Anti-inflammatory polypeptides derived from helminthic glutamate dehydrogenase (HEGDH), methods and uses thereof

Abstract

The present invention relates to novel enzymatically inactive structural polypeptides derived from the N-terminal part of heGDH (helminthic glutamate dehydrogenase), therapeutical methods based thereon and uses thereof.

Description

Anti-inflammatory polypeptides derived from helminthic glutamate dehydrogenase (heGDH), methods and uses thereofCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the right of priority of European patent application EP24173451.6 filed with the European Patent Office on 30 April 2024, the entire content of which is incorporated herein for all purposes.SEQUENCE LISTING

[0002] This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.TECHNICAL FIELD

[0003] Based on their potent immune regulatory capacities, helminth parasites have long been recognized as a potential source of “natural” immunomodulators, which may be particularly efficacious against type 2 inflammatory diseases (Maizels et al. Immunity 49, 801-818 (2018)., Bohnacker et al. Front. Immunol. 11 , 2106 (2020)). Although a large number of anti-inflammatory helminth molecules have been identified and shown efficacy in experimental models of allergy and autoimmunity (Maizels et al. Immunity 49, 801-818 (2018)., Bohnacker et al. Front. Immunol. 11 , 2106 (2020), Cauche et al. Immunology 167, 197-211 (2022), Osbourn et al. Immunity 47, 739-751.e5 (2017), Navarro et al. Sc / . Transl. Med. 8, 362ra143 (2016)), no helminth based therapeutic has been approved or entered the clinic so far.BACKGROUND OF THE INVENTION

[0004] Inhaled and intranasal glucocorticoids (GCs) represent the current standard treatment against asthma and CRSwNP (Chronic Rhinosinusitis with Nasal Polyps). However, a significant number of asthmatic patients experience symptoms and exacerbations despite treatment with inhaled GCs (Henderson et al., Pharmacol. Res. 160, 105189 (2020)) and most nasal polyps are at least partially refractory to intranasal GCs (Riva et al., Am. J. Otolaryngol. 43, 103325 (2022)). When treatment with inhaled / intranasal GCs fails, patients are often subjected to treatment with oral GCs, which however have significant side effects, making them inadequate for long-term treatment. While GCs target a broad range of pro- and anti-inflammatory genes, most other current treatments against asthma and CRSwNP target single molecules. These targets include leukotrienes (LTs), histamine, type 2 cytokines (e.g. IL-4, IL-5) and immunoglobulin E (IgE) as well as their respective receptors. However, like GCs, each of these groups of drugs have significant drawbacks:Leukotriene receptor antagonists (e.g. montelukast / Singulair) have limitedefficacy against asthma and CRSwNP, potentially as a result of their failure to inhibit the synthesis of LTs and due to LT receptor redundancy (Van Gerven et al. The Laryngoscope 128, 1743-1751 (2018)). In addition, the LT synthesis inhibitor Zileuton does not represent a viable long-term treatment option due to its potential hepatotoxic effects (Watkins et al. Drug Saf. 30, 805-815 (2007)). Despite providing relief of acute allergic symptoms, histamine (H1) receptor antagonists are not sufficiently reducing the chronic type 2 inflammation, which drives asthma or CRSwNP. In contrast to these relatively old small molecule inhibitors, the more recently approved monoclonal antibodies (“biologies”) targeting IL-4 receptor alpha, IL-5 or IgE have shown very promising clinical efficacy against CRSwNP and severe “type 2” asthma (Bachert et al., J. Allergy Clin. Immunol. 149, 1711-1721.e6 (2022), Schneier et al., Eur. Respir. J. 2201335 (2022), Akenroye et al., J. Allergy Clin. Immunol. S0091-6749(23)00144-6 (2023)).

[0005] However, nasal polyposis (NP), which often occurs together with asthma, still represents a major unmet clinical need. Traditionally, NP are treated with intranasal glucocorticoids, which however have limited efficacy in most patients. Biological treatments (e.g. monoclonal antibodies targeting IL-4, IL-5 or IgE) have advanced the therapy of NP and severe asthma to certain extent. However, many patients do not benefit from these expensive treatments. In addition, monoclonal antibodies have to be administered systemically by subcutaneous injection and may thus have systemic, currently poorly defined, long-term side effects, which is a clear disadvantage of such therapies.

[0006] Therefore, there is an urgent need for novel, improved local treatments of NP and severe asthma. The invention aims to make use of the unique immune regulatory potential of a helminth enzyme (helminthinc glutamate dehydrogenase, heGDH) and its active N-terminal subunit / s (e.g., polypeptides). Advantageously, the invention relates to the use of the N-terminal peptide / s of heGDH (e.g., in isolation or linked to a suitable protein backbone) as a novel biological treatment against nasal polyposis and / or asthma.SUMMARY OF THE INVENTION

[0007] The present invention relates to H. polygyrus glutamate dehydrogenase (heDGH) that can drive parasite chronicity by e.g., epigenetically and metabolically reprogramming host macrophages, in particularly the present invention relates to identified novel structural and catalytically inactive polypeptides derived from the N-terminal part of the heDGH and methods and uses of said polypeptides (e.g., as a medicament and / or in therapy, preferably for use in a method of treatment / amelioration / prophylaxis of NP, asthma and / or NP comorbid with asthma).

[0008] In one aspect, the present invention relates to a polypeptide, wherein said polypeptide comprising one or more amino acid sequences (e.g., polypeptides) having at least 80% (e.g., at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity toany one of polypeptides (e.g., heDGH-derived, e.g., catalytically inactive, structural polypeptides) selected from the group consisting of:I) SEQ ID NO: 1 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDL); ii) SEQ ID NO: 2 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDLKPMEEQSN); iii) SEQ ID NO: 3 (MLSTLARTSGRLIFRRALSSA); iv) SEQ ID NO: 4 (QMDAHAQVIDDLKP); and v) SEQ ID NO: 5 (MEEQSN).

[0009] In another aspect, the polypeptide of the present of the present invention comprises a fragment or variant of any one of (i)-(v) as defined herein above, preferably said fragment or variant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11 , at least 12, at lest 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 , at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31 , at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39 or at least 40) amino acids long.

[0010] In another aspect, the polypeptide / s of the present of the present invention not have any enzymatic activity and / or is catalytically inactive, e.g., does not have EC 1.4.1.2, EC 1.4.1.3 and / or EC 1.4.1.4 enzymatic activity, preferably does not have EC 1.4.1.3 enzymatic activity), preferably does not have heGDH enzymatic activity (e.g., EC:1.4.1.3).

[0011] In another aspect, the present invention further relates to one or more nucleic acid constructs or expression vectors encoding one or more polypeptides of the present invention.

[0012] In another aspect, the present invention further relates to a host cell (e.g. an isolated and / or recombinant host cell) comprising one or more nucleic acid constructs and / or expression vectors of the present invention and / or expressing one or more polypeptides of the present invention.

[0013] In another aspect, the present invention further relates to a composition or kit comprising one or more of the polypeptides, nucleic acid constructs or expression vectors and / or host cells of the present invention.

[0014] In another aspect, the present invention further relates to a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant airway inflammation, aspirin / NSAID- exacerbated respiratory disease (AERD / N-ERD), nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma,idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; said method comprising administering a therapeutically effective amount of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions of any one of the preceding items, preferably administered locally (e.g., intranasally), further preferably administered to the airways.

[0015] In another aspect, the present invention further relates to one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of of the present invention for use as a medicament and / or in therapy.

[0016] In another aspect, the present invention further relates to one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of of the present invention for use in one or more of the following methods: (i) in a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant airway inflammation, aspirin- exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; preferably administered locally (e.g., intranasally), further preferably administered to the airways; (ii) in a method for modulating the mammalian innate and adaptive immune response, preferably administered locally (e.g., intranasally), further preferably administered to the airways; (iii) in a method for predominantly modulating the mammalian innate immune response over the mammalian adaptive immune response; (iv) in a method for targeting phagocytic cells of the mammalian immune system; (v) in a method for modifying the activation of macrophages and / or granulocytes of the mammalian immune system; (vi) in a method for modifying the activation of one or more of the arachidonic acid pathway of the mammalian immune system; (vii) in a method for decreasing the number of eosinophils and / or inhibiting the migration of granulocytes into tissue of the mammalian immune system; (viii) in a method for eliciting or modulating an immune response in a subject; (ix) in a method for treatment, amelioration, prophylaxis or diagnostics of a steroid-resistant disease; (x) in a method for monitoring development of a disease and / orassessing the efficacy of a therapy of a disease; (xi) in any method of the present invention; (xii) in any combination of methods as in any one of (i)-(xi); (xiii) in any method according to (f)-(xii), wherein said method is an in vitro, in vivo or ex vivo method.

[0017] Accordingly, in the course of the present invention it has been shown that heGDH efficiently suppresses airway inflammation in a mouse model of asthma when administered locally (intranasally) to the airways.

[0018] In another aspect of the present invention it has been shown that the N-terminal part of heGDH (e.g., polypeptides of the present invention) mediates the protein’s unique, broad immune regulatory potential. Furthermore, heGDH simultaneosly targets multiple mechanisms involved in chronic airway inflammation (e.g. suppression of M2 macrophage activation, suppression of leukotrienes, induction of anti-inflammatory IL-10 and prostaglandin E2 (PGE2)). This includes disease-relevant lipid mediators (prostanoids and leukotrienes), which are poorly targeted by standard treatments such as glucocorticoids and current biologies.

[0019] The N-terminal polypeptide sequences of heGDH as described herein are unique to the helminthic protein, thus providing a clear distinction from the mammalian homologues, which lack immune regulatory functions / activities.

[0020] The present application satisfies the demand of providing novel therapeutic entities as described herein by provision of novel and unique polypeptides, compositions, vaccines, kits, isolated host cells, methods and uses described herein below, characterized in the claims and illustrated by the appended Examples.BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1: GDH enables helminth immune evasion by inducing type 2 suppressive macrophages. (A) Experimental model of Hpb infection and treatment with a-heGDH monoclonal antibody (mAb, 4F8) or isotype control antibody (Ab). (B) Worm burden 14 days p.i. in mice treated with a-heGDH mAb or isotype control (n=9 in isotype control group, n=10 in a -heGDH mAb group). (C) Representative immunofluorescence image for heGDH and CD64 in small intestine from mice infected with Hpb (4 days). White arrows: colocalization of heGDH and macrophages in the surrounding of larvae (dashed circle). (D) Representative immunofluorescence image for MDM ± treatment with AF488-labeled heGDH for different time points (scale bar = 20 pm). (E) Volcano plot showing DEGs for MDM ± heGDH (n=3). Significant DEGs were selected by base mean>50, padj<0.1 and logzFOI. Labeled DEGs selected with padj<1e-20 or logzFCM. (F) Heatmap showing top 50 DEGs of MDM treated + heGDH (n=3). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Mann-Whitney test (B) or DESeq2 (E-F). *P < 0.05 (B), *P<0.1 (E-F).

[0022] Figure 2: HeGDH induces a type-2 suppressive switch of the arachidonic acid (AA) metabolism in human macrophages. (A) Left, heatmap of lipid mediators secreted by humanMDM ± treatment with heGDH (LC-MS / MS). Right, levels of major COX metabolites produced by human MDM ± treatment with heGDH or mock vector purification (n=9). (B) Major 5-LOX metabolites (LC-MS / MS) released by human MDM ± treatment with heGDH or mock vector purification (n=9). (C) Secretion of IL-10 (ELISA) by MDM + treatment with heGDH or mock vector purification (n=6). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A, heatmap) and Friedman test (A-C, bar graphs). *P < 0.05; **P < 0.01, ***P < 0.001.

[0023] Figure 3: p300 HAT activation by heGDH mediates the induction of immune- regulatory genes. (A) Gene expression analysis of top DEGs (qPCR) in MDM + heGDH ± p300 HAT inhibitor (n=9). Doted lines indicate expression in control (Ctrl) cells. (B) PGEz, IL-12p and IL-10 secretion (ELISA) from MDM + heGDH ± p300 HAT inhibitor (n=9). (C) Representative immunofluorescence image of p300, DAPI, COX2 and mPGESI in MDM ± heGDH (ctr or p300 knock down), (scale bar = 20 pm). (D) H3K27ac or actin (western blot) in MDM + heGDH + p300 HAT inhibitor. Left, quantification for n=4 donors; right, representative blot for one donor. (E) H3K27ac peaks for MDM ± heGDH (n=3). Colored: FDR £ 0.05; absolute log2FC>1. Peaks of top hits from RNA-seq marked (black dots), bold: most significant peak. Further labeled peaks: overlapping hits from ChlP-and RNA-seq. (F) Average H3K27ac ChlP-seq signal of MDM ± heGDH (n=3) for top differential peaks (grey window; FDR £ 0.05; absolute log2FC > 1 ). (G) Scater plot showing top DEGs from RNA-seq against the top H3K27ac peaks from ChlP-seq. Genes of interest marked black and bold. (H) H3K27ac ChlP-qPCR for IDO1, PTGS2 and ILS in MDM ± heGDH (n=3). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A), Friedman test (B), RM one-way ANOVA (D) or paired t-test (H). *P < 0.05; **P < 0.01; **‘P < 0.001.

[0024] Figure 4: Structural features rather than catalytic activity confer regulatory functions of heGDH. (A) GDH activity of heGDH and heGDHK126A> D204Nin the direction of glutamate utilization and formation (n=4-7). (B) Levels of prostanoids (LC-MS / MS) and IL-10 (ELISA) produced by MDM ± heGDH or heGDHK126A-D204N(n=9). (C) Levels of major COX metabolites (LC-MS / MS) produced by BMDM ± heGDH or heGDHK126A>D204N(n=3). (D) Gene expression analysis of top DEGs (qPCR) in MDM ± heGDH or heGDHK126A> D204N(n=5). (E) Secretion of cysLTs (EIA) from MDM ± heGDH or heGDHK126A'D204N(n=8). (F) Fold change of cysLT secretion (EIA) from MDM + heGDH (Hpb or Ts) or human GDH (n=4-10). (G) Model of heGDH crystal structure fited in the oligomeric cryo-EM reconstruction. N-termini of two neighboring subunits, which are resolved in the X-ray structure are near to a handle-like density in the cryo-EM model. (H) Close up of the handle-like density and the N-termini from the heGDH crystal structure fited in the cryo-EM reconstruction. (I) Secretion of PGE2and IL-10 (EIA, ELISA) from MDM ± heGDH, heGDHc136Sor heGDHAN(n=7). (J) Gene expression analysis of top DEGs (qPCR) in MDM ± heGDH, heGDHc136Sor heGDHAN(n=7). (K) Top three identified proteins bymass spectrometry in MDM (n=4) binding to HA-heGDH. Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by unpaired t-test (A), Friedman test (B-E, l-J) or Kruskal Wallis test (F). *P < 0.05; **P < 0.01 ; ***P < 0.001 , ****P < 0.0001.

[0025] Figure 5: Suppression of leukotrienes is mediated via metabolic reprogramming. (A) Oxygen consumption rate (OCR, left), extracellular acidification rate (ECAR, middle) and basal glycolysis (right) of MDM ± heGDH (n=6). (B) H3K27ac ChlP-seq signal of MDM ± heGDH (n=3) for the PFKFB3 peak (grey window; FDR < 0.05; absolute log2FC > 1 ). (C) Gene expression analysis of PFKFB3 (qPCR) in MDM + heGDH ± p300 HAT inhibitor (n=8). Doted line indicates expression in control (Ctrl) cells. (D) Upper panel: OCR and ECAR of MDM ± heGDH + p300 HAT inhibitor. Lower panel: basal respiration, ATP production and basal glycolysis of MDM ± heGDH ± p300 HAT inhibitor (n=7). (E) Heatmap of targeted metabolomics (LC-MS / MS) in MDM ± heGDH (n =6). (F) Targeted metabolomics (LC-MS / MS) for glutamine, glutamate and 2-hydroxyglutarate (2-HG) in MDM ± heGDH (n=6). (G) Secretion of cysLTs (EIA) by MDM ± L-2-HG (n=6). (H) Leukotriene C4 synthase (LTC4S) activity after incubation with L-2-HG (n=5). (I) LTC4S activity after addition of heGDH (1 or 3 pg) or a specific LTC4S inhibitor (TK05) (n=9). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A, C, E-G), Mann-Whitney test (H), Friedman test (D) or ordinary one-way ANOVA (I). *P < 0.05; **P < 0.01 ; ***P < 0.001 ; ****P < 0.0001.

[0026] Figure 6: heGDH-induced PGEz suppresses type 2 effector functions of macrophages and T cells. (A) Experimental model of 14-days Hpb infection and heGDH treatment. (B) Worm burden, 14 days p.i. in mice treated with PBS or heGDH (n=19 in PBS, n = 22 in heGDH group). (C) Left, representative images of IHC staining for RELMa (scale bar = 150 pm) and right, representative images of IF staining for RELMa, COX2 and DAPI (scale bar = 50 pm) in granulomas of tissues from mice infected with Hpb and treated with PBS or heGDH. (D) Protein amounts of H3K27ac or actin (western blot) in peritoneal macrophages (pMacs) from mice infected with Hpb and treated with PBS or heGDH. Left, quantification for n=7 mice; right, representative blot for four mice. (E) Secretion of PGE2 (EIA) of isolated pMacs (14 days p.i.) from mice infected with Hpb and treated with PBS or heGDH (n=14). Dotted line indicates secretion from pMacs of naive mice. (F) Surface expression of AAM marker CD206 (flow cytometry) in wildtype (WT) or EP2 knock out BMDM ± heGDH (n=8). (G) Experimental model of 28-days primary infection with Hpb and heGDH treatment (H) Worm burden, 28 days p.i. in mice infected with Hpb and treated with PBS or heGDH (left: first- (n = 5); right: second-experiment (n = 5 in PBS, n = 4 in heGDH group)). (I) Secretion of PGE2(EIA) of isolated pMacs (28 days p.i.) from mice infected with Hpb and treated with PBS or heGDH (n=5). (J) TXB2 and 6-keto-PGFiaconcentration (LC-MS / MS) in culture supernatants from isolated intestinal tissue (28 days p.i.) of mice infected with Hpb and treated with PBS or heGDH (n=5). (K) Percentage of Gata3* Th2 cells(flow cytometry) in mesenteric lymph nodes of 28-days infected Hpb-mice treated with PBS or heGDH (n=5). Dotted line indicates percentage in MLN from naive mice. (L) Percentage of IL-4* Th2 cells (flow cytometry) and secretion of IL-4 (ELISA) from human PBMC ± SEA ± heGDH ± mPGESI inhibitor (n=9). (M) Percentage of CD4+CD127*CD25hiFoxP3*(flow cytometry) in human PBMC ± heGDH + mPGESI inhibitor (n=5). Data are pooled from two to five independent experiments and presented as means + SEM. Statistical significance was determined by Mann- Whitney (B, D-E, H-K), Kruskal-Wallis test (F) or Friedman test (M, L). *P < 0.05; **P < 0.01; ***P < 0.001 , ****P < 0.0001.

[0027] Figure 7: Host type 2 immunity limits immune evasion, but not tissue repair driven by heGDH. (A-C) Gene expression analysis of AAM markers (A), top DEGs (B) or PGEz synthesis genes (C) (qPCR) in human MDM ± heGDH or after IL-4, IL-13 pre-treatment (n=6). (D) Secretion of PGEa (EIA) of MDM ± heGDH or after IL-4, IL-13 pre-treatment (n=6). (E) Experimental model of secondary Hpb infection and heGDH treatment. (F) Worm burden, 14 days post secondary Hpb infection in mice treated with PBS or heGDH (n=8 in PBS, n=10 in heGDH group). (G) Left, representative images of IHC staining for RELMa (scale bar = 150 pm) and right, representative images of IF staining for RELMa, COX2 and DAPI (scale bar = 50 pm) in granulomas of tissues from challenge Hpb infected mice treated with PBS or heGDH. (H) Secretion of PGEa (EIA) by peritoneal macrophages (pMacs) (14 days p. challenge infection) of mice treated with PBS or heGDH (n=5). (I) Experimental model of Nb infection treated i.n. with PBS or heGDH. (J) Nb worm burden, 6 days p.i. in mice treated with PBS or heGDH (n=12 in PBS, n=13 in heGDH group). (K) Secretion of PGE2 (EIA) by BAL macrophages from Nb infected mice treated with PBS or heGDH (n=12 in PBS, n=13 in heGDH group). Doted line indicates secretion from BAL macrophages of naive mice treated with PBS, dashed line indicates secretion after heGDH treatment. (L) Levels of LTB4 (LC-MS / MS) per neutrophil counts in BALF from Nb infected mice treated with PBS or heGDH (n=12 in PBS, n=13 in heGDH group). (M) Left, quantification of lung damage as linear means intercept (Lmi) (n=12 in PBS, n=13 in heGDH group). Right, representative H&E staining’s of lung sections from Nb infected mice treated with PBS or heGDH (scale bar = 500 pm). Doted line indicates Lmi of naive mice. Data are pooled from two to four independent experiments and presented as means + SEM. Statistical significance was determined by Friedman test (A-D) or Mann-Whitney (F, H, J-M). *P < 0.05; **P < 0.01.

[0028] Figure 8: HeGDH is present in all stages of Hpb and internalized by macrophages. heGDH in vivo blockade has variable effects on myeloid cell recruitment. (A) Gene expression analysis of heGDH (qPCR) in L3, L4 and L5 stage of Hpb (n=4). Mean of two primer pairs was used to calculate CT values. (B) Representative images of IF staining for 15-LOX, CD64 (rat), Myelodperoxidase (MPO) and DAPI in granulomas of tissues from 14-days primary Hpb infected mice treated intraperitoneal (i.p.) with a-heGDH monoclonal antibody (mAb, 4F8) or isotype control antibody (Ab), (scale bar = 50 pm). (C) Top image:Representative immunofluorescence staining for heGDH and CD64 (rabbit); Middle and bottom image: Representative immunofluorescence staining for heGDH and isotype control antibody (rabbit) for CD64 (rabbit) in small intestine from mice infected with Hpb (4 days). White arrows indicate colocalization of heGDH and macrophages in the surrounding of larvae (dashed circle). (D) Representative immunofluorescence image for heGDH, actin and DAPI in MDM ± heGDH and heGDH in combination with the antigen peptide for the a-heGDH mAb (4F8), (scale bar = 20 pm). (E) Representative immunofluorescence image of actin and DAPI in BMDM ± AF488-labeled heGDH for different time points (30 min, 3h, 24h), (scale bar = 20 pm). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Kruskal-Wallis test (A).

[0029] Figure 9: The maximal dose of LPS found in recombinant heGDH (SEQ ID NO: 27) preparations fails to mimic effects of heGDH. (A) Volcano plot showing DEGs of MDM stimulated with heGDH compared to 1 ng / mL LPS stimulation (n=3) with a padj<0.1, log2FC>1 and a base mean>50. Labeled DEGs are for genes with either padj<1e-20 or with log2FC>2. (B) Heatmap of top 50 DEGs between MDM ± heGDH or 1 ng / mL LPS (n=3), padj<0.1 , log2FC>1; base mean>50. (C) Expression of IL6, PTGS2 and IL1B (RNAseq) upon 1 ng / mL LPS stimulation in MDM (n=3). (D) Secretion of PGE2 (EIA) and IL-10 (ELISA) produced by MDM ± 1 ng / mL LPS (n=6). Statistical significance was determined by DESeq2 (A-B) or Wilcoxon test (C-D). *P<0.1 (A-B), *P < 0.05 (C-D).

[0030] Figure 10: Clone 4F8 mAb attenuates heGDH triggered prostanoid and IL-10 induction but not 5-LOX metabolite suppression. (A) Prostanoids quantified (LC-MS / MS) in supernatants of MDM ± heGDH ± a-heGDH mAb (4F8) (n=5). Dotted lines indicate prostanoid levels in control (Ctrl) cells. (B) Secretion of IL-10 (ELISA) in MDM ± heGDH ± a-heGDH mAb (4F8) (n=5). Doted line indicates mean secretion of IL-10 in control (Ctrl) cells. (C) 5-LOX metabolites quantified (LC-MS / MS) in supernatants of MDM + heGDH ± a-heGDH mAb (4F8) (n=5). Dotted lines indicate mean secretion in control (Ctrl) cells. Data are pooled from two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A-C).

[0031] Figure 11: HeGDH driven p300 HAT activation and H3K27 acetylation specifically Induces type 2 suppressive genes in macrophages. (A) Gene expression analysis of top DEGs (qPCR) in BMDM + heGDH ± p300 HAT inhibitor(A485) (n=7). Doted lines indicate mean expression in control (Ctrl) cells. (B) Gene expression analysis of interferon-induced genes (qPCR) in MDM + heGDH + p300 HAT inhibitor (A485) (n=8). Dotted lines indicate mean expression in control (Ctrl) cells. (C) Cytotoxicity measurement (LDH assay) of MDM (24h) or BMDM (6 h) ± heGDH + p300 HAT inhibitor (A485) (n= 4-6). (D) Quantification of EP300 siRNA knock down (qPCR) in MDM + heGDH ± p300 siRNA (n=8). Doted lines indicate mean expression in control (Ctrl) treated cells. (E) Representative immunofluorescence image of p300,DAPI and COX2 in BMDM + heGDH and heGDH treated cells after p300 siRNA knock down (scale bar = 20 pm). (F) Representative immunofluorescence image of p300, DAPI and CD64 (mouse) in MDM ± heGDH and heGDH treated cells after p300 siRNA knock down (scale bar = 20 pm). (G) Protein amounts of H3K27ac or actin (western blot) in BMDM + heGDH + p300 HAT inhibitor (A485). Top, representative blots for one mouse. Bottom, quantification for n=4 mice. (H) Log2FC of ChlPseq and RNAseq visualized for the eicosanoid pathway. Integration of both data sets into the WP167 (Eicosanoid synthesis) pathway by coloring the background of the genes with the log2FC of the RNA-seq analysis (red - positive log2FC, blue - negative log2FC) and the boarder of the gene boxes according to the log2FC of the ChlP-seq analysis (purple - positive log2FC, green - negative log2FC). Gene names written in grey were filtered from the significant genes due to their low counts and low expression. Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A-B, D), Friedman test (C), or RM one-way ANOVA (G). *P < 0.05; **P < 0.01.

[0032] Figure 12: Cryo-EM structure determination of heGDH and its mutants. (A) Representative micrograph of vitrified heGDH. (B) Cryo-EM image processing workflow for heGDH and its mutants. Picked particles were 2D-classified and used for ab initio reconstruction independently (example shown: heGDHDN). 3D classification and refinement yielded D3-symmetric, hexameric heGDH. (C) Local resolution of cryo-EM reconstruction of the heGDH oligomer (EMDB-ID: 18456).

[0033] Figure 13: Characterization of heGDH activity, structure and cellular targets as well as effects of different heGDH homologues on cysLT formation. (A-C) Activity of recombinant heGDH (SEQ ID NO: 27) in the direction of glutamate utilization and formation with varying concentrations of a-ketoglutarate, glutamate and ammonium (n=4-6). (B) Effects of pH on the activity of recombinant heGDH in the direction of glutamate utilization and formation (n=2). (C) Activity of recombinant heGDH in the direction of glutamate utilization and formation with varying concentrations of cofactors (n=4-6). (D) Effect of GTP or GDH inhibitor bithionol on the activity of heGDH in the direction of glutamate utilization and formation (n=3-7). (E) Effect of a-heGDH mAb (4F8) on the activity of heGDH in the direction of glutamate utilization and formation (n=3). (F) Fold change of cysLT secretion (EIA) from BMDM + heGDH (Hpb or 7s) or human GDH (n=4-8). (G) Overlay of X-Ray structures of human GDH (PDB: 1L1 F) and heGDH (PDB ID: 8QF0). Human GDH is shown in cyan, heGDH is shown in purple. (H) Density map of human GDH simulated from atoms (PDB: 7uzm). (I) Cryo-EM structure of heGDHC136S (PDB ID: 19693). (J) Model of heGDHDN crystal structure (PDB-ID: 8S3G) fitted in the oligomeric Cryo-EM reconstruction of heGDHDN (PDB-ID: 19692). Parts of the handle-like density are still visible in the truncated mutant, while N-termini are lacking. (K) Secretion of PGE2 and IL-10 (EIA, ELISA) from BMDM ± heGDH, heGDHC136S or heGDHDN (n=8). (L) Venn Diagram comparing twoindependent experiments (HA-heGDH added to the lysate of MDM vs. not added to the lysate of MDM before IP) for two timepoints (30 min, 24 h) to identify the overlapping proteins binding to heGDH. Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by unpaired t test (D-E), KruskalWallis test (F) or Friedman test (K). *P < 0.05; **P < 0.01; ***P < 0.001.

[0034] Figure 14: HeGDH reprograms macrophage metabolism via p300. (A) Oxygen consumption rate (OCR, left), extracellular acidification rate (ECAR, middle) and basal glycolysis (right) of BMDM ± heGDH (n=6). (B) Gene expression analysis of Pfkfb3 (qPCR) in BMDM ± heGDH ± p300 HAT inhibitor (A485) (n=7). (C) Heatmap of lipid mediators (LC-MS / MS) produced by human MDM + 1 mM itaconate, a-ketoglutarate (a-KG) or L-2-hydroxyglutarate (L-2-HG) (n=6). (D) Secretion of cysLTs (EIA) by MDM ± D-2- hydroxyglutarate (D-2-HG) (n=3). (E) Gene expression analysis of leukotriene synthesis genes (qPCR) in MDM ± L-2-HG (n=7). (F) Leukotriene C4 synthase activity of MM6 cells ± heGDH or TK05 (n=2-15). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Wilcoxon test (A, D-E), Friedmann test (B, C), or ordinary one-way ANOVA (F). *P < 0.05; **P < 0.01 ; ***P < 0.001.

[0035] Figure 15: HeGDH reduces markers of M2 activation and increases worm fecundity during Hpb Infection, while T-cell differentiation In the MLN and intestinal repair remain largely unaffected. HeGDH fails to Induce Treg cell differentiation in the absence of monocytes. (A) Representative images of IF staining for Ym-1 / 2, iNOS and DAPI (scale bar = 30 pm); (B) for Arg1 , COX-2 and DAPI in granulomas of tissues from 14-days primary Hpb infected mice treated intraperitoneal (i.p.) with PBS or heGDH (scale bar = 50 pm). (C) Egg counts of Hpb infected mice treated i.p. with PBS or heGDH on days 12, 14, 21 and 28 (n = 9 for 12 and 14 days; n = 4-5 for 21 and 28 days). (D) Representative images of IHC staining for a-smooth muscle actin; (E) of Masson Trichrome staining in granulomas of tissues from 14-days primary Hpb infected mice treated i.p. with PBS or heGDH (scale bars = 150 pm). (F) Representative images of Masson Trichrome staining in granulomas of tissues from 28-days primary Hpb infected mice treated i.p. with PBS or heGDH (scale bar = 150 pm). (G) Gating strategy of different T cell poulations (flow cytometry) in mesenteric lymph nodes (MLN) from one mouse infected with Hpb and treated i.p. with PBS. (H) Analysis of different T cell populations (flow cytometry) in MLN from mice treated i.p. with PBS or heGDH (n=4) ± Hpb infection (14 days primary infected) (n=14). (I) Gene expression analysis of type 2 cytokines (qPCR) of 28-days infected Hpb-mice treated i.p. with PBS or heGDH (n=3 in naive group, n = 10 in infected group). (J) Percentage of CD4+CD127-CD25hiFoxP3+(flow cytometry) in human PBMC + heGDH ± monocytes (n=6). Data are pooled from two to four independent experiments and presented as means + SEM. Statistical significance was determined by Mann-Whitney (C), Kruskal-Wallis test (H and I), and Wilcoxon test (J). *P < 0.05; **P < 0.01.

[0036] Figure 16: IL-4 / IL-13 driven macrophage M2 activation is suppressed, while parameters of tissue repair during Hpb challenge Infection remain unaltered by heGDH. (A) Gene expression analysis of AAM markers and 1112b (qPCR) in BMDM ± heGDH or after IL-4, IL-13 pre-treatment (n=8). (B) Representative images of IF staining for Ym-1 / 2, iNOS and DAPI in granulomas of tissues from 14-days challenge Hpb infected mice treated intraperitoneal (i.p.) with PBS or heGDH (scale bar = 30 pm). (C) Analysis of different T cell populations (flow cytometry) in mesenteric lymph nodes (MLN) from mice infected with Hpb (14 days challenge infected) and treated i.p. with PBS or heGDH (n=9 in PBS and n = 10 in heGDH group). (D) Representative images of IF staining for Arg1, COX2 and DAPI (scale bar = 50 pm); (E) of IHC staining for a-smooth muscle actin; (F) of Masson Trichrome staining in granulomas of tissues from 14-days challenge Hpb infected mice treated i.p. with PBS or heGDH (scale bars = 150 pm). Data are pooled from at least two independent experiments and presented as means + SEM. Statistical significance was determined by Friedmanns test (A) or Mann-Whitney (C). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

[0037] Figure 17: Increase of antifibrotic factors and tissue repair despite enhanced IL-17 response and neutrophil recruitment following heGDH treatment during Nb infection. (A) BALF cell counts from mice treated intranasally (i.n.) with PBS or heGDH under naive conditions or after infection with Nb. (B) IL-17A and IL-6 levels in BALF (Legendplex) from mice treated i.n. with PBS or heGDH under naive conditions or after infection with Nb. (C) Representative images of Masson Trichrome staining in lungs from Nb infected mice treatedi.n. with PBS or heGDH (scale bar = 100 pm). (D) Lipid mediators released from intestinal culture supernatants (LC-MS / MS) from mice treated i.n. with PBS or heGDH under naive conditions or after infection with Nb. (E) Analysis of different T cell populations (flow cytometry) in lungs from mice treated i.n. with PBS or heGDH under naive conditions or after infection with Nb. (F) Analysis of different T cell populations (flow cytometry) in mesenteric lymph nodes (MLN) treated i.n. with PBS or heGDH after infection with Nb. (G) Gene expression analysis of Arg1 and 1112b (qPCR) in alveolar macrophages from mice treated i.n. with PBS or heGDH under naive conditions or after infection with Nb. (A-G) Mice treated i.n. with PBS or heGDH under naive conditions (n=3 per group) or after infection with Nb (n=12 in PBS i.n. group and n=13 in heGDH i.n. group). Data are pooled from three independent experiments and presented as means + SEM. Statistical significance was determined by Kruskal-Wallis test (A-B, D-E, G) or Mann-Whitney (F). *P < 0.05; **P < 0.01.

[0038] Figure 18: Sequence alignment of human (SEQ ID NO: 29) and heGDH (SEQ ID NO: 27).

[0039] Figure 19: Recombinant heGDH (SEQ ID NO: 27) increases anti-inflammatory prostaglandin E2 (PGE2) while suppressing pro-inflammatory cysteinyl leukotriene (cysLT) production when administered to nasal polyp tissues from CRSwNP (ChronicRhinosinusitis with Nasal Polyps) patients ex vivo. Suppressive effect is donor dependent and particularly prominent for patients with high cysLT output Nasal polyp (NP) tissues were obtained from n=4 CRSwNP patients by Functional Endoscopic Surgery (FESS) and cultured ex vivo in airway epithelial culture medium (Lonza). NP tissue was incubated for 24h with different concentrations of recombinant heGDH. Tissue culture supernatants were collected and analyzed for lipid mediator (prostanglandin E2 and cysteinyl leukotriene) release by ELISA kits (Cayman Chemicals). Concentrations were normalized to tissue weight (ng / g tissue weight). Data are pooled from three to four independent experiments and presented as means + SEM. Statistical significance was determined by Friedman test. *P < 0.05.OVERVIEW OF THE SEQUENCE LISTING

[0040] As described herein references are made to UniProtKB Accession Numbers (htp: / / www.uniprot.org / , e.g., as available in UniProt release 2024_02 published on March 27, 2024).

[0041] The following structural polypeptides derived from N-terminal part of helminthic glutamate dehydrogenase polypeptide sequence as well as human and heGDH (full length and truncated) are described herein below in Table 1.

[0042] Table 1: Examplary polypeptides of the present invention and full length and truncated human and heGDH polypeptides.

[0043] Table 2 described herein below provides furhter sequence information regarding primer sequences.

[0044] Table 2. Primer sequences for qPCR and cloning:SEQ ID NO: 109 is CAQHSEHRTPTKGG.DETAILED DESCRIPTION OF THE INVENTION

[0045] The molecular mechanisms by which worm parasites evade host immunity are incompletely understood. In a mouse model of chronic intestinal helminth infection, we show that helminthic glutamate dehydrogenase (heGDH) drives parasite chronicity by suppressing macrophage-mediated host defense. Combining RNAseq, ChlPseq and targeted lipidomics, we identify prostaglandin E2 (PGE2) as a major immune regulatory mechanism of heGDH. The induction of PGE2 and further immunoregulatory factors such as IL-12 family cytokines and IDO1 by heGDH depended on the p300-mediated acetylation of histones, whereas the enzyme’s catalytic activity suppressed type-2 promoting leukotrienes by driving 2-hydroxyglutarate production.

[0046] In the course of the present invention it has been shown that heGDH efficiently suppresses airway inflammation in a mouse model of asthma when administered locally (intranasally) to the airways.

[0047] In one aspect of the invention, the induction of immunoregulatory factors surprisingly depended on catalytically inactive heGDH’s N-terminus (e.g., according to polypeptides of the present invention) potentially conferring interaction with cellular targets (e.g., CD64, GPNMB) identified by proteomics. Type-2 cytokines counteracted suppressive effects of heGDH on host defense, showing that type-2 immunity can limit helminth-driven immune evasion. Thus, helminths harness a ubiquitous metabolic enzyme to epigenetically target type-2 macrophage activation and establish chronicity.

[0048] Definitions:

[0049] As referred herein “EC numbers” (Enzyme Commission numbers) may be used to refer to enzymatic activity according to the Enzyme nomenclature database, Release of February 26, 2020 (e.g., available at htps: / / enzyme.expasy.org / ). The EC number refers to Enzyme Nomenclature 1992 from NC-IUBMB, Academic Press, San Diego, Calif., including supplements 1-5 published in Eur. J. Biochem. 1994, 223, 1-5; Eur. J. Biochem. 1995, 232, 1-6; Eur. J.Biochem. 1996, 237, 1-5; Eur. J. Biochem. 1997, 250, 1-6; and Eur. J. Biochem. 1999, 264, 610- 650; respectively.

[0050] The term “polypeptide" is equally used herein with the term "protein". Proteins (including fragments thereof, preferably biologically active fragments, and peptides, usually having less than 30 amino acids) comprise one or more amino acids coupled to each other via a covalent peptide bond (resulting in a chain of amino acids). The term "polypeptide" as used herein describes a group of molecules, which, for example, consist of more than 30 amino acids. Polypeptides may further form multimers such as dimers, trimers and higher oligomers, i.e. consisting of more than one polypeptide molecule. Polypeptide molecules forming such dimers, trimers etc. may be identical or non-identical. The corresponding higher order structures of such multimers are, consequently, termed homo- or heterodimers, homo- or heterotrimers etc. An example for a heteromultimer is an antibody molecule, which, in its naturally occurring form, consists of two identical light polypeptide chains and two identical heavy polypeptide chains. The terms "polypeptide" and "protein" also refer to naturally modified polypeptides / proteins wherein the modification is effected e.g. by post-translational modifications like glycosylation, acetylation, phosphorylation and the like. Such modifications are well known in the art.

[0051] Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter “sequence identity”. For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used may be gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labeled “longest identity” (obtained using the no-brief option) is used as the percent identity and is calculated as follows:

[0052] (Identical Residues* 100) / (Length of Alignment-Total Number of Gaps in Alignment).

[0053] Alternatively, the parameters used may be gap open penalty of 10, gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needle labeled “longest identity” (obtained using the no-brief option) is used as the percent identity and is calculated as follows:

[0054] (Identical Deoxyribonucleotides* 100) / (Length of Alignment-Total Number of Gaps in Alignment).

[0055] Expression: The term "expression" includes any step involved in the production of a variant (polypeptide) including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0056] Expression vector: The term "expression vector” may refer to a linear or circular DNA molecule that comprises a polynucleotide encoding a variant (polypeptide) and is operably linked to control sequences that provide for its expression.

[0057] Fragment: The term "fragment” may refer to a polypeptide having one or more (e.g., several, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10) amino acids absent from the amino and / or carboxyl terminus of a mature polypeptide; wherein the fragment still has an activity and / or structural features (e.g., motifs) as described herein.

[0058] Host cell: The term "host cell" may refer to any cell type that is susceptible to transformation, transfection, transduction, or the like with a nucleic acid construct or expression vector comprising a polynucleotide of the present invention. The term "host cell" encompasses any progeny of a parent cell that is not identical to the parent cell due to mutations that occur during replication.

[0059] Nucleic acid construct: The term "nucleic acid construct" may refer to a nucleic acid molecule, either single- or double-stranded, which is isolated from a naturally occurring gene or is modified to contain segments of nucleic acids in a manner that would not otherwise exist in nature or which is synthetic, which comprises one or more control sequences.

[0060] Operably linked: The term "operably linked" may refer to a configuration in which a control sequence is placed at an appropriate position relative to the coding sequence of a polynucleotide such that the control sequence directs expression of the coding sequence.

[0061] Control sequences: The term "control sequences" as used herein may refer to nucleic acid sequences necessary for expression of a polynucleotide encoding a variant (polynucleotide) of the present invention. Each control sequence may be native (i.e., from the same gene) or foreign (i.e., from a different gene) to the polynucleotide encoding the variant or native or foreign to each other. Such control sequences include, but are not limited to, a leader, polyadenylation sequence, propeptide sequence, promoter, signal peptide sequence, and transcription terminator. At a minimum, the control sequences include a promoter, and transcriptional and translational stop signals. The control sequences may be provided with linkers for the purpose of introducing specific restriction sites facilitating ligation of the control sequences with the coding region of the polynucleotide of the present invention.

[0062] As used herein, the term “corresponding to" may refer to a way of determining the specific amino acid of a sequence wherein reference is made to a specific amino acid sequence (e.g., US2020071638). E.g. for the purposes of the present invention, when references are made to specific amino acid positions, the skilled person would be able to align another amino acid sequence to said amino acid sequence that reference has been made to, in order to determine which specific amino acid may be of interest in said another amino acid sequence. Identification of the corresponding amino acid residue in another polypeptide can be determined by an alignment of multiple polypeptide sequences using several computer programs including, but notlimited to, MUSCLE (multiple sequence comparison by log-expectation; version 3.5 or later; Edgar, 2004, Nucleic Acids Research 32: 1792-1797), MAFFT (version 6.857 or later; Katoh and Kuma, 2002, Nucleic Acids Research 30: 3059-3066; Katoh et al., 2005, Nucleic Acids Research 33: 51 1-518; Katoh and Toh, 2007, Bioinformatics 23: 372-374; Katoh et al., 2009, Methods in Molecular Biology 537: 39-64; Katoh and Toh, 2010, Bioinformatics 26: 1899-1900), and EMBOSS EMMA employing ClustalW (1.83 or later; Thompson et al., 1994, Nucleic Acids Research 22: 4673-4680), using their respective default parameters.

[0063] For purposes of the present invention the polypeptide disclosed in SEQ ID NO: 1 may be used to determine the corresponding amino acid residue in another polypeptide. The amino acid sequence of another polypeptide is aligned with the polypeptide disclosed as SEQ ID NO: 1 , and based on the alignment, the amino acid position number corresponding to any amino acid residue in the polypeptide disclosed as SEQ ID NO: 1 is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

[0064] The term "position" when used in accordance with the present invention may refer to a position of an amino acid within an amino acid sequence depicted herein. The term "corresponding" in this context may include that a position is not only determined by the number of the preceding nucleotides / amino acids.

[0065] As used herein, “silent” mutations mean base substitutions within a nucleic acid sequence which do not change the amino acid sequence encoded by the nucleic acid sequence. “Conservative or equivalent” substitutions (or mutations) mean substitutions as listed in Table 5 herein; “highly conservative" substitutions, are preferred substitutions and are also listed in Table 5 herein.

[0066] Variant: The term "variant" may refer to a polypeptide having specific activity or structural identity as described herein herein comprising an alteration, i.e., a substitution, insertion, and / or deletion, at one or more (e.g., several, e.g., 2, 3, 4 or 5) positions. A substitution means replacement of the amino acid occupying a position with a different amino acid; a deletion means removal of the amino acid occupying a position; and an insertion means adding an amino acid adjacent to and immediately following the amino acid occupying a position.

[0067] In describing the variants of the present invention, the nomenclature described below is adapted for ease of reference. The accepted IUPAC single leter or three tetter amino acid abbreviation is employed.

[0068] Substitutions. For an amino acid substitution, the following nomenclature is used: Original amino acid, position, substituted amino acid. Accordingly, the substitution of Asn (N) atposition 167 with Thr (T) is designated as “N167T” or “Asn167Thr”. Multiple mutations can be separated by addition marks (•’+’') or (",”) e.g., "N167T+F168Y+S174C+F218Y;" or "N167T, F168Y, S174C, F218Y;", representing multiple substitutions at given positions. In the Examples of the present application, multiple mutations can be separated by comma, e.g., N167T, F168Y, S174C, F218Y. Furthermore, „X“ or “Xaa” as used herein may mean any amino acid (e.g., as depicted in Table 1 above). Accordingly, “X167T“ as used herein may mean substitution of any amino acid in position 167 with T (Thr). In case where the original amino acid residue may be any amino acid residue, a short hand notation may also be used indicating only the position and substituted amino acid. Accordingly, “X” or “Xaa” may be omitted in designating substitutions, e.g., “167T“ designation may be used meaning a substitution of any amino acid in position 167 with T (Thr). Furthermore, “X167G,A,S,C,U,I,L,V,T” as used herein may mean substitution of any amino acid in position 167 with any one of G, A, S, C, U, I, L, V or T. In case where the substituting amino acid residue may be any amino acid residue, a short hand notation may also be used indicating only the original amino acid and its position, e.g., “N167”.

[0069] Amino acid motif: The term “amino acid motif’ or “the motif as used herein may refer to a specifically defined amino acid stretch of a polypeptide. Thus, an amino acid motif of the prevent invention may relate to a short sequence of amino acids within a given polypeptide.

[0070] As used herein, the term “larval preparation” may refer to larvae that were prepared, manufactured, compounded, homogenized and / or purified (e.g., to become cell- and / or cell debris free).

[0071] As used herein, the term “immunogenic" may refer to the ability of a substance, such as a polypeptide, to provoke an immune response (e.g., in the body). This response can involve the activation of both the humoral and / or cellular branches of the immune system, leading to the production of antibodies, the activation of T cells, or both.

[0072] As used herein, the term “innate mammalian immune system” may refer to a rapid, non- specific response of a mammalian immune system to pathogens and / or foreign substances through physical barriers (e.g., including, but not limited to skin, mucous membranes, tears, and / or stomach acid), cellular components like macrophages and / or natural killer cells, and chemical mediators such as cytokines and complement proteins.

[0073] As used herein, the term “modulating macrophage- and T-cell mediated immunity" may refer to the process of adjusting the activity of macrophages and T cells to enhance or suppress their roles in the immune response. Such modulation can involve altering the activation, proliferation and / or function of these cells to optimize immune defenses against pathogens or foreign substances, regulate inflammation and / or prevent autoimmune reactions.

[0074] As used herein, the term “epigenetic targeting of prostaglandin synthesis” may refer to the manipulation of epigenetic mechanisms (e.g., introducing epigenetic changes) to influence the production of prostaglandins, which are lipid compounds that perform various roles in inflammation, pain modulation and / or vascular functions. Such epigenetic manipulations (e.g.,changes) including but are not limited to DNA methylation, histone modification and / or RNA- associated silencing, can alter the expression of enzymes involved in the prostaglandin synthesis pathway, thereby modulating their levels and activity in the body.

[0075] As used herein, the term “non-natural amino acids” may refer to non-natural amino acids (nnAAs) that are amino acids that are not found in the genetic code of living organisms and are not typically involved in the natural processes of protein synthesis. Such synthetic or artificially derived amino acids can be incorporated into proteins to introduce new properties and / or functionalities that are not present in natural proteins, such as increased stability, novel enzymatic activities, and / or enhanced therapeutic effects.

[0076] As used herein, the term “immune-therapeutical composition” may refer to a formulation designed to modulate the immune system for therapeutic purposes, e.g., such formulation can be engineered to enhance or suppress immune responses.

[0077] As used herein, the term “disease” may refer to a condition that disrupts the normal structure or function of an organism. In the context of the present invention a disease can be selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant airway inflammation, aspirin-exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy.

[0078] As used herein, the term “Hpb” may refer to Heligmosomoides polygyrus bakeri helminths and can be equally used herein with the term "Heligmosomoides polygyrus bakeri". The nematode Heligmosomoides polygyrus (formerly known as Nematospiroides dubius) is a common parasite found in the duodenum and small intestine of woodmice and other rodents (htps: / / parasite.wormbase.org / Heligmosomoides_polygyrus_prjeb1203 / lnfo / lndex / ).

[0079] The laboratory strain that has been sequenced was originally isolated from Peromyscus in California (Behnke and Harris, 2010), wherein said laboratory strain is named Heligmosomoides polygyrus bakeri. The laboratory strain is typically maintained as described by Camberis et al., 2003 and is often used to model human helminth infection as it can establish chronic infection in different strains of mice.

[0080] Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises" and "comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integer or step. When used herein the term “comprising” can be substituted with the term “containing” or “including” or sometimes when used herein with the term “having”.

[0081] When used herein “consisting of excludes any element, step, or ingredient not specified in the claim element.

[0082] As used herein, the term “consisting essentially” refers to a larval preparation, polypeptide extract or somatic proteins, in which specific further components can be present, namely those not materially affecting the essential characteristics of the corresponding larval preparation, polypeptide extract or somatic proteins ("consists essentially of), e.g., said “further components” can be cofactors of Hpb polypeptides.

[0083] As used herein, the term “cofactors" refers to organic molecules (cf. coenzymes) or ions (usually metal ions) that are required by an enzyme of its activity. They may be atached either loosely or tightly prosthetic group) to the enzyme. A cofactor binds with its associated protein (apoenzymes), which is functionally inactive, to form the active enzyme (holoenzyme).

[0084] As used herein, the terms “nucleic acids” or “nucleotide sequences” refer to DNA molecules (e.g. cDNA or genomic DNA), RNA (mRNA), combinations thereof or hybrid molecules comprised of DNA and RNA. The nucleic acids can be double- or single-stranded and may contain double- and single-stranded fragments at the same time. Most preferred are double stranded DNA molecules.

[0085] The present invention furthermore provides a nucleic acid vector comprising at least one of the nucleic acid sequences as described herein that encode a polypeptide of the present invention. The vector preferably comprises a promoter under the control of which the above nucleic acid sequences are placed. The vector can be prokaryotic or eukaryotic expression vector, where the recombinant nucleic acid is either expressed alone or in fusion to other peptides or proteins.

[0086] The invention also provides a host cell which is transfected with the vector mentioned above. The host cell can be any cell, a prokaryotic cell or a eukaryotic cell and can be used to produce at least parts of a polypeptide of the present invention or fragment or derivative thereof according to the present invention.

[0087] An "adjuvant" is a nonspecific stimulant of the immune response.

[0088] In another aspect the present invention relates to a pharmaceutical composition comprising as an active ingredient a polypeptide of the present invention or fragment or derivative thereof according to the invention. Said pharmaceutical composition may comprise at least one pharmaceutically acceptable carrier or adjuvant or excipient.

[0089] Polypeptides may be provided in pharmaceutically acceptable compositions as known in the art or as listed in a generally recognized pharmacopeia for use in animals, and more particular in humans.

[0090] The composition, if desired, can also contain minor amounts of weting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like.

[0091] The compositions of the invention can be formulated as neutral or salt forms.

[0092] Pharmaceutically acceptable salts include, but are not limited to those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

[0093] The dosage amounts and frequencies of administration are encompassed by the terms therapeutically effective and prophylactically effective. The dosage and frequency of administration further will typically vary according to factors specific for each patient depending on the specific therapeutic or prophylactic agents administered, the type of disease, the route of administration, as well as age, body weight, response, and the past medical history of the patient Suitable regimens can be selected by one skilled in the art. As used herein, the term “therapeutically effective amount” refers to an amount of the therapeutic active component or agent which is sufficient to treat or ameliorate a disease or disorder, to delay the onset of a disease or which provides any therapeutical benefit in the treatment or management of a disease.

[0094] As used herein, the term "treating" and "treatment" refers to administering to a subject a therapeutically effective amount of a pharmaceutical composition according to the invention. A “therapeutically effective amount” refers to an amount of the pharmaceutical composition or the antibody which is sufficient to treat or ameliorate a disease or disorder, to delay the onset of a disease or to provide any therapeutical benefit in the treatment or management of a disease.

[0095] As used herein, the term “prophylaxis” refers to the use of an agent for the prevention of the onset of a disease or disorder. A “prophylacticly effective amount” defines an amount of the active component or pharmaceutical agent sufficient to prevent the onset or recurrence of a disease.

[0096] As used herein, the terms "disorder" and "disease" are used interchangeably to refer to a condition in a subject

[0097] The term "and / or" wherever used herein includes the meaning of "and", "or" and "all or any other combination of the elements connected by said term".

[0098] The term "about" or "approximately" as used herein means within 20%, preferably within 10%, and more preferably within 5% of a given value or range.

[0099] The following detailed description refers to the accompanying Examples that show, by way of illustration, specific details and embodiments, in which the invention may be practised. These embodiments are described in sufficient detail to enable those skilled in the art to practice theinvention. Other embodiments may be utilized such that structural, logical, and eclectic changes may be made without departing from the scope of the invention. Various aspects of the present invention described herein are not necessarily mutually exclusive, as aspects of the present invention can be combined with one or more other aspects to form new embodiments of the present invention.

[0100] Helminth products have been suggested as natural immunoregulators for treating inflammatory diseases. We previously described novel immuneregulatory effects of a larval extract obtained from the L3 stage of the murine intestinal nematode Heligmosomoides polygyrus bakeri (Hpb) and its immunoregulatory candidate protein, glutamate dehydrogenase (GDH) (e.g., W02019193140A1). However, in the course of the present invention it has been surprisingly found that recombinant structural catalytically inactive polypeptides derived from N-terminus of heGDH are capable of suppressing allergic airway inflammation, e.g., when administered locally (intranasally) to the airways of mice in an experimental asthma model.

[0101] In some aspects of the present invention, experiments to further elucidate unique immune regulatory features and therapeutic potential of the helminthic GDH (heGDH) have been carried out. To untangle the role of structure versus catalytic function of heGDH, the structure of heGDH was solved by cryo electron micrscopy (cyro EM) and X-ray crystallography. Moreover, a catalytically inactive mutant (K126A, D204N), a mutant lacking a handle like density (C136S) and a mutant missing the N-terminal sequence (deltaN) were produced recombinantly. Analyzing all the mutants in human macrophage assays showed that the N-terminus is essential for the unique anti-inflammatory and immuneregulatory effects of heGDH. Thus, the N-terminal peptide of heGDH is responsible for its unique therapeutic effects, which overcome the limitations of current biologies and standard treatments such as GCs (such as targeting only single mechanisms and lack of effects / unwanted effects on eicosanoids).

[0102] In further aspects of the present invention, Cryo-EM and X-Ray of heGDH yielded similar structures resembling the overall architecture of mammalian GDH (see Experimental section herein). Unlike its mammalian counterparts or the X-ray model, the cryo-EM density of heGDH contains three “handle”-like densities (**) in the central symmetry plane, each connecting two monomers. These handle-like densities, with the size and shape of a short alpha helix or small unstructured region are connecting to the side chain density of Cys136 and are in close proximity to the N-terminal tails of heGDH as identified by X-ray crystallography. The handles and N-termini are exposed to the exterior solvent and may provide an ideal docking site for interaction partners of heGDH. To decipher the role of these structural features in heGDH-driven immune regulation, effects of a mutant lacking the handle like density (heGDHc136S) or the N-terminus (heGDHAN) were compared to the wildtype protein.

[0103] In some aspects of the present invention, it has been shown that strikingly, all effects on key mediators (PGE2, IL-10) or target genes induced by heGDH were lost in theabsence of the N-terminus, while heGDHc136Sacted similar to the wildtype protein, identifying the N-terminus as the key structural feature responsible for the unique effects of heGDH.

[0104] In some aspects of the present invention, cloning, expression, and purification of heGDH and heGDHANhave been carried out. Accordingly, the heGDH gene was amplified by PCR using Pfu polymerase and a pET-21 a / heGDH construct (GeneArt, Thermo Fisher Scientific) as the template. The obtained PCR product was cloned into a linearized pET TrxA-1a vector, an expression vector containing N-terminal Hise- and thioredoxin-tags followed by a TEV protease cleavage site using the SLiCe method. The truncated protein (AN) was produced by PCR- amplification and cloned into a linearized pET TrxA-1a vector. All reactions used the N-terminal Hiss-tag heGDH construct as the template. All expression constructs were verified by sequencing. The heGDH expression constructs were transformed into E. coli strain BL21 (DE3) CC4 (overexpressing the (co-)chaparones GroEL, GroES, DnaK, DnaJ, GrpE and CIpB) (5) and cultured overnight at 20°C in 2 L flasks containing 500 ml ZYM 5052 auto-induction medium (6) and 100 pg / mL kanamycin, 50 pg / mL spectinomycin and 10 pg / mL chloramphenicol. Cells from 2 L of culture were harvested by centrifugation after reaching saturation, resuspended in 120 mL lysis buffer (50 mM Tris-HCI, 300 mM NaCI, 20 mM imidazole, 10 mM MgCla, 10 pg / mL DNasel, 1 mM AEBSF.HCI, 0,2% (v / v) NP-40, 1 mg / mL lysozyme, 0.01% (v / v) 1 -thioglycerol, pH 8.0), and lysed by sonication. The lysate was clarified by centrifugation (40,000 x g) and filtration (0.2 pm). The supernatant was applied to a 5 mL HiTrap Chelating HP column (Cytiva), equilibrated in buffer A (50 mM Tris-HCI, 300 mM NaCI, 20 mM imidazole, 0.01% (v / v) 1 -thioglycerol, pH 8.0) using an Akta Purifier (Cytiva). The column was washed with buffer A containing 50 mM imidazole until a stable baseline was reached (monitored at 280 nm). Bound proteins were eluted with a linear gradient from 50 to 300 mM imidazole in buffer A. Fractions containing heGDH were pooled and dialyzed overnight at 4°C against 1 L buffer B (50 mM Tris-HCI, 300 mM NaCI, 0.01% (v / v) 1- thioglycerol, pH 8.0). Next, 5 mM ATP (from a 100 mM stock solution at pH 7) and 1 mM MgCb were added, and the solution was incubated overnight at 4°C to detach bound chaperones. The solution was applied to a 5 mL HiTrap Chelating HP column and the protein purified as described above. Fractions containing heGDH were pooled and dialyzed overnight at 4°C against 1 L buffer B in the presence of His-tagged TEV protease in a 1:25 molar ratio (TEV:protein). The cleaved off heGDH was further purified by affinity chromatography as described above and the flow- through and protein containing wash fractions were pooled and concentrated to less than 5 mL. This was subsequently subjected to size exclusion chromatography using a HiLoad 16 / 600 Superdex 200 column (Cytiva), equilibrated in buffer B. The fractions containing heGDH were pooled and dialyzed overnight against 1 L PBS pH 7.4 at 4°C. As the heGDHANwas highly soluble compared to the other constructs, the protein solution was concentrated to 6 mg / mL. Concentrations of the different protein constructs were determined by measuring the absorbance at 280 nm using the specific absorbances for full length heGDH of 1.060 or AN-heGDH of 1.126 mL / mg*cm, resp.

[0105] In some aspects of the present invention, the helminth specific N-terminus rather than the catalytic activity of heGDH confers its immune regulatory functions.

[0106] Accordingly, in further aspects of the present invention in order to discern whether immunoregulation by heGDH depends on the enzyme’s catalytic activity or its structural features, we combined site-directed mutagenesis and structure elucidation by cryo electron microscopy single particle analysis (cryo-EM SPA) and X-ray crystallography (Fig. 12, A, B and C). GDH is a hexameric enzyme that catalyzes the reversible conversion of glutamate to a-ketoglutarate (a- KG) and ammonia while reducing NAD(P)+to NAD(P)H. An enzymatic assay revealed a-KG, glutamate and ammonium as sole substrates and an optimum pH at 8.5 for glutamate utilization and 7.5 for glutamate formation as well as specificity for NAD7NADH as cofactors (Fig. 13, A, B and C). By using the allosteric inhibitor, GTP, and a GDH inhibitor, bithionol, which is also used as an anti-helminthic, heGDH activity was reduced (Fig. 13D). In contrast, clone 4F8 failed to reduce heGDH catalytic activity (Fig. 13E) as well as LT suppression (Fig. 10C), while atenuating the heGDH-mediated induction of the COX pathway (Fig. 10A). This suggested that structural features of heGDH are responsible for the induction of immune regulatory and tissue reparative prostanoids, which is an aspect of the present invention.

[0107] in further aspects of the present invention, to untangle the role of catalytic activity versus structural properties we designed a catalytically inactive mutant of heGDH (heGDHK128A- ago*) (Fig. 4A). This mutant still induced COX-metabolite and IL-10 production by macrophages to an equivalent or higher degree compared to the wildtype protein (Fig. 4, B and C). Gene expression data confirmed a similar or even stronger response of the top DEGs upon stimulation with mutant heGDHK126A-D204N(Fig. 4D). In contrast to its intact effects on prostanoids, heGDHK126A'D204Nfailed to significantly suppress cysLTs (Fig. 4E), supporting the hypothesis that the heGDH-driven induction of the COX pathway via p300 is structure dependent, whereas the catalytic function may be necessary to reduce LT production. A role for the catalytic activity in the suppressive effects on LTs was supported by a similar or even stronger reduction of cysLT formation by distinct helminthic (Ts) or human GDH (Fig. 4F and Fig. 13F).

[0108] In further aspects of the present invention, Cryo-EM and X-Ray crystallography of heGDH yielded similar structures with a D3 symmetry (Fig. 4G) and a resolution at 2.7 and 1.8 A, respectively, resembling the overall architecture of mammalian GDH (Fig. 13G). Unlike its mammalian counterparts or the X-ray model (Fig. 13, G and H), the cryo-EM density of heGDH contains three “handle”-like densities in the central symmetry plane, each connecting two monomers (Fig. 4, G and H). These handle-like densities, with the size and shape of a short alpha helix or small unstructured region are connecting to the side chain density of Cys136 and are in close proximity to the N-terminal tails of heGDH as identified by X-ray crystallography (Fig. 4H). The handles and N-termini are exposed to the exterior solvent and may provide an ideal docking site for interaction partners of heGDH. To decipher the role of these structural features in heGDH- driven immune regulation, effects of a mutant lacking the handle like densities (heGDHc136S) (Fig.131) or the N-terminus (heGDHAN) (Fig. 13J) were compared to the wildtype protein. Strikingly, all effects on key mediators (PGE2, IL-10) (Fig. 4I and Fig. 13K) or target genes (Fig. 4J) induced by heGDH were lost in the absence of the N-terminus, while heGDHc136Sacted similar to the wildtype protein, identifying the N-terminus as the key structural feature responsible for the unique effects of heGDH, which is anspect of the present invention.

[0109] In one proffered aspect, the present invention relates to a polypeptide, wherein said polypeptide comprising one or more amino acid sequences (e.g., polypeptides) having at least 80% (e.g., at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of polypeptides (e.g., heDGH-derived, e.g., catalytically inactive, structural polypeptides) selected from the group consisting of: vi) SEQ ID NO: 1 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDL); vii) SEQ ID NO: 2 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDLKPMEEQSN); viii) SEQ ID NO: 3 (MLSTLARTSGRLIFRRALSSA); ix) SEQ ID NO: 4 (QMDAHAQVIDDLKP); and x) SEQ ID NO: 5 (MEEQSN).

[0110] In another aspect, the polypeptide of the present of the present invention comprises a fragment or variant of any one of (i)-(v) as defined herein above, preferably said fragment or variant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at lest 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 , at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31 , at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39 or at least 40) amino acids long. Further preferably, said fragment or variant comprising an amino acid motif comprising at least 2 (e.g., at least 3, at least 4, at least 5) positively charged amino acids (e.g., arginines (R), lysines (K) and / or histidines (H), preferably said motif comprising at least 2 arginines (RR)), further preferably said amino acid motif is located in the tail-region (e.g., C- or N-terminal end segment / fragment having at least 6 amino acids) of said polypeptide and / or is capable of interacting with one or more negatively charged and / or glycosylated targets / polypeptides, preferably is capable of interacting with CD64 (High affinity immunoglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7)and / or Calcium homeostasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g,, having UniProt accession number: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1, e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT 1 , e.g., having UniProt accession number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having UniProt accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt accession number: Q8NHP8) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4).

[0111] In another preferred aspect, the polypeptide of the present invention consists of a polypeptide selected from SEQ ID NOs: 1-5.

[0112] In another preferred aspect, said heGDH (helminthic glutamate dehydrogenase) having UniProt accession number: A0A183FP08 and / or EC 1.4.1.3 enzymatic activity (e.g., htps: / / enzyme.expasy.Org / EC / 1.4.1.3), e.g., is capable of catalyzing the following reactions: (1) H2O + L-glutamate + NAD(+)<=> 2-oxoglutarate + H(+)+ NADH + NH<(+); (2) H2O + L-glutamate + NADP<+)<=> 2-oxoglutarate + H« + NADPH + NH4W.

[0113] In another preferred aspect, the polypeptide of the present invention does not have enzymatic activity and / or is catalytically inactive.

[0114] In another preferred aspect, the polypeptide of the present invention is capable of interacting with one or more negatively charged and / or glycosylated targets / polypeptides.

[0115] In another preferred aspect, the polypeptide of the present invention is capable of interacting with CD64 polypeptide (High affinity immunoglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11, e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7) and / or Calcium homeostasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt accession number: F8WEL8) and / or Stromal cell- derived factor 2-like protein 1 (SDF2L1 , e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT1, e.g., having UniProt accession number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having UniProt accession number: Q9H5Z1 ) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt accession number: Q8NHP8 ) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4).

[0116] In another preferred aspect, the polypeptide of the present invention is is an immunogenic polypeptide and / or said polypeptide is capable of modulating the innate mammalian immune system, preferably said polypeptide is capable of modulating macrophage-and T-cell mediated immunity, further preferably said polypeptide is capable of regulating macrophage functions and inducing regulatory T cells by epigenetic targeting of prostaglandin synthesis

[0117] In another preferred aspect, the variant of the present invention comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) alterations / modifications at one or more positions (e.g., non-natural amino acid / s), preferably wherein said variant comprises one or more substitutions (e.g., with non-natural amino acid / s), deletions, and / or insertions (e.g., of non-natural amino acid / s) at one or more positions.

[0118] In another preferred aspect, the polypeptide of the present invention further comprises one or more suitable immunoaffinity tags (e.g., for purification purposes).

[0119] In another preferred aspect, the polypeptide / s of the present of the present invention not have any enzymatic activity and / or is catalytically inactive, e.g., does not have EC 1.4.1.2, EC 1.4.1.3 and / or EC 1.4.1.4 enzymatic activity, preferably does not have EC 1.4.1.3 enzymatic activity), preferably does not have heGDH enzymatic activity (e.g., EC:1.4.1.3).

[0120] In another aspect, the present invention further relates to one or more nucleic acid constructs or expression vectors encoding and / or expressing one or more polypeptides of the present invention.

[0121] In another aspect, the present invention further relates to a host cell (e.g. an isolated and / or recombinant host cell) comprising one or more nucleic acid constructs and / or expression vectors of the present invention and / or expressing one or more polypeptides of the present invention.

[0122] In another aspect, the present invention further relates to a composition or kit comprising one or more of the polypeptides, nucleic acid constructs or expression vectors and / or host cells of the present invention.

[0123] In another aspect, the present invention further relates to a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant airway inflammation, aspirin / NSAID- exacerbated respiratory disease (AERD / N-ERD), nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma,idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; said method comprising administering a therapeutically effective amount of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions of any one of the preceding items, preferably administered locally (e.g., intranasally), further preferably administered to the airways.

[0124] In another aspect, the present invention further relates to one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of of the present invention for use as a medicament and / or in therapy.

[0125] In another aspect, the present invention further relates to one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of of the present invention for use in one or more of the following methods: (i) in a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant airway inflammation, aspirin- exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; preferably administered locally (e.g., intranasally), further preferably administered to the airways; (ii) in a method for modulating the mammalian innate and adaptive immune response, preferably administered locally (e.g., intranasally), further preferably administered to the airways; (iii) in a method for predominantly modulating the mammalian innate immune response over the mammalian adaptive immune response; (iv) in a method for targeting phagocytic cells of the mammalian immune system; (v) in a method for modifying the activation of macrophages and / or granulocytes of the mammalian immune system; (vi) in a method for modifying the activation of one or more of the arachidonic acid pathway of the mammalian immune system; (vii) in a method for decreasing the number of eosinophils and / or inhibiting the migration of granulocytes into tissue of the mammalian immune system; (viii) in a method for eliciting or modulating an immune response in a subject; (ix) in a method for treatment, amelioration, prophylaxis or diagnostics of a steroid-resistant disease; (x) in a method for monitoring development of a disease and / orassessing the efficacy of a therapy of a disease; (xi) in any method of the present invention; (xii) in any combination of methods as in any one of (i)-(xi); (xiii) in any method according to (f)-(xii), wherein said method is an in vitro, in vivo or ex vivo method.

[0126] Based on the above at least the following advantages of the present invention are contemplated over molecules known from the prior art and methods based thereon: while current biologies against asthma and / or nasal polyposis target single disease mechanisms (e.g. IL-4 receptor alpha, IL-5, IgE), heGDH and polypeptides of the present invention are capable of acting as a broad immune regulator / s that simultaneosly targets multiple mechanisms involved in chronic airway inflammation. This includes disease-relevant lipid mediators (e.g., prostanoids and leukotrienes), which are poorly targeted by standard treatments such as glucocorticoids and current biologies.

[0127] A key advantage of heGDH and polypeptides of the present invention compared to current biologies (e.g., monoclonal antibodies), which have to be injected and act systemically, is its efficacy when administered to the airways, thus simplifying drug delivery and limiting systemic side effects. The N-terminal sequence of heGDH is unique to the helminthic protein, thus providing a clear distinction from the mammalian homologues, which lack immune regulatory functions.

[0128] The invention is -also characterized by the following items:1. A polypeptide (e.g., a synthetic, recombinant and / or non-naturai polypeptide), wherein said polypeptide comprising one or more (e.g., 2, 3, 4 or 5 etc.) amino acid sequences having (e.g., each having) at least 80% (e.g., at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to a polypeptide selected from the group consisting of: i) SEQ ID NO: 1 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDL); ii) SEQ ID NO: 2 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDLKPMEEQSN); iii) SEQ ID NO: 3 (MLSTLARTSGRLIFRRALSSA); iv) SEQ ID NO: 4 (QMDAHAQVIDDLKP); v) SEQ ID NO: 5 (MEEQSN); and / or vi) optionally, a fragment or variant of any one of (i)-(v), preferably said fragment or vari- ant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11 , at least 12, at test 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 , at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, atleast 39, at least 40, or at least 41 ) amino acids long; further preferably said fragment or variant comprising an amino acid motif comprising at least 2 (e.g., at least 3, at least 4, at least 5) positively charged amino acids (e.g., arginines (R), lysines (K) and / or histidines (H), preferably said motif comprising at least 2 arginines (RR)), fur- ther preferably said amino acid motif is located in the tail-region (e.g., C- or N-terminal end segment / fragment having at least 6 amino acids) of said polypeptide and / or is capable of interacting with one or more negatively charged and / or glycosylated tar- gets / polypeptides, preferably is capable of interacting with CD64 (High affinity immu- noglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession num- ber: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7) and / or Calcium homeostasis endoplasmic re- ticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt accession num- ber: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1, e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT1 , e.g., having UniProt accession number: 075907) and / or Probable ATP-de- pendent RNA helicase DHX35 (DHX35, e.g., having UniProt accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt acces- sion number: Q8NHP8 ) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4), preferably said polypeptide is not a full-length, wild type and / or naturally-occuring heGDH (e.g., having UniProt accession number: A0A183FP08 or SEQ ID NO: 27), further prefera- bly said polypeptide having less than 100% sequence identity (e.g., does not have enzy- matic activity and / or is catalytically inactive) to said full-length heGDH (e.g., wild type heGDH). The polypeptide according to any one of the preceding items, wherein said polypeptide is a fragment or variant of any one of (i)-(v) according to the preceding item 1, preferably said fragment or variant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11 , at least 12, at lest 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 , at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32,at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, or at least 41 ) amino acids long; further preferably said fragment or variant comprising an amino acid motif comprising at least 2 (e.g., at least 3, at least 4, at least 5) positively charged amino acids (e.g., arginines (R), lysines (K) and / or histidines (H), preferably said motif comprising at least 2 arginines (RR)), further preferably said amino acid motif is lo- cated in the tail-region (e.g., C- or N-terminal end segment / fragment having at least 6 amino acids) of said polypeptide and / or is capable of interacting with one or more negatively charged and / or glycosylated targets / polypeptides, preferably is capable of interacting with CD64 (High affinity immunoglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number. Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession num- ber: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt ac- cession number: Q3YEC7) and / or Calcium homeostasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt accession number: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1, e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT1, e.g., having UniProt acces- sion number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having UniProt accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt accession number: Q8NHP8) and / or Tripartite motif-contain- ing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4). The polypeptide according to any one of the preceding items, wherein said fragment or variant is selected from the group consisting of: SEQ ID NOs: 6-26. The polypeptide according to any one of the preceding items, wherein said polypeptide comprises one or more heGDH (helminthic glutamate dehydrogenase)-derived structural motifs having SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and / or SEQ ID NO: 5, preferably SEQ ID NO: 1. The polypeptide according to any one of the preceding items, wherein said polypeptide comprises or consists of SEQ ID NO: 1. The polypeptide according to any one of the preceding items, wherein said polypeptide is isolated and / or purified and / or recombinant and / or synthetic. The polypeptide according to any one of the preceding items, wherein said polypeptide is anti-inflammatory.The polypeptide according to any one of the preceding items, wherein said fragment or variant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11 , at least 12, at lest 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, or at least 41) amino acids long. The polypeptide according to any one of the preceding items, wherein said polypeptide consists of ane or more (e.g., 2, 3, 4 or 5) polypeptides selected from the group consisting of: SEQ ID NOs: 1-5 and / or one or more (e.g., 2, 3, 4 or 5) variant / s thereof, preferably wherein said polypeptide consists of a polypeptide selected from SEQ ID NOs: 1-5. The polypeptide according to any one of the preceding items, wherein said heGDH (helmin- thic glutamate dehydrogenase) having UniProt accession number: A0A183FP08 and / or EC 1.4.1.3 enzymatic activity. The polypeptide according to any one of the preceding items, wherein said polypeptide does not have enzymatic activity and / or is catalytically inactive, e.g., does not have EC 1.4.1.2, EC 1.4.1.3 and / or EC 1.4.1.4 enzymatic activity, preferably not having EC 1.4.1.3 enzymatic activity), preferably does not have heGDH enzymatic activity (e.g., EC:1.4.1.3). The polypeptide according to any one of the preceding items, wherein said polypeptide is capable of interacting with one or more negatively charged and / or glycosylated targets / pol- ypeptides. The polypeptide according to any one of the preceding items, wherein said polypeptide is capable of interacting with (e.g., with one or more of the following): CD64 polypeptide (High affinity immunoglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt ac- cession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7) and / or Calcium homeostasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt accession number: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1 , e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT1 , e.g., having UniProt accession number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having Uni- Prot accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g.,having UniProt accession number: Q8NHP8) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4). The polypeptide according to any one of the preceding items, wherein said polypeptide is an immunogenic polypeptide and / or said polypeptide is capable of modulating the innate mammalian immune system, preferably said polypeptide is capable of modulating macro- phage- and T-cell mediated immunity, further preferably said polypeptide is capable of reg- ulating macrophage functions and inducing regulatory T cells by epigenetic targeting of prostaglandin synthesis. The polypeptide according to any one of the preceding items, wherein said fragment or variant comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30) alterations / modifications at one or more positions (e.g., non-natural amino acid / s), preferably wherein said variant comprises one or more substitutions, deletions and / or insertions at one or more positions. The polypeptide according to any one of the preceding items, wherein the number of sub- stitutions is 1-30, preferably, 1-20, 1-10 or 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 substi- tutions. The polypeptide according to any one of the preceding items, wherein variant comprises one or more substitutions (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions, e.g., compaged to corresponding one or more amino acids in any one of polypeptides having SEQ ID NOs: 1- 5), deletions and / or insertions at one or more positions, preferably said one or more substi- tions are conservative, higly conservative or equivalent substitutions (or mutations) as shown in Table 5 below:The polypeptide according to any one of the preceding items, wherein said fragment or variant comprises or consists of one or more polypeptides selected from the group consist- ing of (Table 6):The polypeptide according to any one of the preceding items, wherein said polypeptide further comprises one or more immunoaffinity tags (e.g., one or more His-tags, Strep-ll tags, ALFA-tags, Avi-tags, C-tags, Calmodulin-tags, polyglutamate-tags, E-tags, FLAG-tags, HA- tags, Myc-tags, NE-tags or any other suitable heGDH (helminthic glutamate dehydrogen- ase )-derived tag known in the art). The polypeptide according to any one of the preceding items, wherein said polypeptide is derived from H. polygyrus (e.g., Heligmosomoides polygyrus bakeri). The polypeptide according to any one of the preceding items, wherein said polypeptide is not longer than 60 amino acids lond, preferably not nongerthan 55 amino acids long, further preferably not longer than 50 amino acids long, most preferably not longer than 45 amino acids long, further most preferably not longer than 41 amino acids long. A nucleic acid construct or expression vector encoding and / or expressing (preferably ex- pressing) one or more polypeptides according to any one of the preceding items. A host cell (e.g. an isolated and / or recombinant host cell) comprising one or more nucleic acid constructs and / or expression vectors according to any one of the preceding items and / or expressing one or more polypeptides according to any one of the preceding items. The host cell according to any one of the preceding items, wherein said host cell is an eucaryotic or prokaryotic host cell. The host cell according to any one of the preceding items, wherein the host cell is selected from a bacterial cell, yeast cell, mammalian cell, insect cell and plant cell, preferably from a bacterial cell and yeast cell. The host cells according to any one of the preceding items, wherein said host cell is non- human. The nucleic acid construct or expression vector according to any one of the preceding items, wherein said nucleic acid construct or expression vector is codon-optimized, preferably for expression in a host cell according to any one of the presediung items. A composition or kit comprising one or more of the polypeptides, nucleic acid constructs or expression vectors and / or host cells of any one of preceding items. The composition or kit of any one of the preceding items, wherein said composition or kit is a diagnostic, pharmaceutical composition and / or vaccine composition or kit. The composition or kit of any one of the preceding items, wherein said composition is an immune-therapeutical composition or kit. A method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant air- way inflammation, aspirin / NSAID-exacerbated respiratory disease (AERD / N-ERD), nasalpolyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune dis- ease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly al- lergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fi- brosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory dis- ease, allergy; said method comprising administering a therapeutically effective amount of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding items, preferably administered locally (e.g., intranasally), further preferably administered to the airways. The method for treatment, amelioration and / or prophylaxis of any one of the preceding items, wherein said administering is carried out locally (e.g., intranasally) or topically, refer- ably to the airways. One or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding items, for use as a medicament and / or in therapy. One or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding items for use in one or more of the following methods: i) in a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory dis- ease, steroid resistant airway inflammation, aspirin-exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic derma- titis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhini- tis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphy- sema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary dis- ease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, ex- trinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy;preferably administered locally (e.g., intranasally), further preferably administered to the airways; ii) in a method for modulating the mammalian innate and adaptive immune response, preferably administered locally (e.g., intranasally), further preferably administered to the airways; iii) in a method for predominantly modulating the mammalian innate immune response over the mammalian adaptive immune response; iv) in a method for targeting phagocytic cells of the mammalian immune system; v) in a method for modifying the activation of macrophages and / or granulocytes of the mammalian immune system; vi) in a method for modifying the activation of one or more of the arachidonic acid path- way of the mammalian immune system; vii) in a method for decreasing the number of eosinophils and / or inhibiting the migration of granulocytes into tissue of the mammalian immune system; viii) in a method for eliciting or modulating an immune response in a subject; ix) in a method for treatment, amelioration, prophylaxis or diagnostics of a steroid-re- sistant disease; x) in a method for monitoring development of a disease and / or assessing the efficacy of a therapy of a disease; xi) in any method according to any one of the preceding items and / or as disclosed in the Examples section herein; xii) in any combination of methods as in any one of (i)-(xi); xiii) in any method according to (i)-(xii), wherein said method is an in vitro, in vivo or ex vivo method. Use of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding items for / in one or more of the following: i) for treatment, amelioration and / or prophylaxis of a disease selected from a group con- sisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid re- sistant airway inflammation, aspirin-exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bron- chitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; preferably administered lo- cally (e.g., intranasally), further preferably administered to the airways; ii) for modulating the mammalian innate and adaptive immune response, preferably ad- ministered locally (e.g., intranasally), further preferably administered to the airways; iii) for predominantly modulating the mammalian innate immune response over the mam- malian adaptive immune response; iv) for targeting phagocytic cells of the mammalian immune system; v) for modifying the activation of macrophages, T-cells and / or granulocytes of the mam- malian immune system; vi) for modifying the activation of one or more of the arachidonic acid pathway of the mammalian immune system; vii) for decreasing the number of eosinophils and / or inhibiting the migration of granulo- cytes into tissue of the mammalian immune system; viii) for eliciting or modulating an immune response in a subject; ix) for treatment, amelioration, prophylaxis or diagnostics of a steroid-resistant disease; x) for monitoring development of a disease and / or assessing the efficacy of a therapy of a disease; xi) for any use according to any one of the preceding items and / or as disclosed in the Examples section herein; xii) in any combination of uses as in any one of (i)-(xi); xiii) in any use according to (i)-(xii), wherein said use is an in vitro, in vivo or ex vivo use.36. One or more polypeptides, nucleic acid constructs or expression vectors, host cells, com- positions, kits, methods, and / or uses of any one of the preceding items, carried out as shown in Examples as disclosed herein (e.g., any one of Figures 1-19 herein) and / or else where herein or herein below.***

[0129] In order that the invention may be readily understood and put into practical effect, some aspects of the invention are described by way of the following non-limiting examples.

[0130] Example 1 : A helminth enzyme subverts macrophage-mediated immunity by epigenetic targeting of prostaglandin synthesis

[0131] INTRODUCTION

[0132] Over 1,000 different parasites can infect humans and around one-third of the human population worldwide are infected by worm parasites (helminths) (1, 2). During evolution, helminths have developed survival strategies to suppress host defense and establish chronic infections. Mechanisms or molecules associated with the evasion of the immune response by parasitic helminths may be exploited for the treatment of type 2 inflammatory disorders (3). However, the mechanisms and molecules by which helminths control antiparasitic immune responses to persist in the host are largely unknown.

[0133] Protective immunity against helminth parasites often relies on the induction of a type 2 immune response characterized by the production of type 2 cytokines that activate host effector cells such as macrophages, granulocytes and T helper 2 (Th2) cells (4-9). Helminths can efficiently suppress this type 2 immune response by targeting cytokines (e.g. IL-33) important for its induction (10) or by inducing regulatory T cells and tolerogenic macrophages (11).

[0134] We have previously shown that larval products containing the glutamate dehydrogenase (GDH) of Heligmosomoides polygyrus bakeri (Hpb) suppress allergic inflammation in a mouse model of asthma and induce a shift in the arachidonic acid (AA) metabolic pathway (12). AA metabolites can act as bioactive derivatives with key roles in infection and inflammation (13). Leukotrienes (LTs) synthesized by 5-lipoxygenase (5-LOX) promote type 2 inflammation and helminth expulsion (14). In contrast, prostaglandins synthesized by the cyclooxygenase (COX) pathway show both type 2 promoting (15) as well as suppressive capacities (16, 17) during infections with helminth parasites. GDH are widely conserved among parasitic helminths, including the human cestode parasite Taenia solium (Ts), in which GDH was shown to drive regulatory ! cells by upregulating the immunoregulatory AA metabolite PGEa (16). However, it remained unclear how helminthic GDH (heGDH) can modulate AA metabolic pathways and whether this would promote immune evasion.

[0135] Here, we demonstrate that heGDH enables parasite chronicity by targeting macrophages. Upon internalization, heGDH regulates macrophage tricarboxylic acid (TCA)- and amino acid metabolism to suppress LTs via its catalytic activity, whereas its structure activates the p300 histone acetyltransferase (HAT) to induce the expression of multiple type 2 suppressive genes, including PGEz synthetic enzymes. The heGDH-mediated induction of myeloid PGEz synthesis suppressed alternative macrophage activation (AAM) and Th2 activation essential for host defense, thus identifying GDH as a key factor of helminthic immune evasion.

[0136] MATERIALS AND METHODS

[0137] Study design

[0138] The aim of this study was to investigate how GDH derived from Hpb can modulate eicosanoid pathways and macrophage activation to regulate the host type 2 immune response. To accomplish this, we used transcriptomic and epigenetic analysis (RNA-seq, ChlP-seq), qPCR, LC-MS / MS and ELISA. To define in vivo effects of heGDH, mice were treated with a monoclonalantibody against heGDH or with recombinant heGDH (SEQ ID NO: 27) during parasite infections. For the human part of our study, healthy volunteers (total n = 47) (Caucasian men and women) were recruited. Sample sizes (determined by Power Analysis by a statistician), replicates, and statistical methods are specified in the figure legends. All blood donors participated in the study after informed written consent. All procedures were approved by the local ethics commitee at the University clinic of the Technical University of Munich (internal reference: 802 / 20S) and in accordance with the declaration of Helsinki. For further details see Supplementary Materials.

[0139] Mice

[0140] C57BL / 6J mice were obtained from Charles River Laboratories (Sulzfeld) and maintained under specific pathogen free conditions at the Helmholtz Munich or at the University of Lausanne. Unless stated otherwise, 6- to 12-week-old mice of both sexes were used. All animal experiments were approved by the local authorities (Regierung von Oberbayern, ROB-55.2- 2532. Vet„02-18-95 and canton de Vaud, VD3809c).

[0141] Treatment with heGDH or mAb (clone 4F8) during helminth infection

[0142] HeGDH treatment (5 pg of heGDH in 100 pL of PBS) was performed intraperitoneally at day 4, 8 and 12 for the 14-days primary infection experiment. When mice were sacrificed at 28 days post primary infection, mice were treated at days 4, 8 and 21. Challenge infected Hpb mice were treated at days 53, 57 and 61. For blocking experiments, 10 pg of a- heGDH mAb (clone 4F8) or isotype control antibody (BioCell) in 100 pL PBS was intraperitoneally given on day 0-2, 4 and 6. Intranasal treatment of Nb infected mice with heGDH was performed at day 0, 2 and 4. For further details see Supplementary Materials.

[0143] Human T-cell analysis

[0144] To assess Treg induction or Th2 cell suppression, human PBMCs were stimulated with 5 pg / mL heGDH alone or in combination with 50 pg / mL of SEA (for Th2 cell induction) and or 10 pM mPGESI inhibitor, 934, for different time points. Cells were analyzed via flow cytometry. A more detailed description for PBMC culture, stimulation and flow cytometry can be found in the Supplementary Materials.

[0145] Macrophage differentiation and culture

[0146] As previously reported (12), CD14+PBMCs were used to generate monocyte- derived macrophages (MDM). Macrophages were cultured in the presence of 10 ng / mL human GM-CSF (Miltenyi Biotec) and 2 ng / mL human TGF-p (Peprotech). Bone marrow derived macrophages (BMDM) from bone marrow of wildtype C57BL / 6 or EP2+mice were isolated and cultured for 6 days in the presence of 20 ng / mL murine recombinant M-CSF (Miltenyi Biotech). Exchange of medium and replenishment of cytokines was performed on the third day. After 6days, cells were harvested and used for further stimulation experiments. More detailed procedures on stimulation and further analysis can be found in the Supplementary Materials.

[0147] RNA sequencing

[0148] Sequencing of RNA from MDM treated with heGDH was performed at the Helmholtz Munich (HMGU) by the Genomics Core Facility. The preparation of samples, library preparation and sequencing was done as previously described (56, 57). More detailed procedures of sequencing and processing of RNA-seq data can be found in the Supplementary Materials.

[0149] Eicosanoid and cytokine analysis

[0150] Eicosanoids in intestinal culture supernatants, BALF and supernatants of cultured macrophages were quantified by liquid chromatography-tandem mass spectrometry (LC-MS / MS) using previously published protocols (58, 59). Cytokines were quantified using commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions. More detailed procedures can be found in the Supplementary Materials.

[0151] Chromatin immunoprecipitation (ChIP)

[0152] For ChIP with formaldehyde crosslinking, macrophages (3-5 x 106cells per condition) were incubated for 30 min at 37°C and 5% CO2 with Accutase (Sigma Aldrich, Merck) to detach the cells. ChIP protocol steps were performed similar as previously described (60). A more detailed procedure of ChIP, ChlP-qPCR, ChlP-seq and data processing can be found in the Supplementary Materials.

[0153] Structure elucidation of heGDH and its mutants

[0154] Crystallization experiments were done on heGDH and heGDHANby sitting drop vapour diffusion methods at 18°C and were performed at the X-ray Crystallography Platform at Helmholtz Munich. At the same time the structure of heGDH, heGDHc136Sand heGDHANwere also determined by cryo-EM SPA on a FEI Titan Krios transmission electron microscope. More detailed procedures of structure elucidation and data processing can be found in the Supplementary Materials.

[0155] Cloning, expression, and purification of heGDH and heGDHAN

[0156] Preparation of the expression constructs

[0157] The heGDH gene was amplified by PCR using Pfu polymerase and a pET- 21a / heGDH construct (GeneArt, Thermo Fisher Scientific) as the template. The obtained PCR product was cloned into a linearized pET TrxA-1a vector, an expression vector containing N- terminal Hise- and thioredoxin-tags followed by a TEV protease cleavage site using the SLiCe method.

[0158] The truncated protein (AN) was produced by PCR-amplification and cloned into a linearized pET TrxA-1a vector. All reactions used the N-terminal Hise-tag heGDH construct as the template. All expression constructs were verified by sequencing.

[0159] Protein expression and purification

[0160] The heGDH expression constructs were transformed into E. coli strain BL21 (DE3) CC4 (overexpressing the (co-)chaparones GroEL, GroES, DnaK, DnaJ, GrpE and CIpB) (5) and cultured overnight at 20°C in 2 L flasks containing 500 ml ZYM 5052 auto-induction medium (6) and 100 pg / mL kanamycin, 50 pg / mL spectinomycin and 10 pg / mL chloramphenicol. Cells from 2 L of culture were harvested by centrifugation after reaching saturation, resuspended in 120 mL lysis buffer (50 mM Tris-HCI, 300 mM NaCI, 20 mM imidazole, 10 mM MgCI2, 10 pg / mL DNasel, 1 mM AEBSF.HCI, 0,2% (v / v) NP-40, 1 mg / mL lysozyme, 0.01% (v / v) 1 -thioglycerol, pH 8.0), and lysed by sonication. The lysate was clarified by centrifugation (40,000 x g) and filtration (0.2 pm). The supernatant was applied to a 5 ml HiTrap Chelating HP column (Cytiva), equilibrated in buffer A (50 mM Tris-HCI, 300 mM NaCI, 20 mM imidazole, 0.01% (v / v) 1 -thioglycerol, pH 8.0) using an Akta Purifier (Cytiva). The column was washed with buffer A containing 50 mM imidazole until a stable baseline was reached (monitored at 280 nm). Bound proteins were eluted with a linear gradient from 50 to 300 mM imidazole in buffer A. Fractions containing heGDH were pooled and dialyzed overnight at 4°C against 1 L buffer B (50 mM Tris-HCI, 300 mM NaCI, 0.01% (v / v) 1- thioglycerol, pH 8.0). Next, 5 mM ATP (from a 100 mM stock solution at pH 7) and 1 mM MgCfe were added, and the solution was incubated overnight at 4°C to detach bound chaperones. The solution was applied to a 5 mL HiTrap Chelating HP column and the protein purified as described above. Fractions containing heGDH were pooled and dialyzed overnight at 4°C against 1 L buffer B in the presence of His-tagged TEV protease in a 1 :25 molar ratio (TEV:protein). The cleaved off heGDH was further purified by affinity chromatography as described above and the flow- through and protein containing wash fractions were pooled and concentrated to less than 5 mL. This was subsequently subjected to size exclusion chromatography using a HiLoad 16 / 600 Superdex 200 column (Cytiva), equilibrated in buffer B. The fractions containing heGDH were pooled and dialyzed overnight against 1 L PBS pH 7.4 at 4°C. As the heGDHANwas highly soluble compared to the other constructs, the protein solution was concentrated to 6 mg / mL. Concentrations of the different protein constructs were determined by measuring the absorbance at 280 nm using the specific absorbances for full length heGDH of 1.060 or AN-heGDH of 1.126 mL / mg*cm, resp.

[0161] Stimulation of MDM for proteomic analysis

[0162] 2.5 x 10® cells were seeded in a 6-well plate. MDM were stimulated with 5 pg / mlHA-heGDH for 30 min or 24 h. For the first experiment and to investigate intracellular interaction partners, additional 5 pg of HA-heGDH were added to the lysate before immunoprecipitation (IP).For the second experiment, no additional HA-heGDH was added. For both experiments, lysates were split in two for IP with anti-HA magnetic beads (Thermo Fisher Scientific) or IP with isotype control magnetic beads (MBL International). After IP, protein solutions were analyzed via mass spectrometry. A more detailed description of sample preparation, proteomics and data processing can be found in the Supplementary Materials.

[0163] Metabolic flux analyses

[0164] A total of 5 x 104cells were plated per well on a Seahorse Miniplate (Agilent Technologies, Santa Clara, Calif). Stimulation of MDM with heGDH was done for 24 hours, while BMDM were treated for 6 hours before mitochondrial stress test (Agilent). A more detailed procedure can be found in the Supplementary Materials.

[0165] Amino acid and TCA cycle intermediate analysis

[0166] A total of 5 x 105MDM was plated and stimulated with heGDH for targeted metabolomics. Metabolite quantification by LC-MS / MS was performed at the Metabolomics Core Facility of the Max Planck Institute for Immunobiology and Epigenetics in Freiburg. A more detailed procedure of targeted metabolite quantification can be found in the Supplementary Materials.

[0167] LTC4S activity assay

[0168] The assay of recombinant LTC4S activity assay was done as previously described (61). To determine the effect of heGDH or L-2-hydroxyglutarate (L-2-HG) on the activity of LTC4S, pre-incubations with 1 pg or 3 pg of heGDH or 1 mM L-2-HG were performed. For LTC4S activity measurements in cell homogenates of differentiated Mono Mac 6 (MM6), 5 pg heGDH was added. A more detailed description for both assays can be found in the Supplementary Materials.

[0169] Statistical Analyses

[0170] Data were analyzed by GraphPad Prism software. For LC-MS / MS (lipid mediator) and ELISA (cytokines) data, missing values below the lower limit of detection were interpolated using % of the minimum value for each metabolite. Statistical analysis of two group comparisons was performed using Mann-Whitney (unpaired), Wilcoxon test (paired) or t test depending on normal distribution. For comparison of more groups, RM one-way ANOVA, Friedmann test (paired) or Kruskal-Wallis test (unpaired) with Dunn correction was used with correction for multiple comparisons. P < 0.05 was considered statistically significant. Details of statistical tests and sample size are provided in the figure legends. Heat maps were generated by R (RNA-seq data) or with the Broad Institute’s Morpheus software.

[0171] Data and materials availability:

[0172] PDB-ID of X-Ray structure (heGDHJull length): 8QF0

[0173] PDB-ID of X-Ray structure (heGDHAN): 8S3G

[0174] PDB-ID of EM-structure (heGDH): 18456

[0175] PDB-ID of EM-structure (heGDHc136S): 19693

[0176] PDB-ID of EM-structure (heGDHAN): 19692

[0177] Proteomics data: is deposited on the databade PRIDE (PXD044102)

[0178] ChIP sequencing data: is deposited on the database GEO (GSE259349).

[0179] RNA sequencing data: is deposited on the database GEO (GSE259350).

[0180] Supplementary Materials

[0181] Supplementary Materials and Methods

[0182] Fig. 8. HeGDH is present in all stages of Hpb and internalized by macrophages. HeGDH in vivo blockade has variable effects on myeloid cell recruitment.

[0183] Fig. 9: The maximal dose of LPS found in recombinant heGDH preparations fails to mimic effects of heGDH.

[0184] Fig. 10: Clone 4F8 mAb atenuates heGDH triggered prostanoid and IL-10 induction but not 5-LOX metabolite suppression.

[0185] Fig. 11: HeGDH driven p300 HAT activation and H3K27 acetylation specifically induces type 2 suppressive genes in macrophages.

[0186] Fig. 12: Cryo-EM structure determination of heGDH and its mutants.

[0187] Fig. 13: Characterization of heGDH activity, structure and cellular targets as well as effects of different heGDH homologues on cysLT formation.

[0188] Fig. 14: HeGDH reprograms macrophage metabolism via p300.

[0189] Fig. 15: HeGDH reduces markers of M2 activation and increases worm fecundity during Hpb infection, while T-cell differentiation in the MLN and intestinal repair remain largely unaffected. HeGDH fails to induce Treg cell differentiation in the absence of monocytes.

[0190] Fig. 16: IL-4 / IL-13 driven macrophage M2 activation is suppressed, while parameters of tissue repair during Hpb challenge infection remain unaltered by heGDH.

[0191] Fig. 17: Increase of antifibrotic factors and tissue repair despite enhanced IL-17 response and neutrophil recruitment following heGDH treatment during Nb infection.

[0192] Table 3. Refinement statistics for X-ray:

[0193] Table 4. Reagents and resources:

[0184] RESULTS

[0195] GOH enables helminth immune evasion by inducing type 2 suppressive macrophages

[0196] We previously identified GDH as a helminth-derived factor able to suppress type 2 inflammation in a mouse model of allergic asthma (12). However, the evolutionary role of GDH during helminth infection remained elusive. Indeed, heGDH is expressed by all stages of the Hpb parasite with highest levels (lowest CT value) found in the infectious L3 stage (Fig. 8A). To investigate whether GDH is essential for parasite immune evasion, we neutralized heGDH by using a specific monoclonal antibody (mAb) (clone 4F8) (12) in mice infected with the nematode parasite Hpb (Fig. 1A). During a primary infestation, Hpb causes a chronic infection with worm counts peaking around two weeks post infection (p.i.). Ab-mediated neutralization of heGDH resulted in lower worm counts 14 days p.i. (Fig. 1B), suggesting that heGDH is a key factor in the Hpb-mediated suppression of type 2 immunity.

[0197] Macrophages are essential players in host defense against parasite infections (18) and recruited bone marrow- / monocyte-derived macrophages are particularly important for anti- helminth immunity (8). 4F8 treatment did not affect the accumulation of CD64* macrophages or Alox15+cells (mostly eosinophils) (19), while neutrophils (MPO+) tended to increase in the granuloma of Hpb infected mice at the peak of infection (Fig. 8B). Immunofluorescence (IF) staining using the 4F8 antibody revealed colocalization of heGDH with CD64+macrophages near Hpb larvae in the small intestinal submucosa early during infection (Fig. 1C and Fig. 8C). In line, stimulation of human monocyte-derived macrophages (MDM) or murine bone marrow-derived macrophages (BMDM) with recombinant heGDH (SEQ ID NO: 27) resulted in binding and its uptake, which was detectable for at least 24 hours in the cytoplasm (Fig. 1D and Fig. 8, D and E).

[0198] To further define the immune regulatory effects of heGDH, RNA sequencing (RNA- seq) was performed, revealing broadly altered transcriptional profiles of heGDH-treated compared to untreated MDM. In particular, heGDH increased the expression of immunoregulatory and type 2 suppressive genes including IDO1, PTGES (mPGES-1), PTGS2 (C0X2), PTGIR, IL12B and EBI3 (Fig. 1, E and F) (20-25). To exclude that the heGDH-triggered induction of regulatory mediators was due to endotoxin contamination, we compared the transcriptional profiles of MDM stimulated with heGDH to MDM stimulated with lipopolysaccharide (LPS) at the concentration present in the preparation of heGDH (0.5- 1 ng / mL) (Fig, 9, A and B). Although low dose (1 ng / mL) LPS upregulated several genes and mediators (Fig. 9, C and D) that were induced by heGDH, MDM stimulated with heGDH showed a much stronger induction of the same top differentially expressed genes (DEGs) (Fig. 9, A and B), confirming that heGDH imprints a type 2 suppressive macrophage phenotype independent of the low amounts of contaminating LPS. In line with the increased expression of genes involved in prostanoid synthesis and signaling, LC- MS / MS analysis revealed a shift from type 2-inducing metabolites (cysLTs) to prostanoids involved in tissue repair and the regulation of type 2 immunity (PGD2, PGE2, PGF2a, TXB2) (26, 27) together with anti-inflammatory IL-10 in heGDH-treated MDM, when compared to control or mock vector treated cells (Fig. 2, A, B and C). Neutralization with clone 4F8 partially abrogated the heGDH-mediated regulation of IL-10 and COX-2, but not of 5-LOX metabolites (Fig. 10, A, B and C), suggesting distinct mechanisms for the modulation of anti-inflammatory, reparative mediators on the one and type 2 promoting mediators (LTs) on the other hand. Taken together, these data suggest that heGDH suppresses anti-helminth host defense by broadly modulating macrophage effector functions.

[0199] p300 HAT activation by heGDH mediates the induction of immune-regulatory genes

[0200] Multiple regulatory genes induced by heGDH, including PGE2synthetic enzymes and IL12B, are regulated via p300 HAT (28-30) (Fig. 1E and Fig. 9A), which suggests an epigenetic mechanism of action. Indeed, addition of a p300 / CBP HAT inhibitor, A485, during treatment with heGDH resulted in a suppression of top DEGs identified by RNA-seq (IL12B, IDO1, PTGS2, EBI3 and PTGES), white the heGDH-triggered induction of interferon stimulated genes (ISG) remained unaffected (Fig. 3Aand Fig. 11, A and B). Decreased gene expression correlated with a strongly diminished secretion of PGE2, IL-12(3 (p40) and IL-10 in GDH-stimulated macrophages treated with the p300 HAT inhibitor (Fig. 3B), which was not due to cellular toxicity (Fig. 11C). Knock-down of p300 during heGDH stimulation by siRNA (Fig. 11D) confirmed the p300 dependent induction of target proteins, including COX-2 and mPGESI in MDM and BMDM (Fig. 3C and Fig. 11E), while CD64 remained unaffected (Fig. 11F). In line with the heGDH- mediated activation of p300, global heGDH-induced H3K27 acetylation (ac) was abrogated by p300 inhibition (Fig. 3D and Fig. 11G). H3K27ac enrichment in enhancer regions of heGDHtreated MDM was confirmed by ChlP-seq analysis, showing a large intersection between H3K27ac peaks and top DEGs from RNA-seq (Fig. 3, E, F and G). Assessment of the linear relationship between ChlP-seq and RNA-seq values across genes revealed a weak, but highly significant positive correlation (r = 0.16, p=2.2e-16), suggesting a robust association between ChlP-seq and RNA-seq values across genes. An overall overlap between both data sets was also apparent for eicosanoid synthesis pathways (Fig. 11 H). Targeted ChlP-qPCR analysis of H3K27ac for IDO1, PTGS2 and IL6 further confirmed this enrichment (Fig. 3H), supporting a key role for p300-mediated H3K27 acetylation in immunoregulation by heGDH.

[0201] The helminth specific N-terminus rather than the catalytic activity of heGDH confers its immune regulatory functions

[0202] To discern whether immunoregulation by heGDH depends on the enzyme’s catalytic activity or its structural features, we combined site-directed mutagenesis and structure elucidation by cryo electron microscopy single particle analysis (cryo-EM SPA) and X-ray crystallography (Fig. 12, A, B and C). GDH is a hexameric enzyme that catalyzes the reversible conversion of glutamate to a-ketoglutarate (a-KG) and ammonia while reducing NAD(P)+to NAD(P)H. An enzymatic assay revealed a-KG, glutamate and ammonium as sole substrates and an optimum pH at 8.5 for glutamate utilization and 7.5 for glutamate formation as well as specificity for NAD7NADH as cofactors (Fig. 13, A, B and C). By using the allosteric inhibitor, GTP, and a GDH inhibitor, bithionol, which is also used as an anti-helminthic, heGDH activity was reduced (Fig. 13D). In contrast, clone 4F8 failed to reduce heGDH catalytic activity (Fig. 13E) as well as LT suppression (Fig. 10C), while atenuating the heGDH-mediated induction of the COX pathway (Fig. 10A). This suggested that structural features of heGDH are responsible for the induction of immune regulatory and tissue reparative prostanoids.

[0203] To untangle the role of catalytic activity versus structural properties we designed a catalytically inactive mutant of heGDH (heGDHK126A< D204N) (31) (Fig. 4A). This mutant still induced COX-metabolite and IL-10 production by macrophages to an equivalent or higher degree compared to the wildtype protein (Fig. 4, B and C). Gene expression data confirmed a similar or even stronger response of the top DEGs upon stimulation with mutant heGDHK126A> D204N(Fig. 4D). In contrast to its intact effects on prostanoids, heGDHK126A-D204Nfailed to significantly suppress cysLTs (Fig. 4E), supporting the hypothesis that the heGDH-driven induction of the COX pathway via p300 is structure dependent, whereas the catalytic function is necessary to reduce LT production. A role for the catalytic activity in the suppressive effects on LTs was supported by a similar or even stronger reduction of cysLT formation by distinct helminthic (Ts) or human GDH (Fig. 4F and Fig. 13F).

[0204] Cryo-EM and X-Ray crystallography of heGDH yielded similar structures with a D3 symmetry (Fig. 4G) and a resolution at 2.7 and 1.8 A, respectively, resembling the overall architecture of mammalian GDH (Fig. 13G), Unlike its mammalian counterparts or the X-raymodel (Fig. 13, G and H), the cryo-EM density of heGDH contains three “handle”-like densities in the central symmetry plane, each connecting two monomers (Fig. 4, G and H). These handle-like densities, with the size and shape of a short alpha helix or small unstructured region are connecting to the side chain density of Cys136 and are in close proximity to the N-terminal tails of heGDH as identified by X-ray crystallography (Fig. 4H). The handles and N-termini are exposed to the exterior solvent and may provide an ideal docking site for interaction partners of heGDH. To decipher the role of these structural features in heGDH-driven immune regulation, effects of a mutant lacking the handle like densities (heGDHc136S) (Fig. 131) or the N-terminus (heGDH'™) (Fig. 13J) were compared to the wildtype protein. Strikingly, all effects on key mediators (PGE2, IL-10) (Fig. 4I and Fig. 13K) or target genes (Fig. 4J) induced by heGDH were lost in the absence of the N-terminus, while heGDHc136Sacted similar to the wildtype protein, identifying the N-terminus as the key structural feature responsible for the unique effects of heGDH.

[0205] To identify cellular targets of heGDH, MDM were stimulated with the HA-tagged protein (SEQ ID NO: 110) for 30 min or 24 h. In one set of experiments, hemagglutinin (HA)- heGDH was again added into the lysate before immunoprecipitation (IP) using anti-HAor isotype control beads. Proteomic analysis revealed 17 overlapping proteins between the two experiments for both timepoints (Fig. 13L and Table S1-not shown). HeGDH was identified as the most abundant protein even when not added to the lysate, supporting efficient uptake into macrophages (Table S1-not shown). Fc gamma receptors (CD64) and GPNMB were identified as the most significantly and abundantly bound proteins by heGDH (relative to isotype control) (Fig. 4K and Table S1-not shown). Thus, structural features of heGDH enable interactions with factors involved in type 2 immunity and anti-helminth host defense (CD64 (32), DGAT 1 (33)) as well as with GPNMB, implicated in tissue repair and AAM activation (34, 35).

[0206] Suppression of leukotrienes is mediated via metabolic reprogramming

[0207] As our data suggested that heGDH can regulate LTs, which promote anti-helminth immunity (14) via its catalytic activity, we elucidated effects of heGDH on macrophage metabolism. In mammalian tissues, oxidative deamination of glutamate via GDH generates a-KG, which can fuel the TCA cycle and further generate ATP in oxidative phosphorylation (Oxphos). Metabolic flux analysis revealed that in line with the suppressive capacity of heGDH on AAM activation the metabolism of macrophages shifted towards increased basal glycolysis, typical for M1 activated macrophages (36) (Fig. 5Aand Fig. 14A). In contrast, AAM have been shown to rely primarily on Oxphos, which tended to be downregulated by heGDH (Fig. 5A and Fig. 14A). The significant p300- and H3K27ac-dependent upregulation of the gene PFKFB3 (Fig. 5B), a positive regulator of glycolysis, after heGDH stimulation suggested a potential link between glycolysis and heGDH-induced epigenetic reprogramming (Fig. 5C and Fig. 14B). Indeed, inhibition of p300 activity in MDM during treatment with heGDH blocked the decrease of basal respiration and ATP production as well as the increase of basal glycolysis (Fig. 5D), suggesting that the metabolic shifttriggered by heGDH is p300 dependent. LC-MS / MS analysis of TCA metabolites revealed higher levels of 2-hydroxyglutarate (2-HG) in heGDH-treated MDM, while the levels of glutamine and glutamate were reduced compared to untreated MDM (Fig. 5, E and F). Furthermore, the aconitate decarboxylase 1 (IRG1) product itaconate, an immunoregulatory byproduct of the TCA cycle was increased in response to heGDH (Fig. 5E). To determine whether the downstream metabolites of heGDH (itaconate, a-ketoglutarate and 2-HG) could affect immune regulatory AA metabolites, we assessed eicosanoid production by MDM following treatment with these metabolites (Fig. 14C). Indeed, we observed that L-2-HG, but not D-2-HG, reduced the production of cysLTs (Fig. 5G and Fig. 14D). To investigate, if L-2-HG directly affects the catalytic activity of leukotriene C< synthase (LTC4S), we performed an LTC4S activity assay. While LTC4S activity was partially inhibited by addition of L-2-HG (Fig. 5H), gene expression of AL0X5 and LTC4S were not affected (Fig. 14E). Interestingly, also heGDH directly affected the enzymatic activity of recombinant LTC4S (Fig. 5I) as well as in a human macrophage cell line (Fig. 14F). Thus, effects of heGDH on the synthesis of key mediators of type 2 immunity are - at least in part - mediated via its downstream metabolites.

[0208] heGDH-induced PGE2suppresses type 2 effector functions of macrophages and T cells100209] To investigate whether recombinant heGDH can modulate host defense in vivo, mice infected with Hpb were treated (i.p.) with the protein during the tissue-dwelling phase of the parasite, associated with AAM-mediated trapping and killing (Fig. 6A). Indeed, our combined RNA-seq, ChlP-seq and proteomics data (Fig. 1-5) suggested that heGDH suppresses multiple key steps in the activation of AAM effector functions, essential for host defense against parasitic nematodes (4, 5, 37). In keeping with these suppressive effects, administration of heGDH led to a significant increase in worm burdens at 14 days p.i. and reduced expression of the AAM markers RELMa and Ym-1 / 2, while the M1 marker INOS was increased (Fig. 6, B and C, and Fig. 15A). In line with our in vitro data (Fig. 3), peritoneal macrophages from heGDH-treated mice showed increased H3K27ac, correlating with enhanced PGEg production and expression of COX-2 in the small intestine (Fig. 6, C, D and E, and Fig. 15B). Reduced downregulation of CD206 in heGDH- treated BMDM from naive mice lacking the PGE2receptor EP2, supported a key role for PGE2in the regulation of AAM activation (Fig. 6F). Assessment of heGDH’s impacton helminth chronicity at a later time point (28 days p.i.) (Fig. 6G), yielded an even stronger increase in worm burdens compared to day 14 (Fig. 6, B and H) and tended to increase egg counts (Fig. 15C), supporting the role of heGDH in promoting helminth chronicity (Fig. 1B). While treatment with heGDH did not induce PGE2in peritoneal macrophages (Fig. 6I) at this later time point, other prostanoids (TXB2and 6-keto-PGFia), associated with tissue repair (26), were increased in the small intestine of mice treated with heGDH (Fig. 6J). However, parameters of intestinal tissue repair (myofibroblast / a-smooth muscle actin (a-SMA) accumulation; collagen deposition)^ 9) remained unaffected byheGDH (Fig. 15, D, E and F). HeGDH treatment also reduced the percentage of Gata3+Th2 cells and type 2 cytokine expression in the mesenteric lymph nodes at 28 days post Hpb infection, while Th2 activation was unaffected at earlier time points (Fig. 6K and Fig. 15, G, H and I). To investigate the importance of heGDH-triggered PGE2 in the regulation of helminth-induced Th2 responses we analyzed the capacity of heGDH to reduce type 2 cytokine production in human peripheral blood mononuclear cells (PBMCs) following stimulation with Schistosoma mansoni soluble egg antigen (SEA), a strong parasitic Th2 trigger. The percentage of SEA-induced IL- 4+CD4+Th2 cells was significantly reduced and SEA-induced IL-4 production was diminished after heGDH treatment (Fig. 6L), while heGDH induced CD4+CD127'CD25hiFoxP3+regulatory T cells in human PBMCs (Fig. 6M). These T cell modulatory functions of heGDH were abrogated by a selective inhibitor of microsomal prostaglandin-E-Synthase-1 (mPGES-1 ) (Fig. 6, L and M) or depletion of monocytes (Fig. 15J), indicating that PGEz acts as a key modulator of macrophage and T cell function during heGDH-induced helminth chronicity.

[0210] Host type 2 immunity limits immune evasion, but not tissue repair driven by heGDH

[0211] To assess how host type 2 immunity may affect heGDH-driven immune regulation, we mimicked a type 2 milieu in vitro by culturing MDM and BMDM in the presence of IL-4 and IL- 13 prior to treatment with heGDH. IL-4 / IL-13-induced genes (AL.OX15 and MRC1 in human MDM; Retnla, Chil3, Mrd in murine BMDM) were or tended to be downregulated by heGDH, confirming the prevention of AAM polarization in a type 2 cytokine milieu (Fig. 7Aand Fig. 16A). While some heGDH-induced genes, including IDO1, IL12B and EBI3 were unaffected by additional treatment with IL-4 and IL-13 (Fig. 7B and Fig. 16A), IL-4 and IL-13 suppressed the induction of PTGS2 and PTGES as well as PGE2(Fig. 7, C and D). In line with these counter-regulatory effects of type 2 cytokines, the capacity of heGDH to trigger immune evasion was impaired during strong type 2 immune responses, i.e. during challenge infection with Hpb or infection with Nippostrongylus brasiliensis (Nb) (Fig. 7, E, F, I and J). However, treatment (i.p.) with heGDH still resulted in a tendency towards increased worm burdens (Fig. 7F) and a reduction of the AAM markers RELMa and Ym-1 / 2 during challenge infection with Hpb (Fig. 7G and Fig. 16B). To study whether heGDH may directly affect macrophage-mediated trapping of Hpb larvae (5), immune serum- activated macrophages were co-incubated with L3 in the presence or absence of heGDH. Indeed, macrophage-mediated larval trapping was impaired by heGDH (Movie S1 ), showing that heGDH directly suppresses a key macrophage effector function during helminth infection. As during primary infection, heGDH treatment during challenge infection did not affect T-cell composition, levels of arginase 1 (Arg1 ), cx-SMA or collagen in the small intestine (Fig. 15B and Fig. 16, C, D, E and F), suggesting that tissue repair remained unaltered. In line with blunted prostanoid induction and immune evasion in the presence of type 2 cytokines (Fig. 7C and F), PGEg secretion by peritoneal macrophages and upregulation of COX-2 by heGDH were atenuated in thegranuloma of challenge-infected as compared to primary infected mice (Fig. 6, C and E; Fig. 7, G and H and Fig. 16D). Intranasal treatment with heGDH during infection with Nb (Fig. 7I), a helminth that triggers a rapid type 2 immune response, similarly failed to significantly affect intestinal worm burdens (Fig. 7J). Despite increasing airway neutrophils, IL-6 and IL-17A production, intranasal heGDH treatment reduced leukotriene B4(LTB4) synthesis, collagen deposition and lung damage in Nb infected mice (Fig. 7, L and M, Fig. 17, A, B and C). Intestinal eicosanoid profiles as well as T-cell responses in the lung and MLN remained largely unaffected by intranasal heGDH treatment except for PGE2, which was increased in the intestine of naive mice (Fig. 17, D, E and F), suggesting that heGDH can indeed influence the lung-gut axis. The decreased type 2 immune response and improved tissue repair in mice treated with heGDH during Nb infection correlated with an increase in anti-fibrotic factors (Arg1 and PGE2) (38, 39) in BAL macrophages (Fig. 7K and Fig. 17G). Together, this suggests that type 2 immune responses have evolved to overcome helminth / GDH-driven immune evasion, while leaving tissue reparative and pro-resolving functions of immune regulatory helminth molecules intact.

[0212] DISCUSSION

[0213] The present study identifies key roles of helminthic GDH in immune evasion, parasite chronicity and tissue repair during helminth infections, which represent a major global health burden (40). HeGDH targets host macrophages, resulting in metabolic, epigenetic, and transcriptional changes that broadly suppress anti-helminth effector functions. Intriguingly, catalytic activity and structural features of heGDH synergize to suppress mediators of type 2 immunity and induce factors that regulate macrophage and T cell activation. Macrophages play crucial roles in anti-helminth immunity by trapping worms, regulating T cell responses, and repairing tissue damage (5, 7, 8, 12, 16, 26), making them prime targets of parasitic immune evasion (18). CD64 positive macrophages co-localize with GDH-containing helminth larvae in the small intestine and our proteomic analysis identified CD64 and GPNMB as potential targets of heGDH. Indeed, the antibody-driven activation of macrophages via CD64 plays an important role in helminth trapping (8, 32), suggesting that heGDH binds CD64 to ensure efficient uptake and evade macrophage-mediated immunity. Although we do not provide experimental evidence that directly links CD64 or GPNMB to p300 activation, a previous study demonstrated that C-reactive protein can bind CD64, resulting in HIF-1a / p300-dependent transcriptional activation (41). Furthermore, GPNMB upregulates H3K27 lysine demethylase JMJD3 (42) which cooperates with p300 to switch towards H3K27ac-mediated transcriptional activation (43). This suggests that heGDH targets multiple factors involved in epigenetic reprogramming and AAM activation to efficiently interfere with anti-helminth immunity (34). In contrast to the suppression of several key IL-4 / IL-13-induced genes (Mrd / CD206, Alox15, Chil3 / Ym1, Fizzll RELMa) by heGDH, the enzyme does not target Arg1, suggesting that Arg 1 -mediated regulation of Th2 responses (38) remains intact in the presence of heGDH.

[0214] While heGDH triggers a broad epigenetic and transcriptional reprogramming, our data support a key role for PGE2 in GDH-driven helminth chronicity. HeGDH-induced PGE2suppresses AAM polarization and Th2 cell activation, two essential mechanisms of anti-helminth immunity (4, 7, 37). In addition, PGE2 may limit ILC2 and mast cell function (44, 45), suggesting that heGDH affects additional cell types involved in type 2 immunity. It will be important to investigate whether heGDH targets additional cell types involved in the initiation of type 2 immunity, including tuft cells, a major early source of host-protective cysLTs (14). Given the roles of eicosanoids in tissue repair (46-49), it will be interesting to investigate whether prostanoids are responsible for tissue-reparative effects of heGDH, e.g. by using mice with deficiencies in PGE2 synthesis (50). Importantly, in contrast to helminthic GDHs, human GDH (Fig. 18) fails to trigger the PGEa-dependent induction of regulatory T-cells (16), suggesting that the N-terminus, which is distinct from mammalian GDHs, allows for interactions of heGDH with its targets (e.g. CD64, GPMNB), thus conferring specific immune regulatory effects to heGDH. Indeed, an N-terminal truncation mutant of heGDH showed a complete loss of all tested immune regulatory effects (including the induction of PGE2and IL-10), suggesting that the exposed N-termini in the heGDH hexamer provide key docking sites for interaction partners.

[0215] While the induction of type-2 suppressive factors (PGE2, IDO1 and IL-12 family cytokines) shared the same structure / p300-dependent upstream mechanism, the suppression of type 2 promoting cysLTs depended on the catalytic activity of GDH. Indeed, L-2-HG induced by heGDH can interfere with LTC4S activity to reduce the synthesis of cysLTs, important for early anti-helminth immunity (14). However, the precise mechanism by which L-2-HG limits LTC4S activity remains to be determined. As LTC4S activity is controlled by phosphorylation via the mTOR / p70S6K pathway (51, 52), which is activated by 2-HG (53), it would be interesting to assess whether heGDH triggers inhibitory phosphorylation of LTC4S.

[0216] A conserved role of GDHs in parasite chronicity is supported by studies identifying GDH as a dominant vaccine target (54) and showing immunomodulatory effects of GDH from the protozoan parasite T. cruzi as well as from the parasitic cestode T solium (16, 55). Future research should thus determine if GDHs from different parasites use the same mechanisms to drive chronicity and tissue repair. The weakened effects of heGDH in the presence of a full-blown type 2 immune response suggest that host type 2 immunity has evolved to counteract helminth- driven immunoregulation. The present study thus identifies an important pathway of host-parasite crosstalk via a ubiquitous metabolic enzyme, which may be harnessed for the therapy and / or prevention of major infectious and inflammatory diseases.

[0217] Example 2: HeGDH increases anti-inflammatory prostaglandin E2 (PGE2) while suppressing pro-inflammatory cysteinyl leukotriene (cysLT) production when administered to nasal polyp tissues from CRSwNP patients ex vivo:

[0218] Methods: Nasal polyp (NP) tissues were obtained from n=4 CRSwNP patients by Functional Endoscopic Surgery (FESS) and cultured ex vivo in airway epithelial culture medium (Lonza). NP tissue was incubated for 24h with different concentrations of recombinant heGDH (SEQ ID NO: 27). Tissue culture supernatants were collected and analyzed for lipid mediator (prostanglandin E2 and cysteinyl leukotriene) release by ELISA kits (Cayman Chemicals). Concentrations were normalized to tissue weight.

[0219] Results: HeGDH increases anti-inflammatory prostaglandin E2 (PGEZ) while suppressing pro-inflammatory cysteinyl leukotriene (cysLT) production when administered to nasal polyp tissues from CRSwNP patients ex vivo. The suppressive effect on cysLTs is donor dependent and particularly prominent for donors with high cysLT output (Figure 19).

[0220] Conclusion: HeGDH increases anti-inflammatory prostaglandin E2 (PGE2) while suppressing pro-inflammatory cysteinyl leukotriene (cysLT) production when administered to nasal polyp tissues from CRSwNP patients ex vivo such that the suppressive effect is donor dependent and particularly prominent for donor with high cysLT output.

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Claims

CLAIMS1. A polypeptide, wherein said polypeptide comprising one or more amino acid sequences having at least 80% (e.g., at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to a heGDH (helminthic glutamate dehydro- genase)-derived polypeptide selected from the group consisting of: i) SEQ ID NO: 1 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDL); ii) SEQ ID NO: 2 (MLSTLARTSGRLIFRRALSSAQMDAHAQVIDDLKPMEEQSN); iii) SEQ ID NO: 3 (MLSTLARTSGRLIFRRALSSA); iv) SEQ ID NO: 4 (QMDAHAQVIDDLKP); and v) SEQ ID NO: 5 (MEEQSN), preferably said polypeptide is not a full-length heGDH (e.g., having UniProt accession num- ber: A0A183FP08 or SEQ ID NO: 27), further preferably said polypeptide having less than 100% sequence identity to said full-length heGDH (e.g., does not have enzymatic activity and / or is catalytically inactive).

2. The polypeptide according to any one of the preceding claims, wherein said polypeptide is a fragment or variant of any one of (i)-(v) according to claim 1 , preferably said fragment or variant is at least 6 (e.g., at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at lest 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31 , at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, or at least 41) amino acids long; further preferably said fragment or variant comprising an amino acid motif comprising at least 2 (e.g., at least 3, at least 4, at least 5) positively charged amino acids (e.g., arginines (R), lysines (K) and / or histidines (H), preferably said motif comprising at least 2 arginines (RR)), further preferably said amino acid motif is located in the tail-region (e.g., C- or N-terminal end segment / fragment having at least 6 amino acids) of said poly- peptide and / or is capable of interacting with one or more negatively charged and / or glyco- sylated targets / polypeptides, preferably is capable of interacting with CD64 (High affinity immunoglobulin gamma Fc receptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt accession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa protein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / orReceptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like protein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7) and / or Calcium homeostasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt accession number: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1 , e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1 (DGAT1, e.g., having UniProt accession number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having Uni- Prot accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt accession number: Q8NHP8) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession number: Q9BYJ4).

3. The polypeptide according to any one of the preceding claims, wherein said polypeptide consists of one or more polypeptide / s selected from SEQ ID NOs: 1-5 and / or one or more variants thereof.

4. The polypeptide according to any one of the preceding claims, wherein said heGDH (hel- minthic glutamate dehydrogenase) having UniProt accession number: A0A183FP08 and / or EC 1.4.1.3 enzymatic activity.

5. The polypeptide according to any one of the preceding claims, wherein said polypeptide does not have enzymatic activity and / or is catalytically inactive, e.g., does not have EC 1.4.1.2, EC 1.4.1.3 and / or EC 1.4.1.4 enzymatic activity, preferably not having EC 1.4.1.3 enzymatic activity), preferably does not have heGDH enzymatic activity (e.g., EC:1.4.1.3).

6. The polypeptide according to any one of the preceding claims, wherein said polypeptide is capable of interacting with one or more negatively charged and / or glycosylated targets / pol- ypeptides.

7. The polypeptide according to any one of the preceding claims, wherein said polypeptide is capable of interacting with CD64 polypeptide (High affinity immunoglobulin gamma Fc re- ceptor I (FCGR1A / FCGR1BP, e.g., having UniProt accession number: P12314 / Q92637) and / or Glycoprotein nonmetastatic melanoma protein B (GPNMB, e.g., having UniProt ac- cession number: Q14956) and / or DnaJ homolog subfamily B member 11 (DNAJB11 , e.g., having UniProt accession number: Q9UBS4) and / or Signal recognition particle 9 kDa pro- tein (SRP9, e.g., having UniProt accession number: P49458) and / or Sideroflexin-3 (SFXN3, e.g., having UniProt accession number: Q9BWM7) and / or Receptor expression-enhancing protein 4 (REEP4, e.g., having UniProt accession number: Q9H6H4) and / or Rab-like pro- tein 6 (RABL6, e.g., having UniProt accession number: Q3YEC7) and / or Calcium homeo- stasis endoplasmic reticulum protein (CHERP, e.g., having UniProt accession number: Q8IWX8) and / or Solute carrier family 25 member40 (SLC25A40, e.g., having UniProt ac- cession number: F8WEL8) and / or Stromal cell-derived factor 2-like protein 1 (SDF2L1, e.g., having UniProt accession number: Q9HCN8) and / or Diacylglycerol O-acyltransferase 1(DGAT 1 , e.g., having UniProt accession number: 075907) and / or Probable ATP-dependent RNA helicase DHX35 (DHX35, e.g., having UniProt accession number: Q9H5Z1) and / or Putative phospholipase B-like 2 (PLBD2, e.g., having UniProt accession number: Q8NHP8) and / or Tripartite motif-containing protein 34 (TRIM34, e.g., having UniProt accession num- ber: Q9BYJ4).

8. The polypeptide according to any one of the preceding claims, wherein said polypeptide is an immunogenic polypeptide and / or said polypeptide is capable of modulating the innate mammalian immune system, preferably said polypeptide is capable of modulating macro- phage-and T-cell mediated immunity, further preferably said polypeptide is capable of reg- ulating macrophage functions and inducing regulatory T cells by epigenetic targeting of prostaglandin synthesis.

9. The polypeptide according to any one of the preceding claims, wherein said variant com- prises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) alterations / modifications at one or more positions (e.g., non-natural amino acid / s), preferably wherein said variant comprises one or more substitutions, deletions, and / or insertions at one or more positions.

10. The polypeptide according to any one of the preceding claims, wherein said polypeptide comprises one or more immunoaffinity tags (e.g., one or more His-tags and / or Strep-I I tags).

11. A nucleic acid construct or expression vector encoding one or more polypeptides according to any one of the preceding claims.

12. A host cell (e.g. an isolated and / or recombinant host cell) comprising one or more nucleic acid constructs and / or expression vectors according to any one of the preceding claims and / or expressing one or more polypeptides according to any one of the preceding claims.

13. A composition or kit comprising one or more of the polypeptides, nucleic acid constructs or expression vectors and / or host cells of any one of preceding claims.

14. The composition or kit of any one of the preceding claims, wherein said composition or kit is a diagnostic, pharmaceutical composition and / or vaccine composition or kit.

15. The composition or kit of any one of the preceding claims, wherein said composition is an immune-therapeutical composition or kit,16. A method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid resistant air- way inflammation, aspirin / NSAID-exacerbated respiratory disease (AERD / N-ERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune dis- ease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantlyallergic asthma, atopic asthma, extrinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fi- brosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory dis- ease, allergy; said method comprising administering a therapeutically effective amount of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions of any one of the preceding claims, preferably administered locally (e.g., in- tranasally), further preferably administered to the airways.

17. One or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kit of any one of the preceding claims for use as a medicament and / or in therapy.

18. One or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding claims for use in one or more of the follow- ing methods: i) in a method for treatment, amelioration and / or prophylaxis of a disease selected from a group consisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory dis- ease, steroid resistant airway inflammation, aspirin-exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic derma- titis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhini- tis, diabetes; bronchitis, chronic bronchitis, mucopurulent chronic bronchitis, emphy- sema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary dis- ease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, ex- trinsic allergic asthma, non-allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino- conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; prefer- ably administered locally (e.g., intranasally), further preferably administered to the air- ways; ii) in a method for modulating the mammalian innate and adaptive immune response, preferably administered locally (e.g., intranasally), further preferably administered to the airways; iii) in a method for predominantly modulating the mammalian innate immune response over the mammalian adaptive immune response; iv) in a method for targeting phagocytic cells of the mammalian immune system;v) in a method for modifying the activation of macrophages and / or granulocytes of the mammalian immune system; vi) in a method for modifying the activation of one or more of the arachidonic acid path- way of the mammalian immune system; vii) in a method for decreasing the number of eosinophils and / or inhibiting the migration of granulocytes into tissue of the mammalian immune system; viii) in a method for eliciting or modulating an immune response in a subject; ix) in a method for treatment, amelioration, prophylaxis or diagnostics of a steroid-re- sistant disease; x) in a method for monitoring development of a disease and / or assessing the efficacy of a therapy of a disease; xi) any method according to any one of the preceding claims; xii) any combination of methods as in any one of (i)-(xi); xiii) any method according to (i)-(xii), wherein said method is an in vitro, in vivo or ex vivo method.

19. Use of one or more polypeptides, nucleic acid constructs or expression vectors, host cells and / or compositions or kits of any one of the preceding claims for / in one or more of the following: i) for treatment, amelioration and / or prophylaxis of a disease selected from a group con- sisting of: nasal polyposis (NP), nasal polyposis (NP) comorbid with asthma, asthma, inflammation, chronic airway inflammation, chronic respiratory disease, steroid re- sistant airway inflammation, aspirin-exacerbated respiratory disease (AERD), nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune disease, inflammatory disease, chronic inflammatory disease, rhinitis, diabetes; bron- chitis, chronic bronchitis, mucopurulent chronic bronchitis, emphysema, MacLeod syndrome, panlobular emphysema, centrilobular emphysema, chronic obstructive pulmonary disease (COPD), chronic obstructive pulmonary disease with acute lower respiratory infection, chronic obstructive pulmonary disease with acute exacerbation, asthma, predominantly allergic asthma, atopic asthma, extrinsic allergic asthma, non- allergic asthma, idiosyncratic asthma, intrinsic nonallergic asthma, mixed asthma, asthmatic bronchitis, late-onset asthma, status asthmaticus, acute severe asthma, bronchiectasis, nasal polyps, cystic fibrosis (CF), allergic rhino-conjunctivitis, atopic dermatitis, autoimmune or inflammatory disease, allergy; preferably administered lo- cally (e.g., intranasally), further preferably administered to the airways; ii) for modulating the mammalian innate and adaptive immune response, preferably ad- ministered locally (e.g., intranasally), further preferably administered to the airways; iii) for predominantly modulating the mammalian innate immune response over the mam- malian adaptive immune response;iv) for targeting phagocytic cells of the mammalian immune system; v) for modifying the activation of macrophages, T-cells and / or granulocytes of the mam- malian immune system; vi) for modifying the activation of one or more of the arachidonic acid pathway of the mammalian immune system; vii) for decreasing the number of eosinophils and / or inhibiting the migration of granulo- cytes into tissue of the mammalian immune system; viii) for eliciting or modulating an immune response in a subject; ix) for treatment, amelioration, prophylaxis or diagnostics of a steroid-resistant disease; x) for monitoring development of a disease and / or assessing the efficacy of a therapy of a disease; xi) for any method according to any one of the preceding claims; xii) any combination of methods as in any one of (i)-(xi); xiii) any use according to (i)-(xii), wherein said use is an in vitro, in vivo or ex vivo use.

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