressing is changed. Frequent dressing changes are necessary to
prevent infection and aid healing. The amount of pain experienced by
patients during wound care remains a worldwide problem for burn victims as
well as a number of other patient populations. When patients are at rest
pain associated with burn can be treated with opioids, which have some
unwanted effects. However, during wound care such as daily bandage
changes, wound cleaning, staple removals etc., opioids are not enough,
with a majority of burn patients reporting severe to excruciating pain
during wound care.
Since both members of the heterodimer for IL-31, i.e., IL-31RA and
OSMRbeta are expressed in dorsal root ganglion cells, an antagonist to
IL-31, such as a neutralizing antibody is useful to prevent, minimize,
limit and/or treat pain, including pain associated with burn or
neuropathy. In vivo models mimicking burn are well known to one skilled in
Persistent pain can provoke hyperplasia such that less than the original
stimulus can cause increased pain, also called allodynia. As both the
IL-31RA and OSMR beta subunits are expressed on dorsal root gangion cells,
an antagonist to IL-31 induced signal transduction in neuronal cells
bearing these subunits can help to mitigate symptoms of allodynia.
Polypeptides of the present invention, such as IL-31, as well as agonists,
fragments, variants and/or chimeras thereof, can also be used to increase
sensitization in mammals. For example, IL-31 polypeptides of the present
invention, including agonists, can be used to increase sensitization
(pain, heat, or mechanical) when delivered locally or topically,
systemically, or centrally and measured in any models or experiments known
to one skilled in the art and/or described herein. Also, the polypeptides
of the present invention can be administered to enhance the sensitivity of
spinal and neuronal cells in order to improve the function of the
surviving neurons to neurotransmitters and therefore might be effective in
Parkinson's or Alzheimers disease, as well as paralysis.
Similarly, where a patient has an increased sensitization to pain,
antagonists to IL-31 can be used to decrease the sensation of pain in a
patient with neuropathy. For example a patient with diabetic neuropathy
and postherpatic neuropathy, have chronic, enhanced pain, the antagonist
to IL-31 may be useful to limit, prevent or decrease the pain.
As a receptor for a protein that is proinflammatory, the presence of
IL-31RA and OSMRbeta in the spinal cord and dorsal root ganglion indicate
that antagonists of IL-31 can be used to reduce inflammation in these
tissues. Thus, conditions such as meningitis may benefit from
administration of the antagonists, including antibodies.
Diseases which involve neurogenic inflammation and stimulation and can
benefit from antagonizing IL-31 induced pain in neuronal tissues,
including dorsal root ganglion cells include: chronic pain, migraines,
arthritis, osteoarthritis, rheumatoid arthritis, polyneuropathy, diabetic
peripheralneuropathy, pain subsequent to nerve severence (eg.
post-surgical pain), inflammatory conditions that involve a neurogenic
pain-producing component, such as inflammatory bowel disease, nephritis,
certain metastic carcinomas, and inflammation of the blood vessels. These
diseases can also be treated by an antagonist of IL-31 induced signal
transduction. In addition, skin conditions, including radiation irritation
and burns, chemical burns, multiple chemical sensitivity, prickly heat,
rhinitis, thermal burns, sunburn, reddening of the skin and chemically
induced lesions, and acute allergic reactions such as acute asthma attack
and inflammation of the lung caused by chemical exposure, and hives as
well as conjunctivitis and gum disease can be treated with IL-31
antagonists. Additionally, scapuloperoneal syndromes are heterogeneous
neuromuscular disorders which are characterized by weakness in the
distribution of shoulder girdle and peroneal muscles. Both neurogenic (scapuloperoneal
spinal muscular atrophy, SPSMA) and myopathic (scapuloperoneal muscular
dystrophy, SPMD) scapuloperoneal syndromes have been described. The
chromosomal locus for SPMD has recently been assigned to chromosome 12q,
which is the same locus as for IL-31. Thus, IL-31 antagonists can be used
to treat these diseases.
In the United States approximately 500,000 people suffer from inflammatory
bowel disease, which can involve either or both the small and large bowel.
Ulcerative colitis and Crohn's disease are the best-known forms of
inflammatory bowel disease, and both are categorized as "idiopathic"
inflammatory bowel disease because the etiology for them is unknown.
Crohn's disease can involve any part of the gastrointestinal tract, but
most frequently involves the distal small bowel and colon. Inflammation
can produce anything from a small ulcer over a lymphoid follicle to a deep
fissuring ulcer to transmural scarring and chronic inflammation. Although
the etiology is unknown, infectious and immunologic mechanisms have been
proposed. Symptoms are variable and can include diarrhea, fever, and pain,
as well as extra-intestinal manifestations of arthritis, uveitis, erythema
nodosum, and ankylosing spondylitis.
The traditional approach to treating inflammatory bowel disease is
immunosuppression with azathioprine (see, for example, Rutgeerts, J.
Gastroenterol. Hepatol. 17(Suppl.):S176-85 (2002)). More recently, the
chimeric monoclonal anti-tumor necrosis factor antibody, infliximab, has
been used to target specific pathogenic disease mechanisms, and allows
thorough suppression of the disease process and healing of the bowel in
the long term. However, this therapy is associated with problems of
immunogenicity. The formation of antibodies to infliximab interferes with
efficacy and is associated with infusion reactions.
Irritable bowel syndrome (IBS) is a chronic functional gastrointestinal
disorder. It is a heterogeneous condition characterized by a variety of
bowel symptoms including abdominal pain and bloating which are usually
associated with altered bowel habit (Collins et al, 2001). It is estimated
that between 12 and 20% of the U.S. population suffer from this condition.
Differing criteria have been proposed for defining IBS, including the
Manning criteria (Manning et al, 1978), the Rome criteria (Thompson et al,
1992), and most recently Rome II (Thompson et al., 1999). Research reports
on IBS frequently classify patients with IBS into the two subtypes of
constipation predominant (CON) and diarrhea predominant (DIA) and
sometimes include a third subtype of alternating pattern (ALT).
Anti-IL-31 molecules, antagonists, antibodies, binding proteins, variants
and fragments, are useful in treating, detecting, and pain associated with
Inflammatory Bowel Disease (IBD) and Irritable Bowel Syndrome (IBS).
Inflammatory Bowel Disease (IBD) can affect the colon and/or rectum
(Ulcerative colitis), or the small and large intestine (Crohn's Disease).
The pathogenesis of these diseases is unclear, but they involve chronic
inflammation of the affected tissues. Potential therapeutics include
anti-IL-31 molecules, including, anti-IL-31 antibodies, other binding
proteins, variants, fragments, chimeras, and other IL-31 antagonists.
These molecules could serve as a valuable therapeutic to reduce
inflammation and pathological effects in IBD and related diseases.
Ulcerative colitis (UC) is an inflammatory disease of the large intestine,
commonly called the colon, characterized by inflammation and ulceration of
the mucosa or innermost lining of the colon. This inflammation causes the
colon to empty frequently, resulting in diarrhea. Symptoms include
loosening of the stool and associated abdominal cramping, fever and weight
loss. Although the exact cause of UC is unknown, recent research suggests
that the body's natural defenses are operating against proteins in the
body which the body thinks are foreign (an "autoimmune reaction"). Perhaps
because they resemble bacterial proteins in the gut, these proteins may
either instigate or stimulate the inflammatory process that begins to
destroy the lining of the colon. As the lining of the colon is destroyed,
ulcers form, releasing mucus, pus and blood. The disease usually begins in
the rectal area and may eventually extend through the entire large bowel.
Repeated episodes of inflammation lead to thickening of the wall of the
intestine and rectum with scar tissue. Death of colon tissue or sepsis may
occur with severe disease. The symptoms of ulcerative colitis vary in
severity and their onset may be gradual or sudden. Attacks may be provoked
by many factors, including respiratory infections or stress. Thus, the
anti-IL-31 molecules of the present invention can be useful to treat and
or detect UC.
Although there is currently no cure for UC available, treatments are
focused on suppressing the abnormal inflammatory process in the colon
lining. Treatments including corticosteroids immunosuppressives (eg.
azathioprine, mercaptopurine, and methotrexate) and aminosalicytates are
available to treat the disease. However, the long-term use of
immunosuppressives such as corticosteroids and azathioprine can result in
serious side effects including thinning of bones, cataracts, infection,
and liver and bone marrow effects. In the patients in whom current
therapies are not successful, surgery is an option. The surgery involves
the removal of the entire colon and the rectum.
There are several animal models that can partially mimic chronic
ulcerative colitis. The most widely used model is the
2,4,6-trinitrobenesulfonic acid/ethanol (TNBS) induced colitis model,
which induces chronic inflammation and ulceration in the colon. When TNBS
is introduced into the colon of susceptible mice via intra-rectal
instillation, it induces T-cell mediated immune response in the colonic
mucosa, in this case leading to a massive mucosal inflammation
characterized by the dense infiltration of T-cells and macrophages
throughout the entire wall of the large bowel. Moreover, this
histopathologic picture is accompanied by the clinical picture of
progressive weight loss (wasting), bloody diarrhea, rectal prolapse, and
large bowel wall thickening (Neurath et al. Intern. Rev. Immunol.
Another colitis model uses dextran sulfate sodium (DSS), which induces an
acute colitis manifested by bloody diarrhea, weight loss, shortening of
the colon and mucosal ulceration with neutrophil infiltration. DSS-induced
colitis is characterized histologically by infiltration of inflammatory
cells into the lamina propria, with lymphoid hyperplasia, focal crypt
damage, and epithelial ulceration. These changes are thought to develop
due to a toxic effect of DSS on the epithelium and by phagocytosis of
lamina propria cells and production of TNF-alpha and IFN-gamma. DSS is
regarded as a T cell-independent model because it is observed in T
cell-deficient animals such as SCID mice.
The administration of IL-31 antagonists or binding partners to these TNBS
or DSS models can be used to measure the amelioration of symptoms and
alter the course of gastrointestinal disease. IL-31 or may play a role in
the inflammatory response and pain associated with colitis, and the
neutralization of IL-31 activity by administrating antagonists is a
potential therapeutic approach for IBD.
Irritable Bowel Syndrome is one of the most common conditions in the
gastrointestinal clinic. Yet, diagnosis and treatment for IBS remain
limited. As the expression of IL-31 and IL-31RA1 have been correlated with
upregulation of Crohn's disease (See Example 5). IL-31 antagonists,
including anti-IL-31 antibodies, other binding proteins, variants,
fragments, chimeras, and other IL-31 antagonists are useful in reducing
symptoms and treatment of the disease.
The administration of IL-31 antagonists or binding partners to a patient
with IBD or IBS can be used to ameliorate symptoms and alter the course of
gastrointestinal disease. IL-31 may play a role in the inflammatory
response in colitis, and the neutralization of IL-31 activity by
administrating antagonists is a potential therapeutic approach for IBD
For disorders related to IBS and IBD, clinical signs of improved function
include, but are not limited to, reduction in pain, cramping and
sensitivity, reduction in diarrhea and improved stool consistency, reduced
abdominal distension, and increased intestinal transit. Improvement can
also be measured by a decrease in mean Crohn's Disease Activity Index (CDAI).
See Best. W. et al., Gastroenterology 70: 439-44, 1976. Additionally,
improved function can be measured by a quality of life assessment as
described by Irvine et al. (Irvine, E. et al., Gastroenterology 106:
Animal models of irritable bowel syndrome are described by Mayer and
Collins. Gastroenterol. 122:2032-2048 (2002). These models can be divided
into those that are mediated primarily by CNS-directed mechanisms ("Stress
Memory" models) and those with primary gut-directed etiologies ("Pain
Memory" and "Immune Memory" models). In one model, animals are surgically
prepared with electrodes implanted on the proximal colon and striated
muscles, and catheters implanted in lateral ventricles of the brain.
Rectal distension is performed by inflation of a balloon rectally
inserted, and the pressure eliciting a characteristic visceromotor
response is measured. A test compound, such as IL-31 antagonist and/or
variants or antagonists, is administered via the appropriate route (p.o.,
i.p., s.c., i.v., or i.m.) and at the appropriate time (i.e. .about.20
min, if i.p. or i.c.v.) prior to distention. Test compound is evaluated
for its ability to affect colonic motility, abdominal contractions, and
Additionally, disorders associated with inflammation of the intestine can
be treated with the IL-31 antagonists such as fragments, agonists and
antagonists thereof described herein. For example, Irritable Bowel
Syndrome (IBS) is characterized by a very broad spectrum of symptoms
(pain; bouts of diarrhea and/or constipation; abnormal gastrointestinal
motility). It is difficult to pinpoint the etiology, and may have
components related to stress, genetics, and/or inflammation. Similarly,
the anti-IL-31 molecules of the present invention, including antibodies
and binding partners, can be used to treat Inflammatory Bowel Disease,
(including colitis and Crohn's disease). IBD is more serious than IBS, and
is characterized by diarrhea, pain, and malnutrition. Patients with IBD
often have increased risk of gastrointestinal cancer.
Gastrointestinal motor activity can be measured in a dog model as follows:
Dogs are anesthetized and the abdominal cavity opened. Extraluminal force
transducers (sensor to measure contraction) are sutured onto five (5)
sites, i.e., the gastric antrum, 3 cm proximal to the pyloric ring, the
duodenum, 5 cm distal to the pyloric ring, the jejunum, 70 cm distal to
the pyloric ring, the ileum, 5 cm proximal to the ileum-colon junction,
and the colon, 5 cm distal to the ileum-colon junction. The lead wires of
these force transducers are taken out of the abdominal cavity and then
brought out through a skin incision made between the scapulae, at which a
connector is connected. After the operation, a jacket protector is placed
on the dog to protect the connector. Measurement of the gastrointestinal
motor activity is started two weeks after the operation. For ad libitum
measurement, a telemeter (electrowave data transmitter) is connected with
the connector to determine the contractive motility at each site of the
gastrointestinal tract. The data is stored in a computer via a telemeter
for analysis. A test compound, such as IL-31 antagonist is administered
via the appropriate route (p.o., i.v., i.p., s.c., i.m.) at the
appropriate time point to assess its ability to affect gastrointestinal
motor activity. This can be performed in normal dogs or dogs in which
gastroparesis/ileus has been induced. The above method is a modification
of those in Yoshida. and Ito. J. Pharmacol. Experiment. Therap. 257,
781-787 (1991) and Furuta et al. Biol. Pharm. Bull. 25:103-1071 (2002).
IL-31 may be a trigger for reactivation of latent viral infections, such
as varicella infection. In primary varicella zoster virus (VZV) infection,
the T cells most likely to be infected by varicella zoster virus are CD4
positive memory T cells expressing CLA and CCR4. These are skin-homing T
cells, which may enhance cell-associated viremia and the transport of
infectious virus to the skin and DRG. These cells are also the primary
producers of IL-31. Thus, IL-31 in primary VZV infection may contribute to
the itch/pain involved in the skin lesions. Reactivation of latent virus
in DRG induces VZV-specific T cell responses, which contribute to the
neurogenic inflammation. Skin-homing T cells are most easily infected with
VZV, and in vivo transfer of virus from T cells to DRG has been observed.
Postherpetic neuralgia is one of the major complications of herpes zoster
caused by the reactivation of varicella-zoster virus and is characterized
by severe pain. See Sato-Takeda, M. et al., Anesthesiology. 2006
104(5):1063-9, herein incorporated by reference. This reference also
teaches a mouse model of postherpetic pain, which corresponds to
postherpetic neuralgia. Briefly, BALB/c mice (MHC haplotype: H-2), C57BL/6
mice (MHC haplotype: H-2), and BALB/b mice, a congenic BALB/c strain with
H-2, are transdermally inoculated on the hind paw with Herpes simplex
virus type I. Unilaterally zosteriform skin lesion and pain-related
responses (acute herpetic pain) are caused, and some mice show
pain-related responses (postherpetic pain) after the cure of skin lesions.
Herpes simplex virus type I antigen and CD3-positive cells are
immunostained in the dorsal root ganglion in the acute phase. See also
Argoff, C. E., et al., J Pain Symptom Manage. 2004 October; 28(4):396-411,
herein incorporated by reference. Thus, antagonists to IL-31 may be useful
to limit or prevent reactivation of viral infections with varicella.
Mouse models for experimental allergic encephalomyelitis (EAE) has been
used as a tool to investigate both the mechanisms of immune-mediated
disease, and methods of potential therapeutic intervention. The model
resembles human multiple sclerosis, and produces demyelination as a result
of T-cell activation to neuroproteins such as myelin basic protein (MBP),
or proteolipid protein (PLP). Inoculation with antigen leads to induction
of CD4+, class II MHC-restricted T-cells (Th1). Changes in the protocol
for EAE can produce acute, chronic-relapsing, or passive-transfer variants
of the model (Weinberg et al., J. Immunol. 162:1818-26, 1999; Mijaba et
al., Cell. Immunol. 186:94-102, 1999; and Glabinski, Meth. Enzym.
288:182-90, 1997). Administration of IL-31 antagonists or other soluble
and fusion proteins may be useful to ameliorate symptoms and alter the
course of disease.
An antagonist to IL-31-induced signal transduction in dorsal root gangion
cells can be useful to treat pruritus uraemicus; pruritis from hepatitis,
hepatic failure, or cholestasis; from scabies or athletes's foot; from
pruritis associated with pregnancy; from pruritis in dualysis patients;
and from pruritis from anaesthasia and psychological disorders as follows.
Pruritus uraemicus or renal itch is an often intolerable symptom of
chronic renal insufficiency (Blachley J D, Blankenship D M, Menter A et
al. Uremic pruritus: skin divalent ion content and response to ultraviolet
phototherapy. Am J Kidney Dis 1985; 5: 237-41.) being present in about 13%
of the cases; secondary skin lesions due to scratching can be seen. It is
even more common in patients undergoing peritoneal dialysis or
hemodialysis (Murphy M, Carmichael A J. Renal itch. Clin Exp Dermatol
2000; 25: 103-6.); it can be localized or generalized. Itching is not
present in acute renal failure. The treatment of renal pruritus is based
on intensive and efficient dialysis to remove pruritogenic substances from
the blood, and on the use of non-complement-activating membranes. One can
also use UV therapy, emollient ointments, activated charcoal,
cholestyramine (4 grams twice a day), phosphate binding agents. Sometimes
parathyroidectomy is necessary.
Pain antagonizes itch. See, for example, Ward, L. et al., Pain 64:129-138,
1996. As such a mediator of pain, such as an IL-31 antagonist can be used
to treat pain associated with itch, thereby ameliorating not only the
itch, or scratching behavior, but also the associated pain.
Pruritus is a well-recognized manifestation among patients with liver
diseases and intrahepatic or posthepatic cholestasis. Hepatic diseases
leading to pruritus include primary biliary cirrhosis, B and C viral
hepatitis, primary sclerosing cholangitis, carcinoma of bile ducts,
alcoholic cirrhosis, autoimmune hepatitis and others. The pruritus is
generalized and more intense on hands, feet and around tight-fitting
clothes, while face, neck and genital area are rarely involved.
Generalized pruritus is present in 1-8% of pregnant women. Pruritus
gravidarum can be differentiated from pruritic dermatoses in pregnancy,
such as pemphigoid gestationis (herpes gestationis), papular and pruritic
dermatosis of pregnancy and others. Pruritus gravidarum manifests without
any rash mostly in the third trimester of pregnancy, but it may also
appear earlier, firstly on the abdomen and then becomes generalized. This
symptom usually tends to be worse at night and disappears after delivery
(within 1-4 weeks). Probably it is associated with intrahepatic
cholestasis, as there is an increase of gamma GT and alkaline phosphatase,
and sometimes also of direct bilirubin level in these patients. Pruritus
is more frequent in multiple pregnancies and can recur in subsequent
pregnancies or during the use of oral contraceptives. Additionally,
pruritic urticarial papulas and plaques of pregnancy (PUPP), the most
common dermatosis associated with pregnancy, does not respond to
antihistamines and often persists beyond parturition.
Some hematological disorders are known to be associated with pruritus. In
polycythemia rubra vera with overproduction of all three hematopoietic
cell lines, patients typically experience severe itch located on the
trunk, but sparing the face, hands and feet, a few minutes after contact
with warm water. Water-induced itching (aquagenic pruritus, or bath itch)
can be present in 70% of the patients. The itch can last for about 15
minutes to one hour, and be so severe that the patients refuse to bathe.
In the last decades pruritus has been described in patients with graft
versus host reactions after bone marrow transplantation.
Chronic delivery of IL-31 induces pruritis and alopecia in mice followed
by the development of skin lesions resembling dermatitis suggesting that
IL-31 may induce itching. See Dillon S. R., et al., Nat Immunol: 5, 752
(2004). The involvement of IL-31 was tested in induction of the itch
response by two methods as shown in Example 2: (i) capsaicin treatment of
IL-31-treated mice and (ii) IL-31 treatment of Tac1 knockout mice, which
have significantly reduced nociceptive pain responses because of lack of
expression of neuropeptides. In addition, whether neutralization of IL-31
in IL-31 treated mice could prevent pruritis and alopecia was tested in
NC/Nga Mice spontaneously develop AD-like lesions that parallel human AD
in many aspects, including clinical course and signs, histophathology and
immunopathology when housed in non-specified pathogen-free (non-SPF)
conditions at around 6-8 weeks of age. In contrast, NC/Nga mice kept under
SPF conditions do not develop skin lesions. However, onset of spontaneous
skin lesions and scratching behaviour can be synchronized in NC/Nga mice
housed in a SPF facility by weekly intradermal injection of crude dust
mite antigen. See Matsuoka H., et al., Allergy: 58, 139 (2003). Therefore,
the development of AD in NC/Nga is a useful model for the evaluation of
novel therapeutics for the treatment of AD.
In addition to the NC/Nga model of spontaneous AD, epicutaneous
sensitization of mice using OVA can also be used as a model to induce
antigen-dependent epidermal and dermal thickening with a mononuclear
infiltrate in skin of sensitized mice. This usually coincides with
elevated serum levels of total and specific IgE, however no skin barrier
dysfunction or pruritus normally occurs in this model. See Spergel J. M.,
et al., J Clin Invest, 101: 1614, (1998). This protocol can be modified in
order to induce skin barrier disregulation and pruritis by sensitizing
DO11.10 OVA TCR transgenic mice with OVA. Increasing the number of
antigen-specific T cells that could recognize the sensitizing antigen may
increase the level of inflammation in the skin to induce visible
scratching behaviour and lichenification/scaling of the skin.
Both the NC/Nga spontaneous AD model and the OVA epicutaneous DO 11.10
model can be used to measure expression of IL-31 and IL-31RA in AD, as
well as the ability of the antagonists described herein to inhibit,
reduce, or neutralize the effects of IL-31. The antagonists described
herein are useful to inhibit scratching associated with dermatitis and
pruritic diseases including atopic dermatitis, prurigo nodularis, and
eczema. In AD, the scratching behavior provoked by intensely itchy skin is
believed to aggravate disease by breaking down skin barrier functions and
activating keratinocytes, leading to chemokine production and increased
inflammation. Many clinicians view AD as a self-propagating cycle, since
lesions formed by frequent scratching are subject to infection and further
antigen stimulation. The fact that patients with near total involvement of
body surface area may have unaffected skin in regions that are hard to
scratch lends credence to this hypothesis. By preventing pruritis,
administration of antagonists of IL-31 or its receptor can be effective in
treating pruritic disease by decreasing IL-31-induced keratinocyte
activation and neurological stimulation, thus breaking the link between
inflammation and pruritis. The reduction in pruritis could also decrease
secretion of neurostimulatory factors and reduce the inflammation and
excoriations associated with constant scratching, leading to an
improvement in disease scores and/or a longer duration between disease
flares. An inhibition, reduction, or prevention of scratching, alone, can
be effective in treating pruritic diseases including, but not limited to,
atopic dermatitis, prurigo nodularis, and eczema, since cessation of
scratching will stop progression of dermatitis, the development of which
is dependent on scratching.
As used herein, the term "antibodies" includes polyclonal antibodies,
affinity-purified polyclonal antibodies, monoclonal antibodies, and
antigen-binding fragments, such as F(ab')2 and Fab proteolytic fragments.
Genetically engineered intact antibodies or fragments, such as chimeric
antibodies, Fv fragments, single chain antibodies and the like, as well as
synthetic antigen-binding peptides and polypeptides, are also included.
Non-human antibodies may be humanized by grafting non-human CDRs onto
human framework and constant regions, or by incorporating the entire
non-human variable domains (optionally "cloaking" them with a human-like
surface by replacement of exposed residues, wherein the result is a
"veneered" antibody). In some instances, humanized antibodies may retain
non-human residues within the human variable region framework domains to
enhance proper binding characteristics. Through humanizing antibodies,
biological half-life may be increased, and the potential for adverse
immune reactions upon administration to humans is reduced. Moreover, human
antibodies can be produced in transgenic, non-human animals that have been
engineered to contain human immunoglobulin genes as disclosed in WIPO
Publication No. WO 98/24893. It is preferred that the endogenous
immunoglobulin genes in these animals be inactivated or eliminated, such
as by homologous recombination.
Antibodies are considered to be specifically binding if: 1) they exhibit a
threshold level of binding activity, and 2) they do not significantly
cross-react with related polypeptide molecules. A threshold level of
binding is determined if anti-IL-31 antibodies herein bind to a IL-31
polypeptide, peptide or epitope with an affinity at least 10-fold greater
than the binding affinity to control (non-IL-31) polypeptide. It is
preferred that the antibodies exhibit a binding affinity (Ka) of 106 M-1
or greater, preferably 107 M-1 or greater, more preferably 108 M-1 or
greater, and most preferably 109 M-1 or greater. The binding affinity of
an antibody can be readily determined by one of ordinary skill in the art,
for example, by Scatchard analysis (Scatchard, G., Ann. NY Acad. Sci. 51:
Whether anti-IL-31 antibodies do not significantly cross-react with
related polypeptide molecules is shown, for example, by the antibody
detecting IL-31 polypeptide but not known related polypeptides using a
standard Western blot analysis (Ausubel et al., ibid.). Examples of known
related polypeptides are those disclosed in the prior art, such as known
orthologs, and paralogs, and similar known members of a protein family.
Screening can also be done using non-human IL-31, and IL-31 mutant
polypeptides. Moreover, antibodies can be "screened against" known related
polypeptides, to isolate a population that specifically binds to the IL-31
polypeptides. For example, antibodies raised to IL-31 are adsorbed to
related polypeptides adhered to insoluble matrix; antibodies specific to
IL-31 will flow through the matrix under the proper buffer conditions.
Screening allows isolation of polyclonal and monoclonal antibodies non-crossreactive
to known closely related polypeptides (Antibodies: A Laboratory Manual,
Harlow and Lane (eds.), Cold Spring Harbor Laboratory Press, 1988; Current
Protocols in Immunology, Cooligan, et al. (eds.), National Institutes of
Health, John. Wiley and Sons, Inc., 1995). Screening and isolation of
specific antibodies is well known in the art. See, Fundamental Immunology,
Paul (eds.), Raven Press, 1993; Getzoff et al., Adv. in Immunol. 43: 1-98,
1988; Monoclonal Antibodies: Principles and Practice, Goding, J. W.
(eds.), Academic Press Ltd., 1996; Benjamin et al., Ann. Rev. Immunol. 2:
67-101, 1984. Specifically binding anti-IL-31 antibodies can be detected
by a number of methods in the art, and disclosed below.
Preparation of monoclonal antibodies is well known to one skilled in the
art. The purified mature recombinant human IL-31 polypeptide (amino acid
residues 27 (Leu) to 167 (Thr) of SEQ ID NO:2) or the mouse ortholog,
produced from expression systems can be used to generate monoclonal
The effect of administering the antagonists of IL-31 mediated signal
transduction can be measured in vivo by a reduction, inhibition,
prevention, minimization, neutralization of inflammation, of skin or
dermal thickening, of recruitment of lymphocytes, and acanthosis, for
example, and other symptoms or composites of symptoms, such as the Eczema
Area and Severity Index (EASI), that are evident to one skilled in the
art. Additional effects could include a change or decrease in the
production of cytokines or chemokines by lesional skin, reduction in an
atopy patch test score, and decrease in release of soluble factors such as
cytokines, chemokines or neuropeptides, as measured by intradermal
microdialysis or other methods. Assessments of degree of itch or pain can
be measured using clinically approved instruments or tools such as the
Visual Analogue Scale. Frequency of scratching can be monitored by limb
movement meters, piezoelectric transducer devices attached to the
fingernails, or time-lapse infrared photography or videography of
nocturnal scratching in patients. Other methods for assessing a decrease
in pain or itch are evident to one skilled in the art.
Monoclonal antibodies purified from tissue culture media are characterized
for their utility in an ELISA for the quantitative determination of
recombinant and native human IL-31. The antibodies are selected and a
quantitative assay is developed.
Monoclonal antibodies purified from tissue culture media are characterized
for their ability to block or reduce the receptor binding activity
("neutralization assay") of purified recombinant huIL-31 on neural cells
expressing the IL-31Ra and OSMRb. A number of "neutralizing" monoclonal
antibodies are identified in this manner. Hybridomas expressing the
neutralizing monoclonal antibodies to human IL-31 described can then be
deposited with the American Type Tissue Culture Collection (ATCC; Manassas
Va.) patent depository as original deposits under the Budapest Treaty.
Five rat anti-mouse hybridomas were generated in a similar fashion and
were given the following clone designations: clone 2220.127.116.11.6, clone
218.104.22.168.1, clone 222.214.171.124.2, clone 2126.96.36.199.1, and clone
2188.8.131.52.5. The monoclonal antibodies produced by these clones were
characterized in a number of ways including binning (i.e, determining if
each antibody could inhibit the binding of any other binding), relative
affinity, and neutralization. The monoclonal antibodies appear to fall
into two separate bins with clone 2184.108.40.206.1 binding to a separate
epitope than the other four.
Monoclonal antibodies in tissue culture media are characterized for their
ability to block or reduce receptor binding when grown in the presence of
the purified recombinant proteins human IL-31.
Binding affinity of the monoclonal antibodies can be generated.
Goat-anti-Rat IgG-Fc gamma specific Antibody (Jackson) is immobilized onto
a CM5 Biacore chip. The assay is optimized to bind each mAb onto the
anti-Rat capture surface and then a concentration series of IL-31 is
injected across the mAb to see association (Ka) and dissociation (Kd).
After each run, the surface is regenerated back to the anti-Rat Antibody
with 2 injections of 20 mM HCl. Data is generated for each and evaluation
software (BIAevaluation software version 3.2, Pharmacia BIAcore, Uppsala,
Sweden) is used to assess the kinetics of the anti-IL-31 antibody binding
to the IL-31 protein
Biochemical confirmation that the target molecule, IL-31, recognized by
the putative anti-IL-31 mAbs is indeed IL-31 are performed by standard
immunoprecipitation followed by SDS-PAGE analysis or western blotting
procedures, both employing soluble membrane preparations from IL-31
transfected versus untransfected Baf3 cells. The mAbs are tested for their
ability to specifically immunoprecipitate or western blot the soluble
Monoclonal antibodies to IL-31 are described in commonly-owned, U.S.
patent application Ser. No. 11/430,066, filed May 8, 2006, U.S. published
patent application number 2006-0275296. These monoclonal antibodies were
purified from tissue culture media were characterized for their ability to
block or inhibit the ability of IL-31 to bind to its receptor in a
neutralization assay. Twenty "neutralizing" monoclonal antibodies were
identified in this manner. The monoclonal antibodies produced by these
clones were characterized in a number of ways including binning (i.e,
determining if each antibody could inhibit the binding of any other
binding), relative affinity, and neutralization. The ten good neutralizing
antibodies appear to be in the same bin, with the other monoclonal
antibodies grouping into three separate bins. In addition, eight of the
good neutralizing antibodies are IgG1 isotype and the other two are IgG2a
isotype. Such monoclonal antibodies can be IgG1 or IgG4 so as to minimize
complement binding and ADCC activity.
Hybridomas expressing the neutralizing monoclonal antibodies to human
IL-31 described above were deposited with the American Type Tissue Culture
Collection (ATCC; Manassas Va.) patent depository as original deposits
under the Budapest Treaty and were given the following ATCC Accession
Nos.: ATCC Patent Deposit Designation PTA-6815, deposited on Jun. 29,
2005; ATCC Patent Deposit Designation PTA-6816, deposited on Jun. 29,
2005; ATCC Patent Deposit Designation PTA-6829, deposited on Jul. 6, 2005;
ATCC Patent Deposit Designation PTA-6830, deposited on Jul. 6, 2005; ATCC
Patent Deposit Designation PTA-6831, deposited on Jul. 6, 2005; ATCC
Patent Deposit Designation PTA-6871, deposited on Jul. 19, 2005; ATCC
Patent Deposit Designation PTA-6872, deposited on Jul. 19, 2005; ATCC
Patent Deposit Designation PTA-6875, deposited on Jul. 19, 2005; and ATCC
Patent Deposit Designation PTA-6873, deposited on Jul. 19, 2005.
A hybridoma expressing the neutralizing monoclonal antibodies to mouse
IL-31 described herein was deposited with the American Type Tissue Culture
Collection (ATCC; Manassas Va.) patent depository as an original deposit
under the Budapest Treaty and was given the following ATCC Accession No.:
ATCC Patent Deposit Designation PTA-6874, deposited on Jul. 19, 2005. The
monoclonal antibodies produced by these hybridoma clones can be cultured
in a growth medium of 90% Iscove's Modified Dulbecco's medium with 2 mM
L-glutamine, 100 .mu.g/mL penicillin, and 100 .mu.g/mL streptomycin
sulfate, and 10% Fetal Clone I Serum (Hyclone Laboratories). The clones
can be propogated by starting cultures at 2.times.105 cells/ml and
maintaining between 1.times.105 and 5.times.105 cell/ml at 37.degree. C.
and 5-6% CO. Cells can be adapted to serum free conditions upon subsequent
transfers. Cells that are frozen are stored in 90% serum, 10% DMSO and
stored in vapor phase of liquid nitrogen freezer.
IL-31 antagonists generated by the methods described herein can be tested
for neutralization, inhibition, reduction, antagonization by a variety of
methods. In addition neutralization can be tested by measuring a decrease
in the production of pro-inflammatory chemokines such as TARC and MDC from
keratinocyte cultures in the presence of ligand and the monoclonal
antibody. Other biomarkers, such as MCP-1, MIP1a, TARC, MCP-1, MDC, IL-6,
IL-8, 1-309, SCYA19, MPIF-1, TECK, MIP-1b, SCYB13, GROa/MGSA, CTACK,
SCCA1/Serpin B3, TSLP, and NT-4 may also be used. Neutralization can also
be measured by the in vivo models described herein.
The bioactive antagonists or antibody conjugates described herein can be
delivered intravenously, intraarterially or intraductally, subcutaneously,
topically, or may be introduced locally at the intended site of action.
The antagonists of the present invention can be measured for their ability
to bind the IL-31 ligand as determined by any of the in vivo models
described herein, including but not limited to the NcNga model, the Ova
epicutaneous model, the chronic hypersensitivity model, the chronic hapten
model, the calcium flux model, the allodynia model.
Additional models to measure the inhibitory effects of the anti-IL-31
antibodies are known to one skilled in the art and described herein are
described by Umeuchi, H. et al., European Journal of Pharmacology, 518:
133-139, 2005; and by Yoo, J. et al., J. Experimental Medicine,
Mouse models to measure neurogenic inflammation are known in the art. See,
for example, Sweitzer, S. M., et al., J. Neuroimmunology 125: 82-93; 2002,
and Honore, P., et al., Neuroscience, (98): 585-598, 2000. See also,
Yonehara, N. and Yoshimura M., Pain, 2001 (9211-2): pp. 259-265).
Within aspects of the invention, the invention provides methods of
treating inflammation in neuronal tissue of a mammal; methods of treating
pain in a mammal; methods of antagonizing IL-31 induced signal
transduction is dorsal root ganglion cells; methods for treating symptoms
associated with burn; methods for treating symptoms associated with viral
infection and for preventing reactivation of viral infection; and methods
of treating pain associated with Inflammatory Bowel Disease. Within an
embodiment, the Inflammatory Bowel Disease is Crohn's Disease.
Within embodiments of these aspects, the invention provides, comprising
admixing neuronal tissue with an IL-31 antagonist, wherein the
inflammation, pain, dorsal root ganglion signal transduction, viral
infection or reactivation, or burn tissue, or pain associated with
Inflammatory Bowel Disease is reduced, limited, prevented, minimized or
Within other embodiments, the IL-31 antagonist binds a polypeptide
comprising an amino acid sequence as shown in SEQ ID NO: 2 from residue 27
to residue 164. Within other embodiments, the antagonist is selected from:
anti-idiotype antibodies; antibody fragments; chimeric antibodies; and
humanized antibodies. Within another embodiment the antagonist an
antibody. Within other embodiments the antibody is a monoclonal antibody.
Within other embodiments the antibody specifically binds a polypeptide
comprising the amino acid sequence of SEQ ID NO: 2 and wherein the
polypeptide is capable of binding the monoclonal antibody produced by the
hybridoma selected from the group consisting of a) ATCC Patent Deposit
Designation PTA-6815; b) ATCC Patent Deposit Designation PTA-6816; c) ATCC
Patent Deposit Designation PTA-6829; d) ATCC Patent Deposit Designation
PTA-6830; e) ATCC Patent Deposit Designation PTA-6831; ATCC Patent Deposit
Designation PTA-6871; g) ATCC Patent Deposit Designation PTA-6872; h) ATCC
Patent Deposit Designation PTA-6875; and i) ATCC Patent Deposit
Designation PTA-6873. Within another embodiment the monoclonal antibody is
selected from a bin of antibodies wherein the hybridoma producing the
antibody is selected from: a) ATCC Patent Deposit Designation PTA-6815; b)
ATCC Patent Deposit Designation PTA-6829; c) ATCC Patent Deposit
Designation PTA-6816; d) ATCC Patent Deposit Designation PTA-6871; and e)
ATCC Patent Deposit Designation PTA-6830. Within another embodiment the
monoclonal antibody is selected from a bin of antibodies wherein the
hybridoma producing the antibody is selected from: a) ATCC Patent Deposit
Designation PTA-6872; b) ATCC Patent Deposit Designation PTA-6873; c) ATCC
Patent Deposit Designation PTA-6875; and d) ATCC Patent Deposit
Claim 1 of 7 Claims
1. A method of treating neuronal tissue
inflammation in a mammal comprising administering to the mammal a
monoclonal antibody or binding fragment thereof which specifically binds
to a polypeptide sequence of amino acid residues 27-164 of SEQ ID NO:2,
and wherein after administration the neuronal tissue inflammation is
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