WO2015131053A1 - Polypeptides dérivés de phl p, procédés et utilisations de ces derniers pour moduler une réponse immunitaire - Google Patents

Polypeptides dérivés de phl p, procédés et utilisations de ces derniers pour moduler une réponse immunitaire Download PDF

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WO2015131053A1
WO2015131053A1 PCT/US2015/018026 US2015018026W WO2015131053A1 WO 2015131053 A1 WO2015131053 A1 WO 2015131053A1 US 2015018026 W US2015018026 W US 2015018026W WO 2015131053 A1 WO2015131053 A1 WO 2015131053A1
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seq
nos
pollen
amino acid
molecule
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PCT/US2015/018026
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English (en)
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Ilka HOOF
Lars Harder CHRISTENSEN
Peters BJOERN
Jason GREENBAUM
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Alk-Abelló A/S
La Jolla Institute For Allergy And Immunology
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Publication of WO2015131053A1 publication Critical patent/WO2015131053A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/35—Allergens
    • A61K39/36—Allergens from pollen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—Inorganic adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/577—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 tolerising response

Definitions

  • the invention relates to pan pollen immunogenic molecules such as polypeptides, proteins and peptides, and methods and uses of such immunogenic molecules for modulating or relieving an immune response in a subject, such as treating a subject for an allergic immune response or inducing or promoting immunological tolerance to the immunogenic molecule or a pollen allergen in a subject.
  • allergen-specific T-cells play an important role in allergic inflammation and that induction of antigen specific T regulatory cells (Tregs) or elimination of allergen-specific T helper type 2 cells (Th2) might be a prerequisite for the induction of specific tolerance. Yet, cross-reactivity among multiple pollen families at the T-cell level is less explored.
  • Allergen-specific immunotherapy SIT is a hyposensitizing immunotherapy introduced in clinical medicine almost a century ago for the treatment of an allergic immune response using the allergens that the subject is sensitized to.
  • An allergic immune response may be mediated by activated allergen-specific Th2 cells, which produce cytokines such as IL-4, IL-5, and IL-13.
  • the allergen-specific T-cell response is mediated predominantly by Th1 cells.
  • SIT may reduce the ratio of Th2:Th1 cells and may alter the cytokine profile, reducing the production of IL-4, IL-5, and IL-13 and increasing the production of IFN-gamma in response to major allergens or allergen extracts.
  • SIT has several limitations, including safety concerns about giving patients allergenic substances. Because most SIT regimens involve the administration of whole, unfractionated, allergen extracts, adverse IgE-mediated events are a considerable risk. Significant efforts have been devoted to developing approaches to modulate allergen-specific T-cell responses without inducing IgE-meditated, immediate-type reactions. These approaches include developing hypoallergens that do not contain IgE- binding epitopes, allergens that are coupled to adjuvants and carriers of bacterial or viral origin or peptides that contain dominant T-cell epitopes and do not react with IgE in allergic individuals.
  • NTGA novel Timothy Grass antigens
  • NTGA's are unrelated to the known allergens of Timothy grass, which mainly are identified based on their high IgE reactivity.
  • International patent application, WO2013/119863 A1 relates to novel antigens (NTGA's) derived from Timothy grass pollen.
  • an immunogenic molecule derived from an allergenic pollen source is able to reduce an allergic immune response caused by an unrelated allergen of the same pollen source via bystander suppression.
  • immunogenic molecules with high sequence conservation/homology to polypeptides identified in several different pollen families.
  • Such immunogenic molecules have potential therapeutical utilization against immune responses triggered by pollen allergens of a broad array of pollen families for example via induction of bystander tolerance towards the offending pollen allergens.
  • immunogenic molecules in short immunogens
  • pan- pollen immunogens which have been detected in Timothy grass pollen (Phleum pratense (in short Phi p) and in at least one non-grass pollen species, which consist of or contain as part of their sequence an amino acid sequence conserved across the Phi p grass pollen species and at least one of the non-grass pollen species investigated.
  • a molecule e.g. an immunogenic molecule
  • a molecule of the invention may consist of or contain as part of its sequence a conserved amino acid sequence detected in several different pollen families, e.g. in a grass pollen species (e.g.
  • Phleum Pratense (Phi p) and at least one of the following non-grass pollen species: Ambrosia psilostachya (Amb p), Ambrosia artemisiifolia, (Amb a), Quercus alba (Que a) and Betula verrucosa, (Bet v).
  • Table 1 shows examples of amino acid sequences of "pan pollen” polypeptides detected and identified in Phi p grass pollen and Table 2 shows amino acid sequences of polypeptides homologous to the polypeptides of Table 1 , which is found in pollen of the species Cyn d, Amb a, Amb p, Que a, and Bet v. As those polypeptides can be detected in pollen, they are referred to herein as wild type polypeptides.
  • GWT Garnier-Weed-Tree
  • Table 3 shows GWT conserved regions identified in the Phi p sequences of Table 1.
  • Table 4 shows the corresponding GWT conserved regions identified in the other pollen species investigated.
  • a conserved region as used herein is defined as the region identified from merging overlapping conserved 15mer peptides in a Phi p sequence, wherein a conserved 15mer peptide is defined as a 15 amino acid peptide containing 0, 1 , or 2 mismatches to a 15mer peptide sequence of a homologous polypeptide in a weed pollen species (Amb a and/or Amb) and in a tree pollen species (Que a and/or Bet v).
  • conserved amino acid sequences of the polypeptides of Table 1 and 2 that are conserved across at least a set of homologous polypeptides found in Phi p grass pollen and in at least one weed pollen species (Amb a or Amp), but not in a tree pollen species (Que a or Bet v).
  • Such sequences are herein named GW (Grass-Weed) conserved stretches of amino acid residues (or alternatively GW conserved regions).
  • Table 5 shows GW conserved regions identified in the Phi p sequences of Table 1.
  • Table 6 shows the corresponding GW conserved regions identified inthe other pollen species.
  • conserved amino acid sequences of the polypeptides of Table 1 and 2 that are conserved across at least a set of homologous polypeptides found in Phi p grass pollen and in at least one tree pollen species (Que a or Bet v), but not in a weed pollen species (Amb a or Amp).
  • Such sequences are herein named GT (Grass- Tree) conserved stretches of amino acid residues (or alternatively GT conserved regions).
  • Table 7 shows GW conserved regions identified in the Phi p sequences of Table 1.
  • Table 8 shows the corresponding GT conserved regions identifiedin the other pollen species.
  • WT Wood-Tree
  • Table 9 shows WT conserved regions identified in polypeptides of Table 2.
  • the GWT, GW, GT or WT conserved sequences may be an immunogenic molecule in itself, or may give rise to additional immunogens comprising the conserved sequences or subsequences thereof.
  • a molecule e.g. an immunogenic molecule
  • an immunogenic molecule of the invention which is identified in Phi p pollen may be able to modify a T cell response in a grass allergic donor as well as in donors allergic to a weed pollen species and/or tree pollen species. This is evidenced using Phi p allergens and a selection of recently identified Phi p antigens as model immunogens (Example 8). It was found that a T cell response of grass allergic donors to a first immunogenic molecule derived from Phi p may also be activated in detectable levels by a second immunogenic molecule containing few mismatches to the first immunogenic molecule, e.g. preferably no more than 2 or 3 mismatches.
  • the presently provided conserved sequences shown in Tables 3 to 8 may be able to induce a T cell response in donors not only sensitized to Phi p pollen allergens, but to allergens of any of the pollen species Cyn d, Amb a, Amb p, Que a and Bet v as well as other pollen species containing polypeptides with similar conserved sequences comprising less than 2 or 3 mismatches to the Phi p sequence in question.
  • the invention relates in a first aspect to a molecule, such as an immunogenic molecule comprising, consisting of or consisting essentially of a) a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 274, 270-273, 212-269 and 275-293 set out in Table 3; SEQ ID NOS: 617-676, 294-616 and 677-767 set out in Table 4; SEQ ID NOS: 768-808 set out in Table 5; SEQ ID NOS: 809-951 set out in Table 6; SEQ ID NOS: 952-1023 set out in Table 7; SEQ ID NOS: 1024-1231 set out in Table 8 and SEQ ID NOS: 1232-1473 set out in Table 9; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NO
  • the invention also relates to the use of said molecule (e.g. said immunogenic molecule) as a medicament, in particularly for use in modulating an immune response, e.g. relieving an immune response, triggered by pollen (i.e. a pollen allergen and/or an immunogenic molecule disclosed herein) in a subject.
  • pollen i.e. a pollen allergen and/or an immunogenic molecule disclosed herein
  • the invention relates to a method for modulating an immune response, e.g. for relieving an immune response (e.g. triggered by pollen or an immunogen present in pollen), in a subject in need thereof, comprising administering an effective amount of a molecule (e.g. an immunogenic molecule) disclosed herein.
  • the invention relates to an immunogenic molecule as disclosed herein for use in modulating an immune response, e.g. for relieving an immune response, in a subject in need thereof, comprising administering an effective amount of a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a molecule e.g. an immunogenic molecule
  • the invention also relates to the use of a molecule (e.g. an immunogenic molecule) as disclosed herein as a medicament, e.g. use of an immunogenic molecule as disclosed herein for the preparation of a medicament for modulating an immune response, e.g. relieving an immune response, in a subject in need thereof.
  • a molecule e.g. an immunogenic molecule
  • an immunogenic molecule as disclosed herein for the preparation of a medicament for modulating an immune response, e.g. relieving an immune response, in a subject in need thereof.
  • the immune response in question may be triggered by pollen, e.g. by pollen of one or more pollen species, pollen genera or pollen families. It follows that the immune response in question may be triggered by one or more pollen allergens from one or more pollen species, pollen genera or pollen families including one or more molecules (e.g.
  • immunogenic molecules disclosed herein, for example the immunogenic molecule itself or a subsequence thereof.
  • a polypeptide of option d) comprises an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 27-30 set out in Table 1 and SEQ ID NOS: 161-172 set out in Table 2;
  • a polypeptide of option a) comprises an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 270-274 set out in Table 3 and SEQ ID NOS: 617-676 set out in Table 4;
  • a polypeptide of option b) comprises an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS
  • the molecule may comprise a combination of two or more polypeptides of option a), option b), option c) or option d), so as to construct molecules with desirable properties e.g. immunogenic molecules with high conservation throughout the entire amino acid sequence.
  • a polypeptide of option a) may contain as part of its sequence a combination of two or more amino acid sequences having at least 65% similarity or identity to a sequence selected from any one of SEQ ID NOS: 212-293 set out in Table 3 and SEQ ID NOS: 294-767 set out in Table 4.
  • a polypeptide of option b) and c) may comprise as part of its sequence a combination of two or more of said subsequences.
  • a cell expresses said molecule.
  • a cell is a eukaryotic or prokaryotic cell and may be a mammalian, insect, fungal or bacterium cell.
  • a molecule (e.g. an immunogenic molecule) of the present invention is suitable as a reagent, for example in immunotherapy against various pollen allergies in a subject.
  • nucleic acid molecules encoding a polypeptide of option a), b), c) or d) or a molecule comprising a polypeptide of option a), b), c) or d).
  • compositions for example pharmaceutical compositions comprising an immunogenic molecule of the invention.
  • a pharmaceutical composition is suitable for immunotherapy (e.g. treatment,
  • a pharmaceutical composition is a vaccine, e.g.
  • FIG. 1 Sensitization pattern of an immunogen of the invention (A0349): It is shown that the in vitro T-cell response towards A0349 is much weaker compared to the response to allergen Phi p 5.
  • FIG. 3 Tolerance induction investigated in mice shows that prophylactic sublingual immunotherapy treatment (SLIT) with A0349 in mice is capable of inducing tolerance towards Phi p extract as shown by the ability of A0349 to reduce the
  • an immunogenic molecule that is weaker than Phi p 5, the major allergen in Phi p extract, is able to reduce an immune response triggered by Phi p extract.
  • FIG 4 Bystander tolerance induction investigated in mice.
  • prophylactic SLIT treatment with A0349 could induce tolerance toward the allergen Phi p 5, which amino acid sequence is quite different from A0349.
  • Phi p 5 an unrelated antigen
  • FIG. 4A Treatment with A0349 resulted in the suppression of an immune response caused by an unrelated antigen, i.e. Phi p 5, possibly via bystander mechanisms, where the tolerance induction towards one immunogen (e.g. A0349) result in suppression of an immune response caused by another unrelated immunogen (e.g. Phi p 5).
  • a or “an” refers to an indefinite number and shall not only be interpreted as “one” but also may be interpreted to mean “some", “several” or one or more.
  • conserved sequence is in the present context meant to include that a given amino acid sequence of one pollen species contains at least 15 contiguous amino acids within the sequence that has less than 3 mismatches compared to an amino acid sequence of 15 amino acid residues from another pollen species. Longer stretches of conserved sequences may contain several stretches of at least 15 contiguous amino acids having less than 3 mismatches compared to another sequence of 15 amino acids from another pollen species, genera or family.
  • the conserved region is usually determined by multiple sequence alignments of three or more homologous pollen polypeptides.
  • polypeptide polypeptide
  • protein protein
  • peptide may be used interchangeably and mean any peptide-linked chain of amino acids regardless of post-translational modification. While polypeptides can be any length, the use of the term “protein” generally means longer polypeptides of at least 100 amino acid residues, whereas the term “peptide” generally means polypeptides shorter than 30 amino acid residues.
  • mismatch is meant to include any substitution of an amino acid residue within the 15mer peptide. Optionally, a mismatch may be a deletion or an addition of an amino acid residue within the 15mer peptide.
  • homologous polypeptides refer to a pollen polypeptide derived from a common ancestor and are typically, although not necessarily, polypeptides having one or more similar functions.
  • homologous pollen polypeptides will have a degree of sequence identity to each other across the entire length of each protein that is at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, 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 greater.
  • homologous pollen polypeptides are found in pollens of different species, genera and families.
  • immunogenic molecule e.g. an allergen or an antigen
  • an immunogenic molecule e.g. an allergen or an antigen
  • the individual's adaptive immune system displays memory to the immunogenic molecule, for example that the immunogenic molecule has induced detectable IgE antibodies against the immunogenic molecule and thus qualifies as an IgE-reactive antigen
  • T-cells stimulated in vitro are able to proliferate under the presence of the immunogenic molecule or fragments of the immunogenic molecule (e.g. linear peptides derived from the immunogenic molecule).
  • allergic immune response is meant to encompass a hypersensitivity immune response, e.g. a type 1 hypersensitivity immune response, such as typically an immune response that is associated with the production of IgE antibodies (i.e. IgE-mediated immune response) and/or production of cytokines usually produced by Th2 cells.
  • An allergic immune response may be associated with an allergic disease, for example atopic dermatitis, urticaria, contact dermatitis, allergic conjunctivitis, allergic rhinitis, allergic asthma, anaphylaxis, food allergy and hay fever.
  • grass pollen is meant to designate pollen of the plant family Poaceae, for example pollen of the plant genus Anthoxanthum, Cynodon, Dactylis, Festuca, Holcus, Hordeum, Lolium, Oryza, Paspalum, Phalaris, Phleum, Poa, Secale, Sorghum, Triticum or Zea.
  • an "immunogenic molecule” refers to a substance, including but not limited to a molecule comprising as part of its structure or exclusively a protein, a polypeptide or a peptide, which optionally modifies, e.g. elicits, induces, stimulates, promotes enhances or decreases, reduces, inhibits, suppresses, relieves an immune response when administered to a subject, for example laboratory mice or modifies, e.g. elicits, induces, stimulates, promotes enhances or decreases, reduces, inhibits, suppresses or relieves a T cell response in vitro in response to the immunogenic molecule.
  • an immunogenic molecule may induce tolerance to itself in a subject.
  • An immune response elicited by an immunogenic molecule may include, but is not limited to, a B cell or a T cell response.
  • An immune response can include a cellular response with a particular pattern of lymphokine/cytokine production (e.g., Th1 , Th2), a humoral response (e.g., antibody production, like IgE, IgG or IgA), or a combination thereof, to a particular immunogenic molecule.
  • an antigen refers to a particular substance to which an immunoglobulin (Ig) isotype may be produced in response to the substance.
  • an immunoglobulin (Ig) refers to an antigen that induces an IgG antibody response.
  • an "IgE antigen” refers to an antigen that induces an IgE antibody response (and thus qualifies as an allergen);
  • an "IgA antigen” refers to a substance that induces an IgA antibody response, and so forth.
  • such an immunoglobulin (Ig) isotype produced in response to an antigen may also elicit production of other isotypes.
  • an IgG antigen may induce an IgG antibody response in combination with one more of an IgE, IgA, IgM or IgD antibody response.
  • an IgG antigen may induce an IgG antibody response without inducing a response to one more of an IgE, IgA, IgM or IgD antibody response.
  • an IgG antigen may induce an IgG antibody response without inducing an IgE, IgA, IgM or IgD antibody response.
  • allergen refers to a particular type of a substance that can elicit production of IgE antibodies, such as in predisposed subjects. For example, if a subject previously exposed to an allergen (i.e.
  • allergic asthma may develop due to a Th2 response characterized by an increased production of type 2 cytokines (e.g., IL-4, IL-5, IL-9, and/or IL-13) secreted by CD4+ T lymphocytes.
  • type 2 cytokines e.g., IL-4, IL-5, IL-9, and/or IL-13
  • the term "immunotherapy” is meant to encompass treatment of a disease by inducing, enhancing, or suppressing an immune response.
  • the therapeutically active agent is an immunogenic molecule, particularly an antigen, more particularly an allergen.
  • An immunogenic molecule may be a protein or a fragment thereof (e.g. immunogenic peptide including a linear peptide).
  • Immunotherapy in connection with allergy usually encompasses repeated administration of a sufficient dose of the immunogenic molecule/antigen/allergen, usually in microgram quantities, over a prolonged period of time, usually for more than 3 months, 6 months, 1 year, such as 2 or 3 years, during which period the immunogenic molecule may be administered daily or less frequently, such as several times a week, weekly, bi-weekly, or monthly, every second month or quarterly.
  • Immunotherapy can be effected by specific immunotherapy or may be effected by bystander tolerance induction.
  • specific immunotherapy in connection with allergy is meant to designate that immunotherapy is conducted with the administration of an immunogenic molecule to which the subject is sensitized to, particularly an immunogenic molecule to which the subject has raised specific IgE antibodies to, e.g. major allergens.
  • immunological tolerance refers to a) a decreased or reduced level of a specific immunological response (thought to be mediated at least in part by antigen-specific effector T lymphocytes, B lymphocytes, antibodies or a combination thereof); b) a delay in the onset or progression of a specific immunological response; or c) a reduced risk of the onset or progression of a specific immunological response to an immunogenic molecule, such as an antigen or an allergen.
  • Specific immunological tolerance occurs when tolerance is preferentially invoked against certain immunogens in comparison with other immunogens. Tolerance is an active immunogenic molecule dependent process and differs from non-specific immunosuppression and immunodeficiency.
  • an immunogenic molecule that elicits, induces, stimulates, promotes enhances or decreases, reduces, inhibits, suppresses, relieves an immune response against another unrelated immunogenic molecule, for example an allergen, e.g. major allergens of pollen.
  • an immunogenic molecule may induce immunological tolerance to itself, and may be able to reactivate T regulatory cells specific to the immunogenic molecule to down-regulate an immune response caused by another unrelated immunogenic molecule, e.g. an allergen.
  • an immunogenic molecule may induce immunological tolerance to another unrelated antigen, e.g. an allergen including a pollen allergen described herein.
  • one immunogenic molecule may provide bystander tolerance induction to another unrelated antigen, e.g. the immunogenic molecule may provide suppression of an immune response triggered by an unrelated antigen (e.g. allergen) via bystander mechanisms.
  • treatment refers to any type of treatment that conveys a benefit to a subject afflicted with allergy or at least sensitized to an allergen , including improvement in the condition of the subject (e.g., in one or more symptoms), delay in the onset of symptoms, slowing the progression of symptoms, or induce disease modification etc.
  • Typical symptoms of an allergic reaction are nasal symptoms, such as itchy nose, sneezing, runny nose, blocked nose; conjunctival symptoms, such asitchy eyes, red eyes, watery eyes; and respiratory symptoms, such as decreased lung function.
  • the treatment may also give the benefit that the patient needs less concomitant treatment with
  • treatment is not necessarily meant to imply cure or complete abolition of symptoms, but refers to any type of treatment that imparts a benefit to a patient.
  • Treatment may be initiated before the subject becomes sensitized to a protein. This may be realized by initiating immunotherapy before the subject has raised detectable serum IgE antibodies capable of binding specifically to the sensitizing protein or before any other biochemical marker indicative of an allergic immune response can be detected in biological samples isolated from the individual.
  • treatment may be initiated before the subject has evolved clinical symptoms of the allergic disease, such as symptoms of allergic rhinitis, allergic asthma or atopic dermatitis.
  • a therapeutically sufficient amount is meant to designate an amount effective to reduce, suppress, relieve or eliminate an allergic immune response, e.g. an amount sufficient to achieve the desirable reduction in clinical relevant symptoms or manifestations of the allergic immune response.
  • a therapeutically sufficient amount may be the accumulated dose of a polypeptide, a set of polypeptides administered during a course of immunotherapy in order to achieve the intended effect or it may be the maximal dose tolerated within a given period.
  • the total dose or accumulated dose may be divided into single doses administered daily, twice a week or more, weekly, every second or fourth week or monthly depending on the route of administration and the pharmaceutical formulation used.
  • the total dose or accumulated dose may vary. It is expected that a single dose is in the microgram range, such as in the range of 5 to 500 microgram dependent on the nature of the polypeptide.
  • Symptoms may be the clinically symptoms of allergic rhinitis, allergic asthma allergic conjunctivitis, atopic dermatitis, food allergy and/or hay fever.
  • the symptoms are the same as experienced with a flu/cold such as sneezing, itching, congestion, coughing, feeling of fatigue, sleepiness and body aches.
  • nasal symptoms may be itchy nose, sneezing, runny nose, blocked nose; conjunctival symptoms may be itchy eyes, red eyes, watery eyes; and respiratory symptoms may be decreased lung function.
  • a responder may also be evaluated by monitoring the patient's reduced need for concomitant treatment with corticosteroids or H1 antihistamines to suppress the clinical symptoms. Symptoms may be subjectively scored or in accordance with official guidelines used in clinical trials of SIT.
  • adjuvant refers to a substance that enhances the immune response to an immunogenic molecule. Depending on the nature of the adjuvant, it can promote either a cell-mediated immune response, humoral immune response or a mixture of the two.
  • an epitope refers to a region or part of an immunogenic molecule that elicits an immune response when administered to a subject.
  • an epitope is a T cell epitope, i.e., an epitope that elicits, stimulates, induces, promotes, increases or enhances a T cell activity, function or response.
  • An immunogenic molecule can be analyzed to determine whether it include at least one T cell epitope using any number of assays (e.g. T cell proliferation assays, lymphokine secretion assays, T cell non-responsiveness studies, etc.).
  • a T-cell epitope refers to an epitope that are MHC Class II binders (i.e. HLA-II binders), for example an epitope able to connect to/ associate with or bind to a HLA-II molecule shown in Tables 11 b or 11c.
  • MHC Class II binders i.e. HLA-II binders
  • immune response includes T cell (cellular) mediated and/or B cell (humoral) mediated immune responses, or both cellular and humoral responses.
  • exemplary immune responses include T cell responses, e.g., lymphokine production, cytokine production and cellular cytotoxicity.
  • T-cell responses include Th1 and/or Th2 responses.
  • immune response includes responses that are indirectly affected by T cell activation, e.g., antibody production (humoral responses) and activation of cytokine responsive cells, e.g., eosinophils, macrophages.
  • Immune cells involved in the immune response include lymphocytes, such as T cells (CD4+, CD8+, Th1 and Th2 cells, memory T cells) and B cells; antigen presenting cells (e.g., professional antigen presenting cells such as dendritic cells, macrophages, B lymphocytes, Langerhans cells, and non-professional antigen presenting cells such as keratinocytes, endothelial cells, astrocytes, fibroblasts, oligodendrocytes); natural killer (NK) cells; myeloid cells, such as macrophages, eosinophils, mast cells, basophils, and granulocytes.
  • lymphocytes such as T cells (CD4+, CD8+, Th1 and Th2 cells, memory T cells) and B cells
  • antigen presenting cells e.g., professional antigen presenting cells such as dendritic cells, macrophages, B lymphocytes, Langerhans cells, and non-professional antigen presenting cells such as
  • sequence means a fragment or part of a longer molecule, e.g. of a full length molecule (e.g. the wild type polypeptides shown in Tables 1 and 2) or of a conserved region thereof (e.g. GWT sequences shown in Tables 3 and 4).
  • subsequence therefore consists of one or more amino acids less than the wild type polypeptide or a conserved region thereof.
  • a molecule e.g. an immunogenic molecule of the invention comprises conserved amino acid sequences detected in a grass pollen species as well as in a weed pollen species and/or a tree pollen species.
  • such molecules can be used to broadly treat a subject with or at risk of developing an immune response to pollen of a variety of pollen species, genera or families, or to broadly induce or promote tolerance of a subject to pollen of a variety of pollen species, genera or families and may include promoting or inducing tolerance to the immunogenic molecule itself or another immunogenic molecule (e.g. an allergen) unrelated to the immunogenic molecule being administered, for example via induction of bystander tolerance towards an offending allergen.
  • an immunogenic molecule e.g. an allergen
  • a multisensitized subject e.g. a subject sensitized to both grass, weed and tree pollen
  • a molecule e.g. an immunogenic molecule
  • the molecule (e.g. the immunogenic molecule) comprises or consists of a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches compared to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 212-293 (e.g. SEQ ID NOS: 274, 270-273, 212-269 and 275-293) set out in Table 3 or SEQ ID NOS: 294-767 (e.g. SEQ ID NOS: 665-676, 617-664 and 677-767) set out in Table 4.
  • SEQ ID NOS: 212-293 e.g. SEQ ID NOS: 274, 270-273, 212-269 and 275-293
  • SEQ ID NOS: 294-767 e.g. SEQ ID NOS: 665-676, 617-664 and 677-767 set out in Table 4.
  • the molecule may comprise one or more T cell epitope(s) optionally a Th-2 cell epitope (i.e. the molecule may comprise one or more T cell epitope-containing amino acid sequence(s)).
  • the polypeptide of option a), option b), option c) and option d) comprises a T cell epitope, optionally a Th-2 cell epitope.
  • the GWT sequences (SEQ ID NOS:212-293 set out in Table 3 and SEQ ID NOS: 294-767 set out in Table 4), the GW sequences (SEQ ID NOS: 768-808 set out in Table 5 and SEQ ID NOS: 809-951 set out in Table 6), GT sequences (SEQ ID NOS: 952-1023 set out in Table 7 and SEQ ID NOS: 1024-1231 set out in Table 8) and WT sequences (SEQ ID NOS: 1232-1473 set out in Table 9) or subsequences thereof may comprise a T cell epitope, optionally a Th-2 cell epitope.
  • a subsequence of a polypeptide of option b) or option c) contains a T cell epitope, optionally a Th-2 cell epitope.
  • sequences selected from any of SEQ ID NOS: 1-37 e.g. SEQ ID NOS: 30, 1-29, 31-37 set out in Table 1
  • SEQ ID NOS: 38- 211 e.g SEQ ID NOS: 161-172, 38-160 and 173-211 set out in Table 2 contains a T cell epitope, optionally a Th-2 cell epitope.
  • immunogens of the present invention may be used in relieving an immune response against phi p grass pollen, non-phi p grass pollen as well as non-grass pollen to the extent that the pollen species contain polypeptides comprising essentially the same GWT, GW, GT or WT conserved region.
  • an immunogenic molecule of the present invention may at least be used in modulating, e.g. relieving, an immune response triggered by pollen of the grass pollen species Phi p and/or Cyn d as well as pollen of other species closely related to Phi p or Cyn d, e.g. pollen species also belonging to the plant family Poales, such as species of the plant genera selected from any of Anthoxanthum, Cynodon, Dactylis, Lolium, Phleum or Poa.
  • immunogenic molecule may also be used in modulating, e.g. relieving, an immune response triggered by pollen of the weed pollen species Amb a and/or Amb p and pollen of species closely related to Amb a and Amp p, e.g. pollen species also belonging to the plant family Asteraceae, such as species of the plant genera selected from any of Ambrosia, Artemisia, Helianthus.
  • the same immunogenic molecule may also be used in modulating, e.g. relieving, an immune response triggered by pollen of the tree pollen species Que a and/or Bet v and pollen of species closely related to Que a and/or Bet v, e.g.
  • pollen species also belonging to the plant families Betulaceae, Fagaceae and Oleaceae, such as species of the plant genera selected from any of Alnus, Betula, Carpinus, Castanea, Corylus, Fagus, Quercus, Fraxinus and Ligustrum.
  • Betulaceae Fagaceae and Oleaceae
  • species of the plant genera selected from any of Alnus, Betula, Carpinus, Castanea, Corylus, Fagus, Quercus, Fraxinus and Ligustrum.
  • a molecule e.g.
  • an immunogenic molecule of the present invention may be used in modulating, e.g., relieving, an immune response triggered by pollen of a plant genus selected from any of Phleum, Cynodon Ambrosia, Quercus and Betula.
  • conserved amino acid sequences disclosed herein have been detected in about five different pollen species and may be found in additional pollen species selected from any of the plant families Poaceae, Asteraceae, Fagaceae, Betulaceae, Oleaceae, and Plantaginaceae, e.g. the plant genera Ambrosia, Artemisia, Helianthus, Alnus, Betula, Carpinus, Castanea, Corylus, Ostrya, Ostryopsis, Fagus, Quercus, Fraxinus, Ligustrum, Lilac, Olea or Plantago using the methodology disclosed herein (see exemplary pollen species below):
  • conserved regions may be found in pollen of other weeds, grasses or tree species, e.g. found in one or more plant species selected from further plant families, like the plant family Chenopidiaceae including the plants Lambs quarters, Russian thistles and Kochias; plant family Amaranthaceae including Pigweeds, plant family Polygonaceae including for example Sheep sorrel, plant family Ulmacea including for example American Elm and Ralphberry, plant family Plantanaceae including for example Sycamore, plant family Salicaceae including for example White poplar and Cottonwood, plant family Aceraceae including for example Box elder and Red maple, plant family Cupressaceae including for example Common juniper and Cedar.
  • Non-limiting examples of species of the genus Ambrosia is Ambrosia artemisiifolia, Ambrosia psilostachya, Ambrosia trifida; a typical species of the genus Artemisia is Artemisia vulgaris; non-limiting examples of species of the genus Betula is Betula verrucosa; non-limiting examples of species of the genus Fagus is Fagus grandifolia or Fagus sylvatica; non-limiting examples of species of the genus Quercus is Quercus alba, non-limiting examples of species of the genus Fraxinus is Fraxinus excelsior; a typical species of the genus Olea is Olea Europaea, a typical species of the genus plantago is Plantago lanceolata, a typical species of the genus plantago is Plantago lanceolata, non- limiting examples of species of
  • a molecule of the invention may modulate an immune response at least triggered by a grass pollen and/or a weed pollen by administering to a subject in need thereof a molecule comprising a GWT conserved sequence set out in Tables 3 or 4 or a subsequence thereof or by administering to a subject in need thereof a molecule comprising a GW conserved sequence set out in Tables 5 or 6 or a
  • the molecule may modulate an immune response triggered by at least a grass pollen and/or a tree pollen by
  • the molecule modulate an immune response triggered by at least a grass pollen, a weed pollen and a tree pollen by administering to a subject in need thereof a molecule comprising a GWT conserved sequence set out in Tables 3 or 4 or a subsequence thereof.
  • a molecule of the invention may modulate, e.g. relieve, an immune response triggered by pollen.
  • Pollen may comprise allergens considered as major and minor allergens according to official guidelines in the art and may in addition comprise additional immunogens, for example a molecule as disclosed herein.
  • a molecule of the invention may be able to relieve an immune response triggered by a major and/or a minor pollen allergen and/or a molecule disclosed herein including the molecule itself.
  • Molecules (e.g. immunogens) eligible for relieving an immune response triggered by an allergen (e.g. a major allergen or minor allergen) unrelated to the molecule is thought, at least in part, to be mediated via induction of bystander tolerance, which mechanism requires, at least in part, co-existence of the triggering allergen and the unrelated molecule at the target organ for the immune response.
  • the co-existence may inherently be obtained by using molecules present in the same pollen source as the allergen.
  • the triggering allergen and the unrelated molecule may be released from the same pollen source within an overlapping period of time (co-release of triggering allergen and unrelated molecule (e,g. immunogenic molecule)) to ensure co-existence at the target organ.
  • Co-release can be examined in vitro using hydrated pollen as described in Example 4 herein.
  • the polypeptide of option a), b), c) or d) may be derived, be a part of or comprise a wild type polypeptide that co-releases/co-elutes from the same pollen source with the major or minor allergens the subject is sensitized to and to which allergens the immune response is sought to be relieved.
  • a molecule with the potential to relieve an immune response triggered by pollen of several different species, genera and families may be identified by investigating the presence of the molecule or a fragment thereof in pollen diffusates of pollen of different species, genera or families including the grass pollen, weed pollen or tree pollen species used for the present investigation within a time period overlapping with the relevant major and/or minor allergen.
  • co-release or “co-elute” may refer to when a molecule starts to release from pollen, e.g. hydrated pollen, within a period overlapping with a major allergen to which the allergic immune response is sought to be relieved.
  • Major allergens start to release from pollen within few minutes after hydration of pollen and continue to be released within the next 30 or 60 minutes.
  • co-release or “co-elute” may refer to when a molecule of the invention starts to release from pollen together with a major allergen, usually within 30 minutes after hydration of the pollen.
  • Co- release/co-elution may be determined by a method comprising extracting pollen in an aqueous solution having pH in the range of 6-8 for a period ranging from 1 to 200 minutes, optionally the aqueous solution comprises at least 60% of water, a buffering agent; a tonicity providing agent.
  • the aqueous solution has an ionic strength corresponding to that of isotonic saline (0.9 g of NaCI in 1 liter of water), e.g. the aqueous solution has an ionic strength in the range of 10 mM to 1000 mM and optionally the aqueous solution has a pH of about 7.
  • the period of extraction is in the range of 1-180, 1-120 minutes, 5-120 minutes, 5 to 90 minutes, for example 5-60 minutes, for example 5-45 minutes, for example 5-30 minutes.
  • some immunogenic molecules may be released from various distinct pollen species, such as from grass pollen species as well as weed and/or tree pollen species.
  • a polypeptide of option a), option b), option c) or option d), as described herein may be derived from a wild type polypeptide that co-releases with a major allergen from grass pollen (e.g. pollen of the genera Phleum and or Cynodon) and from a weed pollen (e.g. pollen of the genera Ambrosia) and from tree pollen (e.g. pollen of the genera Quercus or Betula).
  • grass pollen e.g. pollen of the genera Phleum and or Cynodon
  • a weed pollen e.g. pollen of the genera Ambrosia
  • tree pollen e.g. pollen of the genera Quercus or Betula
  • polypeptides are herein named "GWT polypeptides", such as the set of polypeptides referred to with polypeptide ID NOs: A0349, A0246, A0209, A0211 , A0325, A0357 and A0203.
  • GWT polypeptides such as the set of polypeptides referred to with polypeptide ID NOs: A0349, A0246, A0209, A0211 , A0325, A0357 and A0203.
  • these polypeptides have predicted HLA Class II allele binding sites in their conserved part of their sequence, thereby indicating the polypeptides potentially comprises a T cell epitope in their conserved region.
  • polypeptide A0262 (herein named GT polypeptide) was released from both grass and tree pollen, but not from weed pollen. It was also found that the set of polypeptides having the polypeptide ID NOs A0362,
  • A0316, A0356, A0376, A0336, A0377 and A0366 could be released from grass as well as weed pollen, but not the tree pollen investigated (herein named GW polypeptides).
  • the molecule comprises a conserved region or a subsequence thereof of the "GWT polypeptide" A0349, such as a molecule comprising, consisting of or consisting essentially of a) a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 270-274 set out in Table 3 and SEQ ID NOS: 617-676 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 270-274 set out in Table 3 and SEQ ID NOS: 617-676 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least
  • the molecule comprises a conserved region or a
  • GWT polypeptide such as a molecule comprising, consisting of or consisting essentially of a) a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 233 set out in Table 3 and SEQ ID NOS: 414-418 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 233 set out in Table 3 and SEQ ID NOS: 414-418 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 233 set out in Table 3 and SEQ ID NO NOS: 414-418 set out in Table 4
  • the molecule comprises a conserved region or a
  • GWT polypeptide A0209
  • a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 217-220 set out in Table 3 and SEQ ID NOS: 321-344 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any of SEQ ID NOS: SEQ ID NOS: 217-220 set out in Table 3 and SEQ ID NOS: 321-344 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 217-220 set
  • the molecule comprises a conserved region or a
  • GWT polypeptide A0211
  • a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 221-225 set out in Table 3 and SEQ ID NOS: 345-386 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 221-225 set out in Table 3 and SEQ ID NOS: 345-386 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 221-225 set out in Table 3 and
  • the molecule comprises a conserved region or a
  • GWT polypeptide A0325
  • a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS:256 set out in Table 3 and SEQ ID NOS: 541-547 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS:256 set out in Table 3 and SEQ ID NOS: 541-547 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS:256 set out in Table 3 and SEQ ID NOS: 5
  • GWT polypeptide A0357
  • a molecule comprising or consisting of a) a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 280-281 set out in Table 3 and SEQ ID NOS: 701-709 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 280-281 set out in Table 3 and SEQ ID NOS: 701-709 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 280-281 set out in Table 3 and SEQ ID NO NOS: 701-709 set out
  • the molecule comprises a conserved region or a
  • GWT polypeptide A0203, such as a molecule comprising, consisting of or consisting essentially of a) a polypeptide comprising an amino acid sequence having at least 65% sequence similarity or identity to a sequence selected from any one of SEQ ID NOS: 215, 216 set out in Table 3 and SEQ ID NOS: 309-320 set out in Table 4; or b) a polypeptide comprising an amino acid sequence having at least 65% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 215, 216 set out in Table 3 and SEQ ID NOS: 309-320 set out in Table 4; or c) a polypeptide, which includes at least one amino acid sequence with 0, 1 or 2 mismatches to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 215, 216 set out in Table
  • a molecule of the present invention is an IgE reactive molecule, e.g. able to bind to IgE antibodies specific for the immunogenic molecule.
  • IgE reactivity towards a molecule of the invention may only be conferred by a low fraction of an allergic population.
  • a molecule of the invention may not fall under the usual definitions of a major allergen, which is an allergen with high prevalence in a population of donors allergic to a pollen species, for example the prevalence may be higher than at least 50%.
  • the molecule is able to react with, bind to or induce IgG antibodies in a subject, at least in detectable levels.
  • the molecule does not react with, bind to or induce IgG antibodies, at least in detectable levels. As demonstrated herein, a molecule of the invention seems to be less
  • polypeptides containing GWT, GT, GW or WT conserved regions can be detected in various pollen species and families and share high identity and similarity across the various pollen species. Therefore, a polypeptide of option d) comprises an amino acid sequence having at least 70%, such as at least 75%, 80%,
  • SEQ ID NOS: 1-37 e.g. SEQ ID NOS: 30, 1-29, 31- 37
  • SEQ ID NOS: 38-211 eg. SEQ ID NOS: 161-172, 38-160 and 173-211 set out in Table 2.
  • a polypeptide of option a) comprises an amino acid sequence having at least 70% such as at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% similarity or identity to a sequence selected from any one of SEQ ID NOS: 212- 293 (e.g. SEQ ID NOS: 270-274; 212-269; 275-293) set out in Table 3; SEQ ID NOS: 294-767 (e.g.
  • a polypeptide of option b) comprises an amino acid sequence having at least 70%, such as at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% similarity or identity to a subsequence of at least 15 contiguous amino acid residues of a sequence selected from any one of SEQ ID NOS: 212-293 (e.g. SEQ ID NOS: 270-274; 212-269; 275-293 set out in Table 3; SEQ ID NOS: 294-767 (e.g.
  • the at least 70% such as at least 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% similarity or identity is of a subsequence consisting of at least 16 contiguous amino acid residues, for example at least 17, 18, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 90, 100 or more contiguous amino acids.
  • a subsequence may comprise any number of amino acids, but typically the subsequence has a length of 15 to 50 amino acid residues, for example 15 to 45, 15 to 40, 15 to 35, 15 to 30; 15 to 25; 15 to 20; 15 to 19; 15 to 18; 15 to 17, or 15 to 16 amino acid residues.
  • 15 to 45, 15 to 40, 15 to 35, 15 to 30 15 to 25; 15 to 20; 15 to 19; 15 to 18; 15 to 17, or 15 to 16 amino acid residues.
  • the number of amino acid mismatches may be 0, 1 , 2, or 3, preferably 0, 1 or 2, more preferably 0 or 1.
  • the polypeptide of option b) or option c) has a length of 15-800 or more amino acid residues, for example 15-750, 15-700, 15-650 or 15-600 or more amino acid residues, for example 15-20, 20-25, 25-30, 30-35, 35-40, 45-50, 50-60, 60-70, 70-80, 90-100, 100-125, 125-150, 150-175, 175-200, 200-250, 250-300, 300-350, 350-400, 400- 450, 450-500, 500-550, 550-600, 600-650, 650-700, 700-800 or more amino acid residues.
  • the polypeptide of option a) or of option d) has a length of no more than 800 amino acid residues, for example no more than 750, 700, 650, 600, 550,
  • a polypeptide disclosed herein including a subsequence thereof may contain a T cell epitope, such as a Th2 cell epitope.
  • a subsequence or a polypeptide described herein may have HLA Class II binding properties. HLA Class II binding can be predicted using NetMHCIIpan-3.0 tool (Karosiene, Edita, Michael Rasmussen, Thomas Bö, Ole Lund, S0ren Buus, and Morten Nielsen.
  • a polypeptide of option b) or c) may have different lengths according to the desirable use, for example of about 15-800 or more amino acid residues in length, for example 15-750, 15-700, 15-650, 15-600, 15-500 or more amino acid residues, for example 15-20, 15-25, 15-30, 20-25, 25-30, 30-35, 35-40, 45-50, 50-60, 60-70, 70-80, 90-100, 100-125, 125- 150, 150-175, 175-200, 200-250, 250-300, 300-350, 350-400, 400-450, 450-500, 500- 550, 550-600, 600-650, 650-700, 700-800 or more amino acid residues.
  • a polypeptide of option b) or a polypeptide of option c) has a length in the range of 15 to 30 amino acid residues, for example 15 to 25 amino acid residues.
  • a polypeptide of option a), b), c) or d) is a longer polypeptide which comprises a secondary or tertiary structure, e.g. folded.
  • a polypeptide of option a), b), c) or d) has a length in the range of 30 to 500 amino acid residues or more. It is considered that the length of the amino acid sequence of a polypeptide of option a), b), c) or d) is no more than 800 amino acid residues, for example no more than 750, 700, 650, 600, 550, 500 or 450 amino acid residues.
  • identity and “identical” and grammatical variations thereof, as used herein, mean that two or more referenced entities are the same (e.g. two or more amino acid sequences). Thus, where two polypeptides are identical, they have the same amino acid sequence.
  • the identity can be over a defined area (region or domain) of the sequence, e.g. over the sequence length of a sequence disclosed in Tables 1 to 9 or over a portion thereof e.g. at least 15 contiguous amino acid residues.
  • identity can be over the length of the sequence overlapping the two polypeptides, when aligned with best fit with gaps permitted.
  • the polypeptide may be aligned with a sequence of Tables 1 to 9 and the percent identity be calculated with reference to said sequence. Identity can be determined by comparing each position in aligned sequences. A degree of identity between amino acid sequences is a function of the number of identical or matching amino acids at positions shared by the sequences, i.e. over a specified region.
  • Optimal alignment of sequences for comparisons of identity may be conducted using a variety of algorithms, as are known in the art, including the Clustal Omega program available at http://www.ebi.ac.uk/Tools/msa/clustalo/, the local homology algorithm of Smith and Waterman, 1981 , Adv. Appl. Math 2: 482, the homology alignment algorithm of Needleman and Wunsch, 1970, J. Mol. Biol. 48:443, the search for similarity method of Pearson and Lipman, 1988, Proc. Natl. Acad. Sci.
  • Sequence identity may also be determined using the BLAST algorithm, described in Altschul et al., 1990, J. Mol. Biol. 215:403-10 (using the published default settings).
  • BLAST e.g., BLAST 2.0
  • search algorithm see, e.g., Altschul et al., J. Mol. Biol. 215:403 (1990), publicly available through NCBI
  • mismatch -2 e.g., Altschul et al., J. Mol. Biol. 215:403 (1990), publicly available through NCBI
  • mismatch -2 e.g., Altschul et al., J. Mol. Biol. 215:403 (1990), publicly available through NCBI
  • a BLASTP algorithm is typically used in combination with a scoring matrix, such as
  • FASTA e.g., FASTA2 and FASTA3
  • SSEARCH sequence comparison programs are also used to quantitate the extent of identity (Pearson et al., Proc. Natl. Acad. Sci. USA 85:2444 (1988); Pearson, Methods Mol Biol. 132: 185 (2000); and Smith et al., J. Mol. Biol. 147:195 (1981)).
  • Programs for quantitating protein structural similarity using Delaunay-based topological mapping have also been developed (Bostick et al., Biochem Biophys Res Commun. 304:320 (2003)).
  • a polypeptide sequence is a "homolog” of, or is “homologous” to, another sequence if the two sequences have substantial identity over a specified region and a functional activity of the sequences is preserved or conserved, at least in part. (As used herein, the term 'homologous' does not infer nor exclude evolutionary relatedness).
  • homologous polypeptides include polypeptides with high similarity or identity to Phi P polypeptides and detected in pollen species other than Phi p.
  • a homologous polypeptide to a polypeptide having an amino acid sequence set out in Table 1 or Table 2 may be found in pollen of plant families selected among Asteraceae, Betulaceae, Fagaceae, Oleaceae, or Plantaginaceae, e.g.
  • Two polypeptide sequences are considered to be substantially identical if, when optimally aligned (with gaps permitted), they share at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, etc. identify over a specific region), for example, over all or a part of any amino acid sequence in Tables 1 to 9, or if the sequences share defined functional motifs (e.g., epitopes).
  • the length of the sequence sharing the percent identity is at least 15, 16, 17, 18, 19, 20, etc. contiguous amino acids, e.g. more than 25, 30, 35, 40, 45 or 50 or more contiguous amino acids, including the entire length of a reference sequence of Tables 1 to 9.
  • an "unrelated" or “non-homologous” sequence is considered to share less than 30% identity. More particularly, it may share less than about 25 % identity, with a polypeptide of the invention over a specified region of homology.
  • An amino acid sequence set out in any of Tables 1 to 9 may contain one or more modifications, which optionally may result in greater or less activity or function, for example in the ability to elicit, stimulate, induce, reduce, inhibit, suppress an in vitro immune response (e.g. T cell proliferation or T cell cytokine production); in the ability to bind HLA Class II alleles; in the ability to induce or enhance immunological
  • a modification includes deletions, including truncations and fragments; insertions and additions, substitutions, for example conservative substitutions, site-directed mutants and allelic variants.
  • Non-limiting examples of modifications include one or more amino acid substitutions (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20-25, 25-30, 30-50, 50-100 or more residues), additions and insertions (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or more residues) and deletions (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20-25, 25-30, 30-50, 50-100 or more) of a sequence set out in Tables 1 , 2, 3 and 4.
  • amino acid substitutions e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20-25, 25-30, 30-50, 50-100 or more
  • similarity and “similar” and grammatical variations thereof, as used herein, mean that two or more referenced amino acid sequences contains a limited number of conservative amino acid substitutions of the amino acid sequence.
  • a variety of criteria can be used to indicate whether amino acids at a particular position in a polypeptide are similar.
  • substitutions of like amino acid residues can be made on the basis of relative similarity of side-chain substituents, for example, their size, charge, hydrophobicity, hydrophilicity, and the like, and such substitutions may be assayed for their effect on the function of the peptide by routine testing.
  • a "conservative substitution” is the replacement of one amino acid by a biologically, chemically or structurally similar residue.
  • Biologically similar means that the substitution does not destroy a biological activity.
  • Structurally similar means that the amino acids have side chains with similar length, such as alanine, glycine and serine, or a similar size.
  • Chemical similarity means that the residues have the same charge, or are either hydrophilic or hydrophobic.
  • a conservative amino acid substitution is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain, which include amino acids with basic side chains (e.g., lysine, arginine, histidine); acidic side chains (e.g., aspartic acid, glutamic acid); uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, histidine); nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan); beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan).
  • basic side chains e.g., lysine, arginine, histidine
  • Particular examples include the substitution of one hydrophobic residue, such as isoleucine, valine, leucine or methionine for another or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, serine for threonine, and the like.
  • Proline which is considered more difficult to classify, shares properties with amino acids that have aliphatic side chains (e.g., Leu, Val, lie, and Ala).
  • substitution of glutamine for glutamic acid or asparagine for aspartic acid may be considered a similar substitution in that glutamine and asparagine are amide derivatives of glutamic acid and aspartic acid, respectively.
  • Conservative changes can also include the substitution of a chemically derivatized moiety for a non- derivatized residue, for example, by reaction of a functional side group of an amino acid.
  • Variants and derivatives of polypeptides include forms having a limited number of one or more substituted residues.
  • a polypeptide of option a), b), c) and d) may be longer than the reference sequence set out in Tables 1 to 9.
  • an addition can be one or more additional amino acid residues.
  • a polypeptide of option b) or c) may contain amino acid residues in addition to the subsequence of at least 15 amino acid residues.
  • the additional amino acid residues may be identical to those present in the wild type polypeptide from which the subsequence derives from.
  • the polypeptide of option b) comprises one or more amino acid residues in addition to the subsequence of at least 15 contiguous amino acids, wherein the additional amino acid residue(s) is/are selected from an amino acid residue or an amino acid sequence within the wild type polypeptide that the subsequence is a part of (e.g.
  • wild type polypeptide sequences of Tables 1 or 2 or a GWT sequence of Tables 3 or 4 may be adjacent to, subtended, comprised within, overlapping with or is a part of the subsequence, when present in its natural biological context within the wild type polypeptide.
  • a polypeptide of option a) may contain additional amino acid residues in addition to the GWT sequence set out in Tables 3 and 4.
  • a polypeptide of option a) may comprise one or more amino acid residues in addition to the GWT sequence set out in Tables 3 or 4, wherein the additional amino acid residue(s) is/are selected from an amino acid residue or an amino acid sequence within the wild type polypeptide of which the GWT sequence is a part of (e.g. a wild type polypeptide of Tables 1 or 2).
  • An illustrative example is a GWT sequence of A0349 set out in Table 3 that may be extended with amino acid residues from polypeptide A0349 set out in Tables 1 or 2, such as amino acid residues adjacent to the GWT sequence when aligned with the corresponding wild type polypeptide.
  • the additional amino acid residues may be added to the N- and/or C- terminal end of a sequence set out in Tables 3 to 9, such as additional amino acids selected from amino acids flanking the N- and/or C- terminal ends when sequence is aligned with the source protein it is present in, based upon or derived from.
  • the additional amino acids may be the amino acids flanking the N- and/or C- terminal ends of the sequence when aligned to polypeptide A0349 of Table 1 or 2.
  • a polypeptide of option a), b), c) or d) may include a number of
  • an amino acid sequence (parent sequence) disclosed in any of the Tables or a subsequence thereof may be modified to comprise: a) one or more (e.g. 1 , 2,3, 4, 5, 6, 7, 8, 9 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) amino acid substitutions in a sequence selected from any of the sequences disclosed in Tables 1 to 9, for example a glutamate residue at the N-terminus of an amino acid sequence disclosed in Tables 1 to 9 may be replaced with pyroglutamate and/or one or more cysteine residues in the an amino acid sequence disclosed in Tables 1 to 9 may be replaced with serine or 2-aminobutyric acid; and/or b) one or more amino acid additions (e.g.
  • a polypeptide of option a), b), c) or d) including
  • polypeptide is derivatized.
  • derivatization are covalent or non-covalent attachment of another molecule.
  • Specific examples include glycosylation, acetylation, phosphorylation, amidation, formylation, ubiquitination, and derivatization by protecting/blocking groups and any of numerous chemical modifications.
  • a derivative is formed by reacting a functional side group of an amino acid (e.g. amino, sulfhydryl or carboxy-group) with another molecule to form a covalent or non- covalent attachment of any type of molecule (naturally occurring or designed), such as a sugar moiety.
  • Specific examples of derivatives of a peptide include glycosylation, acylation (e.g.
  • acetylation phosphorylation
  • amidation formylation
  • ubiquitination phosphorylation
  • derivatization by protecting/blocking groups and any of numerous chemical modifications.
  • Additional specific non-limiting examples are tagged peptides, fusion peptides, chimeric peptides including peptides having one or more non-amino acyl groups (q.v., sugar, lipid, etc.) covalently linked to the peptide.
  • a derivative comprises one or more modifications, for example selected from any of: (a) N-terminal acylation (e.g. acetylation or formylation); (b) C-terminal amidation (e.g.
  • a derivative comprises a fusion (chimeric) sequence of peptides, which optionally may contain an amino acid sequence having one or more molecules not normally present in a reference (wild type) sequence covalently attached to the peptide amino acid sequence.
  • chimeric and grammatical variations thereof, when used in reference to a sequence, means that the sequence contains one or more portions that are derived from, obtained or isolated from, or based upon other physical or chemical entities.
  • a derivative is one in which a second heterologous sequence, i.e. a heterologous functional domain, is attached to a peptide disclosed herein, (covalent or non-covalent binding) that may confer a distinct or complementary function to a peptide disclosed herein.
  • Heterologous functional domains are not restricted to amino acid residues.
  • a heterologous functional domain can consist of any of a variety of different types of small or large functional moieties.
  • moieties include nucleic acid, peptide, carbohydrate, lipid or small organic compounds, such as a drug (e.g., an antiviral), a metal (gold, silver), or a radioisotope.
  • Linkers such as amino acid or peptidomimetic sequences, may be inserted between the peptide sequence and the addition (e.g., heterologous functional domain) so that the two entities maintain, at least in part, a distinct function or activity.
  • Linkers may have one or more properties that may include a flexible conformation, an inability to form an ordered secondary structure or a hydrophobic or charged character, which could promote or interact with either domain.
  • Amino acids typically found in flexible protein regions include Gly, Asn and Ser. Other near neutral amino acids, such as Thr and Ala, may also be used in the linker sequence.
  • polypeptides of option a), b), c) or d) or a combination of such polypeptides are not provided as individual polypeptides, but may be fused together or to a carrier molecule to form an isolated molecule (e.g. immunogenic molecule).
  • the polypeptides may be fused to the N- and C-terminus of a surface polypeptide of a virus, e.g. a virus of the hepadnaviridae family as disclosed in international patent application W012168487 A1.
  • a molecule of the invention including a polypeptide of option a), b), c) or d), a subsequence thereof, a modified polypeptide thereof, or a derivatized polypeptide thereof may bind to at least 70% of the group of Class HLA II alleles that the relevant parent polypeptide disclosed in Tables 1 to 9 binds to.
  • a molecule of the invention including a polypeptide of option a), b), c) or d), a modified polypeptide thereof, or a derivatized polypeptide binds to the same, substantially the same or at least 75%, 80%, such as at least 82%, 85%, 88%, 90%, 92%, 95%, 98% or more, of the group of HLA Class II alleles that binds to the relevant parent polypeptide disclosed in Tables 1 to 9, optionally wherein this is determined under same test conditions, either using prediction tools or in-vitro binding assay.
  • a subsequence disclosed herein binds to the same, substantially the same or at least 75%, 80%, such as at least 82%, 85%, 88%, 90%, 92%, 95%, 98% or more, of the group of HLA Class II alleles that binds to the relevant subsequent from which the modified sequence derives, optionally wherein this is determined under same test conditions, either using prediction tools or in-vitro binding assay.
  • the Class HLA II binding is determined with respect to a particular group of Class HLA II alleles, for example one or more or all of the following alleles: DPA1*02:01-DPB1 * 01 :01 ,
  • a polypeptide of option a), b), c) or d), including a modified or derivatized polypeptide thereof may have one or more of the same T cell epitopes or T cell activity (e.g. percent responders) as the amino acid sequence from which it derives (e.g. an amino acid sequence set out in Tables 1 to 9.
  • This may be determined by the ability to induce or stimulate in vitro T cell proliferation using cultured PBMCs (peripheral blood monocytes) in response to the polypeptide of option a), b), c) or d), including a modified or derivatized polypeptide compared to the amino acid sequence set out in Tables 1 to 9, optionally using same test conditions, or by the ability to induce or stimulate production of cytokines, (e.g.
  • T cell assays examples include cytokines, IL-5, IL-13 and/or IL-10) from T cells (obtained from cultured PBMC's) in response to the polypeptide of option a), b), c) or d), including a modified or derivatized polypeptide compared to the amino acid sequence set out in Tables 1 to 9. Examples of T cell assays are described herein in Example 6.
  • a derivative is a fusion (chimeric) sequence, an amino acid sequence having one or more molecules not normally present in the wild type sequence covalently attached to the sequence.
  • chimeric and grammatical variations thereof, when used in reference to a sequence, means that the sequence contains one or more portions that are derived from, obtained or isolated from, or based upon other physical or chemical entities. For example, a chimera of two or more different
  • polypeptides may have one part of a polypeptide, and a second part of the chimera may be from a different sequence, or unrelated protein sequence.
  • heterologous functional domain is attached (covalent or non- covalent binding) that confers a distinct or complementary function.
  • heterologous functional domains are not restricted to amino acid residues.
  • a heterologous functional domain can consist of any of a variety of different types of small or large functional moieties.
  • moieties include nucleic acid, peptide, carbohydrate, lipid or small organic compounds, such as a drug (e.g., an antiviral), a metal (gold, silver), and radioisotope.
  • a tag such as T7 or polyhistidine can be attached in order to facilitate purification or detection of a protein, peptide, etc.
  • a 6-HIS tag may be added to the C- or N-terminal end of a polypeptide of option a), b), c) or d), e.g. the 6- HIS sequence GHHHHHHGSGMLDI, which optionally may remain in the molecule when administered to a subject.
  • a polypeptide linked to a Tag containing histidines may easily be purified by use of a HIS tag affinity column).
  • polypeptides linked to a heterologous domain wherein the heterologous functional domain confers a distinct function on the polypeptide.
  • the polypeptide is derivatized for example to improve solubility, stability, bioavailability or biological activity.
  • tagged polypeptides and fusion proteins and modifications, including peptides having one or more non-amino acyl groups (q.v., sugar, lipid, etc.) covalently linked to the polypeptide and post-translational modifications.
  • Linkers such as amino acid or peptidomimetic sequences may be inserted between the sequence and the addition (e.g., heterologous functional domain) so that the two entities maintain, at least in part, a distinct function or activity.
  • Linkers may have one or more properties that include a flexible conformation, an inability to form an ordered secondary structure or a hydrophobic or charged character, which could promote or interact with either domain.
  • Amino acids typically found in flexible protein regions include Gly, Asn and Ser. Other near neutral amino acids, such as Thr and Ala, may also be used in the linker sequence.
  • the length of the linker sequence may vary without significantly affecting a function or activity of the fusion protein (see, e.g., U.S. Patent No. 6,087,329).
  • Linkers further include chemical moieties and conjugating agents, such as sulfo-succinimidyl derivatives (sulfo-SMCC, sulfo-SMPB), disuccinimidyl suberate (DSS), disuccinimidyl glutarate (DSG) and disuccinimidyl tartrate (DST).
  • sulfo-succinimidyl derivatives sulfo-SMCC, sulfo-SMPB
  • DSS disuccinimidyl suberate
  • DSG disuccinimidyl glutarate
  • DST disuccinimidyl tartrate
  • derivatives are detectable labels.
  • the invention provides polypeptides that are detectably labeled.
  • detectable labels include fluorophores, chromophores, radioactive isotopes (e.g., S 35 , P 32 , I 125 ), electron-dense reagents, enzymes, ligands and receptors.
  • Enzymes are typically detected by their activity.
  • horseradish peroxidase is usually detected by its ability to convert a substrate such as 3,3-',5,5-'-tetramethylbenzidine (TMB) to a blue pigment, which can be quantified.
  • TMB 3,3-',5,5-'-tetramethylbenzidine
  • Modified polypeptides also include one or more D-amino acids substituted for L-amino acids (and mixtures thereof), structural and functional analogues, for example, peptidomimetics having synthetic or non-natural amino acids or amino acid analogues and derivatized forms. Modifications include cyclic structures such as an end-to-end amide bond between the amino and carboxy-terminus of the molecule or intra- or inter- molecular disulfide bond.
  • Polypeptides may be provided in the form of a salt, for example as a pharmaceutically acceptable and/or a physiologically acceptable salt.
  • the salt may be an acid addition salt with an inorganic acid, an acid addition salt with an organic acid, a salt with a basic inorganic acid, a salt with a basic organic acid, a salt with an acidic or basic amino acid or a mixture thereof.
  • a salt, such as a pharmaceutically acceptable salt is an acetate salt.
  • the invention provides polypeptides and molecules in isolated and/or purified form.
  • isolated when used as a modifier of a composition, means that the compositions are made by the hand of man or are separated, completely or at least in part, from their naturally occurring in vivo environment. Generally, isolated compositions are substantially free of one or more materials with which they normally associate with in nature, for example, one or more protein, nucleic acid, lipid, carbohydrate, cell membrane.
  • isolated does not exclude alternative physical forms of the composition, such as fusions/chimeras, multimers/oligomers, modifications (e.g., phosphorylation, glycosylation, lipidation) or derivatized forms, or forms expressed in host cells produced by the hand of man.
  • an “isolated” composition e.g. polypeptides or molecules as defined herein
  • an isolated polypeptide that also is substantially pure or purified does not include polypeptides or polynucleotides present among millions of other sequences, such as polypeptide of a peptide library or nucleic acids in a genomic or cDNA library, for example.
  • compositions can be combined with one or more other molecules.
  • substantially pure or purified does not exclude combinations of compositions, such as combinations of polypeptides other antigens, agents, drugs or therapies.
  • Polypeptides can be prepared recombinant, chemically synthesized, isolated from a biological material or source, and optionally modified, or any combination thereof.
  • a biological material or source would include an organism that produced or possessed any polypeptide or molecule set forth herein.
  • a biological material or source may further refer to a preparation in which the morphological integrity or physical state has been altered, modified or disrupted, for example, by dissection, dissociation, solubilization,
  • polypeptides such as a molecule (e.g. immunogenic molecules) disclosed herein, may be modified by substituting, deleting or adding one or more amino acid residues in the amino acid sequence and screening for biological activity, for example eliciting an immune response.
  • a skilled person will understand how to make such derivatives or variants, using standard molecular biology techniques and methods, described for example in Sambrook et al. (2001) Molecular Cloning: a Laboratory Manual, 3 rd ed., Cold Spring Harbour Laboratory Press).
  • Molecules and polypeptides disclosed herein, including a modified polypeptide thereof, a derivatized polypeptide thereof, or salts thereof may be manufactured synthetically or recombinantly.
  • the molecule, including a polypeptide of option a), b), c), d) or a subsequence thereof is synthetic.
  • the molecule, including a polypeptide of option a), b), c), d) or a subsequence thereof may be isolated and/or purified, e.g. made by the hand of man, such as by synthesis.
  • the polypeptides may be combined after synthesis and freeze-dried or dissolved in aqueous solutions, DMSO, glycerol or the like or mixtures thereof.
  • Molecules e.g. immunogenic molecules
  • Such methods and uses include, for example, administration in vitro and in vivo of one or more molecules disclosed herein.
  • the methods and uses may include modulating an immune response (e.g. modulating an allergic immune response), including, among others, methods and uses for relieving an immune response (e.g. allergic immune response), protecting and treating subjects against a disorder, disease (e.g. allergic disease); and methods and uses of providing immunotherapy, such as specific immunotherapy against an immune response, e.g. an immune response triggered by an allergen (e.g. a pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. a pollen, such as a pollen allergen
  • methods and uses include administration or delivery of a molecule disclosed herein to modulate an immune response in a subject, including, for example, modulating an immune response triggered by an allergen (e.g. triggered by pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. triggered by pollen, such as a pollen allergen
  • a molecule disclosed herein e.g. triggered by pollen, such as a pollen allergen
  • modulate means an alteration or effect on the term modified.
  • modulating involves decreasing, reducing, inhibiting, suppressing, relieving an immune response in a subject triggered by an allergen (e.g. triggered by pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • modulating involves eliciting, stimulating, inducing, promoting, increasing or enhancing an immune response in a subject triggered by an allergen (e.g. triggered by pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • modulate is used to modify the term "immune response triggered by an allergen (e.g.
  • an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein in a subject this means that the immune response in the subject to the allergen, pollen, pollen allergen or molecule is altered or affected (e.g., decreased, reduced, inhibited, suppressed, limited, controlled, prevented, elicited, promoted, stimulated, increased, induced, enhanced, etc.
  • Methods and uses of modulating an immune response triggered by an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein may be used to provide a subject with protection against an immune response or immune reaction to said allergen, pollen, pollen allergen and/or molecule (e.g.
  • methods and uses include administering a molecule (e.g.
  • immunogenic molecule of the invention to protect or treat a subject against an immune response, or one or more symptoms caused by, triggered by or associated with exposure to an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein e.g. a pollen allergen
  • the subject's administration of a therapeutically effective amount of a composition described herein may relieve one or more symptoms of the immune response.
  • the method may comprise relieving one or more symptoms associated with allergic rhinitis, allergic conjunctivitis, allergic asthma and/or allergic eczema (e.g. atopic dermatitis).
  • the one or more symptoms may be associated with allergic rhinitis.
  • the method may comprise reducing one or more of the following symptoms: intensity of itchy nose; number of sneezes within a given period (e.g. daily, weekly, monthly); intensity of blocked nose (e.g. congestion); amount of nasal secretions; eosinophilic count in nasal secretions; specific IgE antibody level (titer) in nasal secretions or in serum; and basophil histamine release of blood.
  • the one or more symptoms may be associated with allergic conjunctivitis.
  • the method may comprise reducing one or more of the following: intensity of itchy eyes, redness in the white of the eyes and/or watery eyes; eosinophilic count in conjunctival tissue scrapings; specific IgE antibody level (titer) in conjunctival tissue scrapings or in serum; and basophil histamine release in blood.
  • the one or more symptoms may be associated with allergic asthma.
  • the method may comprise reducing one or more of the following: number of or frequency of asthma exacerbations (optionally that require hospitalization), intensity and/or number of coughs within a given period (e.g. daily, weekly, monthly); intensity of wheezes; intensity of shortness of breath or congestion (e.g. improvement of being short of breath); reducing Forced Expiratory Volume (FEV1); reducing specific IgE antibody level (titer) in lung fluid or in serum and basophil histamine release in blood; or the method may comprise improving lung function.
  • the one or more symptoms may be symptoms associated with atopic dermatitis.
  • the method may comprise reducing one or more of the following: itch intensity of the skin; eczema score, and number of (peripheral) blood eosinophils.
  • a therapeutic or beneficial effect also includes reducing or eliminating the need, dosage frequency or amount of a second therapeutic method or therapeutically active drug (e.g. anti-inflammatory, decongestants or anti-allergic agent) used for treating a subject having an immune response or one or more symptoms caused by or associated with an allergen.
  • a second therapeutic method or therapeutically active drug e.g. anti-inflammatory, decongestants or anti-allergic agent
  • administration of a peptide combination described herein may reduce the amount of an adjunct therapy administered to a subject, such as reducing the subject's need for concomitant treatment with fast or long-acting p2-agonists, leukotriene modifiers, theophylline corticosteroids or H1 antihistamines (e.g. inhaled or oral) to reduce, relieve, or suppress one or more symptoms of the immune response.
  • an adjunct therapy administered to a subject, such as reducing the subject's need for concomitant treatment with fast or long-acting p2-agonists, leukotriene modifiers, theophylline corticosteroids or H1 antihistamines (e.g. inhaled or oral) to reduce, relieve, or suppress one or more symptoms of the immune response.
  • methods and uses include administering or delivering a molecule (e.g. an immunogenic molecule) of the invention to elicit, stimulate, induce, promote, increase or enhance immunological tolerance of a subject an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • a molecule e.g. an immunogenic molecule
  • an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein include administering or delivering a molecule (e.g. an immunogenic molecule) of the invention to elicit, stimulate, induce, promote, increase or enhance immunological tolerance of a subject an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • a method or use includes administering to the subject an amount of an immunogenic molecule of the invention sufficient to provide the subject with protection against an immune response, or symptoms caused by or associated with exposure to an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • an allergen e.g. pollen, such as a pollen allergen
  • an immunogenic molecule disclosed herein includes administering to the subject an amount of an immunogenic molecule of the invention sufficient to provide the subject with protection against an immune response, or symptoms caused by or associated with exposure to an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • Methods and uses of the invention include providing a subject with protection against an allergen (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein, or symptoms caused by or associated with the subject's exposure to said allergen (e.g. pollen, such as pollen allergen) and/or a molecule disclosed herein, for example, vaccinating the subject to protect against an immune response to said allergen (e.g.
  • an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein e.g. pollen, such as pollen allergen
  • methods and uses include protecting the subject against an immune response triggered by pollen, e.g. a pollen allergen or a molecule disclosed herein by inducing tolerance of the subject (desensitizing) to said allergen, pollen, pollen allergen and/or immunogenic molecule.
  • the terms "protection,” “protect” and grammatical variations thereof, when used in reference to an immune response or symptoms caused by or associated with the exposure to an allergen (e.g. pollen, such as a pollen allergen) and/or an immunogenic molecule disclosed herein, means preventing an immune response or symptoms caused by or associated with the exposure to the allergen, pollen, pollen allergen and/or a molecule disclosed herein, or reducing or decreasing susceptibility to an immune response or one or more symptoms caused by or associated with the exposure to the allergen, pollen, pollen allergen and/or a molecule disclosed herein.
  • an immune response includes but is not limited to an allergic immune response, such as an allergic reaction, hypersensitivity, an inflammatory response or inflammation.
  • an immune response may involve one or more of cell infiltration, production of antibodies, production of cytokines, lymphokines, chemokines, interferons and interleukins, cell growth and maturation factors (e.g., differentiation factors), cell proliferation, cell differentiation, cell accumulation or migration (chemotaxis) and cell, tissue or organ damage or remodeling.
  • an immune response may include an allergic immune response, such as allergic rhinitis; atopic dermatitis; allergic conjunctivitis and asthma. Allergic responses can occur systemically or locally in any region, organ, tissue, or cell.
  • an allergic immune response occurs in the skin, the upper respiratory tract, the lower respiratory tract, pancreas, thymus, kidney, liver, spleen, muscle, nervous system, skeletal joints, eye, mucosal tissue, gut or bowel.
  • Methods and uses herein include relieving a subject of, including treating a subject for, an immune response, or one or more symptoms caused by or associated with an allergen (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. pollen, such as a pollen allergen
  • Such methods and uses include administering to a subject an amount of an immunogenic molecule sufficient to relieve the subject of, such as treat the subject for, the allergic immune response, or one or more symptoms caused by or associated with the allergen, (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • Methods and uses of the invention include treating or administering to a subject previously exposed to an allergen (e.g.
  • methods and uses are for treating or protecting a subject from an allergic immune response, or one or more symptoms caused by or associated with secondary or subsequent exposure to an allergen, (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. pollen, such as a pollen allergen
  • a molecule disclosed herein e.g. a pollen allergen
  • Immunogens described herein may elicit, stimulate, induce, promote, increase or enhance immunological tolerance to an allergen (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • Methods and uses of the invention therefore further include inducing immunological tolerance of a subject to an allergen (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. pollen, such as a pollen allergen
  • molecules described herein can be effective in relieving, such as treating an immune response, including but not limited to an immune response following a secondary or subsequent exposure of a subject to an allergen (e.g. pollen, such as a pollen allergen) and/or a molecule disclosed herein.
  • a method or use includes administering to the subject an amount of a molecule disclosed herein sufficient to induce tolerance in the subject to the allergen (e.g. pollen, such as a pollen allergen) or the molecule itself.
  • the immunological tolerance elicited, stimulated, induced, promoted, increased or enhanced may involve modulation of T cell activity, including but not limited to CD4+ T cells, CD8+ T cells, Th1 cells, Th2 cells and regulatory T cells.
  • immunological tolerance elicited, stimulated, induced, promoted, increased or enhanced from administration of the immunogenic molecule may involve modulation of the production or activity of pro-inflammatory or anti-inflammatory cytokines produced by T cells.
  • a method or use of inducing immunological tolerance in a subject to an allergen (e.g. pollen, such as a pollen allergen) and/or an immunogenic molecule disclosed herein includes a reduction in occurrence, frequency, severity, progression, or duration of physiological conditions, disorders, illnesses, diseases, symptoms or complications caused by or associated an allergic response to the allergen (e.g. pollen, such as a pollen allergen) and/or immunogenic molecule in the subject.
  • inducing immunological tolerance can protect a subject against or treat a subject for an immune response, or one or more symptoms caused by or associated with an allergen (e.g. pollen, such as a pollen allergen) or a molecule disclosed herein.
  • Methods and uses of the invention include treating a subject via immunotherapy, including specific immunotherapy.
  • a method or use includes administering to the subject an amount of a molecule described herein.
  • a molecule administered to a subject during specific immunotherapy to treat the subject is the same immunogenic molecule to which the subject has been sensitized or is hypersensitive (e.g., allergic).
  • a molecule is administered to a subject to treat the subject to a different immunogenic molecule, e.g. an allergen, e.g. a pollen allergen, to which the subject has been sensitized or is hypersensitive (e.g., allergic).
  • the immunotherapeutic mechanism may involve bystander suppression of an immune response caused by an allergen, e.g. a pollen allergen, by administering an unrelated immunogenic molecule, e.g. a molecule disclosed herein.
  • a molecules of the invention may include T cell epitopes, such as Th2 cell epitopes.
  • the subject to be treated has a specific T- cell response to the a molecule before administering the first dose of the a molecule .
  • methods and uses of the invention include administering an amount of an a molecule (e.g., a T cell epitope-containing a molecule ) to a subject sufficient to provide the subject with protection against an immune response disclosed herein, or one or more symptoms disclosed herein.
  • a method includes administering an amount of an a molecule (e.g., a T cell epitope-containing a molecule ) to a subject sufficient to relieve, e.g. treat, vaccinate or immunize the subject against an immune response disclosed herein, or one or more symptoms caused by or associated with an allergen (e.g. pollen, such as a pollen allergen) and/or an a molecule disclosed herein.
  • an allergen e.g. pollen, such
  • the specific T-cell response may be monitored by determining by way of contacting a sample of PBMCs obtained from the subject with a molecule of the invention and measuring the IL-5 secretion or IL-5 mRNA gene expression in response to the molecule.
  • a method or use includes administering to a subject an amount of a polypeptide described herein or derivative thereof including an a molecule described herein, such as a T cell epitope, sufficient to modulate Th2 cell activity in the subject.
  • two or more a molecules may be administered to a subject, e.g. may be administered as a combination composition, or administered separately, such as concurrently or in series or sequentially.
  • methods and uses described herein comprise administration separately or as a combination: at least 2-25 polypeptides defined herein, or separately or as a combination of 3-25, 4-25, 5-25, 6-25, 7-25 polypeptides defined herein, or separately or as a combination of 2-20, 3-20, 4-20, 5-20, 6-20 defined herein, or separately or as a combination of 2-12, 3-12, 4-12, 5-12, 6-12, 7- 12 polypeptides defined herein, or separately or as a combination of 2-10, 3-10, 4-10, 5- 10, 6-10, 7-10 polypeptides defined herein.
  • a further aspect of the invention relates to a composition
  • a composition comprising a polypeptide of option a), b), c) or d) or a combination of two or more polypeptides of option a), b), c) or d).
  • Methods and uses of the invention therefore include any therapeutic or beneficial effect.
  • an immune response, or one or more symptoms caused by or associated with a pollen allergen or an a molecule disclosed herein is reduced, decreased, inhibited, limited, delayed or prevented.
  • Methods and uses of the invention moreover include reducing, decreasing, inhibiting, delaying or preventing onset, progression, frequency, duration, severity, probability or susceptibility of one or more adverse symptoms, disorders, illnesses, diseases or complications caused by or associated with an antigen/allergen.
  • methods and uses include improving, accelerating, facilitating, enhancing, augmenting, or hastening recovery of a subject from an allergic immune response, or one or more physiological conditions, symptoms or complications caused by or associated with a pollen allergen or an a molecule disclosed herein.
  • methods and uses include stabilizing an allergic immune response, or one or more physiological conditions, symptoms or complications caused by or associated with a pollen allergen or an a molecule disclosed herein.
  • a therapeutic or beneficial effect is therefore any objective or subjective measurable or detectable improvement or benefit provided to a particular subject.
  • a therapeutic or beneficial effect can but need not be complete ablation of all or any immune response, or one or more symptoms caused by or associated with a pollen allergen or an a molecule disclosed herein.
  • a satisfactory clinical endpoint is achieved when there is an incremental improvement or a partial reduction in an allergic immune response, or one or more symptoms caused by or associated with an allergen, or an inhibition, decrease, reduction, suppression, prevention, limit or control of worsening or progression of an immune response, or one or more symptoms caused by or associated with a pollen allergen or an a molecule disclosed herein, over a short or long duration (hours, days, weeks, months, etc.).
  • a therapeutic or beneficial effect also includes reducing or eliminating the need, dosage frequency or amount of a second therapeutic protocol or active such as another drug or other agent (e.g., anti-inflammatory) used for treating a subject having or at risk of having an allergic immune response, or one or more symptoms caused by or associated with an allergen.
  • a second therapeutic protocol or active such as another drug or other agent (e.g., anti-inflammatory) used for treating a subject having or at risk of having an allergic immune response, or one or more symptoms caused by or associated with an allergen.
  • reducing an amount of an adjunct therapy such as a reduction or decrease of a treatment for an allergic immune response, or one or more symptoms caused by or associated with an allergen, or a specific immunotherapy, vaccination or immunization protocol is considered a beneficial effect.
  • reducing or decreasing an amount of the a molecule used for specific immunotherapy, vaccination or immunization of a subject to provide protection to the subject is considered a beneficial effect.
  • Methods and uses described herein may relieve one or more symptoms of an allergic immune response or delays the onset of symptoms, slow the progression of symptoms, or induce disease modification.
  • the following symptoms may be decreased or eliminated; nasal symptoms in the form of itchy nose, sneezing, runny nose, blocked nose; conjunctival symptoms in the form of itchy eyes, red eyes, watery eyes; and respiratory symptoms in the form of decreased lung function.
  • the beneficial effect of methods and uses described herein may be observed by the patient's need for less concomitant treatment with corticosteroids or H1 antihistamines to suppress the symptoms.
  • an amount or dose of the immunogenic molecule to be administered, and the period of time required to achieve a desired outcome or result can be determined by one skilled in the art.
  • the immunogenic molecule may be administered to the patient through any route known in the art, including, but not limited to oral, inhalation, sublingual, epicutaneous, intranasal, and/or parenteral routes (intravenous, intramuscular, subcutaneously, intradermal, and intraperitoneal).
  • Methods and uses of the invention include administration of a molecule (e.g. an immunogenic molecule) to a subject prior to contact by or exposure to a pollen allergen or a molecule (e.g. an immunogenic molecule) disclosed herein; administration prior to, substantially contemporaneously with or after a subject has been contacted by or exposed to an allergen; and administration prior to, substantially contemporaneously with or after an allergic immune response, or one or more symptoms caused by or associated with a pollen allergen or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a molecule e.g. an immunogenic molecule
  • a "sufficient amount” or “effective amount” or an “amount sufficient” or an “amount effective” refers to an amount that provides, in single (e.g., primary) or multiple (e.g., booster) doses, a long term or a short term detectable or measurable improvement in a given subject or any objective or subjective benefit to a given subject of any degree or for any time period or duration (e.g., for minutes, hours, days, months, years, or cured).
  • prophylactically effective in each and every subject treated, nor a majority of subjects treated in a given group or population An amount sufficient or an amount effective means sufficiency or effectiveness in a particular subject, not a group of subjects or the general population. As is typical for such methods, different subjects will exhibit varied responses to a method of the invention, such as immunization, vaccination, specific immunotherapy and therapeutic treatments.
  • subject includes but is not limited to a subject at risk of allergen contact or exposure as well as a subject that has been contacted by or exposed to an allergen.
  • a subject also includes those having or at risk of having or developing an immune response to an antigen or an allergen.
  • Such subjects include mammalian animals (mammals), such domestic animal (dogs and cats), a farm animal (poultry such as chickens and ducks, horses, cows, goats, sheep, pigs), experimental animal (mouse, rat, rabbit, guinea pig) and humans.
  • Target subjects and subjects in need of treatment also include those at risk of allergen exposure or contact or at risk of having exposure or contact to an allergen. Accordingly, subjects include those at increased or elevated (high) risk of an allergic reaction; that have, or have previously had or are at risk of developing hypersensitivity to an allergen; and those that have or have previously had or are at risk of developing asthma.
  • the immune response to be treated in a subject in need thereof is against one or more allergens, e.g. one or more pollen, such as one or more pollen allergens.
  • immunological tolerance is desirable to be induced or promoted in a subject in need thereof to one or more allergens, e.g. one or more pollen, such as one or more pollen allergens.
  • the subject may be sensitized to one or more of the pollen allergens.
  • Non-limiting examples of the one or more pollen allergens to which the subject optionally is sensitized or the immune response is triggered by include proteins, polypeptides and peptides including allergens from a plant species selected from any of the plant families /Asferaceae, Betulaceae, Fagaceae, Oleaceae, Poaceae and/or Plantaginaceae, for example plant species selected from any of the plant genera Anthoxanthum, Conydon, Dactylis, Lollium, Phleum, Poa, Ambrosia, Artemisia, Helianthus, AInus, Betula, Carpinus, Castanea, Corylus, Ostrya, Ostryopsis, Fagus, Quercus, Fraxinus, Ligustrum, Lilac, Olea and Plantago; for example plant species selected from any of the plant genera
  • Anthoxanthum Conydon, Dactylis, Lollium, Phleum, Poa, Ambrosia, Artemisia, AInus, Betula, Corylus, Fagus, Quercus, Olea and Plantago; for example plant species selected from the plant genera Ambrosia, Artemisia, Helianthus, AInus, Betula, Carpinus,
  • Quercus such as a plant species selected from any of the plant genera Phleum,
  • the one or more pollen allergens to which the subject optionally is sensitized to include proteins, polypeptides and peptides including allergens from species selected from further plant families, like the plant family Chenopidiaceae including the plants Lambs quarters, Russian thistles and Kochias; plant family Amaranthaceae including Pigweeds, plant family Polygonaceae including for example Sheep sorrel, plant family Ulmacea including for example American Elm and hackberry, plant family
  • Plantanaceae including for example Sycamore, plant family Salicaceae including for example White poplar and Cottonwood, plant family Aceraceae including for example Box elder and Red maple, plant family Cupressaceae including for example Common juniper and Cedar.
  • the subject may be sensitized to one or more pollen species from one or more plant families, for example selected from any of the plant families Poaceae,
  • the subject may be sensitized to a pollen species of the plant family Poaceae and a pollen species of a plant family selected from any one of Asteraceae, Fagaceae and Betulaceae, such as a subject that may be sensitized to a pollen species of the plant genus Phleum and a pollen species of a plant genus selected from any one of the genera Conydon, Ambrosia, Betula and Quercus and combinations thereof.
  • the subject may be sensitized to one or more pollen allergens from one or more plant families, for example selected from the any of the plant families Poaceae, Asteraceae, Fagaceae, Betulaceae, Oleaceae, and Plantaginaceae, preferably Poaceae, Asteraceae, Fagaceae and Betulaceae.
  • the subject may be sensitized to a pollen allergen of the plant family Poaceae and a pollen allergen of a plant family selected from any one of Asteraceae, Fagaceae and Betulaceae and combinations thereof, such as a subject that may be sensitized to a pollen allergen of the plant genus Phleum and a pollen allergen of a plant genus selected from the group consisting of Conydon, Ambrosia, Betula and Quercus and combinations thereof.
  • the immune response may be triggered by pollen from one or more plant families, such as from plant families selected from any of Poaceae, Asteraceae,
  • the immune response is triggered by pollen of the plant family Poaceae and pollen of a plant family selected from any one of Asteraceae, Fagaceae and Betulaceae and combinations thereof, for example the immune response may be triggered by pollen of the plant genus Phleum and pollen of a plant genus selected from the any one of Conydon, Ambrosia, Betula and Quercus or combinations thereof.
  • the immune response may be triggered by one or more pollen allergens, such as one or more pollen allergens from a plant family selected from any of Poaceae, Asteraceae, Fagaceae, Betulaceae, Oleaceae, and Plantaginaceae, preferably Poaceae, Asteraceae, Fagaceae and Betulaceae. Therefore, in some embodiments, the immune response is triggered by one or more pollen allergens from pollen of the plant family Poaceae and one or more pollen allergens from pollen of a plant family selected from any of Asteraceae, Fagaceae and Betulaceae.
  • pollen allergens such as one or more pollen allergens from a plant family selected from any of Poaceae, Asteraceae, Fagaceae, Betulaceae, Oleaceae, and Plantaginaceae, preferably Poaceae, Asteraceae, Fagaceae and Betulaceae. Therefore, in
  • the one or more pollen allergens may be from pollen of the plant genus Phleum and from pollen of a plant genus selected from the group consisting of Conydon, Ambrosia, Betula and Quercus including combinations thereof.
  • the immune responses to be modulated or treated are against pollen allergens of grass species of various plant genera, for example of the genera
  • Phleum and Cynodon thus allowing the treatment or modulation of an immune response caused by a broad range of different grass pollen species. Therefore, in some
  • the one or more pollen allergens are from a grass species selected from any of the plant genera Anthoxanthum, Conydon, Dactylis, Lollium, Phleum and/or Poa, for example from grass species selected from any of the genera Conydon and Phleum.
  • the immune responses to be modulated or treated are against grass pollen allergens as well as weed and/or tree pollen allergens.
  • the immune responses to be modulated or treated are against grass pollen allergens of the genera Phleum and against non-grass pollen of the genera Ambrosia and/or Quercus, thus allowing the treatment or modulation of an immune response caused by a broad range of different grass pollen species and non-grass species.
  • the one or more pollen allergens are from a grass species selected from any of the plant genera Anthoxanthum, Conydon, Dactylis, Lollium, Phleum and Poa and from a non-grass species selected from any of the plant families Asteraceae, Betulaceae, Fagaceae, Oleaceae and Plantaginaceae, e.g.
  • non-grass species are selected from any of the plant genera Ambrosia, Artemisia, Helianthus, Alnus, Betula, Carpinus, Castanea, Corylus, Ostrya, Ostryopsis, Fagus, Quercus, Fraxinus, Ligustrum, Lilac, Olea and Plantago, for example against a non-grass species selected from any of the plant genera Ambrosia, Artemisia, Alnus, Betula, Quercus, Olea and Plantago, such as a non-grass species selected from any of the plant genera Ambrosia, Betula and Quercus.
  • the immune responses to be modulated or treated are against non-grass pollen allergens, such as against weed and/or tree pollen allergens.
  • the immune responses to be modulated or treated are against non-grass pollen allergens of the genera Ambrosia and against non-grass pollen allergens of the genera Quercus, thus allowing the treatment or modulation of an immune response caused by a broad range of different non-grass pollen species.
  • the one or more non-grass pollen allergens are from a plant genus selected from any of
  • Ambrosia Artemisia, Helianthus, Alnus, Betula, Carpinus, Castanea, Corylus, Ostrya, Ostryopsis, Fagus, Quercus, Fraxinus, Ligustrum, Lilac, Olea and/or Plantago.
  • the one or more pollen allergens are from same grass species or alternatively from different grass species, optionally wherein the different grass species are from different plant genera selected from any of Anthoxanthum, Conydon, Dactylis, Lollium, Phleum and Poa, for example Conydon and Phleum.
  • the one or more non-grass pollen allergens are from same non-grass species or alternatively from different non-grass species, optionally wherein the different non-grass species are from different plant genera selected from any of
  • the immune response to be modulated or treated may be at least against one or more pollen allergens of various grass pollen species, e.g. against one or more pollen allergens of the plant genera Phleum. In some embodiments, the immune response to be modulated or treated may be at least against one or more pollen allergens of various weed pollen species, e.g. against one or more pollen allergens of the plant genera Ambrosia.
  • the immune response to be modulated or treated may be at least against one or more pollen allergens of various tree pollen species, e.g. against one or more pollen allergens of the plant genera Quercus. In some embodiments, the immune response to be modulated or treated may be at least against one or more pollen allergens of various tree pollen species, e.g. against one or more pollen allergens of the plant genera Betula.
  • the immune response to be modulated or treated may be against one or more pollen allergens of various grass pollen species and non-grass pollen species, such as two or more, three or more, four or more, five or more pollen allergens of different pollen species, like various grass pollen species and/or non-grass pollen species.
  • Non-limiting examples of the genus Ambrosia may be Ambrosia artemisiifolia, Ambrosia psilostachya, Ambrosia trifida; a typical species of the genus Artemisia may be Artemisia vulgaris; a typical species of the genus Betula may be Betula verrucosa; a typical species of the genus Fagus may be Fagus grandifolia or Fagus sylvatica; a typical species of the genus Quercus may be Quercus alba, a typical species of the genus Fraxinus may be Fraxinus excelsior; a typical species of the genus Olea may be Olea Europaea, a typical species of the genus plantago may be Plantago lanceolata, a typical species of the genus plantago may be Plantago lanceolata, a typical species of the genus
  • Non-limiting examples of non-grass pollen allergens to which a subject optionally may be sensitized to are Aln g 1 , Aln g 4, Amb a 1 , Amb a 2, Amb a 3, Amb a 4, Amb a 5, Amb a 6, Amb a 7, Amb a 8, Amb a 9, Amb a 10, Amb p 5, Amb t 5, Art v 1 , Art v 2, Art v 3, Art v 4, Art v 5, Art v 6, Bet v 1 , Bet v 2, Bet v 3, Bet v 4, Bet v 6, Bet v 7, Car b 1 , Cas s 1 , Cor a 6, Cor a 10, Fag s 1 , Fra e 1 , Hel a 1 , Hel a, Lig v 1 , Ole e 1 , Ole e 2, Ole e 3, Ole e 4, Oie e 5, Ole e 6, Ole e 7, Ole e 8, Ole e 9,
  • Non-limiting examples of grass pollen allergens to which a subject optionally may be sensitized to are Ant o 1 , Cyn d 1 , Cyn d 7, Cyn d 12, Cyn d 15, Cyn d 22w, Cyn d 23, Cyn d 24, Dac g 1 , Dac g 2, Dac g 3, Dac g 4, Dac g 5, Fes p 4, Hoi 1 1 , Hoi I 5, Hor v 1 , Hor v 5, Lol p 1 , Lol p 2, Lol p 3, Lol p 4, Lol p 5, Lol p 11 , Ory s 1 , Pas n 1 , Pha a 1 , Pha a 5, Phi p 1 , Phi p 2, Phi p 4, Phi p 5, Phi p, Phi p 7, Phi p 1 1 , Phi p 12, Phi p 13, Poa p 1 , Poa p 5, Sec c 1 , Sec c 5, Sec
  • the group 1 allergens e.g. Ant o 1 , Cyn d 1 , Dac g 1 , Hoi 1 , Lol p 1 , Pha a 1 , Phi p 1 and Poa p 1
  • group 5 allergens Dac g 5, Lol p 5, Pha a 5, Phi p 5, Poa p 5
  • “Prophylaxis” and grammatical variations thereof mean a method or use in which contact, administration or in vivo delivery to a subject is prior to contact with or exposure to a pollen allergen or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a pollen allergen or a molecule e.g. an immunogenic molecule
  • administration or in vivo delivery to a subject can be performed prior to manifestation of an allergic immune response, or one or more symptoms caused by or associated with an allergen.
  • a subject can be provided protection against an allergic immune response, or one or more symptoms caused by or associated with a pollen allergen or a molecule disclosed herein or provided immunotherapy with a molecule (e.g.
  • an immunogenic molecule of the present invention.
  • a method or use can eliminate, prevent, inhibit, suppress, limit, decrease or reduce the probability of or susceptibility towards an allergic immune response, or one or more physiological conditions, symptoms or complications caused by or associated with an a pollen allergen or a molecule disclosed herein.
  • “Prophylaxis” can also refer to a method or use in which contact, administration or in vivo delivery to a subject is prior to a secondary or subsequent exposure to an allergen (e.g. pollen such as a pollen allergen) and/or a molecule disclosed herein.
  • an allergen e.g. pollen such as a pollen allergen
  • a subject may have had a prior contact or exposure to a pollen allergen or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • an acute allergic reaction may but need not be resolved.
  • Such a subject typically may have developed anti- allergen antibodies due to the prior exposure. Immunization or vaccination, by
  • Such a method or use can eliminate, prevent, inhibit, suppress, limit, decrease or reduce the probability of or susceptibility towards a secondary or subsequent allergic immune response, or one or more symptoms caused by or associated with a pollen allergen or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a method or use includes providing specific immunotherapy to the subject to eliminate, prevent, inhibit, suppress, limit, decrease or reduce the probability of or susceptibility towards a secondary or subsequent allergic immune response, or one or more physiological conditions, symptoms or complications caused by or associated with an a pollen allergen or a molecule disclosed herein.
  • Treatment of an allergic reaction or response can be at any time during the reaction or response.
  • a molecule e.g. an immunogenic molecule
  • a molecule can be administered as a single or multiple dose e.g., one or more times hourly, daily, weekly, monthly or annually or between about 1 to 10 weeks, or for as long as appropriate (e.g. 3 months, 6 months or more, for example, to achieve a reduction in the onset, progression, severity, frequency, duration of one or more symptoms or complications associated with or caused by an allergic immune response, or one or more physiological conditions, symptoms or complications caused by or associated with an antigen/allergen.
  • methods and uses of the invention can be practiced one or more times (e.g., 1-10, 1-5 or 1-3 times) an hour, day, week, month, or year.
  • Doses can be based upon current existing protocols, empirically determined, using animal disease models or optionally in human clinical trials.
  • Initial study doses can be based upon animal studies, e.g. a mouse, and the sufficient amount of immunogenic molecule to be administered for being effective can be determined.
  • Exemplary non-limiting amounts (doses) are in a range of about 0.1 mg/kg to about 100 mg/kg, and any numerical value or range or value within such ranges.
  • doses can be administered, for example, 0.01-500 mg/kg, and any numerical value or range or value within such ranges.
  • the dose can be adjusted according to the mass of a subject, and will generally be in a range from about 1-10 ug/kg, 10-25 ug/kg, 25-50 ug/kg, 50-100 ug/kg, 100-500 ug/kg, 500-1 ,000 ug/kg, 1-5 mg/kg, 5-10 mg/kg, 10-20 mg/kg, 20-50 mg/kg, 50-100 mg/kg, 100-250 mg/kg, 250-500 mg/kg, or more, two, three, four, or more times per hour, day, week, month or annually.
  • a typical range will be from about 0.3 mg/kg to about 50 mg/kg, 0-25 mg/kg, or 1.0-10 mg/kg, or any numerical value or range or value within such ranges.
  • Doses can vary and depend upon whether the treatment is prophylactic or therapeutic, whether a subject has been previously exposed to the antigen/allergen, the onset, progression, severity, frequency, duration, probability of or susceptibility of the symptom, condition, pathology or complication, or vaccination or specific immunotherapy to which treatment is directed, the clinical endpoint desired, previous or simultaneous treatments, the general health, age, gender, race or immunological competency of the subject and other factors that will be appreciated by the skilled artisan. The skilled artisan will appreciate the factors that may influence the dosage and timing required to provide an amount sufficient for providing a therapeutic or prophylactic benefit.
  • Immunogens of the invention can be provided in compositions, and in turn such compositions can be used in accordance with the invention methods and uses.
  • Such compositions, methods and uses include pharmaceutical compositions and formulations.
  • a pharmaceutical composition includes one or more
  • compositions and formulations may be a vaccine, including but not limited to a vaccine to protect against (e.g. modify, relieve or suppress) an immune response disclosed herein, or one or more symptoms caused by or associated with an allergen and/or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a vaccine to protect against (e.g. modify, relieve or suppress) an immune response disclosed herein, or one or more symptoms caused by or associated with an allergen and/or a molecule (e.g. an immunogenic molecule) disclosed herein.
  • a pharmaceutical composition comprises a molecule (e.g. an immunogenic molecule) of the invention and a pharmaceutically acceptable ingredient or carrier.
  • a pharmaceutically acceptable and “physiologically acceptable” mean a biologically acceptable formulation, gaseous, liquid or solid, or mixture thereof, which is suitable for one or more routes of administration, in vivo delivery or contact.
  • Such formulations include solvents (aqueous or non-aqueous), solutions (aqueous or non-aqueous), emulsions (e.g., oil-in-water or water-in-oil), suspensions, syrups, elixirs, dispersion and suspension media, coatings, isotonic and absorption promoting or delaying agents, compatible with pharmaceutical administration or in vivo contact or delivery.
  • Aqueous and non-aqueous solvents, solutions and suspensions may include suspending agents and thickening agents.
  • Such pharmaceutically acceptable carriers include tablets (coated or uncoated), capsules (hard or soft), microbeads, powder, granules and crystals.
  • Supplementary active compounds e.g., preservatives,
  • antibacterial, antiviral and antifungal agents can also be incorporated into the composition.
  • a pharmaceutically acceptable or physiologically acceptable excipient, carrier and/or adjuvants are well-known to the person skilled in the art and may include, but are not limited to, solvents, emulsifiers, wetting agents, plasticizers, solubilizers (e.g. solubility enhancing agents) coloring substances, fillers, preservatives, anti-oxidants, anti-microbial agents, viscosity adjusting agents, buffering agents, pH adjusting agents, isotonicity adjusting agents, mucoadhesive substances, and the like. Examples of formulation strategies are well-known to the person skilled in the art.
  • the pharmaceutical composition may be formulated for parenteral administration, such as formulated for injection, e.g.
  • the pharmaceutical composition may be a liquid (i.e. formulated as a liquid), including a solution, a suspension, a dispersion, and a gelled liquid.
  • a liquid pharmaceutical composition may be formed by dissolving a powder, granulate or lyophilizate of a molecule (e.g. an immunogenic molecule) combination described herein in a suitable solvent and then administering to a subject.
  • Suitable solvents may be any solvent having physiologically acceptable properties and able to dissolve the immunogenic molecule combination in desired concentrations. A desired concentration may depend on the aliquot to be administered (i.e. to be injected) and the desired single dose.
  • a liquid composition comprises each of the immunogenic molecules of the combination in a concentration of 10 to 800 ⁇ , for example 20 to 500 ⁇ or20 to 300 ⁇ .
  • concentration of each immunogenic molecule is the same, such as in an equimolar concentration, but each immunogenic molecule of the composition may be present in different concentrations.
  • the solvent is an aqueous solution, optionally mixed with other solvents.
  • a solvent may comprise at least 60% w/w of water, e.g. at least 65% w/w, 70% w/w, 75% w/w, 80% w/w , 85% w/w, 90% w/w or 95% w/w , 99% w/w of water, such as distilled water, such as sterile water.
  • the solvent is sterile distilled water, e.g. water for injection.
  • An aqueous solution may comprise other solvents than water, for example
  • the aqueous phase of the solvent may be in a physiological acceptable range, typically in the range of 3 to 9, such as in the range of pH 3 to 8, such as in the range of pH 4 to 8, such as in the range of 5 to 8, such as in the range of 6 to 8.
  • the liquid formulation may comprise a pH controlling agent or buffering agent (e.g. citrate buffer, phosphate buffer, acetate buffer), optionally the pH may be adjusted with dilutions of strong base (e.g. sodium hydroxide or the like) and/or dilutions of strong acids (e.g.
  • the liquid formulation is isotonic, and optionally sterile. Therefore, in some embodiments, the formulation comprises saline, such as isotonic saline.
  • the liquid may contain additional excipients, such as another solvent, a solubilizing enhancing agent (e.g. polyoxyethylene (20) sorbitan monolaurate (Tween® 20), ionic and non-ionic emulsifiers (e.g. poloxamers (Kolliphor®)), a dispersant, a thickener, a preservative, an anti-microbial agent, and/or an antioxidant.
  • a solubilizing enhancing agent e.g. polyoxyethylene (20) sorbitan monolaurate (Tween® 20
  • ionic and non-ionic emulsifiers e.g. poloxamers (Kolliphor®)
  • a dispersant e.g. poloxamers (Kolliphor®)
  • a thickener e.g.
  • Non-limiting illustrative examples of solvents include water, saline, DMSO, glycerol, ethanol, acetonitrile, vegetable or synthetic oils.
  • solvents include water, saline, DMSO, glycerol, ethanol, acetonitrile, vegetable or synthetic oils.
  • a pharmaceutical composition may be formulated to contain only a limited amount of water or aqueous solution, e.g. containing less than 10% w/w of water or aqueous solution, such as less than 9, 8, 7, 6, 5, 4, 3, 2, 1 , 0.5% w/w of water or aqueous solution.
  • aqueous solution e.g. containing less than 10% w/w of water or aqueous solution, such as less than 9, 8, 7, 6, 5, 4, 3, 2, 1 , 0.5% w/w of water or aqueous solution.
  • Examples of pharmaceutical compositions with limited levels of water may include granulates, powders, for example lyophilizates, i.e. freeze-dried powders.
  • the freeze-dried composition may be dissolved before use, for example dissolved in an aqueous, optionally sterile, solution, for example a solution having a pH in the range of 3-9, such as pH in the range of 3 to 8, such as pH in the range of 4 to 8.
  • a lyophilizate may contain additional ingredients, e.g. bulking agents and lyoprotectants (e.g. sucrose, lactose, trehalose, mannose, mannitol, sorbitol, glucose, raffinose, glycine, histidine or mixtures thereof), buffering agents (e.g.
  • solubilizers e.g. polyoxyethylene (20) sorbitan monolaurate (Tween® 20)
  • a freeze-dried composition may also be formulated into a solid dosage form that is administered for example by the oral route such as by oral mucosa.
  • the pharmaceutical composition may be formulated for oral administration, for example for sublingual administration. Therefore, the pharmaceutical composition may be a solid dosage form, such as a freeze-dried solid dosage form, typically a tablet, a capsule or sachet, which optionally may be formulated for fast disintegration.
  • compositions, methods and uses of the invention are known in the art (see, e.g., Remington: The Science and Practice of Pharmacy (2003) 20th ed., Mack Publishing Co., Easton, PA; Remington's Pharmaceutical Sciences (1990) 18th ed., Mack Publishing Co., Easton, PA; The Merck Index (1996) 12th ed., Merck Publishing Group, Whitehouse, NJ;
  • compositions can be formulated to be compatible with a particular route of administration, such as by intradermal or by sublingual administration.
  • pharmaceutical compositions may include carriers, diluents, or excipients suitable for administration by various routes.
  • routes of administration for contact or in vivo delivery for which a composition can optionally be formulated include inhalation, intranasal, oral, buccal, sublingual, subcutaneous, intradermal, epicutaneous, rectal, transdermal, or intralymphatic.
  • a composition may take the form of, for example, tablets or capsules, optionally formulated as fast-integrating tablets/capsules or slow-release tablets/capsules.
  • the tablet is freeze-dried, optionally a fast-disintegrating tablet or capsule suitable for being administered under the tongue.
  • the pharmaceutical composition may also be formulated into a "unit dosage form", which used herein refers to physically discrete units, wherein each unit contains a
  • Unit dosage forms also include, for example, ampules and vials, which may include a composition in a freeze-dried or lyophilized state (a lyophilizate) or a sterile liquid carrier, for example that can be added prior to administration or delivery in vivo.
  • Unit dosage forms also include a composition in a freeze-dried or lyophilized state; a sterile liquid carrier, for example, can be added prior to administration or delivery in vivo.
  • Pharmaceutical formulations can be packaged in single or multiple unit dosage form for ease of administration and uniformity of dosage.
  • Immunogenic molecules may be prone to degradation when exposed to oxygen, for example when exposed to air or solvents containing air. Therefore, in some
  • the pharmaceutical composition comprises an inert gas, e.g. argon or nitrogen.
  • Another aspect of the invention relates to a kit comprising a compartment and
  • kits wherein the compartment comprises a pharmaceutical composition as described herein and wherein the instructions are for use in reducing an immune response triggered by pollen, such as a pollen allergen, e.g. of a grass, weed or tree pollen species disclosed herein, such as instructions for use in treating allergy to pollen (e.g. grass, weed and/or tree pollen allergy to a plant family or species disclosed herein.
  • a kit may further comprise packaging material comprising corrugated fiber, glass, plastic, foil, ampules, vials, blister pack, preloaded syringes or tubes, optionally that maintain sterility of the components.
  • a kit may further comprise labels or inserts comprising printed matter or computer readable medium optionally including identifying components, dose amounts, clinical pharmacology and instructions for the clinician or for a subject using one or more of the kit components, prophylactic or therapeutic benefits, adverse side effects or manufacturer information.
  • the kit additionally comprises a container comprising a solvent for dissolving the composition before use. Examples of suitable solvents are described supra.
  • the kit may also comprise a device for use in parenteral injection, e.g. for injecting the composition (e.g. dissolved composition) to a subcutaneous or intradermal tissue.
  • a device may be any suitable device for that purpose, such as a needle or microneedle adapted for intradermal or subcutaneous delivery of the composition.
  • the device may be a microneedle or a device comprising a plurality of microneedles designed for intradermal delivery of liquids, e.g. as described in international patent applications W014064543 A1 , WO05049107 A2, WO06054280 A2, WO07066341 A3 and W014188429 A1.
  • a composition may be lyophilized so as to enhance stability and ease of transportation.
  • the composition may be sterile.
  • compositions can be formulated to be compatible with a particular route of administration.
  • pharmaceutical compositions include carriers, diluents, or excipients suitable for administration by various routes. Exemplary routes of
  • the pharmaceutical composition is aqueous and, in other embodiments, the composition is non-aqueous solutions, suspensions or emulsions of the immunogenic molecule/protein, which compositions are typically sterile and can be isotonic with the biological fluid or organ of the intended recipient.
  • Non-limiting illustrative examples include water, saline, dextrose, fructose, ethanol, vegetable or synthetic oils.
  • a composition can take the form of for example a solid dosage form, e.g. tablets or capsules, optionally formulated as fast- integrating tablets/capsules or slow-release tablets/capsules.
  • the tablet is a freeze-dried, optionally fast-disintegrating tablet suitable for being administered under the tongue.
  • a solid dosage form optionally is sterile, optionally anhydrous.
  • immunogenic molecules can be mixed with adjuvants.
  • Adjuvants include, for example: oil (mineral or organic) emulsion adjuvants such as Freund's complete (CFA) and incomplete adjuvant (I FA) (WO 95/17210; WO 98/56414; WO 99/12565; WO 99/11241 ; and U.S. Patent No.
  • oil mineral or organic
  • I FA incomplete adjuvant
  • metal and metallic salts such as aluminum and aluminum salts, such as aluminum phosphate or aluminum hydroxide, alum (hydrated potassium aluminum sulfate); bacterially derived compounds, such as Monophosphoryl lipid A and derivatives thereof (e.g., 3 De-O-acylated monophosphoryl lipid A, aka 3D-MPL or d3-MPL, to indicate that position 3 of the reducing end glucosamine is de-O-acylated, 3D-MPL consisting of the tri and tetra acyl congeners), and enterobacterial lipopolysaccharides (LPS); plant derived saponins and derivatives thereof, for example Quil A (isolated from the Quilaja Saponaria Molina tree, see, e.g., "Saponin adjuvants", Archiv.
  • Quil A isolated from the Quilaja Saponaria Molina tree, see, e.g., "Saponin adjuvants", Archiv.
  • Cosolvents may be added to the composition.
  • cosolvents contain hydroxyl groups or other polar groups, for example, alcohols, such as isopropyl alcohol; glycols, such as propylene glycol, polyethyleneglycol, polypropylene glycol, glycol ether; glycerol; polyoxyethylene alcohols and polyoxyethylene fatty acid esters.
  • cosolvents contain hydroxyl groups or other polar groups, for example, alcohols, such as isopropyl alcohol; glycols, such as propylene glycol, polyethyleneglycol, polypropylene glycol, glycol ether; glycerol; polyoxyethylene alcohols and polyoxyethylene fatty acid esters.
  • Supplementary compounds can also be incorporated into the compositions.
  • Pharmaceutical compositions may therefore include preservatives, anti-oxidants and antimicrobial agents.
  • Preservatives can be used to inhibit microbial growth or increase stability of ingredients thereby prolonging the shelf life of the pharmaceutical formulation.
  • Suitable preservatives are known in the art and include, for example, EDTA, EGTA, benzalkonium chloride or benzoic acid or benzoates, such as sodium benzoate.
  • Antioxidants include, for example, ascorbic acid, vitamin A, vitamin E, tocopherols, and similar vitamins or provitamins.
  • An antimicrobial agent or compound directly or indirectly inhibits, reduces, delays, halts, eliminates, arrests, suppresses or prevents contamination by or growth, infectivity, replication, proliferation, reproduction, of a pathogenic or non- pathogenic microbial organism.
  • Classes of antimicrobials include antibacterial, antiviral, antifungal and antiparasitics.
  • Antimicrobials include agents and compounds that kill or destroy (-cidal) or inhibit (-static) contamination by or growth, infectivity, replication, proliferation, reproduction of the microbial organism.
  • compositions, methods and uses of the invention are known in the art (see, e.g. Remington: The Science and Practice of Pharmacy (David B. Troy, Paul Beringer Lippincott Williams & Wilkins) 2006).
  • compositions peptides, proteins, antigens, allergens
  • substituents described herein are disclosed by the application to the same extent as if each composition or group of compositions was set forth individually. Thus, selection of particular peptides, proteins, antigens, allergens, etc. is clearly within the scope of the invention.
  • any concentration range, percentage range, ratio range or other integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.
  • numerical values are often presented in a range format throughout this document, a range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention.
  • range expressly includes all possible sub ranges, all individual numerical values within that range, and all numerical values or numerical ranges including integers within such ranges and fractions of the values or the integers within ranges unless the context clearly indicates otherwise.
  • This construction applies regardless of the breadth of the range and in all contexts throughout this patent document.
  • reference to a range of 90-100% includes 91-99%, 92-98%, 93-95%, 91-98%, 91-97%, 91-96%, 91-95%, 91- 94%, 91-93%, and so forth.
  • Reference to a range of 90-100% includes 91%, 92%, 93%, 94%, 95%, 95%, 97%, etc., as well as 91.1%, 91.2%, 91.3%, 91.4%, 91.5%, etc., 92.1%, 92.2%, 92.3%, 92.4%, 92.5%, etc., and so forth.
  • Reference to a range of 5-10, 10-20, 20- 30, 30-40, 40-50, 50-75, 75-100, 100-150, and 150-175, includes ranges such as 5-20, 5- 30, 5-40, 5-50, 5-75, 5-100, 5-150, 5-171 , and 10-30, 10-40, 10-50, 10-75, 10-100, 10- 150, 10-175, and 20-40, 20-50, 20-75, 20-100, 20-150, 20-175, and so forth.
  • reference to a series of ranges of 2-72 hours, 2-48 hours, 4-24 hours, 4-18 hours and 6-12 hours includes ranges of 2-6 hours, 2, 12 hours, 2-18 hours, 2-24 hours, etc., and 4-27 hours, 4-48 hours, 4-6 hours, etc.
  • Bostick D Vaisman II. A new topological method to measure protein structure similarity. Biochem Biophys Res Commun. 304, 320-325, 2003.
  • HLA human leukocyte antigen
  • Needleman SB Wunsch CD. A general method applicable to the search for similarities in the amino acid sequence of two proteins, J. Mol. Biol. 48:443, 1970.
  • This example includes a description of transcriptomic analysis of various pollen species.
  • RNA-sequencing was performed on pollen samples of the following species: Timothy grass (Phleum pratense (Phi p)), Bermuda grass (Cynodon dactylon (Cyn d)), Western ragweed (Ambrosia psilostachya (Amb p)), Short ragweed (Ambrosia artemisiifolia (Amb a)), White oak (Quercus alba (Que a)), and European white birch (Betula verrucosa (Bet v)) (See Table below).
  • Timothy grass Phleum pratense
  • Bermuda grass Cronodon dactylon
  • Western ragweed Ambrosia psilostachya
  • Short ragweed Ambrosia artemisiifolia (Amb a)
  • White oak Quercus alba (Que a)
  • European white birch Betula verrucosa (Bet v)
  • the table below shows the number of reads assembled for each of the different pollens (top), with over 500 million reads over two replicate runs per allergen. Sequences were assembled into transcripts using Trinity (bottom), resulting in over 50 thousand transcripts per allergen with minimum lengths of 200 nucleotides.
  • the transcripts include related variants, such as isoforms, and homologs.
  • RNA sequences of additional species may also be performed on RNA sequences of additional species as shown in the Table below showing pollen RNA-seq reads for the pollen species Kentucky blue grass, Sweet vernal grass, Rye grass and Ash tree, Olive tree and English plantain:
  • This example includes a description of the identification of polypeptides in extracts of Phi p grass pollen and homologous polypeptides thereof in other pollen species.
  • the Phi p grass transcriptome was translated into amino acid sequences in all six reading frames, and they were used to search 17 mass spectrometry samples of Phi p grass pollen extract using the Mascot software. This identified 354 polypeptide sequences. Polypeptide sequences shorter than 50 aa were discarded, resulting in 275 sequences.
  • the closest homologous sequence in the rice (Oryza sativa japonica) proteome via Blast. Rice was chosen since it is a species closely related to Phi p grass with a completely sequenced and annotated genome. Homologous rice sequences were identified for 180 Timothy grass sequences.
  • homologous sequences were identified (via Blast) in the translated transcriptomes of Cyn d, Amb a, Amb p, Que a, and Bet v of all identified sequences, the one(s) sharing the largest number of conserved peptides with the Phi p sequence was selected as homolog. This resulted in 66 sets of homologous polypeptides. In total, there was found evidence of release of the polypeptides from Phi p grass pollen for 52 of these polypeptides upon extracting pollen in a buffered aqueous solution (see Example 4).
  • Table 1 shows amino acid sequences of the polypeptides detected and identified in Phi p grass pollen and Table 2 shows amino acid sequences of polypeptides homologous to the Phi p sequence of Table 1 found in pollen of the species Cyn d, Amb a, Amb p, Que a, and Bet v.

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Abstract

L'invention concerne des polypeptides identifiés dans le pollen de plusieurs familles de plantes différentes. Les polypeptides comprennent des séquences de résidus d'acide aminé conservés parmi différentes familles de plantes, par exemple parmi les familles de pollen de graminées, de mauvaises herbes et d'arbres. L'invention concerne en outre l'utilisation de ces polypeptides ou de leurs résidus d'acide aminé conservés en tant que molécule (par exemple, une molécule immunogène) pour modifier une réponse immunitaire contre ce polypeptide ou un autre antigène non apparenté (par exemple allergène majeur d'une source de pollen) rendue possible par l'induction d'une tolérance "bystander" (de voisinage).
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WO2023242436A1 (fr) * 2022-06-17 2023-12-21 Danmarks Tekniske Universitet Vaccins antiallergiques à base d'allergènes consensus

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Publication number Priority date Publication date Assignee Title
WO2023242436A1 (fr) * 2022-06-17 2023-12-21 Danmarks Tekniske Universitet Vaccins antiallergiques à base d'allergènes consensus

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