Cytokine targeting myeloid cells

Abstract

The invention relates to single chain polypeptides having dual cytokine activities, including CSF1 receptor targeting activity, and to their use in therapy.

Description

[0001] CYTOKINE TARGETING MYELOID CELLS

[0002] Field of the Invention

[0003] The invention relates to single chain polypeptides having dual cytokine activities, including CSF1 receptor targeting activity, and to their use in therapy.

[0004] Background of the Invention

[0005] Cytokines are proteins that are produced in the immune system and act as chemical messengers to communicate between cells of the immune system, and also with other cells of the body. Cytokines control the entire range of immune responses, including their initiation, type, potency, and duration. So much so that its unregulated production triggers a wide variety of inflammatory and autoimmune diseases, allergies, fibrosis, and can even lead to cancer. For this reason, there is great medical interest in the development and use of cytokines as drugs to treat diseases.

[0006] Myeloid cells exclusively express CSF1 receptor, i.e. CSF1R. Stimulation of CSF1R by its ligand, the cytokine CSF1, is essential for the differentiation, proliferation and maintenance of M2-like anti-inflammatory macrophages in the intestine, which play a key role in generating a tolerogenic environment through the production of IL-10. In autoimmune disease, the M2-like anti-inflammatory macrophages of the affected tissues, such as the mucosa of IBD patients, are replaced by inflammatory monocytes that drive disease progression. I L-34 is another ligand of CSF1 R that directs the differentiation of myeloid cells in a tissue-restricted manner, in the skin epidermis and CNS (Wang et al., 2012, Nat. immunol., 24; 13(8): 753-60). IL-34 has been also found to be involved in the development of intestinal fibrosis, a feature of inflammatory bowel disease patients, through CSF1R signaling in fibroblasts (Franze et al., 2020, Journal of Crohn's and Colitis, Volume 14, Issue 10, Pages 1436-1445).

[0007] Monocytes, a subset of circulating immune cells, can differentiate into distinct macrophage subtypes depending on the cytokine environment they encounter. CSF1R signaling is a key driver of monocyte differentiation into macrophages by promoting their survival, proliferation, and functional maturation. However, the presence of additional cytokines during this differentiation process can direct monocytes towards distinct macrophage phenotypes, broadly classified as pro-inflammatory (M1-like) or anti-inflammatory / tissue-repairing (M2-like) macrophages, as well as more specialized subsets depending on the tissue microenvironment.

[0008] When CSF1 is combined with pro-inflammatory cytokines such as interferon-gamma (IFN-y) or tumor necrosis factor-alpha (TNF-a), monocytes differentiate into M1-likemacrophages. These macrophages are characterized by high expression of major histocompatibility complex (MHC) class II molecules, inducible nitric oxide synthase (iNOS), and inflammatory cytokines such as interleukin-12 (IL-12) and IL-6. M1-like macrophages play a crucial role in host defense by producing reactive oxygen species (ROS) and promoting T-helper 1 (Thl)-mediated immune responses.

[0009] Conversely, when CSF1R stimulation is combined with cytokines such as interleukin-4 (IL-4), interleukin-10 (IL-10), or transforming growth factor-beta (TGF-P), monocytes differentiate into M2-like macrophages. These macrophages are characterized by the expression of markers such as CD206 (mannose receptor), arginase-1 (ARG1), and high levels of IL-10. M2-like macrophages promote tissue repair, angiogenesis, and resolution of inflammation by modulating extracellular matrix remodeling and suppressing excessive immune responses. Different M2 subtypes exist, depending on the specific cytokine signals received.

[0010] Interleukin-10 (IL-10) is an anti-inflammatory cytokine that plays a crucial role in regulating immune responses and maintaining immune homeostasis. IL-10 is a homodimeric cytokines that is produced as a monomer by various immune cells, including T cells, B cells, macrophages, and dendritic cells, and it exerts its effects by binding to its receptor complex, which is composed of two subunits, IL-10R1 and IL-10R2. When IL-10 binds to its receptor, IL-10R1 first recognizes and binds to IL-10 with high affinity, forming a complex that is then stabilized by the binding of IL-10R2. The binding of IL- 10 to its receptor complex leads to the activation of downstream signaling pathways.

[0011] When binds to its receptor on myeloid cells, IL-10 exerts potent anti-inflammatory activities, including inhibition of the production of pro-inflammatory mediators, expression of MHCII and costimulatory molecules. Simultaneously, activation of myeloid cells by IL-10 leads to the expression of anti-inflammatory mediators and promotes the development of tolerogenic phenotype, like M2-like anti-inflammatory macrophages and tolerogenic dendritic cells. In contrast to the strong anti-inflammatory activities on myeloid cells, IL- 10 is a growth factor for B cells and promotes plasma cell differentiation and antibody production. Similarly, IL-10 has stimulatory effects on CD8+ T cells, inducing their proliferation, IFNY production, and cytotoxicity.

[0012] While in preclinical studies IL-10 has shown promising results in ameliorating inflammation and tissue damage in models of colitis, clinical trials investigating the therapeutic potential of IL-10 in patients with colitis have not been successful. One possible explanation is that the delivery of IL- 10 to the site of inflammation may be a limiting factor. However, some strategies aimed to enhance the delivery of IL-10 at the site of inflammation did not show clinical benefit (AMT-101).Another possible explanation is because the anti-inflammatory effects of IL-10 on myeloid cells are counteracted by pro-inflammatory tissue-damaging effects, including activation, differentiation, and induction of antibodies by B cells and induction of cytotoxic activities by CD8+ T cells.

[0013] To address this limitation, the inventors developed IL-10-based therapeutics that can selectively activate myeloid cells while avoiding activation of other immune cell subsets. More specifically, they constructed single chain polypeptides having IL-10 and CSF1R targeting activities, i.e. single chain polypeptides combining IL-10 and CSF1 activities (herein ‘ORK1’) and single chain polypeptides combining IL-10 and IL-34 activities (herein ‘ORK6’), and that inhibit activation of myeloid cells and induce differentiation of monocyte-derived macrophages that have a regulatory phenotype. Furthermore, the dual CSF1R tageting-IL-10 cytokine has specificity for monocytes and does not activate or has reduced activation properties on CD8 T cells and B cells.

[0014] The inventors further constructed single chain polypeptides having IFN-y and CSF1R targeting activities, i.e. single chain polypeptides combining IFN-y and CSF1 activities (herein “ORK9’). The single chain polypeptides activate IFN-yR signaling in monocytes and induce differentiation of monocyte-derived macrophages that have a pro-inflammatory phenotype. Furthermore, the induced macrophages promote T cell activation and cytotoxic cytokine secretion. The single chain polypeptides having IFN-y and CSF1R targeting activities are thus suitable for reprogramming the tumor microenvironment and enable antitumor immunity.

[0015] Single chain polypeptides having dual CSF1 and IL-2 activities (herein “ORK10’), dual CSF1 and IL-4 activities (herein “ORK11’), and dual CSF1 and GM-CSF activities (herein “ORK12’) were also designed. These single chain polypeptides were shown to binding to monocytes in a CSF1-dependent manner. ORK11 was additionally demonstrated to preferentially activate monocytes when compared to T cells, indicating selectivity towards myeloid cells compared to lymphoid cells, in contrast with IL-4.

[0016] These results thus support the development of a dual cytokine platform in the form of a single chain polypeptide combining a CSF1R targeting cytokine and another cytokine of interest. The dual cytokine platform enables to fine-tune differentiation of monocytes into pro-inflammatory or regulatory macrophages depending on the context of use thereof.

[0017] Summary of the Invention

[0018] The invention is as defined by the claims.

[0019] It is disclosed a single chain polypeptide having dual activities of a cytokine targeting CSF1 R and of a cytokine of interest.In some aspects, the single chain polypeptide which comprises in the N-terminus to C-terminus direction, (i) a cytokine domain targeting CSF1R, a peptide linker L1, and a cytokine of interest, or (ii) a cytokine of interest, a peptide linker L1 , and a cytokine domain targeting CSF1R.

[0020] In some aspects, in the single chain polypeptide, the cytokine domain targeting CSF1R is a CSF1 monomer, a dimeric single chain CSF1 polypeptide, or an IL-34 monomer.

[0021] In some aspects, the cytokine domain targeting CSF1R is:

[0022] (i) an IL-34 monomer comprising sequence SEQ ID NO: 3, or a sequence at least 80% identical thereto and that retains IL-34 activity,

[0023] (ii) a CSF1 monomer comprising sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 101, or a sequence at least 80% identical thereto and that retains CSF1 activity,

[0024] (iii) a CSF1 monomer comprising sequence SEQ ID NO: 207, or a sequence at least 80% identical thereto and that retains CSF1 activity.

[0025] In some aspects, the cytokine monomer targeting CSF1R is a CSF1 monomer that comprises a sequence that differs from SEQ I D NO: 1 , SEQ I D NO: 2, or SEQ I D NO: 101: (i) by a substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132, and 140 in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 101, or

[0026] (ii) by substitution M65Y, S18I, F54G, L85I, Q58E, T64H, M65Y, R66Y, A74D, V78Y, Q81Y, N122Q, L56Y V120F, E5R, H9Q, R86Y, K93E, K100Y, W132N, K51Q, S18L, F54N, or Q58F.

[0027] In some aspects, the cytokine of interest is selected from the group consisting of IL-10, GM-CSF, IL-2, IL-4, IFN-a, IFN-p, IFN-y, IL-27, IL-33, IL-13, IL-24, IL-ip, IL-36, TNFa, TGFp, IL-6 and IL-35.

[0028] In some aspects, the single chain polypeptide is a dual CSF1-IFN-y cytokine, a dual IFN-y-CSF1 cytokine, a dual CSF1-IL-2 cytokine, a dual IL-2-CSF1 cytokine, a dual CSF1-IL-4 cytokine, a dual IL-4-CSF1 cytokine, a dual CSF1 -GM-CSF cytokine, a dual GM-CSF-CSF1 cytokine.

[0029] In some aspects, the single chain polypeptide comprises, from the N-terminus to the C-terminus direction, (i) a CSF1 domain, the cytokine of interest, and a Fc fragment, or (ii) a Fc fragment, the cytokine of interest, and the CSF1 domain

[0030] In some aspects, in the single chain polypeptide, one or more of the following conditions are met:

[0031] (i) the peptide linker L1 comprises from 5 to 50 amino acid residues;(ii) the peptide linker L1 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;

[0032] (iii) the peptide linker L1 comprises sequence SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, or SEQ ID NO: 16;

[0033] (iv) the peptide linker L1 comprises sequence (SEQ ID NO: 17)n, n being an integer from 1 to 10;

[0034] (v) the peptide linker L1 is a rigid peptide sequence composed of Glu, Ala, and Lys residues; or

[0035] (vi) the peptide linker L1 comprises sequence EAAAK (SEQ ID NO: 118), or (EAAAK; SEQ ID NO: 118)nwith n being a integer from 2 to 10, such as sequence SEQ ID NO: 119.

[0036] In some aspects, the single chain polypeptide is fused to a human immunoglobulin Fc fragment, or to albumin or albumin fragment, or to an anti-albumin antibody or a fragment thereof, or conjugated to a poly(ethylene glycol) (PEG) molecule.

[0037] It is further disclosed a protein comprising a single chain polypeptide as herein defined fused to an immunoglobulin Fc fragment, preferably a human immunoglobulin Fc fragment.

[0038] In some aspects, the protein comprises one single chain polypeptide having dualcytokine activities and one immunoglobulin Fc fragment.

[0039] In some aspects, the protein comprises two single chain polypeptides having dualcytokine activities and one immunoglobulin Fc fragment.

[0040] In some embodiments, the single chain polypeptide(s) having dual-cytokine activities and the immunoglobulin Fc fragment are fused through a peptide linker L2.

[0041] In some aspects, in the single chain polypeptide, one or more of the following conditions are met:

[0042] (i) the peptide linker L2 comprises from 3 to 45 amino acid residues;

[0043] (ii) the peptide linker L2 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;

[0044] (iii) the peptide linker L2 comprises sequence any one of sequences SEQ ID NO:

[0045] 15, SEQ ID NO: 17 to SEQ ID NO: 28, SEQ ID NO: 99, SEQ ID NO: 130, SEQ ID NO: 134;

[0046] (iv) the peptide linker L2 comprises sequence (SEQ ID NO: 17)p, p being an integer from 1 to 8;

[0047] (v) the peptide linker L2 is a rigid peptide sequence composed of Glu, Ala, and Lys residues; or(vi) the peptide linker L2 comprises sequence EAAAK (SEQ ID NO: 118), or (EAAAK; SEQ ID NO: 118)nwith n being a integer from 2 to 10, such as sequence SEQ ID NO: 119.

[0048] In some aspects of the protein, the Fc fragment or domain is from lgG1, lgG2, lgG3 or I gG4, silent or non-silent.

[0049] In some aspects, the protein comprises a first polypeptide chain comprising any one of sequence:

[0050] (i) SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or a sequence at least 80%% identical thereto and that retains CSF1 and IFN-y activities;

[0051] (ii) SEQ ID NO: 144, SEQ ID NO: 145, or a sequence at least 80% identical thereto wherein said protein retains CSF1 and IL-2 activities;

[0052] (iii) SEQ ID NO: 146, SEQ ID NO: 147, or a sequence at least 80% identical thereto and that retains CSF1 and IL-4 activities;

[0053] (iv) SEQ ID NO: 148 to SEQ ID NO: 151, or a sequence at least 80% identical thereto and that retains CSF1 and GM-CSF activities.

[0054] In some aspects, the protein further comprises a second polypeptide chain comprising SEQ ID NO: 105, or a sequence at least 80% identical thereto, wherein the first and second polypeptides dimerise.

[0055] It is further disclosed a pharmaceutical composition comprising the single chain polypeptide or the protein as herein defined, and a pharmaceutically acceptable carrier.

[0056] It is further disclosed the single chain polypeptide, the protein, or the pharmaceutical composition as herein defined, for use as a medicament.

[0057] It is further disclosed the single chain polypeptide, the protein, or the pharmaceutical composition as herein defined, for use for treating a disease comprising cancers, infectious diseases, inflammatory diseases, auto-immune diseases, neuro-inflammatory diseases, infectious diseases and tissue repair.

[0058] Detailed description of the invention

[0059] Main definitions

[0060] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, “a ribonucleotide” is understood to represent one or more ribonucleotides. As such, the terms “a,” “an,” “one or more,” and “at least one” can be used interchangeably hereinThroughout this specification and embodiments, the words “have” and “comprise,” or variations such as “has,” “having,” “comprises,” or “comprising” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. It is further understood that wherever embodiments are described herein with the language “comprising” or “having,” or grammatical equivalents thereof, otherwise analogous embodiments described in terms of “consisting of” and / or “consisting essentially of” are also provided.

[0061] As used herein “IL-34 activity” or “IL-34 activities” denotes one or more biological activities mediated by activation of CFS1-R by IL-34. These comprise or consist of (i) the activation of CFS1-R at the surface of monocytes or progenitors thereof, and / or (ii) the differentiation of monocytes into macrophages, preferentially both.

[0062] As used herein “CSF1 activity” or “CSF1 activities” denotes one or more biological activities mediated by activation of CFS1-R by CSF1. These comprise or consist of (i) the activation of CFS1-R at the surface of monocytes or progenitors thereof, and / or (ii) the differentiation of monocytes into macrophages, preferentially both

[0063] As used herein ‘IL-10 activity” or “IL-10 activities” denotes one or more biological activities mediated by IL-10 through binding to its receptor, IL-10R. These comprise or consist of (i) the binding to IL-1 OR on myeloid cells, in particular monocytes, or (ii) antiinflammatory activities, or preferentially both. Anti-inflammatory activities include (a) the inhibition of production of pro-inflammatory mediators (e.g. pro-inflammatory cytokines such as tumour necrosis factor-a (TNF-a), IL-1, IL-12, IL-6 and granulocyte-macrophage colonystimulating factor, inflammatory enzymes such as cyclo-oxygenase 2 and inducible nitric oxide synthase, chemokines such as RANTES, membrane inflammatory protein-1 a (MIP1a), IL-8, and eotaxin), and / or (b) the inhibition of expression of MHCII and costimulatory molecules by myeloid cells, and / or (c) the induction of differentiation of monocytes into tolerogenic macrophages. In some embodiments, anti-inflammatory activities comprise inhibition of production of TNF-a and / or IL-6 by activated monocytes. In some embodiments, anti-inflammatory activities comprise induction of differentiation of monocytes into tolerogenic macrophages. Tolerogenic macrophages have M2-like phenotype and can be identified by their regulatory phenotype and high levels of expression of CD206 and CD163. In some embodiments, IL-10 activities comprise or consist of binding to IL-10R on myeloid cells, in particular monocytes, inhibition of production of TNF-a and / or IL-6 by activated monocytes, and differentiation of monocytes into tolerogenic macrophages.

[0064] As used herein “GM-CSF activity” or “GM-CSF activities” denotes one or more biological activities mediated by activation of GM-CSF receptor, also known as CD116, byGM-CSF. CD116 is primarily present in the immune cells like granulocytes, endothelial cells and macrophages. These comprise or consists of the stimulation of growth and differentiation for cells in the granulocyte, monocyte / macrophage and eosinophil lineage.

[0065] As used herein “IL-2 activity” or “IL-2 activities” denotes one or more biological activities mediated by activation of trimeric IL-2 receptor (IL-2R), consisting of IL-2Ra (CD25), IL-2RP (CD122), and the common gamma chain (CD132), or dimeric IL-2Rs, made of CD122 and CD132, by IL-2. Trimeric IL-2R is mainly expressed on immunosuppressive regulatory T (Treg) cells. Trimeric IL-2R is mainly present on resting, antigen-experienced (memory) effector T (Teff) and natural killer (NK) cells. IL-2 has pleiotropic effects on the immune system: IL-2 activities include (i) stimulation of proliferation and activation of regulatory cells (essentially at low dose II-2), and (ii) stimulation of proliferation and activation of effector cells (essentially at high dose IL-2).

[0066] As used herein “IL-4 activity” or “IL-4 activities” denotes one or more biological activities mediated by activation of IL-4 type I and / or type II receptor by IL-4. IL-4 orchestrates both regulatory and stimulatory immune effects. As used herein, IL-4 activity / activities comprises or consists of triggering the differentiation of naive T cells into T helper 2 (TH2) cells and suppressing type 1 inflammation.

[0067] As used herein, “IFN-a activity” or “IFN-a activities” denotes one or more biological activities mediated by activation of IFN-a / p receptor subunit (IFNAR)1 / IFNAR2 heterodimer by IFN-a. These comprise or consist of (i) inducing antiviral innate immune responses, and (ii) antitumoral activity.

[0068] As used herein, “IFN- activity” or “IFN- activities” denotes one or more biological activities mediated by activation of IFN-a / p receptor subunit (IFNAR)1 / IFNAR2 heterodimer by IFN-a. IFN-a and -p bind to the same receptor. IFN-exerts immunomodulatory effects that include reduction of pathogenic Th1 and Th17 CD4+ T cells, and increase in IL-10 producing Treg cells.

[0069] As used herein, “IFN-y activity” or “IFN-y activities” denotes one or more biological activities mediated by activation of its receptor composed of two subunits, IFNGR1 and IFNGR2, by IFN-y. These comprise or consists of (i) promoting antiviral activity, (ii) facilitating macrophage activation, (iii) controlling Th1 / Th2 balance, or (iv) regulating cellular apoptosis and proliferation.

[0070] As used herein, “IL-27 activity” or “IL-27 activities” denotes one or more biological activities mediated by activation of its receptor by IL-27. IL-27 is a heterodimeric, cytokine composed of EBI3 (IL-27B) and p28 (IL-27A) subunits, and interacts with a heterodimeric receptor composed of WSX-1 and gp130. The activities comprise or consist of promoting naive CD4 T cell proliferation and Th1 differentiation.As used herein, “IL-35 activity” or “IL-35 activities” denotes one or more biological activities mediated by activation of its receptor by IL-35. IL-35 is a heterodimeric, cytokine composed of EBI3 (IL-27B) and p35 (IL-12A) subunits, and can signal through at least 4 receptors (2 homodimers: I L-12Rp2 / l L-12Rp2 and gp130 / gp130, and 2 heterodimers: IL-12Rp2 / gp130 and IL-12RP2 / WSX-1). IL-35 activities comprise or consist of immunosuppressive functions, including (i) reducing proliferation of TH1, TH2 and TH17 CD4+ T cells, (ii) reducing secretion of IFN-y, IL-17 IL-4 and / or IL-12 by CD4+ T cells, (iii) reducing proliferation of CD8+ T cells, (iv) reducing secretion of TH1 cytokines and expression of CD28 by CD8+ T cells, (v) reducing proliferation of B cells, or (vi) reducing IgG production by B cells.

[0071] As used herein, “IL-33 activity” or “IL-33 activities” denotes one or more biological activities mediated by activation of its receptor by IL-33. IL-33 is a member of the IL-1 family and interacts with the ST2 (IL1RL1) receptor and IL-1 receptor accessory protein (IL-1 RAcP). The activities comprise or consist of promoting type 2 immune responses, inducing M2 macrophage polarization, and amplifying IL-4 and IL-13 signaling.

[0072] As used herein, “IL-13 activity” or “IL-13 activities” denotes one or more biological activities mediated by activation of its receptor by IL- 13. IL- 13 is a type 2 cytokine that interacts with the IL-13 receptor alpha-1 (IL-13Ra1) and IL-4 receptor alpha (IL-4Ra). The activities comprise or consist of inducing M2 macrophage polarization, promoting tissue remodeling, and suppressing excessive inflammatory responses.

[0073] As used herein, “TGF- activity” or “TGF- activities” denotes one or more biological activities mediated by activation of its receptor by TGF-p. TGF-p is a multifunctional cytokine that interacts with the TGF-p receptor complex, composed of type I and type II serine / threonine kinase receptors. The activities comprise or consist of promoting M2 macrophage differentiation, suppressing pro-inflammatory cytokine production, modulating immune tolerance, and regulating tissue repair and fibrosis

[0074] As used herein, “IL-i activity” or “IL-i activities” denotes one or more biological activities mediated by activation of its receptor by IL-1 p. IL-1 p is a pro-inflammatory cytokine of the IL-1 family and interacts with the IL-1 receptor type 1 (IL-1R1) and IL-1 receptor accessory protein (IL-1RAcP). The activities comprise or consist of inducing M1 macrophage polarization, stimulating IL-6 and TNF-a production, promoting dendritic cell activation, and amplifying inflammatory responses.

[0075] As used herein, “IL-6 activity” or “IL-6 activities” denotes one or more biological activities mediated by activation of its receptor by IL-6. IL-6 is a pleiotropic cytokine that interacts with the IL-6 receptor (IL-6R) and the signal-transducing gp130 receptor. The activities comprise or consist of inducing acute-phase responses, regulating monocytedifferentiation, promoting pro-inflammatory cytokine production, and enhancing antigen presentation by dendritic cells.

[0076] As used herein, “TNF-a activity” or “TNF-a activities” denotes one or more biological activities mediated by activation of its receptor by TNF-a. TNF-a is a pro-inflammatory cytokine that interacts with TNF receptors TNFR1 (p55) and TNFR2 (p75), activating NF-KB and apoptosis pathways. The activities comprise or consist of inducing M1 macrophage activation, promoting inflammatory cytokine secretion, stimulating endothelial cell activation, and enhancing monocyte-to-dendritic cell differentiation.

[0077] As used herein, “IL-36 activity” or “IL-36 activities” denotes one or more biological activities mediated by activation of its receptor by IL-36. IL-36 cytokines belong to the IL-1 family and interact with IL-36 receptor (IL-36R, also known as IL-1R6) and IL-1 receptor accessory protein (IL-1RAcP). The activities comprise or consist of promoting M1 macrophage polarization, enhancing dendritic cell maturation, inducing pro-inflammatory cytokine secretion, and amplifying innate immune responses.

[0078] As used herein, “IL-24 activity” or “IL-24 activities” denotes one or more biological activities mediated by activation of its receptor by IL-24. IL-24 is a member of the IL-10 cytokine family and interacts with IL-20R1 / IL-20R2 and IL-22R1 / IL-20R2 receptor complexes. The activities comprise or consist of regulating monocyte differentiation, promoting macrophage anti-inflammatory responses, inducing apoptosis in tumor cells, and modulating wound healing processes.

[0079] An “immunoglobulin Fc fragment” or “Fc fragment” consists of two polypeptides linked by disulfide bonds, each polypeptide, from the N-terminal to C-terminal, being composed of a hinge region, a CH2 domain and a CH3 domain. The structure of the Fc fragment is nearly identical across all subtypes of human immunoglobulin. As used herein “an immunoglobulin Fc domain” or “Fc domain” denotes the monomeric polypeptide comprising immunoglobulin hinge and CH2, CH3 immunoglobulin heavy chain constant domains. Fc domain as used herein encompasses native Fc and Fc variants. The term "native Fc" as used herein refers to a molecule comprising the sequence of a non-antigen-binding fragment resulting from digestion of an antibody, or produced by other means, and that contain the hinge region. The original immunoglobulin source of the native Fc is preferably of human origin and can be any of the immunoglobulins, of isotype as IgM, I g D, IgG, IgA, or IgE, IgG has several subclasses, including, but not limited to, lgG1, lgG2, lgG3, and lgG4. The term "Fc variant" as used herein refers to a molecule or sequence that is modified from a native Fc but still comprises a binding site for the salvage receptor, FcRn (neonatal Fc receptor). Exemplary Fc variants, and their interaction with the salvage receptor, are known in the art. Thus, the term "Fc variant" can comprise a molecule orsequence that is humanized from a non-human native Fc. Furthermore, a native Fc comprises regions that can be removed because they provide structural features or biological activity that are not required for the antibody-like binding proteins of the invention. Thus, the term "Fc variant" comprises a molecule or sequence that lacks one or more native Fc sites or residues, or in which one or more Fc sites or residues has be modified, that affect or are involved in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC).

[0080] By ‘similar affinity’ it is meant herein the affinity preferably does not vary (i.e. increase or decrease) by more than a 5-fold, preferably 4-fold, 3-fold, 2-fold, or 1.5-fold factor compared to the reference level of affinity of the wild-type cytokine (IL-34).

[0081] By ‘higher’ or ‘reduced’ it is meant herein the affinity preferably varies (i.e. increase or decrease) by at least a 5-fold, preferably, 7-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 200-fold factor compared to the reference level of affinity of the wild-type cytokine (IL-34 or IL-10 as appropriate).

[0082] Determining cytokine activities can be readily performed by the skilled person using conventional methods in the art, such as HEK cell reported assays, as disclosed in the examples.

[0083] Single chain polypeptide having dual activities of a cytokine targeting CSF1R and of a cytokine of interest

[0084] A single chain polypeptide having dual activities of a cytokine targeting CSF1 R and of a cytokine of interest is provided.

[0085] In some embodiments, the single chain polypeptide comprises, in the N-terminus to C-terminus direction, a cytokine domain targeting CSF1R, a peptide linker L1, and a cytokine of interest.

[0086] In some embodiments, the single chain polypeptide comprises, in the N-terminus to C-terminus direction, a cytokine of interest, a peptide linker L1, and a cytokine domain targeting CSF1R.

[0087] In some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1 R and cytokine of interest comprises an additional cytokine domain targeting CSF1R.Methods for designing homo- or hetero-dimeric single chain polypeptide comprising helix-bundle cytokines of type I or Type II have been disclosed in the international patent application WO2023144393 A1, which content is incorporated by reference.

[0088] Cytokine domain targeting CSF1R

[0089] As used herein, a cytokine targeting CSF1R, or cytokine domain targeting CSF1R, denotes a CSF1 monomer or IL-34 monomer, or dimeric single chain CSF1 or IL-34 polypeptide.

[0090] In some embodiments, the cytokine domain targeting CSF1R targeting is a CSF1 monomer.

[0091] The CSF1 monomer incorporated into the single chain polypeptide having dual cytokine activities is a wild-type CSF1 monomer, a fragment, or a mutant thereof, or a circular perm utant thereof. Preferably, the CSF1 monomer is a human CSF1 monomer.

[0092] A canonical sequence of human CSF1 is available from the database UniProtKB under accession number P09603-1 (entry version 234 of 27 November 2024). The sequence P09603-1 of human CSF1 includes a signal peptide of 32 amino acids (SEQ ID NO: 42) and a 522 amino acid long mature polypeptide.

[0093] In some embodiments, the CSF1 monomer is a polypeptide sequence that comprises or consists of SEQ ID NO: 1 (amino acids E1-D150 of mature human CSF1 protein, also referred to as “short version ending in QD”), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity. In some embodiments, a polypeptide sequence of wild-type human CSF1 comprises or consists of SEQ ID NO: 2 or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0094] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of SEQ ID NO: 101 (amino acids E1-V151 of mature human CSF1 protein, also referred to as “short version ending in QDV” in Table 1), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0095] In some embodiments, the CSF1 monomer is a polypeptide that comprises or consists of SEQ ID NO: 2 (amino acids E1-R223 of mature human CSF1 protein, also referred to as “full length”) or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0096] In some embodiments, the CSF1 monomer is polypeptide that comprises or consists of a sequence that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101 by oneor more mutations (and is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101). In some embodiments, the one or more mutations in SEQ ID NO: 1 , SEQ ID NO: 2, or SEQ ID NO: 101, comprise or consist of (positions indicated based on mature human CSF1 protein sequence, or by reference to any one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101):

[0097] a) A substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132 and 140;

[0098] b) S18I;

[0099] c) F54G;

[0100] d) L85I;

[0101] e) Q58E;

[0102] f) T64H;

[0103] g) M65Y;

[0104] h) R66Y;

[0105] i) A74D;

[0106] j) V78Y;

[0107] k) Q81Y

[0108] l) N122Q;

[0109] m) L56Y;

[0110] n) V120F;

[0111] o) E5R;

[0112] p) H9Q;

[0113] q) R86Y;

[0114] r) K93E;

[0115] s) K100Y;

[0116] t) W132N;

[0117] u) K51Q;

[0118] v) F54N;

[0119] w) Q58F;

[0120] x) or any combination of mutations listed in b) to w).

[0121] In some embodiments the one or more mutations in SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101 comprise or consist of substitution M65Y, as present e.g. in variant ORK1-P-532.In some embodiments, the one or more mutations in SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101 comprise or consist of (positions indicated based on mature human CSF1 protein sequence SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101):

[0122] a) A substitution at positions i) 18, 64, 65, 66, 85, and 120; ii) 5, 18, 66, 78, 85, 86, and 120; iii) 18, 64, 66, 78, 85, 86, and 120; iv) 18, 64, 74, 85, 86, and 120; v) 18, 64, 66, 85, and 86; vi) 18, 64, 66, 74, and 85; vii) 5, 64, 66, 85, and 86; viii) 18, 64, 66, 74, 85, 86, and 120; or ix) 18, 64, 66, 74, 86, and 120;

[0123] b) S18I, T64H, M65F, R66V, L85I, R86Y, V120F;

[0124] c) E5R, S18I, T64H, R66L, L85V, R86Y, V120F;

[0125] d) S18I, T64H, R66L, V78Y, L85I, R86Y, V120F;

[0126] e) S18I, T64H, R66L, L85I, R86Y, V120F;

[0127] f) S18I, T64H, R66Y, L85I, R86Y, V120F;

[0128] g) S18I, T64H, A74D, L85I, R86Y, V120F;

[0129] h) S18I, T64H, R66I, L85I, R86Y;

[0130] i) S18I, T64H, R66V, A74D, L85I;

[0131] j) E5R, T64H, R66L, L85I, R86Y;

[0132] k) S18I, T64H, R66V, A74D, L85V, R86Y, V120F;

[0133] l) S18I, T64H, R66I, A74D, L85V, R86Y, V120F;

[0134] m) S18I, T64H, R66V, A74D, L85I, R86Y, V120F;

[0135] n) S18I, T64H, R66I, A74D, L85I, R86Y, V120F;

[0136] o) S18I, T64H, R66V, A74D, R86Y, V120F, or

[0137] p) S18I, T64H, R66V, A74D, L85Y, R86Y, V120F.

[0138] In some embodiments, the one or more mutations in SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101 comprise or consist of substitutions:

[0139] S18I, T64H, R66V, A74D, L85V, R86Y, and V120F, as present e.g. in variant ORK1-P-576;

[0140] S18I, T64H, R66V, A74D, L85I, R86Y, and V120F, as present e.g. in variant ORK1-P-578; or

[0141] S18I, T64H, R66I, A74D, L85I, R86Y, and V120F, as present e.g. in variant ORK1-P-579.

[0142] In some embodiment the CSF1 monomer comprises or consists of a sequence that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101 by no other mutation or substitution than the one defined above.

[0143] In some embodiments, the CSF1 monomer is a CSF1 circular permutant comprising or consisting of sequence SEQ ID NO: 207, or a sequence at least 80%, at least 85%, atleast 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0144] The dimeric single chain CSF1 polypeptide may comprise a substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132, and 140 of SEQ ID NO: 207, or any specific substitution or combination of substitutions as described above in respect to SEQ ID NO: 1 , SEQ ID NO: 2, or SEQ ID NO: 101.

[0145] In some embodiments, the cytokine domain targeting CSF1R is a dimeric single chain CSF1 polypeptide.

[0146] In some embodiments, the dimeric single chain CSF1 polypeptide comprise or consists of sequence SEQ ID NO: 201, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 activity.

[0147] In some embodiments, the cytokine domain targeting CSF1R is an IL-34 monomer.

[0148] The IL-34 monomer incorporated into the single chain polypeptide having IL- 10 and IL-34 activities may be a wild-type IL-34 monomer, a fragment, or a mutant thereof.

[0149] A canonical sequence of human IL-34 is available from the database UniProtKB under accession number Q6ZMJ4-1 (entry version 144 of 27 November 2024). The sequence Q6ZMJ4-1 of human IL-34 includes a signal peptide of 20 amino acids (SEQ ID NO: 71) and a 222 amino acid long mature polypeptide.

[0150] In some embodiments, a polypeptide sequence of IL-34 thus comprises or consists of sequence SEQ ID NO: 3 (amino acids N1-P222 of mature human IL-34 protein), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-34 activity.

[0151] In some embodiments, the IL-34 monomer is a mutant IL-34 that has similar or higher affinity to CSF1R compared to wild-type IL-34.

[0152] Cytokine of interest

[0153] In some embodiments, a “cytokine of interest” denotes a cytokine exercising a biological activity of interest on myeloid cells.

[0154] In some embodiments, the cytokine of interest is selected from the group consisting of IL-10, GM-CSF, IL-2, IL-4, IFN-a, IFN-p, IFN-y, IL-27, IL-35, TNF-a, IL-ip, IL-6, IL-36, IL-13, TGF-p, IL-24 and IL-33. In some embodiments, the cytokine of interest is selected from the group consisting of IL-10, GM-CSF, IL-2, IL-4, and IFN-y.In some embodiments, the cytokine of interest is selected from the group consisting of GM-CSF, IL-2, IL-4, IFN-a, IFN- , IFN-y, IL-27, IL-35, TNF-a, IL-ip, IL-6, IL-36, IL-13, TGF-p, IL-24 and IL-33. In some embodiments, the cytokine of interest is selected from the group consisting of GM-CSF, IL-2, IL-4, and IFN-y.

[0155] In some embodiments, the cytokine of interest is IFN-y. In some embodiments, the cytokine of interest is GM-CSF. In some embodiments, the cytokine of interest is IL-2. In some embodiments, the cytokine of interest is IL-4.

[0156] In some embodiments, the cytokine of interest is IL-10.

[0157] According to an embodiment, the cytokine of interest is different from IL-10. According to this embodiment, a single chain polypeptide having dual activities of a cytokine targeting CSF1R and IL- 10, in particular dual CSF1-IL-10 activities or dual IL-34-IL-10 activities, is excluded.

[0158] In some embodiments, the cytokine of interest is a cytokine monomer. In some embodiments, the cytokine of interest is an homodimeric single chain cytokine.

[0159] In some embodiments, the cytokine of interest is IL-10.

[0160] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL- 10 is provided

[0161] The single chain polypeptide comprises an IL-10 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-10 domain is an IL-10 monomer or a single chain dimeric IL-10.

[0162] The IL-10 may be an IL-10 monomer as shown in SEQ ID NO: 4, or a mutant thereof, or a polypeptide comprising the sequence of mature IL-10 and on the N-terminal end 1, 2, 3 or more additional amino acids residues consecutively present in the signal peptide of IL-10 (i.e. A, RA or VRA).

[0163] The IL-10 may be a single chain dimeric IL-10, such as described in the international patent application WO2023144393 A1. In some embodiments, single chain dimeric IL-10 comprises sequence SEQ ID NO: 5, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL-10 receptor. In some embodiments, single chain dimeric IL-10 comprises sequence SEQ ID NO: 6, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL- 10 receptor.

[0164] In some embodiments, the cytokine of interest is GM-CSF.A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and GM-CSF is provided.

[0165] The single chain polypeptide comprises a GM-CSF domain fused to an IL-34 monomer or CSF1 monomer, wherein the GM-CSF domain is a GM-CSF monomer or a single chain dimeric GM-CSF.

[0166] In some embodiments, the GM-CSF domain is a GM-CSF monomer, in particular human GM-CSF. A reference sequence of human GM-CSF is available in Uniprot database, under accession number P04141 (entry version 219 of 27 November 2024). In some embodiments, the GM-CSF monomer is a polypeptide that comprises or consists of sequence SEQ ID NO: 199, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains GM-CSF activity.

[0167] In some embodiments, the GM-CSF domain is a dimeric single chain GM-CSF, such as disclosed in the international patent application WO2023144393 A1.

[0168] In some embodiments, the cytokine of interest is IL-2.

[0169] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-2 is provided.

[0170] The single chain polypeptide comprises an IL-2 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-2 domain is an IL-2 monomer or a single chain dimeric IL-2.

[0171] In some embodiments, the IL-2 domain is an IL-2 monomer, in particular human IL-2. A reference sequence of human IL-2 is available in Uniprot database, under accession number P60568 (entry version 201 of 27 November 2024). In some embodiments, the IL-2 monomer is a polypeptide that comprises or consists of sequence SEQ ID NO: 200, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-2 activity.

[0172] In some embodiments, the IL-2 domain is a dimeric single chain IL-2, such as disclosed in the international patent application WO2023144393 A1.

[0173] In some embodiments, the cytokine of interest is IL-4.

[0174] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-4 is provided.

[0175] The single chain polypeptide comprises an IL-4 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-4 domain is an IL-4 monomer or a single chain dimeric IL-4.In some embodiments, the IL-4 domain is an IL-4 monomer, in particular human IL-4. A reference sequence of human IL-4 is available in Uniprot database, under accession number P05112 (entry version 249 of 27 November 2024, SEQ ID NO: 135). The sequence of human IL-4 includes a signal peptide of 24 amino acids and a 129 amino acid long mature polypeptide. In some embodiments, the IL-4 monomer is a polypeptide that comprises or consists of sequence SEQ ID NO: 136 (amino acids H1-S129 of mature human IFN-y protein), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IFN-y activity. In some embodiments, the IL-4 monomer is a mutated IL-4 with substitution T13D, i.e. a polypeptide that comprises or consists of sequence SEQ ID NO: 137.

[0176] In some embodiments, the IL-4 domain is a dimeric single chain IL-4, such as disclosed in the international patent application WO2023144393 A1.

[0177] In some embodiments, the cytokine of interest is IFN-a.

[0178] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IFN-a is provided.

[0179] The single chain polypeptide comprises an IFN-a domain fused to an IL-34 monomer or CSF1 monomer, wherein the IFN-a domain is an IFN-a monomer or a single chain dimeric IFN-a.

[0180] In some embodiments, the IFN-a domain is an IFN-a monomer, in particular human IFN-a. In some embodiments, IFN-a is selected from the group consisting of IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17 and IFNA21.

[0181] In some embodiments, the IFN-a domain is a dimeric single chain IFNa1 / 13 or dimeric single chain IFNa2, such as disclosed in the international patent application WO2023144393 A1.

[0182] In some embodiments, the cytokine of interest is IFN- .

[0183] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IFN-p is provided.

[0184] The single chain polypeptide comprises an IFN-p domain fused to an IL-34 monomer or CSF1 monomer, wherein the IFN-p domain is an IFN-p monomer or a single chain dimeric IFN-p.In some embodiments, the I FN-p domain is an I FN- monomer, in particular human IFN-p. A reference sequence of human I FN-p is available in Uniprot database, under accession number P01574 (entry version 235 of 27 November 2024).

[0185] In some embodiments, the IFN-p domain is a dimeric single chain IFN-p, such as disclosed in the international patent application WO2023144393 A1.

[0186] In some embodiments, the cytokine of interest is IFN-y.

[0187] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IFN-y is provided.

[0188] The single chain polypeptide comprises an IFN-y domain fused to an IL-34 monomer or CSF1 monomer, wherein the IFN-y domain is an IFN-y monomer or a single chain dimeric IFN-y.

[0189] In some embodiments, the IFN-y domain is an IFN-y monomer, e.g. a wild-type IFN-y monomer, a fragment or mutant thereof. Preferably, the IFN-y monomer is a human IFN-y monomer, a fragment or mutant thereof.. A reference sequence of human IFN-y is available in Uniprot database, under accession number P01579 (entry version 250 of 27 November 2024, SEQ ID NO: 120). The sequence of human IFN-y includes a signal peptide of 23 amino acids and a 143 amino acid long mature polypeptide.

[0190] In some embodiments, the IFN-y monomer is a polypeptide that comprises or consists of sequence SEQ ID NO: 121 (amino acids Q1-G138 of mature human IFN-y protein), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IFN-y activity.

[0191] In some embodiments, the IFN-y domain is a dimeric single chain IFN-y, such as disclosed in the international patent application WO2023144393 A1.

[0192] In some embodiments the dimeric single chain IFN-y polypeptide comprises or consists of two IFN-y monomers (i.e. two wild-type IFN-y monomers (in particular human IFN-y),afragment or mutant thereof) linked by a peptide linker (herein called L1.1 linker).

[0193] In some embodiments the dimeric single chain IFN-y comprises or consists of two IFN-y monomers of sequence SEQ ID NO: 121, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IFN-y activity, linked by a peptide linker (linker L1.1). In some embodiments the dimeric single chain IFNy comprises or consists one IFNy monomer of sequence SEQ ID NO: 13 linked by a peptide linker (linker L1.1) to an IFNy monomer consisting of amino acids amino acids Q1-R137 of SEQ ID NO: 121.

[0194] The L1.1 linker sequence bridging the C-terminus of one IFN-y monomer and the N-terminus of the other IFN-y monomer may have any suitable sequence. In someembodiments the L1.1 linker is a (flexible) peptide sequence composed of Gly and Ser residues, in different proportions. The L1.1 linker comprises from (or consists of) 10 to 45 amino acid residues, preferably 10 to 30 amino acids, still preferably 10 to 15 amino acids. According to some embodiments, the L1.1 linker comprises or consists of a sequence (GGGGS, SEQ ID NO: 17)n, n being a integer from 1 to 10, e.g. from 3 to 6. According to an embodiment, the L1.1 linker comprises or consists of GGGGSGGGGS (SEQ ID NO: 24).

[0195] In an embodiment the dimeric single chain IFN-y comprises or consists of sequence SEQ ID NO: 122, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IFN-y activity.

[0196] In some embodiments, the cytokine of interest is IL-27.

[0197] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-27 is provided.

[0198] The single chain polypeptide comprises an IL-27 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-27 domain is an IL-27 monomer (e.g., IL-27A or IL-27B) or a single chain dimeric IL-27. A reference sequence of human IL-27A is available in Uniprot database, under accession number Q8NEV9 (entry version 136 of 27 November 2024). A reference sequence of human IL-27B is available in Uniprot database, under accession number Q14213 (entry version 191 of 27 November 2024).

[0199] In some embodiments, the IL-27 domain is a dimeric single chain IL-27, such as disclosed in the international patent application WO2023144393 A1.

[0200] In some embodiments, the cytokine of interest is IL-35.

[0201] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-35 is provided.

[0202] The single chain polypeptide comprises an IL-35 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-35 domain is an IL-35 monomer (e.g., IL-12A or IL-27B) or a single chain dimeric IL-35. A reference sequence of human IL-127A is available in Uniprot database, under accession number P29459 (entry version 202 of 27 November 2024).

[0203] In some embodiments, the cytokine of interest is TNF-a.

[0204] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and TNF-a is provided. The single chain polypeptide comprises a TNF-a domainfused to an IL-34 monomer or CSF1 monomer, wherein the TNF-a domain is a TNF-a monomer or a single chain dimeric TNF-a.

[0205] In some embodiments, the TNF-a domain is a TNF-a monomer, in particular human TNF-a. A reference sequence of human TNF-a is available in the Uniprot database, under accession number P01375 (entry version 277 of 23 January 2025). In some embodiments, the TNF-a domain is a dimeric single chain TNF-a, such as disclosed in the international patent application WO2023144393 A1.

[0206] In some embodiments, the cytokine of interest is IL-1 .

[0207] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-10 is provided.

[0208] The single chain polypeptide comprises an IL-10 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-1 domain is an IL-1 monomer or a single chain dimeric IL-10.

[0209] In some embodiments, the IL-10 domain is an IL-10 monomer, in particular human IL-10. A reference sequence of human IL-10 is available in the Uniprot database, under accession number P01584 (entry version 235 of 10 December 2024). In some embodiments, the IL-10 domain is a dimeric single chain IL-10, such as disclosed in the international patent application WO2023144393 A1.

[0210] In some embodiments, the cytokine of interest is IL-6.

[0211] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-6 is provided.

[0212] The single chain polypeptide comprises an IL-6 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-6 domain is an IL-6 monomer or a single chain dimeric IL-6.

[0213] In some embodiments, the IL-6 domain is an IL-6 monomer, in particular human IL-6. A reference sequence of human IL-6 is available in the Uniprot database, under accession number P05231 (entry version 273 of 17 January 2025). In some embodiments, the IL-6 domain is a dimeric single chain IL-6, such as disclosed in the international patent application WO2023144393 A1.

[0214] In some embodiments, the cytokine of interest is IL-36 (IL-36a, IL-360, IL-36y). A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-36 is provided.The single chain polypeptide comprises an IL-36 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-36 domain is an IL-36 monomer or a single chain dimeric IL-36.

[0215] In some embodiments, the IL-36 domain is an IL-36 monomer, in particular human IL-36. A reference sequence of human IL-36a, IL-36P, and I L-36y is available in the Uniprot database under accession numbers Q9UHA7, Q9UBH0, and Q9NZH8, respectively (entry version 190 of 22 February 2025).

[0216] In some embodiments, the IL-36 domain is a dimeric single chain IL-36, such as disclosed in the international patent application WO2023144393 A1.

[0217] In some embodiments, the cytokine of interest is IL-13.

[0218] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-13 is provided.

[0219] The single chain polypeptide comprises an IL-13 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-13 domain is an IL-13 monomer or a single chain dimeric IL-13.

[0220] In some embodiments, the IL-13 domain is an IL-13 monomer, in particular human IL-13. A reference sequence of human IL-13 is available in the Uniprot database, under accession number P35225 (entry version 248 of 12 March 2025).

[0221] In some embodiments, the IL-13 domain is a dimeric single chain IL-13, such as disclosed in the international patent application WO2023144393 A1.

[0222] In some embodiments, the cytokine of interest is TGF- .

[0223] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and TGF-p is provided.

[0224] The single chain polypeptide comprises a TGF-p domain fused to an IL-34 monomer or CSF1 monomer, wherein the TGF-p domain is a TGF-p monomer or a single chain dimeric TGF-p.

[0225] In some embodiments, the TGF-p domain is a TGF-p monomer, in particular human TGF-pi. A reference sequence of human TGF-pi is available in the Uniprot database, under accession number P01137 (entry version 306 of 5 February 2025). In some embodiments, the TGF-p domain is a dimeric single chain TGF-p, such as disclosed in the international patent application WO2023144393 A1.

[0226] In some embodiments, the cytokine of interest is IL-33.A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-33 is provided.

[0227] The single chain polypeptide comprises an IL-33 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-33 domain is an IL-33 monomer or a single chain dimeric IL-33. In some embodiments, the IL-33 domain is an IL-33 monomer, in particular human IL-33. A reference sequence of human IL-33 is available in the Uniprot database, under accession number 095760 (entry version 210 of 14 January 2025).

[0228] In some embodiments, the IL-33 domain is a dimeric single chain IL-33, such as disclosed in the international patent application WO2023144393 A1.

[0229] In some embodiments, the cytokine of interest is IL-24.

[0230] A single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1R and IL-24 is provided.

[0231] The single chain polypeptide comprises an IL-24 domain fused to an IL-34 monomer or CSF1 monomer, wherein the IL-24 domain is an IL-24 monomer or a single chain dimeric IL-24. In some embodiments, the IL-24 domain is an IL-24 monomer, in particular human IL-24. A reference sequence of human IL-24 is available in the Uniprot database, under accession number Q13007 (entry version 201 of 18 March 2025).

[0232] In some embodiments, the IL-24 domain is a dimeric single chain IL-24, such as disclosed in the international patent application WO2023144393 A1.

[0233] Exemplary sequences are disclosed in the appended sequences for TNF-a, I L- 1 p, IL-6, IL-36, IL-13, TGF-p, IL-24, and IL-33.

[0234] Peptide linker L1

[0235] In some embodiments the cytokine targeting CSF1R and a cytokine of interest are linked through a linker L1 that comprises from (or consists of) 5 to 50, or from 10 to 45 amino acid residues, preferably 12 to 40 amino acids, still preferably 15 to 40 amino acids, still preferably 20 to 35 amino acids, even more preferably 25 to 35 amino acids.

[0236] Preferably, the linker L1 bridging the cytokine targeting CSF1R to the N-terminus of the cytokine of interest, or bridging the N-terminus of the cytokine of interest to the C-terminus of the cytokine targeting CSF1R, is a (flexible) peptide sequence composed of Gly and Ser residues, in different proportions. Examples of suitable linkers comprise or consist of GGSGGSGGSGGSGGG (15 amino acid long, SEQ ID NO: 13), GGGSGGSGGSGGSGGSGGSGGSGGG (25 amino acid long, SEQ ID NO: 14), GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (30 amino acid long, SEQ ID NO: 15),or GGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGG (35 amino acid long, SEQ ID NO: 16). In some embodiments the L1 linker comprises or consists of a sequence (GGGGS, SEQ ID NO: 17)n, n being a integer from 1 to 10, such as from 2 to 10, or from 3 to 6.

[0237] In some embodiments, the linker L1 is a (rigid) peptide sequence composed of Glu, Ala, and Lys residues. Examples of suitable linkers comprise or consist of sequence EAAAK (SEQ ID NO: 118), or (EAAAK; SEQ ID NO: 118)nwith n being an integer from 2 to 10, e.g. from 2 to 3 or from 3 to 6, such as n = 6 (i.e. sequence SEQ ID NO: 119).

[0238] In some embodiments, the linker L1 is a short linker, such as 5-15 amino acid long, or 5-10 amino acid long, or 5 amino acid long, either flexible or rigid, as described above. Short L1 linkers improve the selectivity of the single chain polypeptides for monocytes over T cells. Either to induce M2 a regulatory phenotype of macrophages (the case of ORK1) or to induce a M1 pro-inflammatory phenotype (in the case of ORK9).

[0239] Resulting single chain polypeptide having dual cytokine activities

[0240] CSF1R and IFN-y

[0241] In some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1R and IFN-y incorporates a combination of any of the cytokine targeting CSF1R, IFN-y cytokine domain, and peptide linker L1 as defined above. In some embodiments the single chain polypeptide has dual CSF1 and IFN-y activities. In some embodiments the single chain polypeptide has dual IL-34 and IFN-y activities.

[0242] The single chain polypeptide having dual CSF1 and IFN-y activities comprises a CSF1 monomer or dimeric single chain CSF1 polypeptide according to any of the embodiments described above. The single chain polypeptide having dual CSF1 and IFN-y activities further comprises an IFN-y monomer or dimeric single chain IFN-y polypeptide according to any of the embodiments described above.

[0243] According to some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1 R and IFN-y comprises or consists of:

[0244] i) a CSF1 monomer-L1-dimeric single chain IFNy polypeptide, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 123, ii) dimeric single chain IFNy polypeptide-L1-CSF1 monomer, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 124, iii) a CSF1 monomer-L1-IFNy monomer, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 7 or SEQ ID NO: 208,

[0245] iv) a IFNy monomer-L1-CSF1 monomer, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 8, or SEQ ID NO: 209,

[0246] v) a CSF1 monomer-L1-dimeric single chain IFNy-L1-CSF1 monomer, whereinthe two L1 linkers may be identical or different, preferably identical, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 210, vi) a CSF1 monomer-L1-CSF1 circular permutant-L1 -dimeric single chain I FNy, wherein the two L1 linkers may be identical or different, preferably identical, such as a polypeptide comprising or consisting of sequence SEQ ID NO: 211 , vii) or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 and IFNy activities.

[0247] CSF1R and IL-2

[0248] In some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1R and IL-2 incorporates a combination of any of the cytokine targeting CSF1R, IL-2 cytokine domain, and peptide linker L1 as defined above. In some embodiments the single chain polypeptide has dual CSF1 and IL-2 activities. In some embodiments the single chain polypeptide has dual IL-34 and IL-2 activities.

[0249] The single chain polypeptide having dual CSF1 and IL-2 activities comprises a CSF1 monomer according to any of the embodiments described above. The single chain polypeptide having dual CSF1 and IL-2 activities further comprises an IL-2 monomer as described above.

[0250] In some embodiments, the single chain polypeptide having dual cytokine activities comprises or consists of a sequence SEQ ID NO: 9 (CSF1JL-2 in QRK10_P_01), SEQ ID NO: 10 (IL-2_CSF1 in QRK10_P_02), or a sequence at least at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain the dual CSF1 and IL-2 activities.

[0251] CSF1R and IL-4

[0252] In some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1R and IL-4 incorporates a combination of any of the cytokine targeting CSF1R, IL-4 cytokine domain, and peptide linker L1 as defined above. In some embodiments the single chain polypeptide has dual CSF1 and IL-4 activities. In some embodiments the single chain polypeptide has dual IL-34 and IL-2 activities.

[0253] The single chain polypeptide having dual CSF1 and IL-4 activities comprises a CSF1 monomer according to any of the embodiments described above. The single chain polypeptide having dual CSF1 and IL-4 activities further comprises an IL-4 monomer as described above.In some embodiments, the single chain polypeptide having dual cytokine activities comprises or consists of a sequence SEQ ID NO: 11 (CSF1-IL-4 in QRK11_P_01), SEQ ID NO: 12 (IL-4-CSF1 in ORK11_P_02), or a sequence at least at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain the dual CSF1 and IL-4 activities.

[0254] CSF1R and GM-CSF

[0255] In some embodiments, the single chain polypeptide having dual activities of a cytokine targeting CSF1R and GM-CSF incorporates a combination of any of the cytokine targeting CSF1R, GM-CSF cytokine domain, and peptide linker L1 as defined above. In some embodiments the single chain polypeptide has dual CSF1 and GM-CSF activities. In some embodiments the single chain polypeptide has dual IL-34 and GM-CSF activities.

[0256] The single chain polypeptide having dual CSF1 and GM-CSF activities comprises a CSF1 monomer according to any of the embodiments described above. The single chain polypeptide having dual CSF1 and GM-CSF activities further comprises a GM-CSF monomer as described above.

[0257] In some embodiments, the single chain polypeptide having dual cytokine activities comprises or consists of a sequence SEQ I D NO: 152 (CSF 1 -GMCSF in ORK12_P_01 and ORK12_P_03), SEQ ID NO: 153 (GMCSF-CSF1 in QRK12_P_02 and QRK12_P_04), ora sequence at least at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain the dual CSF1 and GM-CSF activities.

[0258] CSF1R and IL10

[0259] In some embodiments, the single chain polypeptide having dual cytokine activities comprises a sequence SEQ ID NO: 38 (CSF1-IL-10 of ORK1_455_P) or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 and IL- 10 activities.

[0260] In some embodiments, the single chain polypeptide having dual cytokine activities comprises a sequence SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 91, or SEQ ID NO: 92, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-34 and IL- 10 activities.

[0261] Modified dual-cytokine single chain polypeptide having dual cytokine activities and increased half-life

[0262] According to some embodiments, the single chain polypeptide having dual-cytokine activities is modified to increase its half-life. In some embodiments, said modificationcomprises one or more alterations selected from the group consisting of a fusion to an immunoglobulin Fc fragment, albumin, or any albumin-binding domain; conjugation with poly(ethylene glycol) (PEG) or other polymer-based moieties; or any other chemical or genetic modification known to extend protein stability and circulation time in vivo.

[0263] Accordingly, the invention further relates to a protein comprising the single chain polypeptide having dual-cytokine activities as herein defined fused to an immunoglobulin Fc fragment or to albumin, or albumin fragment, or to an anti-albumin antibody or a fragment thereof, or conjugated to a poly(ethylene glycol) (PEG) molecule.

[0264] In some embodiments, the protein comprises a single chain polypeptide having dualcytokine activities and an immunoglobulin Fc fragment, preferably a human immunoglobulin Fc fragment. In some embodiments, the immunoglobulin Fc fragment is a human lgG1 or lgG4 Fc fragment.

[0265] In some embodiments, the protein comprises a single chain having dual-cytokine activities and albumin or a fragment thereof, preferably human albumin. In some embodiments, the protein comprises a single chain having dual-cytokine activities and an anti-albumin antibody or a fragment thereof, preferably an anti-human albumin antibody or a fragment thereof.

[0266] In some embodiments, the protein comprises a single chain polypeptide having dualcytokine activities conjugated to PEG. Process for protein pegylation has been reviewed for instance by Pfister and Morbidelli (Journal of Controlled Release, Volume 180, 2014, Pages 134-149).

[0267] Protein comprising one single chain polypeptide having dual-cytokine activities and one immunoglobulin Fc fragment

[0268] In some embodiments, the protein comprises one SC polypeptide having dualcytokine activities and one immunoglobulin Fc fragment. In some embodiments, the SC polypeptide having dual-cytokine activities and the immunoglobulin Fc fragment are fused through a peptide linker L2. In this aspect, the protein comprises a first polypeptide comprising the SC polypeptide having dual activities fused to a first immunoglobulin Fc fragment (Fc1) through a peptide linker L2, and a second polypeptide comprising a second immunoglobulin Fc fragment (Fc2).

[0269] In some embodiments, the protein comprises one SC polypeptide having dualcytokine activities and one immunoglobulin Fc fragment, wherein the SC polypeptide having dual-cytokine activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting the C-terminus of the SC polypeptide having dual-cytokine activities with the N-terminus of one hinge region of the Fc fragment.According to some aspects the protein then comprises two polypeptide chains comprising or consisting of formula (I):

[0270] (i) cytokine domain targeting CSF1R monomer-L1 -cytokine of interest-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0271] (ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0272] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. According to some aspects, the protein also comprises two polypeptide chains comprising or consisting of formula (II):

[0273] (i) cytokine of interest-L1 -cytokine domain targeting CSF1R monomer -L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0274] (ii) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0275] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0276] In some embodiments, the protein comprises one SC polypeptide having dualcytokine activities and one immunoglobulin Fc fragment, wherein the SC polypeptide having dual-cytokine activities is fused to the immunoglobulin Fc fragment through a peptide linker L2 connecting the C-terminus of the one of the CH3 domains of the Fc fragment with the N-terminus of the SC polypeptide having dual-cytokine activities.

[0277] According to some aspects, the protein then comprises two polypeptide chains comprising or consisting of formula (III):

[0278] (i) Fc1-L2- cytokine domain targeting CSF1R monomer-L1 -cytokine of interest, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0279] (i) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0280] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. According to some aspects, the protein then comprises two polypeptide chains comprising or consisting of formula (IV):

[0281] (i) Fc1-L2-cytokine of interest -L1-cytokine domain targeting CSF1R monomer, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0282] (i) Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0283] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0284] In some embodiments, the linker L2 connecting the single chain polypeptide having dual-cytokine activities and the immunoglobulin Fc fragment is preferably a peptide sequence that comprises from, or consists of, 3 to 45 amino acid residues, preferably 5 to 40 amino acid residues, preferably 5 to 25 amino acids, still preferably 10 to 25 amino acids.The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility, but linkers comprising amino acids randomly selected from the group consisting of valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartate, glutamate, asparagine, glutamine, glycine, and proline may also be suitable. A well-suited linker L2 according to the present disclosure contains or consists of glycine and serine residues and is for example of the format (GGGGS, SEQ ID NO: 17)p, wherein p is an integer from 1 to 8, notably from 1 to 4, advantageously 2, 3 or 4. In some embodiments, the linker L2 is (GGGGS)3(SEQ ID NO: 18). In some embodiments, the linker L2 is a (rigid) peptide sequence composed of Glu, Ala, and Lys residues. Examples of suitable linkers comprise or consist of sequence EAAAK (SEQ ID NO: 118), or (EAAAK; SEQ ID NO: 118)nwith n being an integer from 2 to 10, e.g. from 2 to from 3 to 6, such as n = 6 (i.e. sequence SEQ ID NO: 119).

[0285] In some embodiments, the linker is selected from the group consisting of the amino acid sequences SGGGGSGGGGS (SEQ ID NO: 19), SGGGGSGGGGSAP (SEQ ID NO: 20), NFSQP (SEQ ID NO: 21), KRTVA (SEQ ID NO: 22), GGGSGGGG (SEQ ID NO: 23), GGGGSGGGGS (SEQ ID NO: 24), GGGGSGGGGSGGGGS (SEQ ID NO: 18), THTCPPCPEPKSSDK (SEQ ID NO: 25), GGGS (SEQ ID NO: 26), EAAKEAAKGGGGS (SEQ ID NO: 27), EAAKEAAK (SEQ ID NO: 28), GGGGS (SEQ ID NO: 17), (SG)m where m is an integer comprised between 1 and 7, GGSSGSGSGSTGTSSSGTGTSAGTTGTSASTSGSGSGGGGGSGGGGSAGG (SEQ ID NO: 130), GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 99), GGGGGSGGGGSGGGGS (SEQ ID NO: 134), EAAAK (SEQ ID NO: 118), or (EAAA K; SEQ ID NO: 118)nwith n being a integer from 2 to 10, e.g. from 2 to 3 or from 3 to 6, such as n = 6 (i.e. sequence SEQ ID NO: 119).

[0286] The linker L1 is as defined above.

[0287] In some embodiments, the linker L1 comprises or consists of sequence SEQ ID NO: 15, and the linker L2 comprises or consists of sequence SEQ ID NO: 18.

[0288] The cytokine domain targeting CSF1R monomer and cytokine of interest are as defined above.

[0289] In some embodiments, the protein comprises a single chain polypeptide having dual activities of a cytokine targeting CSF1R and IFN-y. In some embodiments, the linker L1 comprises or consists of sequence SEQ ID NO: 15, and the linker L2 comprises or consists of sequence SEQ ID NO: 18. In some embodiments, the protein comprises SEQ ID NO: 125 (ORK9_P_01), SEQ ID NO: 126 (ORK9_P_02), SEQ ID NO: 127 (ORK9_P_03), SEQID NO: 128 (ORK9_P_04), SEQ ID NO: 202 (QRK9_P_05), SEQ ID NO: 203 (QRK9_P_06), SEQ ID NO: 204 (ORK9_P_07), SEQ ID NO: 205 (ORK9_P_08), SEQ ID NO: 206 (ORK9_P_09), or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains CSF1 and IFN-Y activities. In some embodiments, the protein comprises a Fc2 domain comprising or consisting of SEQ ID NO: 105.

[0290] In some embodiments, the protein comprises:

[0291] a) a first polypeptide chain comprising any one of SEQ ID NO: 125 to SEQ ID NO:

[0292] 128, and SEQ ID NO: 202 to SEQ ID NO: 206, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, and

[0293] b) a second polypeptide chain comprising or consisting of SEQ ID NO: 105, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto,

[0294] wherein the first and second polypeptides dimerise, and

[0295] wherein said protein retains CSF1 and IFN-y activities.

[0296] In some embodiments, the protein comprises a single chain polypeptide having dual activities of a cytokine targeting CSF1R and IL-2. In some embodiments, the linker L1 comprises or consists of sequence SEQ ID NO: 15, and the linker L2 comprises or consists of sequence SEQ ID NO: 18. In some embodiments, the protein comprises a first polypeptide chain comprising SEQ ID NO: 144, SEQ ID NO: 145, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto wherein said protein retains CSF1 and IL-2 activities. In some embodiments the protein further comprises a second polypeptide chain comprising or consisting of SEQ ID NO: 105, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, wherein the first and second polypeptides dimerise.

[0297] In some embodiments, the protein comprises a single chain polypeptide having dual activities of a cytokine targeting CSF1R and IL-4. In some embodiments, the linker L1 comprises or consists of sequence SEQ ID NO: 15, and the linker L2 comprises or consists of sequence SEQ ID NO: 18. In some embodiments, the protein comprises a first polypeptide chain comprising SEQ ID NO: 146, SEQ ID NO: 147, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto wherein said protein retains CSF1 and IL-4 activities. In some embodiments theprotein further comprises a second polypeptide chain comprising or consisting of SEQ ID NO: 105, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, wherein the first and second polypeptides dimerise.

[0298] In some embodiments, the protein comprises a single chain polypeptide having dual activities of a cytokine targeting CSF1R and GM-CSF. In some embodiments, the linker L1 comprises or consists of sequence SEQ ID NO: 15, and the linker L2 comprises or consists of sequence SEQ ID NO: 18. In some embodiments, the protein comprises a first polypeptide chain comprising any one of SEQ ID NO: 148 to SEQ ID NO: 151, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto wherein said protein retains CSF1 and GM-CSF activities. In some embodiments the protein further comprises a second polypeptide chain comprising or consisting of SEQ ID NO: 105, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto, wherein the first and second polypeptides dimerise.

[0299] Protein comprising two single chain polypeptides having dual-cytokine activities and one immunoglobulin Fc fragment

[0300] In some embodiments, the protein comprises two SC polypeptides having dualcytokine activities and one immunoglobulin Fc fragment. In some embodiments, the SC polypeptides having dual-cytokine activities and the immunoglobulin Fc fragment are fused through a peptide linker L2. According to this aspect, the protein comprises two polypeptide chains comprising the SC polypeptide having dual activities of a cytokine targeting CSF1R and cytokine of interest, fused to a Fc fragment through a peptide linker L2.

[0301] Preferably, the two SC polypeptides having dual-cytokine activities are identical. In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a polypeptide having dual-cytokine activities, a peptide linker L2, and a Fc domain, wherein the two polypeptide chains dimerise through their Fc domains, thereby forming a Fc fragment.

[0302] In some embodiments the peptide linker L2 connects the C-terminus of the cytokine of interest with the hinge of the Fc domain; the protein then comprises two polypeptide chains comprising or consisting of formula (V):

[0303] (i) cytokine targeting CSF1R-L1 -cytokine of interest-L2-Fc1 wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and(i) cytokine targeting CSF1R-L1 -cytokine of interest-L2-Fc2 wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0304] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some embodiments the peptide linker L2 connects the C-terminus of the cytokine targeting CSF1R with the hinge of the Fc domain; the protein then comprises two polypeptide chains comprising or consisting of formula (VI):

[0305] (i) cytokine of interest-L1-cytokine targeting CSF1R-L2-Fc1 wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0306] (i) cytokine of interest-L1- cytokine targeting CSF1R -L2-Fc2 wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0307] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0308] In some embodiments, the protein comprises two polypeptide chains that each comprise, in the N-terminus to C-terminus direction, a Fc domain, a peptide linker L2, a polypeptide having dual-cytokine activities, wherein the two polypeptide chains dimerise through their Fc domains, thereby forming a Fc fragment.

[0309] In some embodiments the peptide linker L2 connects the C-terminus of the Fc fragment with the N-terminus of the cytokine targeting CSF1R; the protein then comprises two polypeptide chains comprising or consisting of formula (VII):

[0310] (i) Fc1-L2- cytokine targeting CSF1R-L1 -cytokine of interest, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0311] (i) Fc2-L2- cytokine targeting CSF1R-L1 -cytokine of interest, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3

[0312] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. In some embodiments the peptide linker L2 connects the C-terminus of the Fc fragment with the N-terminus of the cytokine targeting CSF1R; the protein then comprises two polypeptide chains comprising or consisting of formula (VIII):

[0313] (i) Fc1-L2-cytokine of interest-L1 -cytokine targeting CSF1 R, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0314] (i) Fc2-L2-cytokine of interest-L1 -cytokine targeting CSF1 R, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3

[0315] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0316] The linkers L1 and L2 are as defined above.

[0317] In some embodiments, the linker L1 comprises or consist of sequence SEQ ID NO: 15, and the linker L2 comprises or consist of sequence SEQ ID NO: 18.Fc fragment or Fc domain

[0318] In some embodiments, the immunoglobulin Fc fragment or Fc domain is of the isotype IgG, such as lgG1, lgG2, lgG3, and lgG4, preferably lgG1 or lgG4. In some embodiments, the immunoglobulin Fc fragment or Fc domain is an engineered Fc fragment or Fc domain.

[0319] Accordingly, in some embodiments the protein comprises a Fc fragment that comprises two polypeptides that comprise a hinge region, an IgG CH2 domain and an IgG CH3 domain. Accordingly, in some embodiments the protein comprises two polypeptide chains that each comprise a Fc domain that comprise a hinge region, an IgG CH2 domain and an IgG CH3 domain.

[0320] Preferably the Fc fragment is silenced, i.e. the Fc fragment is mutated to reduce or eliminate binding to Fc gamma receptors and / or complement protein C1 q and thereby reduce or abolish immune effector functions.

[0321] In certain embodiments, a human IgG heavy chain Fc fragment or domain extends from Cys226 to the carboxyl-terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc fragment or domain may or may not be present, without affecting the structure or stability of the Fc fragment. Unless otherwise specified herein, numbering of amino acid residues in the IgG or Fc region is according to the EU numbering system for antibodies, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0322] In certain embodiments, Fc fragment or domain refers to an immunoglobulin IgG heavy chain constant region comprising a hinge region (starting at Cys226), an IgG CH2 domain and CH3 domain. The term “hinge region” or “hinge sequence” as used herein refers to the amino acid sequence that is, in some instance, located between the linker L2 and the CH2 domain. In certain embodiments, the hinge region comprises the amino acid sequence CPPCP (SEQ ID NO: 35), a sequence found in the native lgG1 or lgG2 hinge region at positions 239-243, sequence CPRCP (SEQ ID NO: 36), a sequence found in the native lgG3 hinge region at positions 239-241 B, sequence CPSCP (SEQ ID NO: 37), a sequence found in the native lgG4 hinge region at positions 239-243, to facilitate dimerization. In certain other embodiments, the Fc fragment or domain starts at the hinge region and extends to the C-terminus of the IgG heavy chain. In certain particular embodiments, the Fc fragment or domain comprises the Fc fragment or domain of human IgG, lgG2, lgG3 or lgG4. In certain particular embodiments, the Fc fragment or domain comprises the CH2 and CH3 domain of lgG4. In certain other particular embodiments, the Fc fragment or domain comprises the CH2 and CH3 domain of lgG1. In certainembodiments, the IgG CH2 domain starts at Ala 231. In certain other embodiments, the CH3 domain starts at Gly 341. It is understood that the C-terminus Lys residue of human IgG can be optionally absent. In some embodiments, the Fc fragment or domain is from human lgG1 heavy chain as shown in SEQ ID NO: 102. The lgG1 Fc fragment typically comprises or consists of sequence SEQ ID NO: 103, or amino acids 1-226 or 1-225 of SEQ ID NO: 103 (Fc with absent C-terminal Lys, or Gly-Lys), or a silenced variant thereof.

[0323] In certain embodiments, the Fc fragment is a human lgG4 Fc comprising substitution S228P in the hinge region and R409K in the CH3 region. In certain embodiments, the Fc fragment is engineered to modulate Fc mediated antibody functions. In some embodiments, the Fc fragment of the present disclosure does not induce antibody dependent cellular cytotoxicity (ADCC) and / or is an “Fc silent” antibody. Examples of silent lgG4 Fc fragments comprise mutations at positions 234, 235, 236, 237, 238, 265 and 329 in the lgG4 Fc amino acid sequence (Ell numbering). For example, lgG4 Fc silent fragment may include any one of the following mutations or combinations of mutations: F234A / L235A (FALA); L235E; F234A / L235A / G237A / P238S; and F234S / L235T / G236R (STR).

[0324] Examples of silent I gG 1 Fc fragments comprise mutations at positions 234, 235, 236, 239, 265, 297, 329 and / or 331 in the lgG1 Fc amino acid sequence (Ell numbering) corresponding to positions 14, 15, 16, 19, 45, 77, 109 and / or 112 of SEQ ID NO: 103. Another silent lgG1 Fc fragment comprises the N297A mutation, which results in aglycosylated or non-glycosylated Fc fragments. For example, lgG1 Fc silent fragment may include any one of the following mutations or combinations of mutations: D265A / P329A; L234F / L235Q / K322Q (FQQ); L234A / L235A (LALA); L234A / L235A / K322A (LALAKA); N297Q (aglycosyl); L234F / L235E / P331S (FES); L234A / L235A / G237A (LALAGA); L234A / L235E; L234A / L235A / G237A / P238S / H268A / A330S / P331 S; L234A / L235A / P329G (LALAPG); F234S / L235T / G236R (STR); L234A / L235A / P329S (LALAPS) L234A / L235A / D265 (LALADS) and G236R / L328R.

[0325] For certain proteins whose Fc Region-containing first and second polypeptide chains are not identical, it is desirable to reduce or prevent homodimerization from occurring between the CH2-CH3 Domains of two first polypeptide chains or between the CH2-CH3 Domains of two third polypeptide chains. The CH2 and / or CH3 Domains of such polypeptide chains need not to be identical in sequence, and advantageously are modified to foster complexing between the two polypeptide chains. For example, an amino acid substitution (preferably a substitution with an amino acid comprising a bulky side group forming a "knob", e.g., tryptophan) can be introduced into the CH2 or CH3 Domain such that steric interference will prevent interaction with a similarly mutated domain and will obligate the mutated domain to pair with a domain into which a complementary, or accommodatingmutation has been engineered, i.e., "the hole" (e.g., a substitution with glycine). Such sets of mutations can be engineered into any pair of polypeptides comprising CH2-CH3 Domains that forms a Fc Region to foster heterodimerization. Methods of protein engineering to favor heterodimerization over homodimerization are well known in the art, in particular with respect to the engineering of immunoglobulin-like molecules, and are encompassed herein (see e.g., Ridgway et al. (1996), Protein Engr. 9:617-621, Atwell et al. (1997), J. Mol. Biol.

[0326] 270: 26-35, and Xie etal. (2005), J. Immunol. Methods 296:95-101; each of which is hereby incorporated herein by reference in its entirety).

[0327] A preferred knob is created by modifying an IgG Fc Region to contain the modification T366W. A preferred hole may also be created by modifying an IgG Fc Region to contain the modification T366S, L368A and Y407V.

[0328] According to some embodiments of the invention, the first Fc fragment comprises an S354C or Y349C mutation and the second heavy chain comprises a Y349C or S354C mutation.

[0329] In some embodiments, the Fc fragment or Fc pair comprises or consists of sequence SEQ ID NO: 104 (lgG4 Fc (knob)) and / or SEQ ID NO: 105 (lgG4 Fc FALA (hole)), or SEQ ID NO: 106 (lgG4 silent FALA (R409K mutation, knob) and / or SEQ ID NO: 107 (lgG4 silent FALA (R409K mutation, hole)).

[0330] In some embodiments, the Fc fragment or Fc pair comprises or consist of sequence SEQ ID NO: 108 (lgG1 murine silent (knob)) and / or SEQ ID NO: 109 (lgG1 murine silent (hole)).

[0331] In some embodiments, the lgG1 Fc fragment or Fc pair comprises or consist of sequence SEQ ID NO: 110 (lgG1 silent STR (knob)) and / or SEQ ID NO: 111 (lgG1 silent STR (hole)); SEQ ID NO: 112 (lgG1 silent LALAPG (knob)) and / or SEQ ID NO: 113 (lgG1 silent LALAPG (hole)); SEQ ID NO: 114 (lgG1 silent LALAPS (knob)) and / or SEQ ID NO: 115 (lgG1 silent LALAPS (hole)); or SEQ ID NO: 116 (lgG1 silent LALADS (knob)) and / or SEQ ID NO: 117 (lgG1 silent LALADS (hole)).

[0332] Pharmaceutical composition

[0333] The invention further relates to a pharmaceutical composition comprising single chain polypeptides having dual cytokine activities, including CSF1 receptor targeting activity, or the protein comprising it, and a pharmaceutically acceptable carrier.

[0334] A suitable pharmaceutically acceptable ‘carrier’ (such as diluents, adjuvants, excipients or vehicles) is described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Pharmaceutical compositions of the invention are formulated to conform toregulatory standards and can be administered orally, intravenously, topically, or via other standard routes. As used herein, the term ‘carrier’ includes any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. The phrase ‘pharmaceutically acceptable’ or ‘pharmacologically-acceptable’ refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human or a non-human mammal.

[0335] Acceptable pharmaceutical vehicles can be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The pharmaceutical vehicles can be saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliary, stabilising, thickening, lubricating and colouring agents may be used. When administered to a subject, the pharmaceutically acceptable vehicles are preferably sterile. Water is a suitable vehicle particularly when the compound of the invention is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid vehicles, particularly for injectable solutions. Suitable pharmaceutical vehicles also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The present compositions, if desired, can also contain minor amounts of wetting or emulsifying agents, or buffering agents.

[0336] The medicaments and pharmaceutical compositions of the invention can take the form of liquids, solutions, suspensions, lotions, gels, tablets, pills, pellets, powders, modified-release formulations (such as slow or sustained-release), suppositories, emulsions, aerosols, sprays, capsules (for example, capsules containing liquids or powders), liposomes, microparticles or any other suitable formulations known in the art. Other examples of suitable pharmaceutical vehicles are described in Remington's Pharmaceutical Sciences, Alfonso R. Gennaro ed., Mack Publishing Co. Easton, Pa., 19th ed., 1995, see for example pages 1447-1676.

[0337] In some embodiments, the therapeutic compositions or medicaments of the invention are formulated in accordance with routine procedures as a pharmaceutical composition adapted for oral administration (more suitably for human beings). Compositions for oraldelivery may be in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs, for example. Thus, in various embodiments, the pharmaceutically acceptable vehicle may be a capsule, tablet or pill.

[0338] Orally administered compositions may contain one or more agents, for example, sweetening agents such as fructose, aspartame or saccharin; flavouring agents such as peppermint, oil of Wintergreen, or cherry; colouring agents; and preserving agents, to provide a pharmaceutically palatable preparation. When the composition is in the form of a tablet or pill, the compositions may be coated to delay disintegration and absorption in the gastrointestinal tract, so as to provide a sustained release of active agent over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving compound are also suitable for orally administered compositions. In these dosage forms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace the agent or agent composition through an aperture. These dosage forms can provide an essentially zero order delivery profile as opposed to the spiked profiles of immediate release formulations. A time delay material such as glycerol monostearate or glycerol stearate may also be used. Oral compositions can include standard vehicles such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Such vehicles are preferably of pharmaceutical grade. For oral formulations, the location of release may be the stomach, the small intestine (the duodenum, thejejunem, or the ileum), or the large intestine. The person skilled in the art is able to prepare formulations that will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine. Suitably, the release will avoid the deleterious effects of the stomach environment, either by protection of the peptide (or derivative) or by release of the peptide (or derivative) beyond the stomach environment, such as in the intestine. To ensure full gastric resistance a coating impermeable to at least pH 5.0 would be essential. Examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and Shellac, which may be used as mixed films.

[0339] To aid dissolution of the therapeutic agent(s) into the aqueous environment a surfactant might be added as a wetting agent. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents might be used and could include benzalkonium chloride or benzethomium chloride. Potential nonionic detergents that could be included in the formulation as surfactants include: lauromacrogol 400, polyoxyl 40 stearate,polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 20, 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants, when used, could be present in the formulation of the peptide or nucleic acid or derivative either alone or as a mixture in different ratios.

[0340] Typically, compositions for intravenous administration comprise sterile isotonic aqueous buffer. Where necessary, the compositions may also include a solubilising agent.

[0341] Mixtures of the polypeptides as described herein may be prepared in water suitably mixed with one or more excipients, carriers, or diluents. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form may be sterile and may be sufficiently fluid to enable injection by an appropriate syringe. Suitably, the composition is stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and / or vegetable oils. Proper fluidity may be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of a dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0342] For parenteral administration in an aqueous solution, for example, the solution may be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, subcutaneous, and intracerebroventricular administration. In this connection, sterile aqueous media that can be employed will be known to those of skill in the art.

[0343] Another suitable route of administration for the pharmaceutical compositions of the invention is via pulmonary or nasal delivery.

[0344] Additives may be included to enhance cellular uptake of a therapeutic agent of the invention, such as the fatty acids, oleic acid, linoleic acid and linolenic acid.Therapeutic applications

[0345] The single chain polypeptide having dual cytokine activities of the invention, the protein, or pharmaceutical composition comprising it, is for use as a medicament.

[0346] Accordingly, the invention further relates to a method of treatment which comprises administering the single chain polypeptide having dual activities of the invention, or the protein, to a subject in need thereof.

[0347] The subject may be a human or non-human mammal such as a rodent (mouse or rat), a primate (e.g. a monkey), a pig, etc.

[0348] The single chain polypeptide having dual cytokine activities of the invention, the protein, or pharmaceutical composition comprising it, is further for use for treating a disease. Accordingly, the invention further relates to a method of treating a disease which comprises administering the single chain polypeptide having dual activities of the invention, or the protein, to a subject in need thereof. The nature of the disease depends on the cytokine of interest comprised in the single chain polypeptide having dual cytokine activities or the protein containing it.

[0349] In some embodiments, the diseases to be treated comprise cancers, infectious diseases, inflammatory diseases, auto-immune diseases, neuro-inflammatory diseases, infectious diseases or tissue repair.

[0350] According to some embodiments, the cytokine of interest and corresponding disease to be treated are as follows:

[0351] i) the cytokine of interest is IL-10 and the disease is an inflammatory disease, an auto-immune disease, a neuro-inflammatory disease;

[0352] ii) the cytokine of interest is IFN-y and the disease is cancer or an infectious disease;

[0353] iii) the cytokine of interest is IL-35 and the disease is an inflammatory disease or an auto-immune disease;

[0354] iv) the cytokine of interest is IL-27 and the disease is an inflammatory disease or an infectious disease;

[0355] v) the cytokine of interest is GM-CSF and the disease is cancer or an infectious disease;

[0356] vi) the cytokine of interest is IL-2 and the disease is cancer or an infectious disease;

[0357] vii) the cytokine of interest is IL-4 and the disease is an inflammatory disease or IL-4 is for tissue repair;viii) the cytokine of interest is IFN-a and the disease is cancer or an infectious disease;

[0358] ix) the cytokine of interest is IFN-p and the disease is cancer or an infectious disease;

[0359] x) the cytokine of interest is IL-33 and the disease is an inflammatory disease or an auto-immune disease;

[0360] xi) the cytokine of interest is IL-13 and the disease is an inflammatory disease or an auto-immune disease;

[0361] xii) the cytokine of interest is TGF and the disease is an inflammatory disease or an auto-immune disease;

[0362] xiii) the cytokine of interest is IL-6 and the disease is cancer or an infectious disease;

[0363] xiv) the cytokine of interest is IL-36 and the disease is cancer or an infectious disease;

[0364] xv) the cytokine of interest is TN Fa and the disease is cancer or an infectious disease;

[0365] xvi) the cytokine of interest is IL-1 and the disease is cancer or an infectious disease;

[0366] xvii) the cytokine of interest is IL-24 and the disease is cancer or an infectious disease.

[0367] Miscellaneous embodiments

[0368] The application further discloses:

[0369] Embodiment 1. A single chain polypeptide having dual activities of a cytokine targeting CSF1R and of a cytokine of interest.

[0370] Embodiment 2. The single chain polypeptide according to embodiment 1, which comprises in the N-terminus to C-terminus direction, a cytokine monomer targeting CSF1R, a peptide linker L1, and a cytokine of interest.

[0371] Embodiment 3. The single chain polypeptide according to embodiment 1, which comprises, in the N-terminus to C-terminus direction, a cytokine of interest, a peptide linker L1, and a cytokine monomer targeting CSF1R.

[0372] Embodiment 4. The single chain polypeptide according to any one of embodiments 1 to 3, wherein one or more of the following conditions are met:

[0373] (i) the peptide linker L1 comprises from 10 to 45 amino acid residues;(ii) the peptide linker L1 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;

[0374] (iii) the peptide linker L1 comprises sequence SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36;

[0375] (iv) the peptide linker L1 comprises sequence (SEQ ID NO:17)n, n being an integer from 2 to 10, preferably from 3 to 6.

[0376] Embodiment 5. The single chain polypeptide according to any one of embodiments 1 to 4, wherein the cytokine monomer targeting CSF1R is a CSF1 monomer.

[0377] Embodiment 6. The single chain polypeptide according to any one of embodiments 1 to 5, wherein the cytokine monomer targeting CSF1R is a CSF1 monomer comprising sequence SEQ ID NO: 1, SEQ ID NO: 2, or a sequence at least 80% identical thereto and that retains CSF1 activity.

[0378] Embodiment 7. The single chain polypeptide according to any one of embodiments 1 to 4, wherein the cytokine monomer targeting CSF1R is an IL-34 monomer.

[0379] Embodiment 8. The single chain polypeptide according to any one of embodiments 1 to 4 and 7, wherein the cytokine monomer targeting CSF1R is an IL-34 monomer comprising sequence SEQ ID NO: 3, or a sequence at least 80% identical thereto and that retains IL-34 activity.

[0380] Embodiment 9. The single chain polypeptide according to any one of embodiments 1 to 8, wherein the cytokine of interest is selected from the group consisting of IL-10, GM-CSF, IL-2, IL-4, IFN-a, IFN-p, IFNy, IL-27, IL-33, IL-13, IL-24, IL-1 p, IL-36, TNFa, TGFp, IL-6 and IL-35.

[0381] Embodiment 10. The single chain polypeptide according to any one of embodiments 1 to 9, wherein the cytokine of interest is IL-10.

[0382] Embodiment 11. The single chain polypeptide according to embodiment 10, wherein IL-10 consists of an IL-10 monomer comprising sequence SEQ ID NO: 4, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL- 10 receptor.

[0383] Embodiment 12. The single chain polypeptide according to embodiment 10, wherein IL-10 consists of a single chain dimeric IL-10 that comprises sequence SEQ ID NO: 5, or SEQ ID NO: 6, or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain at least the same stability, and / or at least the same level of interaction with IL-10 receptor.

[0384] Embodiment 13. The single chain polypeptide according to embodiment 10, which is a dual CSF1 -IL-10 cytokine comprising sequence SEQ ID NO: 38, or a sequenceat least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain CSF1 and IL- 10 activities.

[0385] Embodiment 14. The single chain polypeptide according to embodiment 10, which is a dual I L-34-I L-10 cytokine comprising sequence SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 91, or SEQ ID NO: 92 or a sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retains IL-34 and IL-10 activities.

[0386] Embodiment 15. The single chain polypeptide according to any one of embodiments 1 to 9, wherein the cytokine of interest is selected from the group consisting of GM-CSF, IL-2, IL-4, IFN-a, IFN- , IFNy, IL-27, IL-33, IL-13, IL-24, IL-ip, IL-36, TNFa, TGFp, IL-6 and IL-35.

[0387] Embodiment 16. The single chain polypeptide according to embodiment 15, wherein the cytokine of interest is a monomer or an homodimeric single chain cytokine.

[0388] Embodiment 17. The single chain polypeptide according to embodiment 15 or 16, which is a dual CSF1-IFN-y cytokine, a dual IFNy-CSF1 cytokine, a dual CSF1-IL2 cytokine, a dual IL2-CSF1 cytokine, a dual CSF1-IL4cytokine, ora dual IL4-CSF1 cytokine.

[0389] Embodiment 18. The single chain polypeptide according to any one of embodiments 15 to 17, which comprises a sequence selected from the group consisting of SEQ ID NO: 7 to SEQ ID NO: 12, or a sequence at least at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical thereto and that retain the respective dual cytokine activities.

[0390] Embodiment 19. The single chain polypeptide according to any one of embodiments 1 to 18, which is modified to increase its half-life, optionally fused to a human immunoglobulin Fc fragment or to albumin or any albumin-binding domain, or conjugated to a poly(ethylene glycol) (PEG) molecule or other polymer-based moieties; or any other chemical or genetic modification known to extend protein stability and circulation time in vivo.

[0391] Embodiment 20. A protein comprising a single chain polypeptide according to any one of embodiments 1 to 19 fused to an immunoglobulin Fc fragment, preferably a human immunoglobulin Fc fragment.

[0392] Embodiment 21. The protein according to embodiment 20, which comprises one single chain polypeptide having dual-cytokine activities and one immunoglobulin Fc fragment.

[0393] Embodiment 22. The protein according to embodiment 21, which comprises two polypeptide chains comprising formula (I):cytokine targeting CSF1R monomer-L1 -cytokine of interest-L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0394] Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3, wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains. Embodiment 23. The protein according to embodiment 21, which comprises two polypeptide chains comprising formula (II):

[0395] cytokine of interest-L1-cytokine targeting CSF1R monomer -L2-Fc1, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0396] Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3, wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0397] Embodiment 24. The protein according to embodiment 21, which comprises two polypeptide chains comprising formula (III):

[0398] Fc1-L2-cytokine targeting CSF1R monomer-L1 -cytokine of interest, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0399] Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3, wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0400] Embodiment 25. The protein according to embodiment 21, which comprises two polypeptide chains comprising formula (IV):

[0401] Fc1-L2-cytokine of interest -L1 -cytokine targeting CSF1 R monomer, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0402] Fc2, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3, wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0403] Embodiment 26. The protein according to embodiment 20, which comprises two single chain polypeptides having dual-cytokine activities and one immunoglobulin Fc fragment.

[0404] Embodiment 27. The protein according to embodiment 26, which comprises two polypeptide chains comprising formula (V):

[0405] cytokine targeting CSF1R-L1 -cytokine of interest-L2-Fc1 wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0406] cytokine targeting CSF1R-L1 -cytokine of interest- L2-Fc2 wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0407] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.Embodiment 28. The protein according to embodiment 26, which comprises two polypeptide chains comprising formula (VI):

[0408] cytokine of interest-L1-cytokine targeting CSF1R-L2-Fc1 wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0409] cytokine of interest-L1- cytokine targeting CSF1R -L2-Fc2 wherein Fc2 is a Fc domain comprising hinge-CH2-CH3,

[0410] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0411] Embodiment 29. The protein according to embodiment 26, which comprises two polypeptide chains comprising formula (VII):

[0412] Fc1-L2- cytokine targeting CSF1R-L1 -cytokine of interest, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0413] Fc2-L2- cytokine targeting CSF1R-L1 -cytokine of interest, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3

[0414] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0415] Embodiment 30. The protein according to embodiment 26, which comprises two polypeptide chains comprising formula (VIII):

[0416] Fc1-L2-cytokine of interest-L1-cytokine targeting CSF1R, wherein Fc1 is a Fc domain comprising hinge-CH2-CH3, and

[0417] Fc2-L2-cytokine of interest-L1-cytokine targeting CSF1R, wherein Fc2 is a Fc domain comprising hinge-CH2-CH3

[0418] wherein the two polypeptide chains dimerise through the Fc1 and Fc2 domains.

[0419] Embodiment 31. The protein according to any one of embodiments 20 to 30, wherein one or more of the following conditions are met:

[0420] (i) the peptide linker L2 comprises from 3 to 45 amino acid residues;

[0421] (ii) the peptide linker L2 is a flexible peptide sequence composed of Gly and Ser residues;

[0422] (iii) the peptide linker L2 comprises sequence (GGGGS, SEQ ID NO: 17)p, wherein p is an integer from 1 to 8;

[0423] (iv) the peptide linker L2 comprises any one of sequence SEQ ID NO: 17 to SEQ ID NO: 33.

[0424] Embodiment 32. The protein according to any one of embodiments 20 to 31, wherein the Fc fragment or domain is from lgG1, lgG2, lgG3 or lgG4, silent or non- silent.Embodiment 33. The protein according to any one of embodiments 20 to 32, wherein the Fc fragment or domain comprises sequence SEQ ID NO: 34.

[0425] Embodiment 34. A pharmaceutical composition comprising the single chain polypeptide according to any one of embodiments 1 to 19, or the protein according to any one of embodiments 20 to 33, and a pharmaceutically acceptable carrier.

[0426] Embodiment 35. A single chain polypeptide according to any one of embodiments 1 to 19, or the protein according to any one of embodiments 20 to 33, or the pharmaceutical composition according to embodiment 34, for use as a medicament.

[0427] Embodiment 36. The single chain polypeptide according to any one of embodiments 1 to 19, or the protein according to any one of embodiments 20 to 33 for use for treating a disease comprising cancers, infectious diseases, inflammatory diseases, autoimmune diseases, neuro-inflammatory diseases, infectious diseases and tissue repair.

[0428] The invention will be further illustrated in view of the following figures and examples.

[0429] FIGURES

[0430] Figure 1. Structural organisation of different proteins comprising a single chain polypeptide having IL-10 and CSF1 activities and an immunoglobulin Fc fragment.

[0431] Symetric molecules comprising two single chain polypeptides having IL-10 and CSF1 activities and one immunoglobulin Fc fragment (ORK1_Fc_01, ORK1_Fc_02, ORK1_Fc_03, ORK1_Fc_04) and asymmetric molecules comprising one single chain polypeptide having IL-10 and CSF1 activities and one immunoglobulin Fc fragment (ORK1_Fc_05, ORK1_Fc_06). Fc fragment is from lgG4. In asymmetric molecules, Fc fragment comprises stabilization mutation S228P and silent FALA.

[0432] Figure 2. Spatial Orientation and Structure to Optimize Function. Tested N-to-C and C-to-N orientations of CSF1 and IL-10 together with two CSF1 molecules linked and hybrid human-viral IL-10 to assess impact on monocyte binding and T cell activity. Asymmetric formats (Fc-05 to Fc-12) were evaluated to assess how the positioning of IL-10, CSF-1, and the Fc region affects monocyte binding and T-cell activity. Linker 1 (L1) is defined as the linker connecting the IL-10 domain to the CSF-1 domain. Linker 1.1 (L1.1) is defined as intrapolypeptide linker connecting one CSF-1 domain to another CSF-1 domain. Linker 2 (L2) is defined as the linker connecting the cytokine domain (either IL-10 or CSF-1) to the FC domain.

[0433] Figure 3. Structure-Activity Relationship: IL-10 Signaling in Monocytes is CSF1-Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL-10 (not shown), ORK1-068 or ORK1-341 for 20 min at 37°C. Cells werestained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) PBMC from healthy donors were plated o / n in 96-well plates and treated next day when specified with saturating concentrations of rCSF1 (37°C, 5 min) then stimulated with IL-10 (not shown) or ORK1-068 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h.

[0434] Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM, similar results were obtained from 2 different donors. Each biological replicate was normalized by assigning the highest IL-10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1-068 and ORK1-341 were normalized accordingly.

[0435] Figure 4. ORK1-494 Binding to Monocytes is CSF1 -Dependent. (A) Total PBMC from healthy donors were treated when specified with saturating concentrations of rCSF1, rlL-10 or both together. After 5 min at 37°C, cells were washed and resuspend in buffer containing sodium azide 0,1%. After that, cells were treated with ORK1-494 for 20 min on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted on graphs in comparison with control sample with no blocking conditions.

[0436] Figure 5. ORK1 Binding to Monocytes is CSF1 -Dependent and drives IL-10 signaling. (A) Total PBMC from healthy donors were treated with ORK1-494, ORK1-513 and ORK1-532 for 20 min at 37°C. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647 and anti-human lgG4-FC PE. Samples were analyzed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted on graphs. (B) Total PBMC from healthy donors were treated with or without saturating concentrations of rCSF1. After that, samples were stimulated with ORK1-532 or IL-10 as a control. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. ORK1-532 improves selective binding to monocytes and increases IL-10 signaling.Figure 6. ORK1 optimization show increased ratio between T cells and monocytes in STAT3 activation. (A) Total PBMC from healthy donors were treated with IL-10, ORK1-494, ORK1-513, ORK1-532, ORK1-555 and ORK1-587. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-10 % of positive cells value of the top concentration as 100% and the lowest value of an untreated control as 0%. The samples treated with ORK1 variants were normalized accordingly. Ratio between values from T cells vs monocytes is shown in the graph (interpolated at 50% maximum stimulation). Doseresponse curves showing the % of positive cells from STAT3 activation in monocytes and T cells are shown in graph.

[0437] Figure 7. ORK1-532 variant induces regulatory macrophages without activating T cells. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-513 or ORK1-532. Cytokines were added every 2 days. Macrophages were stimulated overnight with LPS+IFNg. TNFa was measured by ELISA. Data from 4 different donors is shown. (B) CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with a-CD3 and a-CD28 and treated with IL- 10, ORK1-532 or ORK1-513 with IL-2, after 2 days supernatants were collected. GzmB was determined by ELISA. Data from 4-8 different donors is shown.

[0438] Figure 8. Short L1 Linkers Improve Monocyte Potency and Restore IL-10-Like Vmax. Total PBMC from healthy donors were treated with IL-10, ORK1-513 and linker variants designed. Cells were stained with antibodies specific for CD14 and CD3, followed by intracellular staining for phosphorylated STAT3 (pSTAT3). Samples were analyzed with LSRII cytometer and mean fluorescence intensity (MFI) values corresponding to STAT3 activation were quantified using GraphPad Prism software. (A) Graph to illustrate Vmax from ORK1-555 compared to ORK1-513 and IL-10 as a control. (B) Graph correlation between Vmax values (relative to IL-10) and EC50 in monocytes from pSTAT3 activation, all linker variants were included and ORK1-513 was used as a control.

[0439] Figure 9. ORK1 optimization. Description of molecule structure, number of the molecules and description of changes applied to improve function. ORK1 Fc was optimized followed a systematic, multi-parametric approach. Initial format ORK1-068, then ORK1-405 adding lgG4 and later sequence optimization in ORK1-494. Screening was performed to determine the optimal spatial orientation of the cytokine domains (CSF1 and IL-10) and the Fc subunit within the molecular construct, ORK1-513. In parallel, multiple CSF1 affinityvariants were evaluated (ORK1-532), together with systematic optimization of linker length and rigidity between the individual domains (ORK1-555, ORK1-563, ORK1-564 and ORK1-566). Later, affinity mutation was synergized with linker modification (ORK1-587).

[0440] Figure 10. Structural organisation of different proteins comprising a single chain polypeptide having IL-10 and IL-34 activities and an immunoglobulin Fc fragment. Symmetric molecules comprising two single chain polypeptides having IL-10 and IL-34 activities and one immunoglobulin Fc fragment: Format ORK6-Fc-O1 from ORK6_P_011, Format ORK6-FC-02 from ORK6_P_012 and asymmetric molecules comprising one single chain polypeptide having IL-10 and IL-34 activities and one immunoglobulin Fc fragment: Format ORK6-Fc-03 from ORK6_P_013, Format ORK6-Fc-04 from ORK6_P_014. Fc fragment is from lgG4. In asymmetric molecules, Fc fragment comprises stabilization mutation S228P and silent FALA.

[0441] Figure 11. ORK6-13 Binding to Monocytes is CSF1 -Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK6-013 is added and incubated 20’ on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647, CD3 APC / Cy7 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer. (B) Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted using GraphPad Prism software. Flow cytometry analysis shows dose-dependent binding of ORK6-013 to CD14+ monocytes. Binding is reduced upon CSF1R blocking, confirming specificity.

[0442] Figure 12. ORK6 shows selectivity towards myeloid cells compared to lymphoid cells. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL10, ORK6-004, ORK6-013 or ORK6-014 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. (B) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 and ORK6 was normalized accordingly.

[0443] Figure 13. ORK6 induces regulatory macrophages without activating T and B cells. (A) PBMCs were isolated from human buffy coats and CD14+ cells were purified bypositive selection (Miltenyi). Monocytes were cultured for 7 days in the presence or absence of: CSF1, IL-34, ORK6-004 or ORK6-013. Cytokines were added every 2 days. At day 7 cells were stimulated overnight with LPS±IFN-y and TNFa levels in the supernatants were determined by ELISA. (B) CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The lymphocytes were stimulated with soluble anti-CD3 and anti-CD28 for 72h. Then, cells were counted, replated and treated with IL- 10, ORK6-004 or ORK6-013 with IL-2 for 48h. Cell supernatants were obtained and GzmB was detected by ELISA. (C) B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes were stimulated with CpG (T-independent conditions) and IL-10, ORK6-004 or ORK6-013 at dO and d2. After 7 days cells were harvested for the analysis of plasma cells by flow cytometry. Similar results were obtained when stimulating with CD40L (T-dependent conditions).

[0444] Figure 14. Structural organisation of different proteins comprising a single chain polypeptide having IFNy and CSF1 activities and an immunoglobulin Fc fragment. GRK9-P-001, GRK9-P-002, GRK9-P-003, and GRK9-P-004, GRK9-P-005, ORK9-P-006, ORK9-P-007. ORK9 variants were designed by varying the orientation and positioning of CSF1 and IFN-y to tune biological potency and receptor engagement This modular approach enables optimization of: IFNy signaling strength, myeloid cell targeting via CSF1 and balance between efficacy and safety.

[0445] Figure 15. ORK9 Binding to Monocytes is CSF1 -Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, GRK9-001, ORK9-002, ORK9-003 and ORK9004 were added and incubated 20’ on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted using GraphPad Prism software.

[0446] Figure 16. ORK9 shows selectivity towards myeloid cells compared to lymphoid cells. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IFN-y or GRK9-001, GRK9-002, GRK9-003 and GRK9-004 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTATI (pY701) (BD) for 1h. (B-C) Samples were analyzed with LSRII cytometer and MFI values obtained from STAT1 activation were plotted usingGraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM.

[0447] Figure 17. ORK9-differentiated macrophages have a pro-inflammatory profile. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence of CSF1, CSF1 + IFN-y or ORK9 variants at 0.56 nM. Cytokines were added every 2 days. On day 7, macrophages were stimulated with LPS for 17 hours. Following this period, the culture supernatants were collected and analyzed to quantify protein levels (TNFa and IL-10) by ELISA.

[0448] Figure 18. ORK9-induced macrophages promote pro-inflammatory T cell responses. (A) CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 5 days in the presence CSF 1 or ORK9 variants at 0.56 nM . Cytokines were added every 2 days. At day 5 cells were counted and replated to a 96 round well plate (40.000 cells / well) and last dose of cytokines was added. After 2 days, media was washed and refreshed, macrophages were activated with LPS and CD3 T cells from an allogenic donor were purified and added over the macrophages (120.000 CD3 T cells / well, ratio of 1:3 macrophages:? cells). Supernatants were harvested at 48h and T cells were counted on day 5 of co-culture. . (B) Cell supernatants were obtained and GzmB, IFN-y, TNFa and IL-23 were measured by ELISA at 48h of co-culture.

[0449] Figure 19. Structural organisation of different proteins comprising a single chain polypeptide having IL-2 and CSF1 activities and an immunoglobulin Fc fragment. GRK10-P-001 and GRK10-P-002. ORK10 variants were designed by varying the orientation and positioning of CSF1 and IL-2 to tune biological potency and receptor engagement This modular approach enables optimization of IL-2 signaling strength, myeloid cell targeting via CSF1 and balance between efficacy and safety.

[0450] Figure 20. ORK10 Binding to Monocytes is CSF1 -Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK10-001 or ORK10-002 were added and incubated 20’ on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted using GraphPad Prism software.

[0451] Figure 21. Structural organisation of different proteins comprising a single chain polypeptide having IL-4 and CSF1 activities and an immunoglobulin Fc fragment. GRK11-P-001 and GRK11-P-002. ORK11 variants were designed by varying the orientation and positioning of CSF1 and IL-4 to tune biological potency and receptorengagement This modular approach enables optimization of IL-4 signaling strength, myeloid cell targeting via CSF1 and balance between efficacy and safety.

[0452] Figure 22. ORK11 Binding to Monocytes is CSF1 -Dependent. (A) PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, GRK11-001 or GRK11-002 were added and incubated 20’ on ice. Cells were washed and resuspended in stain buffer containing the mix of antibodies: CD14 Alexa 647 and anti-human lgG4-FC PE. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC PE were plotted using GraphPad Prism software.

[0453] Figure 23. ORK11 shows selectivity towards myeloid cells compared to lymphoid cells. (A) PBMC from healthy donors were plated o / n in 96-well plates and stimulated next day with IL-4, ORK11-001 or ORK11-002 for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT6 (pY641) (BD) for 1h. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT6 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM. Each biological replicate was normalized by assigning the highest IL-4 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK11 was normalized accordingly.

[0454] Figure 24: Structural organisation of different proteins comprising a single chain polypeptide having GM-CSF and CSF1 activities and an immunoglobulin Fc fragment. Design of ORK12 variants illustrating molecule structure, composition (lgG4-Fc, CSF1 and GM-CSF subunits), spatial orientation and number of molecules.

[0455] Figure 25: ORK12 Binding to Monocytes is CSF1 -Dependent. (A) Total PBMC from healthy donors were treated when specified with saturating concentrations of rCSF1 (5’ at 37°C). After this, cells were washed and resuspend in buffer containing PBS + 1% FBS + sodium azide 0,1%. Cells were then resuspended and treated with ORK12 variants for 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647 and anti-human lgG4-FC PE to detect ORK12. Samples were analysed with LSRII cytometer and percentage of lgG4-Fc PE positive cells were plotted on graphs (grey lines) in comparison with control sample with no blocking conditions (black lines). Data show the variants individually plotted.EXAMPLES

[0456] Example 1: Production of CSF1-IL10 combined bioactivity in a single-chain protein

[0457] Our goal is to create a dual molecule that can activate IL- 10 signaling exclusively in cells expressing CSF1R, which are predominantly myeloid cells and no other cells such as CD8+ T cells. To achieve this, we have engineered variants of IL-10 and / or Foldikine-10 with reduced affinity to I L1 ORA and I L1 ORB linked to a CSF 1 or I L-34 monomer. The CSF1 R taregting component acts as a targeting tool to circulating monocytes, facilitating preferential binding of IL-10 to these cells.

[0458] Example 2: Generation of single-chain (SC) molecules fused to Fc fragment The principles of protein expression are as follows.

[0459] In bold is highlighted the signal peptide (SP). Signal peptide is NOT included in final ORF as it is processed.

[0460] • In italic is included the tag used for purification (TEV-his-hibit). Tag is included in the ORF.

[0461] • Some proteins are the result of the co-transfection of two vectors. The two vectors are displayed. Knob chain or hole chain is specified.

[0462] • For context. Specified origin of the signal peptide at the end of the document

[0463] >ORK1-68_P MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQM ETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLK SCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDI KAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYI EAYMTM N NGGLDYLPNM LRDLRDAFSRVKTFFQM KDQLDN LLLKESLLEDFKGYLG CQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKA VEQVKNAFN KLQEKGIYKAMSEFDI Fl NYI EAYMTM KI RN GENL YFQSGGHHHHHHGGGV SGWRLFKKIS*

[0464] (SEQ ID NO: 97)

[0465] >ORK1-455_P MTAPGAAGRCPPTTWLGSLLLLVCLLASRSITEEVSEYCSHMIGSGHLQSLQRLIDSQM ETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLK SCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYI EAYMTM KIRNGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSR VKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVN SLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTM KI RN GENL YFQSGGHHHHHHGGG VSG WRLFKKIS*(SEQ ID NO: 98)

[0466] Signal peptide context:

[0467] MEWSWVFLFFLSVTTGVHS (SEQ ID NO : 41) - Taken from antibodies. Used as universal in different constructs.

[0468] MTAPGAAGRCPPTTWLGSLLLLVCLLASRSIT (SEQ ID NO : 42) (From CSF1 human).

[0469] Tables 1 and 2 disclose the different polypeptides constituting the protein comprising single chain polypeptide having IL- 10 and CSF1 activities fused to a Fc fragment.Table 1 : Description of CSF1-IL10 proteins: description of Polypeptides for Protein 1 and Protein 2

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[0482] Table 2: Description of CSF1-IL10 proteins including mutation(s) in CSF1 or linker variation

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[0517] "Protein quantification

[0518] Since we generated different point mutations to improve or decrease affinity to the receptors as well as we have broken loops in some of the constructs or made new linkers there is a danger that the resulting molecules will not be recognized by the antibody used in commercial ELISA kits. To avoid this issue, we have used the H I BIT approach to quantify all of our constructs (https: / / www.promega.es / en / resources / technologies / hibit-protein-tagging-system / ). Essentially this involves fusing a peptide tag spaced by a random linker to the Nt or CT of aThe Nano-Gio® Hi BiT Extracellular Detection System is a method that can detect HiBiT-tagged proteins secreted into the extracellular medium. The detection is done by adding a nonlytic detection reagent containing a substrate and Large BiT (LgBiT). HiBiT binds tightly to LgBiT, promoting complex formation and generating a bright, luminescent enzyme. The amount of luminescence generated is proportional to the amount of HiBiT-tagged protein present, and the system can detect protein amounts over seven orders of magnitude. The detection process is simple and requires only an add-mix-read assay protocol.). By having a peptide reference of know concentrations it is extremely easy to quantify the expression of any protein.

[0519] Plasmid generation

[0520] All plasmids generated in this work were assembled following the Gibson method (Gibson DG et al. Nat Methods. 2009 May;6(5):343-5). When required, Integrated DNA Technologies (IDT) Corporation performed gene synthesis (gBIock double-stranded fragments) and oligonucleotides synthesis. Gene amplification was carried out with Phusion DNA polymerase (Thermo Fisher Scientific) and transformed in Escherichia coli DH5-alpha competent cells (NEB).

[0521] For mammalian protein expression in ExpiCHO cells, the vector used was pcDNA3.1 (V790-20, Invitrogen). All the plasmids were verified by Sanger sequencing (Eurofins Genomics).

[0522] Expression in CHO cells

[0523] ExpiCHO protein expression kit cells were purchased from Thermofisher (A29133). For a low-scale production (2.5 mL), 24 deep well plates were used (AXYPDW10ML24CS, Merck) and covered by gas-permeable film (ThermoFisher). The day -1 of production, ExpiCHO-cells were split to a final density of 3 x 106viable cells / mL. Grown at 37C, 8% CO2 and shaking at 110rpm. The Day 0 of production, ExpiCHO cells were split to a final density of 3 x 106viable cells / mL per sample aliquot 2.5mL of the cell suspension into one well of the plate. Per sample, 2ug of DNA to 200ul of cold OptiPRO SFM & 9.0 pLExpiFectamine CHO Reagent was added. Then, 200 pL of the complexation mixture was added to each relevant well. All the plate was covered with a gas permeable seal and placed at 37°C, 8% CO2 and 225rpm. The day 1 after 18-22 hours post-transfection, ExpiFectamine CHO Enhancer and ExpiCHO Feed was added to each well. For an entire plate of 24 wells, 400 pL ExpiFectamine CHO Enhancer and 16 mL ExpiCHO Feed in a conical tube were mix. From this mixture, 600 pLwas added to each well of the plate. Place at 37C, 8% CO2 and 225rpm. After 4 days the samples were harvested. Plates were centrifuged 5 minutes at 4600 rpm at room temperature. Supernatants were aliquoted, snap frozen liquid nitrogen and stored at -80°C.

[0524] Protein purification

[0525] Affinity purification of Fc proteins was performed with HiTrap MabSelect PrismA. Binding buffer was PBS, 850 mM NaCI. Elution was performed with the following buffer: 0.1 M Na-citrate pH 3.3. Samples were neutralized right after elution with 1 M Tris-HCI, pH 9 (10% v / v). Purity, as defined by the fraction of Fc proteins, was determined by HPLC-SEC. SDS-PAGE of purified products was performed to verify the fragmentation and / or aggregation status of the final material.

[0526] Inferring EC-50 data

[0527] For the inference of an EC-50, we measured a dose-response analysis by following up absorbance (HEKBIuelO) or fluorescence (HEK293-CSF1 reporter).

[0528] The changes in absorbance at 630 nm due to different IL-10 tested concentrations were fitted to a saturation binding model using the following equation:

[0529] Y = Baseline+( B Max -Baseline) / (1+(X / EC50)A-Hill-slope).

[0530] In this equation, EC-50 is the apparent dissociation constant (as the number of active receptors per cell is unknown), h is the Hill-slope, and B Max is the saturation signal. This equation assumes specific binding only; all non-specific signals were subtracted. For EC- 50 calculated the equation used was log(agonist) vs. response -variable slope (Y=Bottom + (Top-Bottom) / (1+10A((LogEC50-X)*HillSlope)) from Prism 9 software.

[0531] For the CSF1 analysis we measured changes in the ratio of firefly / renilla luminescence.

[0532] The different experiments were analysed independently and fitted using GraphPad Prism 9 software. For IL-10 and FoldikinelO variants in those experiments were the basal or final lines of some mutants were not defined we used the average values from those mutants with lower EC-50 (final value at saturation) or high EC-50 (initial value with no ligand). In the case of CSF1 since we cannot be sure that the transformation efficiency withthe CSFR1 kit was the same in all cases, we fix the initial value as indicated above and we fix the Hill slope to the average of the mutants with lower EC-50. In each Figure we indicated how the fitting was done.

[0533] Example 3: in v / Yro functional evaluation of the dual CSF1-IL10 cytokine (ORK1) PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with saturating concentrations of rCSF1 for 5 min when indicated. Then samples were treated with ORK1-068, ORK1-341 and rlL-10 as a control (data not shown) (10 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 and CD3, fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with pSTAT3 (pY705) (BD) for 1h (Figure 3). Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK1 variants was normalized accordingly. In Figure 3A, ORK1-341 contain a mutation in CSF1 that cannot bind to CSF1R. In Figure 3B, an excess of soluble rCSF1 was added to saturate CSF1 receptors. In both examples ORK1-068 activates STAT3 preferentially in monocytes but this preferential signal is lost when ORK1 is unable to bind to CSF1R.

[0534] ORK1-494 selectively target monocytes via CSF1R. PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK1-494 (15nM per treatment, 7 dilutions 1 / 3) is added and incubated 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14, CD3 and anti-human lgG4-FC. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from anti-human lgG4-FC were plotted using GraphPad Prism software (Figure 4). The results show a preferential binding to monocytes (CD14+). Binding of ORK1-494to human PBMCs is predominantly restricted to CD14+monocytes and is inhibited by CSF1R blockade, by not by IL-10R blockade, confirming CSF1R-dependent targeting. Minimal binding to CD3+T cells suggest low off-target activity.

[0535] ORK1 was optimized followed a systematic, multi-parametric approach. Initial format ORK1-068, then ORK1-405 adding lgG4 (data not shown) and later sequence optimization in ORK1-494. Screening was performed to determine the optimal spatial orientation of the cytokine domains (CSF1 and IL- 10) and the Fc subunit within the molecular construct, ORK1-513. In parallel, multiple CSF1 affinity variants were evaluated(ORK1-532), together with systematic optimization of linker length and rigidity between the individual domains (ORK1-555). Later, affinity mutation was synergized with linker modification (ORK1-587).

[0536] Total PBMC from healthy donors were treated with IL-10, ORK1-494, ORK1-513, ORK1-532, ORK1-555 and ORK1-587. Cells were stained with CD14 and CD3 antibodies and intracellular staining for pSTAT3. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from STAT3 activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-10 % of positive cells value of the top concentration as 100% and the lowest value of an untreated control as 0%. The samples treated with ORK1 variants were normalized accordingly. Data was interpolated to 50% of maximum stimulation. Ratio between values from T cells vs monocytes is shown in the graph. Dose-response curves of % of positive cells from STAT3 activation in monocytes and T cells from samples treated with ORK1 variants are shown in the graph (Figure 6).

[0537] ORK1-532 improves selective binding to monocytes and increases IL-10 signaling. Total PBMC from healthy donors were stimulated with ORK1-532, ORK1-513, ORK1-494 or IL-10 as a control. Cells were stained with CD14 anti-human lgG4. Samples were analyzed with LSRII cytometer and values obtained from % of positive cells from lgG4 were plotted using GraphPad Prism software. (Figure 5A)

[0538] Previous data reported demonstrated that increase binding translates in increased IL-10 signaling in monocytes compared to T cells. The increased IL-10 signaling could be mediated by CSF1-CSF1R interaction (Figure 5B). The optimization strategy in this case was based on increase selectivity towards monocytes by increasing CSF1 affinity to CSF1 R based on a high-throughput mutational screening.

[0539] ORK1-532 variant induces regulatory macrophages without activating T cells.

[0540] CD14+ cells were purified from total PBMC of healthy donors. Monocytes were cultured for 7 days in the presence or absence of: CSF1, ORK1-513 or ORK1-532. Cytokines were added every 2 days. Macrophages were stimulated overnight with LPS+IFN-y. TNF-a was measured by ELISA. In monocyte-derived macrophages, ORK1-532 outperforms previous formats and maintain the highest IL-10 anti-inflammatory activity (Figure 7A).

[0541] CD8 T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). Cells were activated for 3 days with a-CD3 and O-CD28 and treated with IL-10, ORK1-532 orORK1-513 with IL-2 and after 2 days supernatants were collected. GzmB wasdetermined by ELISA. ORK1-532 is similar at ORK1-513, both reducing activation of CD8 T cells (Figure 7B).

[0542] Total PBMC from healthy donors were treated with IL-10, ORK1-513 and linker variants designed. Cells were stained with antibodies specific for CD14 and CD3, followed by intracellular staining for pSTAT3. MFI values corresponding to STAT3 activation were indicated in Figure 8 A. Graph to illustrate Vmax restore in ORK1-555 compared to ORK1-513 and IL-10 as a control.

[0543] In Figure 8B correlation between Vmax values (relative to IL-10) and EC50 in monocytes from pSTAT3 activation, all linker variants were included and ORK1-513 was used as a control. 5 variants were selected as best performers with higher Vmax and low EC50 in monocytes (ORK1-564, ORK1-563, ORK1-555, ORK1-556 and ORK1-566), all of them with short L1 linkers. Short L1 linkers improve monocyte potency and restore IL-10-like Vmax.

[0544] Example 4: Dual IL34-IL10 cytokine (ORK6)

[0545] Our goal was to create a dual molecule that can activate IL- 10 signaling exclusively in cells expressing CSF1R, which are predominantly myeloid cells and no other cells such as CD8+ T cells. To achieve this, we have engineered variants of IL-10 and / or Foldikine-10 with reduced affinity to I L1 ORA and I L1 ORB linked to a CSF 1 or I L-34 monomer. The CSF1 R targeting component acts as a targeting tool to circulating monocytes, facilitating preferential binding of IL-10 to these cells.

[0546] Plasmid generation

[0547] All plasmids generated in this work were assembled following the Gibson method (Gibson DG et al. Nat Methods. 2009 May;6(5):343-5). When required, Integrated DNA Technologies (IDT) Corporation performed gene synthesis (gBIock double-stranded fragments) and oligonucleotides synthesis. Gene amplification was carried out with Phusion DNA polymerase (Thermo Fisher Scientific) and transformed in Escherichia coli DH5-alpha competent cells (NEB).

[0548] For mammalian protein expression in ExpiCHO cells, the vector used was pcDNA3.1 (V790-20, Invitrogen). All the plasmids were verified by Sanger sequencing (Eurofins Genomics).

[0549] Expression in CHO cells

[0550] ExpiCHO protein expression kit cells were purchased from Thermofisher (A29133). For a low-scale production (2.5 mL), 24 deep well plates were used (AXYPDW10ML24CS,Merck) and covered by gas-permeable film (ThermoFisher). The day -1 of production, ExpiCHO-cells were split to a final density of 3 x 106viable cells / mL. Grown at 37C, 8% CO2 and shaking at 110rpm. The Day 0 of production, ExpiCHO cells were split to a final density of 3 x 106viable cells / mL per sample aliquot 2.5mL of the cell suspension into one well of the plate. Per sample, 2ug of DNA to 200ul of cold OptiPRO SFM & 9.0 pL ExpiFectamine CHO Reagent was added. Then, 200 pL of the complexation mixture was added to each relevant well. All the plate was covered with a gas permeable seal and placed at 37°C, 8% CO2 and 225rpm. The day 1 after 18-22 hours post-transfection, ExpiFectamine CHO Enhancer and ExpiCHO Feed was added to each well. For an entire plate of 24 wells, 400 pL ExpiFectamine CHO Enhancer and 16 mL ExpiCHO Feed in a conical tube were mix. From this mixture, 600 pLwas added to each well of the plate. Place at 37C, 8% CO2 and 225rpm. After 4 days the samples were harvested. Plates were centrifuged 5 minutes at 4600 rpm at room temperature. Supernatants were aliquoted, snap frozen liquid nitrogen and stored at -80°C.

[0551] Protein purification

[0552] Affinity purification of Fc proteins was performed with HiTrap MabSelect PrismA. Binding buffer was PBS, 850 mM NaCI. Elution was performed with the following buffer: 0.1 M Na-citrate pH 3.3. Samples were neutralized right after elution with 1 M Tris-HCI, pH 9 (10% v / v). Purity, as defined by the fraction of Fc proteins, was determined by HPLC-SEC. SDS-PAGE of purified products was performed to verify the fragmentation and / or aggregation status of the final material.

[0553] The principles of protein expression are as follows.

[0554] In bold is highlighted the signal peptide (SP). Signal peptide is NOT included in final

[0555]

[0556] In italic is included the tag used for purification (TEV-his-hibit). Tag is included in the ORF. Some proteins are the result of the co-transfection of two vectors. The two vectors are dis =played. Knob chain or hole chain is specified. For context. Specified origin of the signal peptide at the end of the document.

[0557] > SEQ I D NO: 93 ORK6_1 (I L10-I L34) -tev-h is- h i b it MEWSWVFLFFLSVTTGVHSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGEN LKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKI RNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGNEPLEMWPLTQNEECTVTGFLRD KLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATE SVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPK ALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSP SSPPHSTGS RP RFQGEGLLPGENLYFQSGGHHHHHHGGGVSGWRLFKKIS*> SEQ ID NO: 94 ORK6_5- tev-his-hibit MPRGFTWLRYLGIFLGVALGNEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYF PINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKY LQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCS CCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRA QGEGLLPGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFK GYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCEN KSKAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM K\RNGENLYFQSGGHHHHHH GGGVSGWRLFKKIS*

[0558] >ORK6_14_P = co-transfection of two vectors (ORK6_10_V I ORK1_287_V)

[0559] > SEQ I D NO: 95 ORK6_10_V (knob chain) MEWSWVFLFFLSVTTGVHSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRT PEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSC SVMHEALHNHYTQKSLSLSLGGGGGGSGGGGSGGGGSNEPLEMWPLTQNEECTVT GFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVL VSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPG PNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAA TQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSGGSGGSGGS GGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESL LEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQA ENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDI Fl NYI EAYMTM KI RN*

[0560] > SEQ ID NO: 96 ORK1_287_V (hole chain) MEWSWVFLFFLSVTTGVHSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRT PEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVCTLPPSQEEMTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSRLTVDKSRWQEGNVFSC SVM H EALH N HYTQKSLSLSLG*

[0561] Signal peptide context:

[0562] MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 70) (Taken from antibodies. Used as universal in different constructs).

[0563] MPRGFTWLRYLGIFLGVALG (SEQ ID NO: 71) (From IL34 human).

[0564] The dual cytokine single chain I L34-IL10 polypeptides as disclosed in Table 3 were constructed. Dual cytokine single chain I L34-IL10 polypeptides fused to an immunoglobulin Fc fragment are further disclosed in Table 4.Table 3: details of the dual cytokine single chain IL34-IL10 polypeptides (0RK6)

[0565]

[0566] Table 4: details of the dual cytokine single chain I L34-IL10 polypeptides (0RK6) fused o Fc

[0567]

[0568] In vitro functional evaluation of the dual IL34-IL10 cytokine

[0569] Flow cytometry analyses were conducted with ORK6 showing dose-dependent binding of ORK6-013 to CD14+ monocytes, and reduction of binding upon CSF1 R blocking, thereby confirming that ORK6 binding to monocytes is CSF1 -dependent, indicating functional CSF1R engagement (Figure 11). PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1, rlL-10 or both (5 min, 37°C), after that cells were washed and resuspend in buffer containing sodium azide 0,1%. Then, ORK6-013 (15nM per treatment, 8 dilutions 1 / 3) is added and incubated 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14, CD3 and anti-human lgG4-FC. Finally, cells were resuspended in stain buffer. Samples were analysed with LSRII cytometer and MFI values obtained from antihuman lgG4-FC were plotted using GraphPad Prism software. The results show a preferential binding to monocytes (CD14+).

[0570] Further characterization of receptor binding and selectivity of the dual cytokine single chain IL34-IL10 polypeptide, with or without an Fc

[0571] PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with rl L10, ORK6-004, ORK6-013 or ORK6-014 (15 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT3 (pY705) (BD) for 1h. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT3 activation were plotted using GraphPad Prism software. One representative result with two technical replicates is shown as mean with error bar depicting the SEM. Each biological replicate was normalized by assigning the highest IL10 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK6 variants was normalized accordingly. These results show that ORK6 variants show selectivity towards myeloid cells compared to lymphoid cells, in contrast with IL-10 (Figure 12).

[0572] Induction of regulatory macrophages was analyzed by cultivating CD14+ monocytes treated with CSF1, IL-34, ORK6-004 orORK6-013 over 7 days. Cytokine production (TNFa) was analyzed by ELISA. The phenotype of the ORK6induced macrophages is consistent with the phenotype of regulatory macrophages. Differentiated macrophages were further stimulated by LPS and IFN-y: TNFa production was found decreased in ORK6-004 and ORK6-013 induced macrophages, compared to IL-34 or CSF-1 induced macrophages (Figure 13A).T cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The lymphocytes (1.5 million / ml) were stimulated with anti-CD3 (ThermoFisher) (1 pg / ml) and anti-CD28 (ThermoFisher) (3 pg / ml) for 72h. Then, cells were splited, replated and treated with IL10, ORK6-004, ORK6-013 10 nM per treatment, 6 dilutions 1 / 4) with IL-2 (5 ng / ml) for 48h.. GzmB production measured by ELISA in aTCR-stimulated CD8+ T cells treated with ORK6 (ORK6-004 or ORK6-013) demonstrates attenuated CD8 activation (Figure 13B).

[0573] B cells were isolated from human PBMCs using positive microbeads selection (Miltenyi). The isolated lymphocytes (50,000 cells / well) were stimulated with CpG (InvivoGene) (1 pg / ml) and IL10, ORK6-004 orORK6-013 (10 nM per treatment, 8 dilutions 1 / 3) at dO and d2. Similar results were obtained when stimulating with CD40L. Results shown that ORK6 is less potent than IL-10 in promoting plasma cell differentiation and therefore induce reduced B cell activation compared to IL-10 (Figure 13C).

[0574] Example 5: Design of dual CSF1 and IFN-y cytokines (ORK9)

[0575] ORK9 was designed in order to achieve targeted IFN-y delivery to induce myeloid-derived macrophages with pro-inflammatory phenotype, with application in treating e.g. cancer and infectious diseases.

[0576] ORK9 variants were designed by varying the orientation and positioning of CSF1 and IFN-y to tune biological potency and receptor engagement.

[0577] This modular approach enables optimization of:

[0578] i. IFN-y signaling strength

[0579] ii. Myeloid cell targeting via CSF1

[0580] iii. Balance between efficacy and safety.

[0581] The structural organization of GRK9-P-001, ORK9-P-002, ORK9-P-003, ORK9-P-004, ORK9-P-005, ORK9-P-006 and ORK9-P-007 is shown in Figure 14. The constitutive domains of the ORK9 variants are detailed in Table 5.Table 5: details of the dual cytokine single chain CSF1-IFN-y polypeptides (0RK9) fused to Fc

[0582] <

[0583] <

[0584] <

[0585] <

[0586] <

[0587]

[0588] For binding assays, PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5’, 37°C); after that, cells were washed and resuspend in stain buffer containing PBS + FBS 1% + sodium azide 0,1%. Then, ORK9 variants were added and incubated 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647, and anti-human lgG4-Fc PE to detect ORK9. Samples were analyzed with LSRII cytometer and percentage of lgG4-Fc positive cells as plotted using GraphPad Prism software (Fig. 15). ORK9-004 shows preferential binding for monocytes (CD14+).

[0589] PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with IFN-y or ORK9 variants (5 nM per treatment, 8 dilutions 1 / 3) for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT 1 (pY701) (BD) for 1 h. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT1 activation were plotted using GraphPad Prism software. One representative result with two technical replicates (line graphs) is shown as mean with error bar depicting the SEM ORK9 show preferential signaling for monocytes compared to T cells. (Fig. 16)

[0590] PBMCs were isolated from human buffy coats and CD14+cells were purified by positive microbead selection (Miltenyi). Isolated monocytes (0.5 million / ml) were cultured for 7 days in the presence or absence of: rCSF1, rCSF1+rlFN-y or ORK9 variants (0.56 nM per treatment) or untreated condition. Cytokines were added every 2 days. At day 7, macrophages were stimulated with LPS for 17h and cytokine levels were measured in the supernatant. The results show that ORK9 induces the differentiation of pro-inflammatory monocyte-derived macrophages, as identified by the increase on TNFa production and the decrease on IL- 10 production (Fig. 17).

[0591] PBMCs were isolated from human buffy coats and CD14+monocytes or CD3+T cells were purified by positive selection (Miltenyi). Monocytes (0.5 million / ml) were cultured for 7 days in the presence or absence of: rCSF1, rCSF1+rlFN-y or ORK9 variants (1.08 nM). Cytokines were added every 2 days. At day 5, cells were harvested and replated in 96 U bottom well / plates (40.000 cells / well in 150 pL) and last dose of cytokines was added. At day 7, cells were washed and stimulated with LPS during 4h. After that, CD3+T cells (120.000 cells in 50 pL, ratio 3:1 respect macrophages) were added to the culture. At day 2 of MLR co-culture, supernatant for some wells was harvested for measuring cytokine production (TNFa, IFNy, GzmB and IL-23). At day 5, total T cell number was counted. Co-culture with 0RK9-differentiated macrophages increases T cell proliferation and production of TNFa, IFNy, GzmB and IL-23 similar as co-culture with CSFI+IFNy-differentated macrophages (Fig. 18).

[0592] Example 6: Design of dual CSF1 and IL-2 cytokines (ORK1Q)

[0593] The structural organization of GRK10-P-001 and ORK10-P-002 is shown in Figure 19. The constitutive domains of the ORK10 variants are detailed in Table 6.

[0594] in vitro functional evaluation of the dual CSF1-IL-2-Fc cytokine

[0595] For binding assays, PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5’, 37°C); after that, cells were washed and resuspend in stain buffer containing PBS + FBS 1% + sodium azide 0,1%. Then, ORK10 variants were added and incubated 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647and anti-human lgG4-Fc PE to detect ORK10. Samples were analyzed with LSRII cytometer and percentage of lgG4-Fc positive cells as plotted using GraphPad Prism software (Fig. 20). GRK10-002 is the variant with preferential binding affinity to monocytes (CD14+).Table 6: details of the dual cytokine single chain CSF1-IL-2, or CSF1-IL-4, or CSF1-GMCSF polypeptides fused to Fc

[0596]

[0597] Example 7: Design of dual CSF1 and IL-4 cytokines (ORK11)

[0598] The structural organization of GRK11-P-001 and GRK11-P-002 is shown in Figure 21. The constitutive domains of the ORK11 variants are detailed in Table 6.

[0599] in vitro functional evaluation of the dual CSF1-IL-4-Fc cytokine

[0600] For binding assays, PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5’, 37°C); after that, cells were washed and resuspend in stain buffer containing PBS + FBS 1% + sodium azide 0,1%. Then, ORK11 variants were added and incubated 20’ on ice. Cells were washed and resuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647and anti-human lgG4-Fc PE to detect ORK11. Samples were analyzed with LSRII cytometer and percentage of lgG4-Fc positive cells was plotted using GraphPad Prism software (Fig. 22). ORK11-002 is the variant with preferential binding affinity to monocytes (CD14+).

[0601] For pSTAT6 signaling, PBMC (150,000 cells / well) from healthy donors were plated on in 96-well plates and stimulated next day with rlL-4 or ORK11 variants (starting at 5 nM, 8serial dilutions 1 / 4) for 20 min at 37°C. Cells were stained with CD14 Alexa647 (Biolegend) and CD3 APC / Cy7 (Biolegend), fixed with 4% PFA and permeabilized for intracellular staining with 80% MetOH. Cells were stained with AlexaFluor488 pSTAT6 (pY641) (BD) for 1 h. Samples were analyzed with LSRII cytometer and MFI values obtained from STAT6 activation were plotted using GraphPad Prism software. Each biological replicate was normalized by assigning the highest IL-4 MFI value of the top concentration as 100% and the lowest MFI value of an untreated control as 0%. The MFI from the samples treated with ORK11 variants was normalized accordingly. ORK11 preferentially activates monocytes when compared to T cells. These results show that ORK11 variants have selectivity towards myeloid cells compared to lymphoid cells, in contrast with IL-4 (Fig. 23).

[0602] Example 8: Design of dual CSF1 and GM-CSF cytokines (ORK12)

[0603] The structural organization of GRK12-P-001, ORK12-P-002, ORK12-P-003 and ORK12-P-004 is shown in Figure 24. The constitutive domains of the ORK12 variants are detailed in Table 6.

[0604] in vitro functional evaluation of the dual CSF1-GM-CSF-Fc cytokine

[0605] For binding assays, PBMC from healthy donors were plated o / n in 96-well plates and treated when specified with saturating concentrations of rCSF1 (5’, 37°C); after that, cells were washed and resuspend in stain buffer containing PBS + FBS 1% + sodium azide 0,1%. Then, ORK12variants were added and incubated 20’ on ice. Cells were washed andresuspend in stain buffer containing the mix of antibodies: CD14 Alexa 647and anti-human lgG4-Fc PE to detect 0RK12. Samples were analyzed with LSRII cytometer and percentage of lgG4-Fc positive cells as plotted using GraphPad Prism software (Fig. 25). ORK12-003 is the variant with preferential binding affinity to monocytes (CD14+).

[0606] Sequences

[0607] >SEQ ID NO: 1 CSF1 short version ending in QD EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQD

[0608] >SEQ ID NO: 2 CSF1 full-length EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVVTKPDCNCLYPKAIPSSDPASVSPHQPL APSMAPVAGLTWEDSEGTEGSSLLPGEQPLHTVDPGSAKQRPPR

[0609] >SEQ ID NO: 3 IL-34 (UniProtKB Q6ZMJ4-1 without signal peptide) NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVS PKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLP

[0610] >SEQ ID NO: 4 IL-10 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM KI RN

[0611] >SEQ ID NO: 5 Foldikine 10 (without Methionine, only C108N (2nddomain) mutation, and N-terminal completed) SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRV KTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNS LGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRN

[0612] >SEQ ID NO: 6 single chain dimeric IL-10 domain in ORK1-455 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM KI RNGGGSGGGGSSPGQGTQS ENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSE MIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKN AFNKLQEKGIYKAMSEFDIFINYI EAYMTM KIRN

[0613] >SEQ ID NO: 7 CSF1_IFN-y (ORK9_P_03) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGQDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQSQIVSFYF KLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNYSVTDLNVQRKAIHELIQ VM AELSPAAKTGKRKRSQM LFRG

[0614] >SEQ ID NO: 8 IFN-Y_CSF1 (ORK9_P_04) QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQSQIVSFYFKLFKNF KDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNYSVTDLNVQRKAIHELIQVMAELS PAAKTGKRKRSQMLFRGGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCS HMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFR DNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLD KDWNIFSKNCNNSFAECSSQDV

[0615] >SEQ ID NO: 9 CSF1JL-2 (ORK10_P_01) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKH LQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEF LNRWITFCQSIISTLT

[0616] >SEQ ID NO: 10 IL-2_CSF1 (ORK10_P_02) APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEE ELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITF CQSIISTLTGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGHLQS LQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQ LQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNC NNSFAECSSQDV

[0617] >SEQ ID NO: 11 CSF1JL-4 (ORK11_P_01) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGHKCDITLQEIIKDLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRAATVLRQFYS HHEKDTRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQSTLENFLERL KTIMREKYSKCSS

[0618] >SEQ ID NO: 12 IL-4_CSF1 (ORK11_P_02) HKCDITLQEIIKDLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRAATVLRQFYSHHEKD TRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQSTLENFLERLKTIMRE KYSKCSSGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGHLQSL QRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQL QELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCN NSFAECSSQDV

[0619] >SEQ ID NO: 13

[0620] GGSGGSGGSGGSGGG

[0621] >SEQ ID NO: 14

[0622] GGGSGGSGGSGGSGGSGGSGGSGGG

[0623] >SEQ ID NO: 15

[0624] GGGGGSGGSGGSGGSGGSGGSGGSGGSGGG

[0625] >SEQ ID NO: 16GGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGG

[0626] >SEQ ID NO: 17

[0627] GGGGS

[0628] >SEQ ID NO: 18

[0629] GGGGSGGGGSGGGGS

[0630] >SEQ ID NO: 19

[0631] SGGGGSGGGGS

[0632] >SEQ ID NO: 20

[0633] SGGGGSGGGGSAP

[0634] >SEQ ID NO: 21

[0635] NFSQP

[0636] >SEQ ID NO: 22

[0637] KRTVA

[0638] >SEQ ID NO: 23

[0639] GGGSGGGG

[0640] >SEQ ID NO: 24

[0641] GGGGSGGGGS

[0642] >SEQ ID NO: 25

[0643] THTCPPCPEPKSSDK

[0644] >SEQ ID NO: 26

[0645] GGGS

[0646] >SEQ ID NO: 27

[0647] EAAKEAAKGGGGS

[0648] >SEQ ID NO: 28

[0649] EAAKEAAK

[0650] >SEQ ID NO: 29 GGSSGSGSGSTGTSSSGTGTSAGTTGTSASTSGSGSGGGGGSGGGGSAGG

[0651] >SEQ ID NO: 30

[0652] NGGLDY

[0653] >SEQ ID NO: 31

[0654] FGGLDY

[0655] >SEQ ID NO: 32

[0656] YKTIT

[0657] >SEQ ID NO: 33

[0658] DKDIRDGD

[0659] >SEQ ID NO: 34 Fc of lgG4 silent FALA allotypeESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL

[0660] >SEQ ID NO: 35

[0661] CPPCP

[0662] >SEQ ID NO: 36

[0663] CPRCP

[0664] >SEQ ID NO: 37

[0665] CPSCP

[0666] >SEQ ID NO: 38 CSF1-IL-10 ORK1_P_455 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLE DFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPG QGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGC QALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVE QVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN

[0667] > SEQ ID NO: 39 ORK1_P-068 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRV KTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNS LGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRN

[0668] > SEQ ID NO: 40 ORK1-P-455 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLE DFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPG QGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGC QALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVE QVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRN

[0669] > SEQ ID NO: 41 signal peptide

[0670] MEWSWVFLFFLSVTTGVHS

[0671] > SEQ ID NO: 42 signal peptide

[0672] MTAPGAAGRCPPTTWLGSLLLLVCLLASRSIT

[0673] > SEQ ID NO: 43 ORKI-Fc-01 (ORK1_P-400)ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYL KKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPL QLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGS GGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLK ESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLR DLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQ DPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEF DIFINYI EAYMTM KIRN

[0674] >SEQ ID NO: 44 ORKI-Fc-02 (ORK1_P_401) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRV KTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNS LGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRNGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA LHNHYTQKSLSLSLG

[0675] >SEQ ID NO: 45 ORKI-Fc-03 (ORK1_P_402) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSGGGGSGGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQI TFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKD YEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGG GGSGGSGGSGGSGGSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSR VKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVN SLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSE MIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKN AFN KLQEKGI YKAMSEFDI Fl N Yl EAYMTM KI RN*

[0676] >SEQ ID NO: 46 ORKI-Fc-04 (ORK1_P_403) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRV KTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNS LGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRNGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQ EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDK SRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG*

[0677] >SEQ ID NO: 47 ORKI-Fc-05 (ORK1_P_404) EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKG YLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRV KTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNS LGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYM TMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMIS RTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLG*

[0678] >SEQ ID NO: 48 ORKI-Fc-06 (ORK1_P_405) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRN*

[0679] >SEQ ID NO: 49 ORK1_P_438 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRN*

[0680] >SEQ ID NO: 50 ORK1_P-439 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRN*

[0681] >SEQ ID NO: 51 ORK1_440_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS EFDIFINYIEAYMTMKIRN*

[0682] >SEQ ID NO: 52 ORK1_P_441 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGMTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYIEAYMTMKIRN*

[0683] >SEQ ID NO: 53 ORK1_473_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLL KESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYIEAYMTMKIRN*

[0684] >SEQ ID NO: 54 ORK1_474_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLR LRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLD YLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVM PQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDI Fl NYI EAYMTM KI RN*

[0685] >SEQ ID NO: 55 ORK1_475_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLL KESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QDPDIKAHVNSLGENLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYIEAYMTMKIRN*

[0686] >SEQ ID NO: 56 ORK1_476_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLR LRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLD YLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVM PQAENQDPDIKAHVNSLGENLKTLRLRLRRAHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMKIRN*

[0687] >SEQ ID NO: 57 ORK1_482_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLR LRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYI EAYMTM KIRNGG GSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKES LLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRF LPCEN KSKAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM KI RN*

[0688] >SEQ ID NO: 58 ORK1_485_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLL KESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNML RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN QDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSE FDIFINYI EAYMTM KIRN*

[0689] >SEQ ID NO: 59 ORK1_486_P ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSG GSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQ LDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLR LRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLD YLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVM PQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMKIRN*

[0690] >SEQ ID NO: 60 ORK1_P_493 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGS GGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDIFINYI EAYMTM KIRN*>SEQ ID NO: 601 ORK1_P_494 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGS GGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKD QLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTL RLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGL DYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEV MPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKG IYKAMSEFDI FINYI EAYMTM KI RN*

[0691] >SEQ ID NO: 62 ORK1_P_495 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGS GGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLL LKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRR CHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNM LRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAE NQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAM SEFDI FINYI EAYMTM KIRN*

[0692] >SEQ ID NO: 63 ORK1_P_496 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGS GGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKD QLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTL RLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGL DYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEV MPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKG IYKAMSEFDIFINYI EAYMTM KIRN*

[0693] >SEQ ID NO: 64 ORK1_P_497 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVC YLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYET PLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVVTKPDCNCLYPKAIPSSD PASVSPHQPLAPSMAPVAGLTWEDSEGTEGSSLLPGEQPLHTVDPGSAKQRPPRGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVK TFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSL GENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMT MNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMI QFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAF NKLQEKGIYKAMSEFDIFINYI EAYMTM KIRN*

[0694] >SEQ ID NO: 65 ORK1_P_455 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLE DFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGSGGGGSSPG QGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGC QALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVE QVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN

[0695] >SEQ ID NO: 66 ORK6_P_001 (IL10-IL34) SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM KI RNGGGGGSGGSGGSGGSGG SGGSGGSGGSGGGNEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKIS VPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVET LLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSS VLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLP

[0696] >SEQ ID NO: 70 signal peptide

[0697] MEWSWVFLFFLSVTTGVHS

[0698] >SEQ ID NO: 71 signal peptide

[0699] MPRGFTWLRYLGIFLGVALG

[0700] >SEQ ID NO: 72 IL10-IL34 ORK6_P_001 SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGY LGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKS KAVEQVKNAFN KLQEKGI YKAMSEFDI Fl NYI EAYMTM KI RNGGGGGSGGSGGSGGSGG SGGSGGSGGSGGGNEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKIS VPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVET LLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSS VLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLP

[0701] >SEQ ID NO: 73 IL34-IL10 ORK6_P_002 NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVS PKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSG GSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLK TLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 74 I L34-Foldikine10 ORK6_P_004 NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSG GSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLD NLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYL PNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMP QAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYI EAYMTM KIRN

[0702] >SEQ ID NO: 75 ORK6_P_013 sequence encoded by plasmid 1 NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVS PKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSG GSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLD NLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYL PNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMP QAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYI EAYMTM KIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFN STYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQE EMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKS RWQEG N VFSCSVM H EALH N H YTQKSLSLSLG*

[0703] >SEQ ID NO: 77 ORK6_P_014 sequence encoded by plasmid 1 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGGS GGGGSGGGGSNEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPY EGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLL NVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVL NWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPG GGGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFS RVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHV NSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEA YMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALS EMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVK NAFNKLQEKGIYKAMSEFDIFINYI EAYMTM KIRN*

[0704] > SEQ ID NO: 80 IL36a KIDTPQQGSIQDINHRVWVLQDQTLIAVPRKDRMSPVTIALISCRHVETLEKDRGNPIYLGL NGLNLCLMCAKVGDQPTLQLKEKDIMDLYNQPEPVKSFLFYHSQSGRNSTFESVAFPG WFIAVSSEGGCPLILTQELGKANTTDFGLTMLF

[0705] > SEQ ID NO: 81 IL36b REAAPKSYAIRDSRQMVWVLSGNSLIAAPLSRSIKPVTLHLIACRDTEFSDKEKGNMVYL GIKGKDLCLFCAEIQGKPTLQLKLQGSQDNIGKDTCWKLVGIHTCINLDVRESCFMGTLD QWGIGVGRKKWKSSFQHHHLRKKDKDFSSMRTNIGMPGRM

[0706] > SEQ ID NO: 82 IL1bAPVRSLNCTLRDSQQKSLVMSGPYELKALHLQGQDMEQQVVFSMSFVQGEESNDKIPV ALGLKEKNLYLSCVLKDDKPTLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPN WYISTSQAENMPVFLGGTKGGQDITDFTMQFVSS

[0707] > SEQ ID NO: 83 IL6 VPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENN LNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKV LIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRALR QM

[0708] > SEQ ID NO: 84 IL13 LTCLGGFASPGPVPPSTALRELIEELVNITQNQKAPLCNGSMVWSINLTAGMYCAALESLI NVSGCSAIEKTQRMLSGFCPHKVSAGQFSSLHVRDTKIEVAQFVKDLLLHLKKLFREGQF

[0709] N

[0710] > SEQ ID NO: 85 IL-24 QEFHFGPCQVKGVVPQKLWEAFWAVKDTMQAQDNITSARLLQQEVLQNVSDAESCYLV HTLLEFYLKTVFKNYHNRTVEVRTLKSFSTLANNFVLIVSQLQPSQENEMFSIRDSAHRRF LLFRRAFKQLDVEAALTKALGEVDILLTWMQKFYKL

[0711] > SEQ ID NO: 86 IL33 MKPKMKYSTNKISTAKWKNTASKALCFKLGKSQQKAKEVCPMYFMKLRSGLMIKKEACY FRRETTKRPSLKTGRKHKRHLVLAACQQQSTVECFAFGISGVQKYTRALHDSSITGISPIT EYLASLSTYNDQSITFALEDESYEIYVEDLKKDEKKDKVLLSYYESQHPSNESGDGVDGK MLMVTLSPTKDFWLHANNKEHSVELHKCEKPLPDQAFFVLHNMHSNCVSFECKTDPGV FIGVKDNHLALIKVDSSENLCTENILFKLSET

[0712] > SEQ ID NO: 87 IL36a KIDTPQQGSIQDINHRVWVLQDQTLIAVPRKDRMSPVTIALISCRHVETLEKDRGNPIYLGL NGLNLCLMCAKVGDQPTLQLKEKDIMDLYNQPEPVKSFLFYHSQSGRNSTFESVAFPG WFIAVSSEGGCPLILTQELGKANTTDFGLTMLF

[0713] > SEQ ID NO: 88 IL36b REAAPKSYAIRDSRQMVWVLSGNSLIAAPLSRSIKPVTLHLIACRDTEFSDKEKGNMVYL GIKGKDLCLFCAEIQGKPTLQLKLQGSQDNIGKDTCWKLVGIHTCINLDVRESCFMGTLD QWGIGVGRKKWKSSFQH H H LRKKDKDFSSM RTN IGM PGRM

[0714] > SEQ ID NO: 89 TNFa MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVIGPQRE EFPRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELR DNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRET PEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL

[0715] > SEQ ID NO: 90 TGFb MPPSGLRLLPLLLPLLWLLVLTPGRPAAGLSTCKTIDMELVKRKRIEAIRGQILSKLRLASP PSQGEVPPGPLPEAVLALYNSTRDRVAGESAEPEPEPEADYYAKEVTRVLMVETHNEIY DKFKQSTHSIYMFFNTSELREAVPEPVLLSRAELRLLRLKLKVEQHVELYQKYSNNSWRY LSNRLLAPSDSPEWLSFDVTGVVRQWLSRGGEIEGFRLSAHCSCDSRDNTLQVDINGFT TGRRGDLATIHGMNRPFLLLMATPLERAQHLQSSRHRRALDTNYCFSSTEKNCCVRQLY IDFRKDLGWKWIHEPKGYHANFCLGPCPYIWSLDTQYSKVLALYNQHNPGASAAPCCVP QALEPLPIVYYVGRKPKVEQLSNMIVRSCKCS

[0716] > SEQ ID NO: 91 Foldikinel 0-1 L34 ORK6_P_003TQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQA LSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQV KNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLK TLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN GGGGGSGGSGGSGGSGGSGGSGGSGGSGGGNEPLEMWPLTQNEECTVTGFLRDKL QYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESV QDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALL DNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSP PHSTGSVRPVRAQGEGLLP

[0717] > SEQ ID NO: 92 IL34-sclL10 NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVS PKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSG GSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLK TLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN GGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLL KESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN SEQ ID NO: 101 CSF1 short version ending in QDV EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDV

[0718] SEQ ID NO: 105 lgG4 silent FALA allotype (KIH) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVCTLPPSQEEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLVSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 118

[0719] EAAAK SEQ ID NO: 119

[0720] EAAAKEAAAKEAAAKEAAAKEAAAKEAAAK SEQ ID NO: 122 single chain dimeric IFNy QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQSQIVSFYFKLFKNF KDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNYSVTDLNVQRKAIHELIQVMAELS PAAKTGKRKRSQMLFRGGGGGSGGGGSQDPYVKEAENLKKYFNAGHSDVADNGTLFL GILKNWKEESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRD DFEKLTNYSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFR

[0721] >SEQ ID NO: 129 ORK6_P_011 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSNEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLN VQQGLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLN WQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGG GGGSGGSGGSGGSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSR VKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVN SLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAY MTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSE MIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKN AFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN

[0722] >SEQ ID NO: 130 ORK6_P_012 NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRL QRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQGLTDVEVS PKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQS CSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLPGGGGGSGGSGGSG GSGGSGGSGGSGGSGGGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLD NLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQEPDIKAHVNSLGENLKTLRLR LRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYL PNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMP QAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPS VFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSR WQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 144 Protein 1 of ORK10_P_01 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYL KKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPL QLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGG SGGSGGSGGSGGGAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYM PKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYA DETATIVEFLNRWITFCQSIISTLT SEQ ID NO: 145 Protein 1 of ORK10_P_02 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKAT ELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETAT IVEFLNRWITFCQSIISTLTGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCS HMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFR DNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLD KDWNIFSKNCNNSFAECSSQDV SEQ ID NO: 146 Protein 1 of ORK11_P_01 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYL KKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPL QLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGG SGGSGGSGGSGGGHKCDITLQEIIKDLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRA ATVLRQFYSHHEKDTRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQS TLENFLERLKTIMREKYSKCSS SEQ ID NO: 147 Protein 1 of ORK11_P_02 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSHKCDITLQEIIKDLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRAATVL RQFYSHHEKDTRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQSTLEN FLERLKTIMREKYSKCSSGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCS HMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFR DNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLD KDWNIFSKNCNNSFAECSSQDV SEQ ID NO: 148 Protein 1 of ORK12_P_01 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSEEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYL KKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPL QLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGS GGSGGSGGSGGGAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEM FDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESF KENLKDFLLVIPFDCWEPVQE SEQ ID NO: 149 Protein 1 of QRK12_P_02 ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTI SKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSG GGGSGGGGSAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDL QEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKEN LKDFLLVIPFDCWEPVQEGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCS HMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFR DNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLD KDWNIFSKNCNNSFAECSSQD SEQ ID NO: 150 Protein 1 of QRK12_P_03 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCL QTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLV IPFDCWEPVQEGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVM H EALH N H YTQKSLSLSLGSEQ ID NO: 151 Protein 1 of ORK12_P_04 APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRL ELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFD CWEPVQEGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGHLQS LQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQ LQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNC NNSFAECSSQDGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVM H EALH N H YTQKSLSLSLG

[0723] SEQ ID NO: 152 CSF1-GMCSF in ORK12_P_01 and ORK12_P_03 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCL QTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLV IPFDCWEPVQE

[0724] SEQ ID NO: 153 GMCSF-CSF1 in QRK12_P_02 and QRK12_P_04 APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRL ELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFD CWEPVQEGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGEEVSEYCSHMIGSGHLQS LQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQ LQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNC NNSFAECSSQD SEQ ID NO: 154 Protein 1 of ORK1_P_533 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMIFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVF NETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGG SGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLE DFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRD AFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIK AHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFIN YIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKP KDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSL WCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVF SCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 155 Protein 1 of ORK1_P_534 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNDIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVM H EALH N H YTQKSLSLSLG SEQ ID NO: 156 Protein 1 of ORK1_P_535 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIYQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVM H EALH N H YTQKSLSLSLG SEQ ID NO: 157 Protein 1 of ORK1_P_536 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLYELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVM H EALH N H YTQKSLSLSLG SEQ ID NO: 158 Protein 1 of ORK1_P_537 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FQETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVM H EALH N H YTQKSLSLSLG SEQ ID NO: 159 Protein 1 of ORK1_P_538EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDI MEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNV FNETKNLLDKDWNIFSKNCQNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSG GSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLL EDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLR DAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDI KAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFI NYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVM H EALH N H YTQKSLSLSLG SEQ ID NO: 160 Protein 1 of ORK1_P_539 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLYVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 161: Protein 1 of ORK1_P_540 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 162: Protein 1 of ORK1_P_541 EEVSRYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLGSEQ ID NO: 163: Protein 1 of ORK1_P_542 EEVSEYCSQMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 164: Protein 1 of ORK1_P_543 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG

[0725] SEQ ID NO: 165: Protein 1 of ORK1_P_544 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTEDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 166: Protein 1 of ORK1_P_545 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDYACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 167: Protein 1 of ORK1_P_546 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDNNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 168: Protein 1 of ORK1_P_547 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLQKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 169: Protein 1 of ORK1_P_548 EEVSEYCSHMIGSGHLQLLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 170: Protein 1 of ORK1_P_549 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKANLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 171: Protein 1 of ORK1_P_550 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVFDIMEDTMRFRDNTPNA IAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQ DVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTF FQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRF LPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 172: Protein 1 of ORK1_P_551 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSG GSGGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGV EVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQE EMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLG SEQ ID NO: 173: Protein 1 of ORK1_P_552 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 174: Protein 1 of ORK1_P_553 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKE AAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKN QVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLG SEQ ID NO: 176: Protein 1 of ORK1_P_554 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 176: Protein 1 of ORK1_P_555 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 177: Protein 1 of ORK1_P_556 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDV SQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKA KGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 178: Protein 1 of ORK1_P_557 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCPAPEAAGGPSVF LFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGSEQ ID NO: 179: Protein 1 of ORK1_P_558 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGSSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 180: Protein 1 of ORK1_P_559 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMK DQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGS GGSGGGESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEE MTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLG SEQ ID NO: 181: Protein 1 of QRK1_P_560 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMK DQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGGPSV FLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLnTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0726] SEQ ID NO: 182: Protein 1 of ORK1_P_561 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMK DQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAA AKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN HYTQKSLSLSLGSEQ ID NO: 183: Protein 1 of ORK1_P_562 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQM KDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFL PCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMK DQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLP CENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGPSVF LFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 184: Protein 1 of ORK1_P_563 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGESKYGPPCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 185: Protein 1 of ORK1_P_564 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNGGGGSESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDV SQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKA KGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 186: Protein 1 of ORK1_P_565 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNEAAAKEAAAKEAAAKEAAAKEAAAKEAAAKESKYGPPCPPCPAPEAAGGPSVF LFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 187: Protein 1 of ORK1_P_566 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQDVEAAAKSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQ ALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIY KAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQAL SEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRFLPCENKSKAVEQVKNAFNKLQEKGIYK AMSEFDIFINYIEAYMTMKIRNEAAAKESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPCQEEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 188: Protein 1 of ORK1_P_567 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHFVFRDNTPNA IAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSSQ DVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTF FQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRF LPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 189: Protein 1 of ORK1_P_568 EEVSRYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMLFRDNTPN AIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 190: Protein 1 of ORK1_P_569 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMLFRDNTPN AIAIYQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 191:

[0727] EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMLFRDNTPN AIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSSQDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 192: Protein 1 of ORK1_P_571 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMYFRDNTPN AIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 193: Protein 1 of ORK1_P_572 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMRFRDNTPN DIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 194: Protein 1 of ORK1_P_573 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMIFRDNTPNA IAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSSQ DVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTF FQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRF LPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 195: Protein 1 of ORK1_P_574EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMVFRDNTPN DIAIVQLQELSIRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 196: Protein 1 of ORK1_P_575 EEVSRYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMLFRDNTPN AIAIVQLQELSIYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 197: Protein 1 of ORK1_P_576 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMVFRDNTPN DIAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKT FFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRC HRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFF QMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHR FLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 198: Protein 1 of ORK1_P_577 EEVSEYCSHMIGSGHLQILQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDHMIFRDNTPND IAIVQLQELSVYLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNFFNETKNLLDKDWNIFSKNCNNSFAECSSQ DVGGGGGSGGSGGSGGSGGSGGSGGSGGSGGGSPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTF FQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCH RFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMNNGGLDYLPNMLRDLRDAFSRVKTFFQ MKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRNHRF LPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRNGGGGGSGGGGSGGGGSESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPCQEEMTKNQVSLWCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLGSEQ ID NO: 201: single chain CSF1 EEVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQEQLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPN AIAIVQLQELSLRLKSCFTKDYEEHDKACVRTFYETPLQLLEKVKNVFNETKNLLDKDWNIFSKNCNNSFAECSS QDVGGGGGSGGGGSGGSGGSGGSGGSGGSGGSGGSGGGHDKACVRTFYETPLQLLEKVKNVFNETKNLL DKDWNIFSKNCNNSFNECEDQDRRQPGTKTTGKVSEYCSHMIGSGHLQSLQRLIDSQMETSCQITFEFVDQE QLKDPVCYLKKAFLLVQDIMEDTMRFRDNTPNAIAIVQLQELSLRLKSCFTKDYEE

Claims

CLAIMS1. A single chain polypeptide having dual activities of a cytokine targeting CSF1 R and of a cytokine of interest.

2. The single chain polypeptide according to claim 1 , which comprises in the N-terminus to C-terminus direction, (i) a cytokine domain targeting CSF1R, a peptide linker L1 , and a cytokine of interest, or (ii) a cytokine of interest, a peptide linker L1 , and a cytokine domain targeting CSF1R.

3. The single chain polypeptide according to claim 1 or 2, wherein the cytokine domain targeting CSF1R is a CSF1 monomer, a dimeric single chain CSF1 polypeptide, or an IL-34 monomer.

4. The single chain polypeptide according to any one of claims 1 to 3, wherein the cytokine domain targeting CSF1R is:(i) an IL-34 monomer comprising sequence SEQ ID NO: 3, or a sequence at least 80% identical thereto and that retains IL-34 activity,(ii) a CSF1 monomer comprising sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 101, or a sequence at least 80% identical thereto and that retains CSF1 activity,(iii) a CSF1 monomer comprising sequence SEQ ID NO: 207, or a sequence at least 80% identical thereto and that retains CSF1 activity.(iv) The single chain CSF1 polypeptide comprising SEQ I D NO: 201 , or a sequence at least 80% identical thereto and that retains CSF1 activity.

5. The single chain polypeptide according to claim 4, wherein the cytokine monomer targeting CSF1R is a CSF1 monomer that comprises a sequence that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 101:(i) by a substitution at one or more of positions 5, 9, 18, 51, 54, 56, 58, 64, 65, 66, 74, 78, 81, 85, 86, 93, 100, 120, 122, 132, and 140 in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 101, or(ii) by substitution M65Y, S18I, F54G, L85I, Q58E, T64H, M65Y, R66Y, A74D, V78Y, Q81Y, N122Q, L56Y V120F, E5R, H9Q, R86Y, K93E, K100Y, W132N, K51Q, S18L, F54N, or Q58F.

6. The single chain polypeptide according to any one of claims 1 to 4, wherein the cytokine of interest is selected from the group consisting of IL- 10, GM-CSF, IL-2, IL-4, IFN-a, IFN-p, IFN-y, IL-27, IL-33, IL-13, IL-24, IL-1 p, IL-36, TNFa, TGFp, IL-6 and IL-35.

7. The single chain polypeptide according to claim 6, comprising from the N-terminus to the C-terminus direction, (i) a CSF1 domain, the cytokine of interest, and a Fc fragment, or (ii) a Fc fragment, the cytokine of interest, and the CSF1 domain.

8. The single chain polypeptide according to any one of claims 2 to 7, wherein one or more of the following conditions are met:(i) the peptide linker L1 comprises from 5 to 50 amino acid residues;(ii) the peptide linker L1 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;(iii) the peptide linker L1 comprises sequence SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, or SEQ ID NO: 16;(iv) the peptide linker L1 comprises sequence (SEQ ID NO: 17)n, n being an integer from 1 to 10;(v) the peptide linker L1 is a rigid peptide sequence composed of Glu, Ala, and Lys residues; or(vi) the peptide linker L1 comprises sequence EAAAK (SEQ ID NO: 118), or (EAAAK; SEQ ID NO: 118)nwith n being a integer from 2 to 10;(vii) the peptide linker L1 comprises sequence SEQ ID NO: 119.

9. The single chain polypeptide according to any one of claims 1 to 8, which is a single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1 R and GM-CSF.

10. The single chain polypeptide according to claim 9, which comprises sequence SEQ ID NO: 152, SEQ ID NO: 153, or a sequence at least at least 80% identical thereto and that retain the dual CSF1 and GM-CSF activities.

11. The single chain polypeptide according to any one of claims 1 to 8, which is a single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1 R and IL-4.

12. The single chain polypeptide according to claim 11, which comprises sequence SEQ ID NO: 11, SEQ ID NO: 12, or a sequence at least at least 80% identical thereto and that retain the dual CSF1 and IL-4 activities.

13. The single chain polypeptide according to any one of claims 1 to 8, which is a single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1 R and IL-2.

14. The single chain polypeptide according to claim 13, which comprises sequence SEQ ID NO: 9, SEQ ID NO: 10, or a sequence at least at least 80% identical thereto and that retain the dual CSF1 and IL-2 activities.10815. The single chain polypeptide according to any one of claims 1 to 8, which is a single chain polypeptide having dual cytokine activities of a cytokine targeting CSF1 R and IFN-y.

16. The single chain polypeptide according to claim 15, which comprises sequence SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 7, SEQ ID NO: 208, SEQ ID NO: 8, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 211, or a sequence at least 80% identical thereto and that retain the dual CSF1 and IFN-y activities.

17. The single chain polypeptide according to any one of claims 1 to 16, which is fused to a human immunoglobulin Fc fragment, to albumin or an albumin fragment, to an anti-albumin antibody or fragment thereof, or which is conjugated to a poly(ethylene glycol) (PEG) molecule.

18. A protein comprising a single chain polypeptide according to any one of claims 1 to 17 fused to an immunoglobulin Fc fragment.

19. The protein according to claim 18, which comprises a first polypeptide comprising the single chain polypeptide having dual activities of a cytokine targeting CSF1R and cytokine of interest according to any one of claims 1 to 15 fused to a first immunoglobulin Fc fragment through a peptide linker L2, and a second polypeptide comprising a second immunoglobulin Fc fragment.

20. The protein according to claim 18, which comprises two polypeptide chains comprising the single chain polypeptide having dual activities of a cytokine targeting CSF1R and cytokine of interest according to any one of claims 1 to 15 fused to a Fc fragment through a peptide linker L2.

21. The protein according to claim 19 or 20 wherein one or more of the following conditions are met:(i) the peptide linker L2.comprises from 3 to 45 amino acid residues;(ii) the peptide linker L2 is a flexible peptide sequence composed of Gly and Ser residues, in different proportion;(iii) the peptide linker L2 comprises any one of sequences SEQ ID NO: 15, SEQ ID NO: 17 to SEQ ID NO: 28, SEQ ID NO: 99, SEQ ID NO: 130, SEQ ID NO: 134;(iv) the peptide linker L2 comprises sequence (SEQ ID NO: 17)p, n being an integer from 1 to 8,(v) the peptide linker L2 is a rigid peptide sequence composed of Glu, Ala, and Lys residues,(vi) the peptide linker L2 comprises sequence SEQ ID NO: 118 or (SEQ ID NO:118)n with n being a integer from 1 to 10,109(vii) the peptide linker L2 comprises sequence SEQ ID NO: 119.

22. The protein according to any one of claims 18 to 21, wherein the Fc fragment or domain is from lgG1, lgG2, lgG3 or lgG-4, silent or non-silent.

23. The protein according to any one of claims 18 to 22, which comprises a first polypeptide chain comprising any one of sequence:(i) SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or a sequence at least 80%% identical thereto and that retains CSF1 and IFN-y activities;(ii) SEQ ID NO: 144, SEQ ID NO: 145, or a sequence at least 80% identical thereto wherein said protein retains CSF1 and IL-2 activities;(iii) SEQ ID NO: 146, SEQ ID NO: 147, or a sequence at least 80% identical thereto and that retains CSF1 and IL-4 activities;(iv) SEQ ID NO: 148 to SEQ ID NO: 151, or a sequence at least 80% identical thereto and that retains CSF1 and GM-CSF activities.

24. The protein according to claim 23, which further comprises a second polypeptide chain comprising SEQ I D NO: 105, or a sequence at least 80% identical thereto, wherein the first and second polypeptides dimerise.

25. A pharmaceutical composition comprising the single chain polypeptide according to any one of claims 1 to 17, or the protein according to any one of claims 18 to 24, and a pharmaceutically acceptable carrier.

26. A single chain polypeptide according to any one of claims 1 to 16, the protein according to any one of claims 18 to 24, or the pharmaceutical composition according to claim 25, for use as a medicament.

27. A single chain polypeptide according to any one of claims 1 to 17, the protein according to any one of claims 18 to 24, or the pharmaceutical composition according to claim 26, for use for treating a disease comprising cancers, infectious diseases, inflammatory diseases, auto-immune diseases, neuro-inflammatory diseases, infectious diseases and tissue repair.

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