COMPOUNDS AND METHODS TARGETING INTERLEUKIN-34
The present invention relates to compounds, pharmaceutical compositions, and
methods, which include antibodies directed against human interleukin-34 (IL-34), which
5 are expected to be useful in the field of neuroinflammation and acute or chronic
inflammatory diseases. In particular, the embodiments are expected to be useful in
treatment and/or diagnostic applications relating to Alzheimer's Disease, as well as other
tauopathi es.
Alzheimer's disease (AD), a leading cause of dementia, develops in one percent of
10 the population between the ages 65 and 69, and increases to 40-50% in those 95 years and
older. AD patients exhibit telltale clinical symptoms that include cognitive impairment
and deficits in memory function. In these patients, the presence of AD is confirmed by
heavy senile plaque burden and neurofibrillary tangles (NFT) found in the cerebral cortex
upon post-mortem histopathological examination. The mature senile plaques consist of
15 extracellular ~-amyloid peptides derived from enzymatic processing of amyloid precursor
protein and intracellular neurofibrillary tangles (NFT), which are derived from filaments
of hyperphosphorylated tau proteins. Aggregates of hyperphosphorylated tau, such as
neurofibrillary tangles, are linked to the degree of cognitive impairment in Alzheimer's
disease. In AD and various other tauopathies, tau aggregates appear in specific brain
20 regions and patterns that are linked to disease risk, onset, and or progression, and these
regions and patterns are known to skilled artisans.
Cytokines regulate normal homeostatic tissue functions, and dysregulation of
these cytokine networks is associated with pathological conditions. The central nervous
system (CNS), where few blood-borne immune cells circulate, seems to be particularly
25 vulnerable to dysregulated cytokine networks. In neurodegenerative diseases, CNSresident
cells are the predominant producers of pro-inflammatory cytokines and can
contribute to dysregulated cytokine networks and neuroinflammation. Damage to the
CNS may involve recruitment of circulating immune cells resulting in an innate immune
response consisting of resident microglia, peripherally derived monocytes, macrophages
30 and dendritic cells. The activation states of microglia and macrophages are not strictly pro
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or anti-inflammatory and instead may have a spectrum of functional states. Microglia
and/or peripherally derived monocytes and macrophages may acquire an antiinflammatory
phenotype, in which they remove debris and promote regeneration and
homeostasis. Neuronal dysfunction or damage can also activate microglia to produce pro-
S inflammatory cytokines and recruit leukocytes from the bloodstream. In
neurodegenerative conditions, such as Alzheimer's disease (AD), microglia activation is a
frequent finding and reflects the tissue response to accumulation of extracellular betaamyloid
plaques and hyperphosphorylated tau aggregates. Neuroinflammation is an
important component of neurodegenerative diseases and is characterized by elevated
10 production of pro-inflammatory cytokines by CNS cells (Becher, B., Spath, S. &
Goverman, J. Cytokine networks in neuroinjlammation. Nat Rev Immunol 17, 49-59
(2017)). Neuroinflammation and microgliosis are believed to be mechanisms underlying
neurodegenerative diseases such as plaque accumulation in Alzheimer's disease, and
neuronal death and dysfunction in Parkinson's disease and Huntington's disease.
15 Microgliosis involves the abnormal proliferation and/or hypertrophy of microglia
in response to inflammatory signals. Broadly, IL-34 acts as a potent and pleiotropic
cytokine in the regulation of inflammatory and immune processes and is a key regulatory
cytokine for the growth of CNS-resident microglia in normal tissue homeostasis. IL-34 is
expressed by neurons in the cortex, the anterior olfactory nucleus and the hippocampus.
20 IL-34 is closely related to colony-stimulating factor 1 (CSF1; also known as M-CSF), and
both cytokines bind the CSF 1 receptor. IL-34 is a secreted homodimeric cytokine that
acts as one of two activating ligands for CSF 1R, and triggers receptor
autophosphorylation and dimerization with subsequent activation of multiple signaling
pathways (See, for example, Structural basis for the dual recognition of helical cytokines
25 IL-34 and CSF-1 by CSF-JR. Structure 20, 676-687, and Felix J, De Munck S, Verstraete
K, Meuris L, Callewaert N, Elegheert J. et al.). Human and mouse IL-34 polypeptides are
disclosed for example in US Patent No. 9, 770,486. IL-34 is a protein of 242 amino acids
in humans (SEQ ID NO: 41), and 235 amino acids in mouse (SEQ ID NO: 42).
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Anti-IL-34 antibodies have been described in the art, and for example, WO
2016/196679 recites various anti-IL-34 antibodies and potential uses thereof However,
to date, no antibody targeting IL-34 has been approved for therapeutic use.
Thus, there remains an unmet need for alternative and/or improved anti-IL-34
5 antibodies, pharmaceutical compositions thereof, and methods of using the same for
therapeutic and/or in diagnostic applications relating to immune-mediated diseases
involving IL-34, and/or diseases treatable with an anti-IL-34 antibody, such as
neuroinflammatory disorders, and/or Alzheimer's Disease. Further, there remains an
unmet need for alternative and/or improved anti-IL-34 antibodies which have a
10 combination of particularly advantageous properties over prior art anti-IL-34 antibodies,
in view of at least one or more properties from the following: 1) desirable association and
dissociation rates, 2) potency in neutralization of human IL-34 to achieve an antineuroinflammatory
response and in vivo efficacy, 3) sufficiently potent as a monotherapy
for the treatment and/or prevention of immune-mediated and/or inflammatory disorders;
15 4) a sustained duration of action; 5) sufficiently limited induction ofundesirable cytokine
release, 6) acceptably low immunogenicity (i.e., sufficiently non-immunogenic in
humans); 7) avoidance ofuntoward immunocompromise; and/or 8) desirable in vivo
stability, physical and chemical stability including, but not limited to, thermal stability,
solubility, low self-association, and pharmacokinetic characteristics which are acceptable
20 for development and/or use in the treatment of inflammatory or neuroinflammatory
disorders, for example AD.
Summary of Invention:
Embodiments of the present invention provide novel anti-human IL-34 and anti-
25 murine IL-34 antibodies. According to some embodiments, the present invention provides
antibodies which comprise a light chain variable region (LCVR) and a heavy chain
variable region (HCVR), wherein the LCVR comprises complementarity determining
regions (CDRs) LCDR1, LCDR2 and LCDR3 and the HCVR comprises CDRs HCDR1,
HCDR2 and HCDR3 are selected from the groupings of CDR combinations provided in
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Table 1. The sequence identifiers used herein are listed in Table 1, and the sequences are
provided in the amino acid and nucleotide sequence listing provided herein.
Table 1: Amino Acid and Nucleotide Sequences
Antibody 1 Antibody 2 Antibody 3
SEQIDHC 1 13 19
SEQIDLC 2 2 20
SEQIDHCVR 3 14 21
SEQIDLCVR 4 4 22
SEQIDHCDR1 5 15 23
SEQIDHCDR2 6 16 24
SEQIDHCDR3 7 17 25
SEQID LCDR1 8 8 26
SEQIDLCDR2 9 9 27
SEQIDLCDR3 10 10 28
SEQ ID: DNA HC 11 18 29
SEQIDDNALC 12 12 30
5 Accordingly, embodiments of the present invention also provide antibodies
comprising a LCVR and a HCVR selected from:
a. the LCVR having the amino acid sequence of SEQ ID NO: 4 and the HCVR having
the amino acid sequence of SEQ ID NO: 3;
b. the LCVR having the amino acid sequence of SEQ ID NO: 4 and the HCVR having
10 the amino acid sequence of SEQ ID NO: 14;
c. the LCVR having the amino acid sequence of SEQ ID NO: 22 and the HCVR
having the amino acid sequence of SEQ ID NO: 21.
According to other embodiments, the present invention also provides antibodies
comprising a combination ofLCVR and HCVR as described above in a-c, with a hinge
15 region and Fe region selected from SEQ ID NO: 51 and SEQ ID NO: 52.
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According to other embodiments, the present invention also provides antibodies
comprising a LC and a HC selected from, or having amino acid sequences with at least
95% homology to the amino acid sequences of:
a. the LC having the amino acid sequence of SEQ ID NO: 2 and the HC having the
5 amino acid sequence of SEQ ID NO: 1;
b. the LC having the amino acid sequence of SEQ ID NO: 2 and the HC having the
amino acid sequence of SEQ ID NO: 13; and
c. the LC having the amino acid sequence of SEQ ID NO: 20 and the HC having the
amino acid sequence of SEQ ID NO: 19.
10 As used herein "Antibody 1" refers to an antibody having the HCDR1 amino acid
sequence of SEQ ID NO: 5, the HCDR2 amino acid sequence of SEQ ID NO: 6, the
HCDR3 amino acid sequence of SEQ ID NO: 7, the LCDR1 amino acid sequence of SEQ
ID NO: 8, the LCDR2 amino acid sequence of SEQ ID NO: 9, the LCDR3 amino acid
sequence of SEQ ID NO: 10, the HCVR amino acid sequence of SEQ ID NO: 3, the
15 LCVR amino acid sequence of SEQ ID NO: 4, the HC amino acid sequence of SEQ ID
NO: 1, the LC amino acid sequence of SEQ ID NO: 2, the HC DNA sequence of SEQ ID
NO: 11, and the LC DNA sequence of SEQ ID NO: 12. The framework and CDR
sequences in each of the antibodies for which sequences are set forth herein are annotated
using annotation rules in agreement with the method ofNorth, et al.. J Mol. Bioi. 2011:
20 406: 228-256 unless otherwise specified.
As used herein "Antibody 2" refers to an antibody having the HCDR1 amino acid
sequence of SEQ ID NO: 15, the HCDR2 amino acid sequence of SEQ ID NO: 16, the
HCDR3 amino acid sequence of SEQ ID NO: 17, the LCDR1 amino acid sequence of
SEQ ID NO: 8, the LCDR2 amino acid sequence of SEQ ID NO: 9, the LCDR3 amino
25 acid sequence of SEQ ID NO: 10, the HCVR amino acid sequence of SEQ ID NO: 14,
the LCVR amino acid sequence of SEQ ID NO: 4, the HC amino acid sequence of SEQ
ID NO: 13, the LC amino acid sequence of SEQ ID NO: 2, the HC DNA sequence of
SEQ ID NO: 18, and the LC DNA sequence of SEQ ID NO: 12. The framework and
CDR sequences in each of the antibodies for which sequences are set forth herein are
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annotated using annotation rules in agreement with the method of North, et al.. J Mol.
Bioi. 2011: 406: 228-256 unless otherwise specified.
As used herein "Antibody 3" refers to an antibody having the HCDR1 amino acid
sequence of SEQ ID NO: 23, the HCDR2 amino acid sequence of SEQ ID NO: 24, the
5 HCDR3 amino acid sequence of SEQ ID NO: 25, the LCDR1 amino acid sequence of
SEQ ID NO: 26, the LCDR2 amino acid sequence of SEQ ID NO: 27, the LCDR3 amino
acid sequence of SEQ ID NO: 28, the HCVR amino acid sequence of SEQ ID NO: 21,
the LCVR amino acid sequence of SEQ ID NO: 22, the HC amino acid sequence of SEQ
ID NO: 19, the LC amino acid sequence of SEQ ID NO: 20, the HC DNA sequence of
10 SEQ ID NO: 29, and the LC DNA sequence of SEQ ID NO: 30. The framework and
CDR sequences in each of the antibodies for which sequences are set forth herein are
annotated using annotation rules in agreement with the method of North, et al.. J Mol.
Bioi. 2011: 406: 228-256 unless otherwise specified.
The carboxy-terminal portion of each HC defines a constant region primarily
15 responsible for effector functions, and in some embodiments of the present invention the
antibodies have one or more modifications in the constant region of each HC that reduce
effector functions. Preferably, embodiments of the present invention are IgG4 antibodies,
and thus contain an IgG4 Fe region, or an Fe region derived from human IgG4, e.g., a
modified IgG4 Fe region.
20 According to some embodiments, modifications in the constant region of both
HCs which reduce effector functions, and amino acid substitutions are introduced into the
IgG4 hinge and Fe regions. Thus, some embodiments have modifications in the constant
region of both HCs which include the amino acid alanine at both residues 230 and 231
(exemplified in HC of Antibody 1, HC of Antibody 2, and SEQ ID NO: 52, respectively),
25 and further modifications in the constant region of both HCs promoting stability,
including the amino acid proline at residue 224 (exemplified in HC of Antibody 1, HC of
Antibody 2, and for example in SEQ ID NO: 51), and the deletion of the amino acid
lysine at residue 443 (exemplified HC of SEQ ID N0.1).
The antibodies of the present invention are believed to have a combination of
30 particularly advantageous properties over prior art anti-IL-34 antibodies, including but
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not limited to, one or more of the following properties: 1) desirable association and
dissociation rates, 2) potency in neutralization of human IL-34 to achieve an antineuroinflammatory
response and in vivo efficacy, 3) sufficiently potent as a monotherapy
for the treatment and/or prevention of immune-mediated and/or inflammatory disorders;
5 4) a sustained duration of action; 5) sufficiently limited induction of undesirable cytokine
release, 6) acceptably low immunogenicity (i.e., sufficiently non-immunogenic in
humans); 7) avoidance ofuntoward immunocompromise; and/or 8) desirable in vivo
stability, physical and chemical stability including, but not limited to, thermal stability,
solubility, low self-association, and pharmacokinetic characteristics which are acceptable
10 for development and/or use in the treatment of inflammatory or neuroinflammatory
disorders, for example AD.
Embodiments of the present invention provide a significant advance over the prior
art by providing compositions and methods useful in the prevention, downregulation, or
amelioration of inflammatory and/or neuroinflammatory related disorders, through IL-34
15 neutralization, using a pharmacologically advantageous anti-human IL-34 antibody as
provided in the embodiments described herein. Anti-human IL-34 antibodies of the
present invention are capable of improving immune and/or inflammatory pathology, or
restoring immune homeostasis, preferably, through inhibition of the innate arm of the
immune response, and/or abrogation of microgliosis or other monocyte/macrophage
20 lineage cellular activation and or proliferation, thereby directly modifying underlying
disease pathology. The use of such antibodies clinically may lead to durable long-term
improvement of the disease(s) being treated.
Further, there is a need for diagnostic anti-human IL-34 antibodies that are
specific for human IL-34, and possess improved binding affinity, and demonstrate
25 enhanced sensitivity in human IL-34 determinations, and improved enzyme-linked
immunosorbent assay (ELISA) assay conditions that result in minimal interference and
broad dilutionallinearity. According to some aspects of the present invention, anti-human
IL-34 antibodies, including human IL-34 neutralizing antibodies, are provided which bind
human IL-34 given by SEQ ID NO: 41. Interleukin 34 (IL-34; also known as
30 uncharacterized protein C16orf77) is secreted as a homodimer consisting of39 kDa
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monomers. It belongs to no known cytokine family. Human IL-34 is synthesized as a 242
amino acid (AA) precursor that contains a 20 AA signal sequence, and results in a 222
AA mature chain. As used herein IL-34 refers to the mature chain. The mature chain
contains one potential site ofN-linked glycosylation. Human IL-34 is 71% identical to
5 mouse IL-34 on the amino acid level. IL-34 is expressed in various tissues, including the
heart, brain, liver, kidney, spleen, thymus, testes, ovary, small intestine, prostate, and
colon, and is most abundant in the spleen. "h IL-34" or "human IL-34" when used herein
in reference to an IL-34 polypeptide, unless otherwise stated, refers to wild-type human
IL-34, and preferably has the amino acid sequence set forth in SEQ ID NO: 41, which is
10 mature IL-34 having the leader sequence removed. (See, for example, Lin et.al., Science
(2008) Vol. 320, Issue 5877, pp. 807-811).
An exemplary human IL-34 (including the 20 AA signal peptide sequence which
is removed to yield the mature polypeptide) is:
15 MPRGFTWLRYLGIFLGV ALGNEPLEMWPL TQNEECTVTGFLRDKLQYRS
RLQYMKHYFPI NYKISVPYEGVFRIANVTRLQRAQVSERELR YL WVL VSLSATES
VQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKL
VRPKALLDNCFRVMELL YCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQL
YPPPPWSPSSPPHSTGSVRPVRAQGEGLLP
20 (SEQ ID NO: 41, see NCBI Ref Seq. No. NP 689669.2, March 1, 2018).
According to some aspects of the present invention, anti-murine IL-34 antibodies,
including murine IL-34 neutralizing antibodies, are provided which bind human IL-34
given by SEQ ID NO. 42. An exemplary murine IL-34 (including the 20 AA signal
peptide sequence which is removed to yield the mature polypeptide) is:
25 MPWGLAWL YCLGILLDV ALGNENLEIWTLTQDKECDLTGYLRGKLQYK
NRLQYMKHYFPINYRIA VPYEGVLRV ANITRLQKAHVSERELRYL WVL VSLNAT
ESVMDVLLEGHPSWKYLQEVQTLLENVQRSLMDVEIGPHVEA VLSLLSTPGLSL
KL VRPKALLDNCFRVMELL YCSCCKQSPILKWQDCELPRLHPHSPGSLMQCTAT
NVYPLSRQTPTSLPGSPSSSHGSLP
30 (SEQ ID NO: 42, see NCBI Ref Seq. No. NP 001128572.1, March 1, 2018).
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As used herein, "human anti-IL34 antibody" or "anti-human IL-34 antibody"
refers to an antibody that binds to human IL-34, and when administered in vitro or in
vivo, results in an IL-34 activity-neutralizing and/or blocking response, such as at least
one significantly lessened desired activity, for example a desired reduction in IL-34
5 signaling as evidenced by a change in an IL-34 responsive molecular or cellular endpoint.
For instance, microglia number, density, or phenotype in the CNS, is an example of an
IL-34 responsive molecular or cellular effect. As used herein, the terms "signaling" and
"signal transduction" and "IL-34-mediated", as they relate to IL-34, refer to cellular
and/or intercellular responses which result from the activity ofiL-34.
IO The term "antibody," as used herein, refers to an immunoglobulin molecule that
binds an antigen. Embodiments of an antibody include a monoclonal antibody, polyclonal
antibody, human antibody, humanized antibody, chimeric antibody or conjugated
antibody. The antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any
subclass (e.g., IgG I, IgG2, IgG3, IgG4). An exemplary antibody is an immunoglobulin G
I5 (IgG) type antibody comprised of four polypeptide chains: two heavy chains (HC) and
two light chains (LC) that are cross-linked via inter-chain disulfide bonds. LCs are
classified as kappa or lambda, which are each characterized by a specific constant region.
Embodiments of the present invention may comprise an IgGI or IgG4 antibody, and
further comprise kappa light chains or lambda light chains. Preferably antibodies of the
20 present invention comprise light chain constant regions which are kappa constant regions.
HCs are classified as gamma, mu, alpha, delta, or epsilon, and define the isotype
of an antibody as IgG, IgM, IgA, IgD, or IgE, respectively. The amino-terminal portion of
each of the four polypeptide chains includes a variable region of about IOO-I25 or more
amino acids primarily responsible for antigen recognition. The carboxyl-terminal portion
25 of each of the four polypeptide chains contains a constant region primarily responsible for
effector functions. Each heavy chain is comprised of a heavy chain variable region (VH)
and a heavy chain constant region. The constant region of the heavy chains contains CHI,
CH2, and CH3 domains. CHI comes after the HCVR; the CHI and HCVR form the
heavy chain portion of an antigen-binding (Fab) fragment, which is the part of an
30 antibody that binds antigen(s). CH2 comes after the hinge region and before CH3. CH3
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comes after CH2 and is at the carboxy-terminal end of the heavy chain. The constant
region of the light chains contains one domain, CL. CL comes after the LCVR; the CL
and LCVR form the light chain portion of a Fab.
The antibodies of the present invention include IgG HCs which can be further
5 divided into subclasses, e.g., IgG1, IgG2, IgG3, IgG4, and embodiments ofthe present
invention may include one or more modifications in the constant region of each HC, for
example that enhance or reduce effector function. The term "Fe region" as used herein
refers to a region of an antibody, which comprises the CH2 and CH3 domains of the
antibody heavy chain. Optionally, the Fe region may include a portion of the hinge
10 region or the entire hinge region of the antibody heavy chain. IgG 1 is known to induce
antibody-dependent cell cytotoxicity (ADCC) and complement-dependent cytotoxicity
(CDC), and Fe mutations described herein may reduce aggregation, reduce or enhance
ADCC or CDC activities, (or other functions), and/or modify the pharmacokinetics of the
antibodies. Embodiments of anti-human IL-34 antibodies described herein have reduced
15 binding to the F cy R and C 1 q receptors, thereby reducing or eliminating the cytotoxicity
which may be induced by antibodies with wild type IgG Fe regions. Thus, according to
some embodiments, mutations are introduced in the Fe region at positions as described
herein. Patient safety can be improved with sufficiently reduced or eliminated effector
functions of such anti-human IL-34 antibodies comprising a modified Fe region, and in
20 combination with other properties described herein, provide therapeutic agents with an
improved profile of useful activities while avoiding undesirable activities.

We Claim:
1. An antibody that binds human IL-34 wherein the antibody comprises a heavy chain
variable region (VH) and a light chain variable region (VL ), wherein the VH comprises
5 heavy chain complementarity determining regions (HCDR) HCDR1, HCDR2, and
HCDR3, and the VL comprises light chain complementarity determining regions (LCDR)
LCDR1, LCDR2, and LCDR3, wherein
the HCDR1 comprises SEQ ID NO: 5,
the HCDR2 comprises SEQ ID NO: 6,
10 the HCDR3 comprises SEQ ID NO: 7,
the LCDR1 comprises SEQ ID NO: 8,
the LCDR2 comprises SEQ ID NO: 9, and
the LCDR3 comprises SEQ ID NO: 10.
15 2. The antibody of claim 1, wherein the VH comprises SEQ ID NO: 3 and the VL
comprises SEQ ID NO: 4.
20
3. The antibody of claim 1 or 2, wherein the antibody comprises a heavy chain (HC)
comprising SEQ ID NO: 1 and a light chain (LC) comprising SEQ ID NO: 2.
4. An antibody that binds human IL-34 comprising a VH and a VL, wherein the VH
comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and
LCDR3, wherein
the HCDR1 comprises SEQ ID NO: 15,
25 the HCDR2 comprises SEQ ID NO: 16,
the HCDR3 comprises SEQ ID NO: 17,
the LCDR1 comprises SEQ ID NO: 8,
30
the LCDR2 comprises SEQ ID NO: 9, and
the LCDR3 comprises SEQ ID NO: 10.
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5. The antibody of claim 4, wherein the VH comprises SEQ ID NO: 14 and the VL
comprises SEQ ID NO: 4.
6. The antibody of claim 4 or 5, wherein the antibody comprises a HC comprising SEQ
5 ID NO: 13 and a LC comprising SEQ ID NO: 2.
10
15
7. An antibody that binds murine IL-34 wherein the antibody comprises a VH and a VL,
wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises
LCDR1, LCDR2, and LCDR3, wherein
the HCDR1 comprises SEQ ID NO: 23,
the HCDR2 comprises SEQ ID NO: 24,
the HCDR3 comprises SEQ ID NO: 25,
the LCDR1 comprises SEQ ID NO: 26,
the LCDR2 comprises SEQ ID NO: 27, and
the LCDR3 comprises SEQ ID NO: 28.
8. The antibody of claim 7, wherein the VH comprises SEQ ID NO: 21 and the VL
comprises SEQ ID NO: 22.
20 9. The antibody of claim 7 or 8, wherein the antibody comprises a HC comprising SEQ
ID NO: 19 and a LC comprising SEQ ID NO: 20.
25
10. A nucleic acid comprising a sequence encoding a SEQ ID NO selected from one or
more of the group consisting of: 11, 12, 18, 29, and 30.
11. A vector comprising the nucleic acid of claim 10.
12. The vector of claim 11, wherein the vector comprises a first nucleic acid sequence
encoding SEQ ID NO: 11 or 18, and a second nucleic acid sequence encoding SEQ ID
30 NO: 12.
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13. The vector of claim 11, wherein the vector comprises a first nucleic acid sequence
encoding SEQ ID NO: 29, and a second nucleic acid sequence encoding SEQ ID NO: 30.
5 14. A composition comprising a first vector comprising a nucleic acid sequence encoding
SEQ ID NO: 11 or 18, and a second vector comprising a nucleic acid sequence encoding
SEQ ID NO: 12.
15. A composition comprising a first vector comprising a nucleic acid sequence encoding
10 SEQ ID NO: 29, and a second vector comprising a nucleic acid sequence encoding SEQ
IDNO: 30.
16. A cell comprising the vector of claim 12 or 13.
15 17. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID
NO: 11 or 18, and a second vector comprising a nucleic acid sequence encoding SEQ ID
NO: 12.
18. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID
20 NO: 29, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:
30.
19. The cell of any one of claims 16-18, wherein the cell is a mammalian cell.
25 20. A process of producing an antibody comprising culturing the cell of any one of
claims 16-19 under conditions such that the antibody is expressed and recovering the
expressed antibody from the culture medium.
21. An antibody produced by the process of claim 20.
30
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22. A pharmaceutical composition comprising the antibody of any one of claims 1-9 or
21, and a pharmaceutically acceptable excipient, diluent or carrier.
23. A method of treating an immune-mediated disease in a subject in need thereof,
5 comprising administering to the subject a therapeutically effective amount of the antibody
of any one of claims 1-9 or 21, or the pharmaceutical composition of claim 22.
24. The method of claim 23, wherein the immune-mediated disease is selected from the
group consisting of Alzheimer's Disease; a tauopathy disease; Sjogren's syndrome (SS);
10 Rheumatoid arthritis (RA); inflammatory bowel disease (IBD), atopic dermatitis, kidney
disease, sepsis, and/or non-alcoholic fatty liver disease (NAFLD).
15
20
25. The method of claim 23, wherein the immune-mediated disease is Alzheimer's
Disease.
26. The anti body of any one of claims 1-9 or 21 for use in therapy.
27. The antibody of any one of claims 1-9 or 21, or the pharmaceutical composition of
claim 22, for use in the treatment of an immune-mediated disease.
28. The antibody or pharmaceutical composition of claim 27, wherein the an immunemediated
disease is selected from the group consisting of Alzheimer's Disease; a
tauopathy disease; Sjogren's syndrome (SS); Rheumatoid arthritis (RA); inflammatory
bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and/or non-alcoholic fatty
25 liver disease (NAFLD).
29. The antibody or pharmaceutical composition of claim 27, wherein the immunemediated
disease is Alzheimer's Disease.
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30. Use of the antibody of any one of claims 1-9 or 21, in the manufacture of a
medicament for the treatment of an immune-mediated disease.
31. The use of claim 30, wherein the immune-mediated disease is selected from the
5 group consisting of Alzheimer's Disease; a tauopathy disease; Sjogren's syndrome (SS);
Rheumatoid arthritis (RA); inflammatory bowel disease (IBD), atopic dermatitis, kidney
disease, sepsis, and/or non-alcoholic fatty liver disease (NAFLD).
10
15
20
32. The use of claim 30, wherein the immune-mediated disease is Alzheimer's Disease.
33. A method of determining the human IL-34level in a bodily fluid comprising:
(a) contacting the bodily fluid with an anti-human IL-34 diagnostic
monoclonal antibody, or antigen-binding fragment thereof, that specifically
binds to human IL-34 consisting of the amino acid sequence as in SEQ ID
NO: 41, the antibody, or antigen-binding fragment thereof, comprising: light
chain complementarity determining regions LCDRI, LCDR2, and LCDR3
comprising the amino acid sequences (SEQ ID NO: 8), (SEQ ID NO: 9), and
(SEQ ID NO: 10), respectively, and heavy chain complementarity determining
regions HCDRI, HCDR2, and HCDR3 comprising the amino acid sequences
(SEQ ID NO: 15), (SEQ ID NO: 16), and (SEQ ID NO: 17), respectively;
(b) optionally, removing any non-specifically bound monoclonal antibody
or, antigen-binding fragment thereof; and
(c) detecting and/or quantifying the amount of monoclonal antibody, or
antigen-binding fragment thereof, which is specifically bound to human IL-34.
25 34. The method of claim 33, wherein said bodily fluid is blood, serum or plasma, or
cerebrospinal fluid, and said contacting occurs ex vivo.