WO2017148950A1 - Gene signature for the detection of vernal keratoconjunctivitis - Google Patents

Gene signature for the detection of vernal keratoconjunctivitis Download PDF

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WO2017148950A1
WO2017148950A1 PCT/EP2017/054652 EP2017054652W WO2017148950A1 WO 2017148950 A1 WO2017148950 A1 WO 2017148950A1 EP 2017054652 W EP2017054652 W EP 2017054652W WO 2017148950 A1 WO2017148950 A1 WO 2017148950A1
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signature
markers
invention comprises
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another embodiment
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PCT/EP2017/054652
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French (fr)
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Andrea Leonardi
Philippe Daull
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Santen Sas
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Definitions

  • the present invention relates to the field of vernal keratoconjunctivitis detection and prognosis. More specifically, the present invention relates to a signature based on differential gene expression in different conditions of vernal keratoconjunctivitis, for the detection and/or prognosis of the disease in a subject.
  • the main types of ocular allergy are seasonal allergic conjunctivitis (SAC), perennial allergic conjunctivitis (PAC), vernal keratoconjunctivitis (VKC), atopic keratoconjunctivitis (AKC), and giant papillary conjunctivitis (GPC).
  • SAC seasonal allergic conjunctivitis
  • PAC perennial allergic conjunctivitis
  • VKC vernal keratoconjunctivitis
  • ATC atopic keratoconjunctivitis
  • GPC giant papillary conjunctivitis
  • Diagnosis of allergic conjunctivitis is generally made by thorough history and careful clinical observation. Type I hypersensitivity reactions are important in these diseases, although they are not the only pathophysiologic mechanism.
  • VKC is a chronic, bilateral, and severe form of allergic inflammation affecting the ocular surface and can cause severe damage to the ocular surface, leading to corneal scarring and vision loss if not treated properly.
  • VKC Main clinical symptoms of VKC are ocular itching, watery eye, photophobia, and blurred vision, while signs are essentially stringy white mucous exudate, palpebral, limbal and corneal manifestations. Contrary to SAC, VKC involves various different types of immune reactions, with raised IgE levels in tears and serum, and mast cells and eosinophils in the conjunctival epithelium. It is therefore important to diagnose properly the allergic background responsible for VKC. For that purpose it is important to assess the biomarker profile of VKC patients. Indeed, markers may help in the diagnosis of VKC and in the prescription of the most appropriate treatment option as well as in a better follow-up of patient’s response to the treatment.
  • WO2012038574 discloses a method or diagnosing ocular allergy in a child by analyzing histamine, ECF, MBP, ECP, EDN and/or E-selectin markers.
  • this set of markers is used to diagnose several types of allergic conjunctivitis, i.e. seasonal conjunctivitis (SAC), perennial conjunctivitis (PAC) and vernal keratoconjunctivitis (VKC).
  • SAC seasonal conjunctivitis
  • PAC perennial conjunctivitis
  • VKC vernal keratoconjunctivitis
  • VKC have to be distinguished from other allergic conjunctivitis, in particular from AKC with which it shares the main symptoms and signs, ⁇ while the underlying mechanism might be different.
  • complications may arise without therapy to control symptoms.
  • the present invention relates to a signature of VKC of particular clinical relevance for the detection of this disease in a subject, as well as for the graduation of its severity for providing the appropriate treatment to patients.
  • the present invention relates to a method for the detection of vernal keratoconjunctivitis (VKC) in a subject, wherein the method comprises assessing the expression of markers of a signature comprising at least two VKC markers in a sample from said subject, wherein the at least two VKC markers are markers whose expression is different between a VKC patient and a healthy person.
  • the signature comprises at least 2 markers, preferably at least 3, preferably at least 4, more preferably at least 5 markers.
  • the VKC markers are selected from the list of markers of Table 1, fragments, variants and equivalents thereof.
  • the VKC markers are selected from the list of markers of Table 2, fragments, variants and equivalents thereof.
  • the VKC markers are selected from the list of markers of Table 3, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 4, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 5, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 6, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 7, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 8, fragments, variants and equivalents thereof.
  • the VKC markers are selected from the list of markers of Table 9, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 10, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 11, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 12, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 13, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 14, fragments, variants and equivalents thereof. In one embodiment, the sample is conjunctival superficial cells of said subject. According to an embodiment, the said subject is a human, preferably a child. In one embodiment, the method of the invention further comprises a step of comparing the expression of VKC markers with a reference expression profile. In one embodiment, the said method comprises the steps of:
  • the present invention also relates to a genechip specific for vernal keratoconjunctivitis (VKC), comprising at least 3 of the genes selected from the group comprising markers of the lists of Table 1, preferably of Table 2, more preferably of Table 3, even more preferably of Table 4, Table 5, Table 6, Table 7 Table 8, Table 9, Table 10, Table 11, Table 12, Table 13 or Table 14, fragments, variants and equivalents thereof.
  • VKC vernal keratoconjunctivitis
  • the present invention further relates to a kit for implementing the method according to the invention, wherein the kit comprises means for determining the expression of the markers of the signature.
  • Prognosis refers to the likelihood of vernal keratoconjunctivitis progression during the natural history of the disease, or to the likelihood of a beneficial response to a specific treatment, wherein a beneficial response means an improvement in any measure of patient status including, but not limited to, ocular itching, watery eye, photophobia, and blurred vision.
  • a“prognostic signature” refers to a signature that may be used for the prognosis of a subject.
  • the term “prognostic signature” also includes“predictive signature”, wherein said term refers to a signature that may be used for anticipating the response of a subject to a specific treatment.
  • Signature refers to a group of markers (i.e. at least 2, preferably at least 3, more preferably at least 5, and even more preferably at least 10 markers) whose combined expression profile is indicative of a biological condition, or of a particular prognosis or of a particular response of a subject to a treatment.
  • the signature of the invention comprises or consists of one marker.
  • A“marker” corresponds to a nucleotide sequence isolated from the genome, preferably to a gene in the genome, i.e. each marker is identifiable as all or a portion of a gene.
  • a marker may thus correspond to an entire gene, or to an EST (wherein EST stands for Expressed Sequence Tag) derived from this gene.
  • Expression refers interchangeably to expression of a marker, including the encoded polypeptide or protein.
  • Expression of a marker may be determined, for example, by immunoassay using one or more antibody(ies) that bind(s) with the polypeptide.
  • expression of a marker may be determined by measurement of mRNA levels, for example, by RT-PCR, RT-qPCR (wherein qPCR stands for quantitative PCR), or using a microarray, or using sequencing methods.
  • the term“expression” of a marker may also refer to modification of a protein or peptide, preferably to post-translational modification of a protein or peptide.
  • Subject or“patient” refers to an animal, preferably a mammal, more preferably a human.
  • the subject is a patient, i.e. a recipient of health care services.
  • the subject is a VKC patient or the subject is susceptible or suspected of having VKC.
  • the subject is a child, i.e. a person under the age of 18, preferably under the age of 14.
  • the present invention relates to a signature for VKC disease, wherein said signature comprises markers whose expression is different between a VKC patient and a“normal” patient, i.e. a patient who does not have VKC.
  • the term“normal patient” may be replaced by the term“healthy patient” or“healthy person”.
  • a VKC patient is a patient suffering from VKC.
  • the signature of the invention comprises at least 2 markers, preferably at least 3 markers, more preferably at least 5 markers, and even more preferably at least 8 markers.
  • Methods for determining markers are well-known from the skilled artisan, and include, without limitation, comparing the transcriptome (in an embodiment wherein expression relates to transcription of a marker) or proteome (in an embodiment wherein expression relates to translation of a marker) in a VKC patient and a“normal” patient.
  • An example of such a method, based on the comparison of transcriptomes, is presented in the Examples.
  • the genes are identified as differentially expressed in VKC, when there is at least about a 2 fold difference in expression from normal, at least about 2.5 fold, 3 fold, 5 fold and 10 fold.
  • the genes are identified as differentially expressed in VKC, when there is at least about a 20 fold difference in expression from normal, including at least about 25 fold, 30 fold, 35 fold, 40 fold, 45 fold, 50 fold, 60 fold, 70 fold, 80 fold and 90 fold.
  • the genes are identified as differentially expressed in VKC, when there is at least about a 100 fold difference in expression from normal, including at least about 125 fold, 150 fold, 200 fold, 300 fold, 400 fold, and 500 fold.
  • some genes can show a higher difference in expression than others. These genes can be more involved or alternatively, equally involved in the manifestation of disease as a gene that is less differentially expressed.
  • the difference in expression is positive or negative.
  • genes may be induced or down-regulated, respectively.
  • Markers whose expression is different between a VKC patient and a“normal” patient are hereinafter referred as“VKC markers”.
  • the present invention thus also relates to“VKC markers”.
  • VKC markers are selected from the list of the 335 VKC markers of Table 1 below, as well as their variants, fragments or equivalents.
  • Table 1 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 2.
  • a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence comprising at least 25 contiguous nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 contiguous nucleotides of said nucleotide sequence SEQ ID NO: X.
  • a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence comprising the nucleotide sequence SEQ ID NO: X and additional nucleic acids in 3’ and/or 5’ of SEQ ID NO: X, wherein the number of additional nucleic acids ranges from 1 to 500, preferably from 1 to 200, more preferably from 1 to 100 nucleotides.
  • a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence that typically differs from said nucleotide sequence SEQ ID NO: X in one or more substitutions, deletions, additions and/or insertions.
  • a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence of at least 25, preferably of at least 50, 100, 150, 200, 300, 400, 500, 1000, 1500, 2000 or 3000 nucleotides having at least 75%, 80%, 90%, 95%, or at least 96%, 97%, 98%, 99% identity with the nucleotide sequence SEQ ID NO: X.
  • identity refers to the degree of sequence relatedness between polypeptides, as determined by the number of matches between strings of two or more amino acid residues.“Identity” measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (i.e.,“algorithms”). Identity of related polypeptides can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.
  • Preferred computer program methods for determining identity between two sequences include the GCG program package, including GAP (Devereux et al., Nucl. Acid. Res. ⁇ 2, 387 (1984); Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul et al., J. MoI. Biol.215, 403- 410 (1990)).
  • the BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB/NLM/NIH Bethesda, Md. 20894; Altschul et al., supra).
  • NCBI National Center for Biotechnology Information
  • the well-known Smith Waterman algorithm may also be used to determine identity.
  • a fragment is a nucleotide sequence of at least 25 nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 nucleotides.
  • a fragment of a sequence SEQ ID NO: X is a sequence of at least 25 contiguous nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 contiguous nucleotides of SEQ ID NO: X.
  • an equivalent of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence, preferably a gene involved in the same pathway than the nucleotide sequence SEQ ID NO: X.
  • VKC markers are selected from the list of the 315 VKC markers of Table 2 below, as well as their variants, fragments or equivalents. Table 2 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 2.
  • VKC markers are selected from the list of the 206 VKC markers of Table 3 below, as well as their variants, fragments or equivalents.
  • Table 3 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 5.
  • VKC markers are selected from the list of the 193 VKC markers of Table 4 below, as well as their variants, fragments or equivalents.
  • Table 4 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 5.
  • VKC markers are selected from the list of the 149 VKC markers of Table 5 below, as well as their variants, fragments or equivalents.
  • Table 5 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 10.
  • VKC markers are selected from the list of the 138 VKC markers of Table 6 below, as well as their variants, fragments or equivalents.
  • Table 6 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 10.
  • VKC markers are selected from the list of the 95 VKC markers of Table 7 below, as well as their variants, fragments or equivalents.
  • Table 7 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 25.
  • VKC markers are selected from the list of the 86 VKC markers of Table 8 below, as well as their variants, fragments or equivalents.
  • Table 8 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 25.
  • VKC markers are selected from the list of the 63 VKC markers of Table 9 below, as well as their variants, fragments or equivalents.
  • Table 9 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 50.
  • VKC markers are selected from the list of the 57 VKC markers of Table 10 below, as well as their variants, fragments or equivalents.
  • Table 10 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 50.
  • VKC markers are selected from the list of the 36 VKC markers of Table 11 below, as well as their variants, fragments or equivalents.
  • Table 11 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 100.
  • VKC markers are selected from the list of the 32 VKC markers of Table 12 below, as well as their variants, fragments or equivalents.
  • Table 12 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 100.
  • VKC markers are selected from the list of the 14 VKC markers of Table 13 below, as well as their variants, fragments or equivalents.
  • Table 13 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 500.
  • VKC markers are selected from the list of the 12 VKC markers of Table 14 below, as well as their variants, fragments or equivalents.
  • Table 14 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 500.
  • the signature of the invention comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 VKC markers.
  • the signature of the invention comprises at least 1 marker, preferably at least 2 or 3 markers, selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, even more preferably from the list of Table 5, even more preferably from the list of Table 6, even more preferably from the list of Table 7.
  • the signature of the invention comprises at least 1 marker, preferably at least 2 or 3 markers, selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7, even more preferably from the list of Table 8, even more preferably from the list of Table 9, even more preferably from the list of Table 10, even more preferably from the list of Table 11, even more preferably from the list of Table 12, even more preferably from the list of Table 13, even more preferably from the list of Table 14.
  • the signature of the invention comprises or consists of 2 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises or consists of 2 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises or consists of 3 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises at least 3 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises or consists of 4 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises at least 4 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises or consists of 5 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises at least 5 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4.
  • the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14.
  • the signature of the invention comprises or consists of 1, 2, 3 markers or more selected from the list of Table 7 and 1, 2, 3 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6.
  • the signature of the invention comprises or consists of 1 marker selected from the list of Table 7, and 1, 2, 3, 4, 5 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6.
  • the signature of the invention comprises or consists of 2 markers selected from the list of Table 7, and 1, 2, 3, 4 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6.
  • the signature of the invention comprises or consists of 3 markers selected from the list of Table 7, and 1, 2, 3 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6.
  • the signature of the invention comprises or consists of 1, 2, 3 markers or more selected from the list of Table 14 and 1, 2, 3 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.
  • the signature of the invention comprises or consists of 1 marker selected from the list of Table 14, and 1, 2, 3, 4, 5 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.
  • the signature of the invention comprises or consists of 2 markers selected from the list of Table 14, and 1, 2, 3, 4 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.
  • the signature of the invention comprises or consists of 3 markers selected from the list of Table 14, and 1, 2, 3 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.
  • the VKC markers of the invention are selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the VKC markers of the invention are selected from the group comprising CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the VKC markers of the invention are selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention comprises at least 2 markers selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention comprises or consists of 2 markers selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises at least 3 markers selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention comprises or consists of 3 markers selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention comprises the markers CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention consists of the markers CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
  • the signature of the invention comprises or consists of CCL18 and CCL2.
  • the signature of the invention comprises or consists of CCL18 and CCL24.
  • the signature of the invention comprises or consists of CCL18 and CCL3.
  • the signature of the invention comprises or consists of CCL18 and CCL4.
  • the signature of the invention comprises or consists of CCL18 and CSF3R.
  • the signature of the invention comprises or consists of CCL18 and CXCR1.
  • the signature of the invention comprises or consists of CCL18 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2 and CCL24. In another embodiment, the signature of the invention comprises or consists of CCL2 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL2 and CCL4.
  • the signature of the invention comprises or consists of CCL2 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24 and CCL3.
  • the signature of the invention comprises or consists of CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL24 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24 and NLRP3.
  • the signature of the invention comprises or consists of CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3 and NLRP3.
  • the signature of the invention comprises or consists of CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R and CXCR1.
  • the signature of the invention comprises or consists of CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CXCR1 and LILRB2.
  • the signature of the invention comprises or consists of CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of LILRA5 and LILRB3.
  • the signature of the invention comprises or consists of LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL24. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL4.
  • the signature of the invention comprises or consists of CCL18, CCL2 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and NLRP3.
  • the signature of the invention comprises or consists of CCL18, CCL24 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and LILRB2.
  • the signature of the invention comprises or consists of CCL18, CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and LILRA5.
  • the signature of the invention comprises or consists of CCL18, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and LILRA5.
  • the signature of the invention comprises or consists of CCL18, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and LILRB2.
  • the signature of the invention comprises or consists of CCL18, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and NLRP3.
  • the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, LILRA5 and LILRB3.
  • the signature of the invention comprises or consists of CCL18, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CSF3R.
  • the signature of the invention comprises or consists of CCL2, CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and CCL4.
  • the signature of the invention comprises or consists of CCL2, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and NLRP3.
  • the signature of the invention comprises or consists of CCL2, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and NLRP3.
  • the signature of the invention comprises or consists of CCL2, CSF3R and CXCR1. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and IL1RL1. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRA5. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRB2. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRB3. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and IL1RL1.
  • the signature of the invention comprises or consists of CCL2, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRB3.
  • the signature of the invention comprises or consists of CCL2, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRB3 and NLRP3.
  • the signature of the invention comprises or consists of CCL24, CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRB3.
  • the signature of the invention comprises or consists of CCL24, CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRB3.
  • the signature of the invention comprises or consists of CCL24, CCL4 and NLRP3.
  • the signature of the invention comprises or consists of CCL24, CSF3R and CXCR1.
  • the signature of the invention comprises or consists of CCL24, CSF3R and IL1RL1.
  • the signature of the invention comprises or consists of CCL24, CSF3R and LILRA5.
  • the signature of the invention comprises or consists of CCL24, CSF3R and LILRB2.
  • the signature of the invention comprises or consists of CCL24, CSF3R and LILRB3.
  • the signature of the invention comprises or consists of CCL24, CSF3R and NLRP3.
  • the signature of the invention comprises or consists of CCL24, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRB2.
  • the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRB2 and NLRP3.
  • the signature of the invention comprises or consists of CCL24, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRB3.
  • the signature of the invention comprises or consists of CCL3, CCL4 and NLRP3.
  • the signature of the invention comprises or consists of CCL3, CSF3R and CXCR1.
  • the signature of the invention comprises or consists of CCL3, CSF3R and IL1RL1.
  • the signature of the invention comprises or consists of CCL3, CSF3R and LILRA5.
  • the signature of the invention comprises or consists of CCL3, CSF3R and LILRB2.
  • the signature of the invention comprises or consists of CCL3, CSF3R and LILRB3.
  • the signature of the invention comprises or consists of CCL3, CSF3R and NLRP3.
  • the signature of the invention comprises or consists of CCL3, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRB2.
  • the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRB2 and NLRP3.
  • the signature of the invention comprises or consists of CCL3, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and NLRP3.
  • the signature of the invention comprises or consists of CCL4, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRB2.
  • the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRB2 and NLRP3.
  • the signature of the invention comprises or consists of CCL4, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and NLRP3.
  • the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB3.
  • the signature of the invention comprises or consists of CSF3R, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRB2.
  • the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRB2 and NLRP3.
  • the signature of the invention comprises or consists of CXCR1, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRB2 and NLRP3.
  • the signature of the invention comprises or consists of IL1RL1, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of LILRA5, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB2, LILRB3 and NLRP3. In a particular embodiment of the invention, the signature of the invention comprises or consists of one, two or three of CCL4, CCL3 and CSF3R.
  • the signature of the invention comprises at least the three markers CCL4, CCL3 and CSF3R.
  • the signature of the invention consists of CCL4, CCL3 and CSF3R.
  • the signature of the invention comprises or consists of two, three or four of CCL4, CCL3, CSF3R and NLRP3.
  • the signature of the invention comprises at least the four markers CCL4, CCL3, CSF3R and NLRP3.
  • the signature of the invention consists of CCL4, CCL3, CSF3R and NLRP3.
  • the signature of the invention comprises or consists of two, three, four or five of CCL4, CCL3, CSF3R, NLRP3 and LILRB3. In a particular embodiment of the invention, the signature of the invention comprises at least the five markers CCL4, CCL3, CSF3R, NLRP3 and LILRB3. In a particular embodiment of the invention, the signature of the invention consists of CCL4, CCL3, CSF3R, NLRP3 and LILRB3.
  • the VKC marker of the invention is not TGFB. In one embodiment, the VKC marker of the invention is not PDGF. In one embodiment, the VKC marker of the invention is not TNF. In one embodiment, the VKC marker of the invention is not IL2R.
  • the VKC marker of the invention is not IL4. In one embodiment, the VKC marker of the invention is not IL6. In one embodiment, the VKC marker of the invention is not IL6R or IL6sR. In one embodiment, the VKC marker of the invention is not IL7. In one embodiment, the VKC marker of the invention is not IL8. In one embodiment, the VKC marker of the invention is not IFNG. In one embodiment, the VKC marker of the invention is not IL1B. In one embodiment, the VKC marker of the invention is not IL13. In one embodiment, the VKC marker of the invention is not IL5. In one embodiment, the VKC marker of the invention is not CD1A. In one embodiment, the VKC marker of the invention is not CD4.
  • the VKC marker of the invention is not CD45R0. In one embodiment, the VKC marker of the invention is not CD45RA. In one embodiment, the VKC marker of the invention is not HLA-DR. In one embodiment, the VKC marker of the invention is not MMP9. In one embodiment, the VKC marker of the invention is not MMP1. In one embodiment, the VKC marker of the invention is not TIMP-1. In one embodiment, the VKC marker of the invention is not IL10. In one embodiment, the signature of the invention is an array or a genechip that includes the genes that are identified as differentially expressed in VKC, which can be referred to as a“human VKC genechip”. In one embodiment, the array or the genechip of the invention is specific for VKC.
  • a genechip can be produced that includes only those genes that are expressed in mild disease or in severe disease.
  • a genechip can be produced with only those genes that are identified as possessing key roles in each aspect of the disease.
  • the array or the genechip of the invention comprises at least 2 of the genes selected from the group comprising markers of Table 1, preferably of Table 2, more preferably of Table 3, even more preferably of Table 4, Table 5, Table 6, Table 7, Table 8, Table 9, Table 10, Table 11, Table 12, Table 13 or Table 14, fragments, variants and equivalents thereof.
  • the array or the genechip of the invention comprises at least 2 of the genes according to the invention, preferably at least 3, 4, 5, 6, 7, 8, 9 or 10 of the genes according to the invention.
  • the present invention also relates to a signature as hereinabove described, for the identification of VKC in a subject.
  • the term“identification of VKC in a subject” means identification of a subject suffering from VKC.
  • the term“identification” may be replaced by“determination”,“detection” or“diagnostic”.
  • the present invention further relates to a signature as hereinabove described, for the prognosis of VKC in a subject.
  • the present invention further relates to a method for the detection of VKC in a subject, wherein said method comprises assessing the expression of markers in a sample of said subject, whose expressions are different between a VKC patient or a“normal” patient.
  • the method of the invention is a method to identify a subject suffering from VKC.
  • the present invention also relates to a method for the prognosis of the VKC severity in a subject.
  • the method of the invention is a non-invasive method.
  • the method of the invention is for determining a personalized course of treatment of the subject. Indeed, according to the prognosis obtained, a personalized treatment may be administered to the subject.
  • a personalized treatment may be administered to the subject.
  • the expression of at least 2, preferably of at least 3, more preferably of at least 4, even more preferably 5 markers is assessed.
  • the expression of at least 6 markers is assessed.
  • the subject is susceptible or suspected of having VKC.
  • the subject suffer from at least one of the following symptoms: ocular itching, watery eye, photophobia, and blurred vision.
  • the subject suffer from at least one of the following signs: stringy white mucous exudate, palpebral, limbal and corneal manifestations.
  • the subject is at risk of developing VKC.
  • the risks of developing VKC are family or medical history of allergies or atopic diseases, such as for example asthma, rhinitis, and eczema.
  • the subject is a VKC patient.
  • the severity of the disease is not yet determined.
  • the signature or the method may be for identifying the severity of the disease and thereby identifying the appropriate treatment, such as, for example, lubricating eye drops, mast cell stabilizers, corticosteroids, steroids, theophylline or antihistamines.
  • the signature or the method may be for assessing the likelihood of a beneficial response of the patient to a specific anti-VKC treatment.
  • the subject of the invention is young.
  • the term “young” means that the subject is at most 20 years old, at most 15 or 10 years old.
  • the subject is a child.
  • the term“child” refers to a human being (person) during the period between birth and puberty. By“puberty” it means the time in which sexual and physical characteristics mature person because of hormonal changes.
  • the present invention child is considered a person of up to 14 years (inclusive).
  • the subject is a man.
  • the subject is a woman.
  • the method of the invention for the detection of VKC or the prognosis of VKC severity in a subject comprises determining the expression profile of markers of a signature of the invention in a sample of said subject.
  • the sample was previously taken from the subject, i.e. the method of the invention does not comprise a step of recovering a sample from the subject. Consequently, according to this embodiment, the method of the invention is a non-invasive method.
  • sample includes fluid, cells or tissue can be obtained from the eye of a subject, such as, for example, conjunctiva, cornea, tears, aqueous humor, and the like. Such samples may be obtained by conventional methods known to those skilled in the art.
  • said biological sample is a sample of tears.
  • the term “tear” refers to aqueous fluid secreted by the lacrimal glands to wet and clean the conjunctiva of the eye through the eyelid movement.
  • the tear sample can be obtained by conventional methods known to those skilled in the art, for example by an absorbent, a capillary, one absorbing sponge, paper or similar device.
  • the sample can be obtained from subjects previously diagnosed or undiagnosed, as well as subjects undergoing treatment, or who have been previously treated for this disease.
  • the sample of the invention is a biological sample.
  • the sample is conjunctival superficial cells or tears.
  • the sample is conjunctival superficial cells.
  • the term“conjunctival superficial cells” means cells from superficial layers of the ocular surface epithelium.
  • the ocular surface epithelium englobes two, three or more layers of cells.
  • conjunctival superficial cells include conjunctival epithelial cells and inflammatory cells integrated in the tissue, i.e. the ocular surface epithelium.
  • the method of the invention comprises a step of comparing the expression profile of the markers of the signature of the invention measured in the sample of the subject with a reference expression profile, measured in a reference sample.
  • a reference expression profile can be relative to an expression profile derived from population studies, including without limitation, such subjects having similar age range, subjects in the same or similar ethnic group, similar VKC history and the like.
  • the reference expression profile is constructed using algorithms and other methods of statistical and structural classification.
  • the reference expression profile is derived from the measurement of the expression profile of markers of a signature of the invention in a control sample derived from one or more substantially healthy subjects, also called “normal” patients.
  • a“substantially healthy subject” has not been previously diagnosed or identified as having or suffering from VKC.
  • the reference expression profile is derived from the previous measurement of the expression profile of markers of a signature of the invention in a reference sample derived from the same subject, such as, for example, the expression profile measured one month before, preferably six months before, more preferably one year before or more.
  • the reference expression profile is derived from the measurement of the expression profile of markers of a signature of the invention in a reference population.
  • the reference sample is thus derived from a reference population.
  • the reference population comprises substantially healthy subjects, preferably at least 5, more preferably at least 10, more preferably at least 20 and even more preferably at least 50 substantially healthy subjects (or“normal” patients).
  • the reference expression profile corresponds to the mean expression profile of the markers of the signature of the invention measured in the reference population. In one embodiment of the invention, the reference expression profile corresponds to the median expression profile of the markers of the genetic signature of the invention measured in the reference population. In one embodiment of the invention, the expression of the VKC markers of the invention corresponds to the transcription level (i.e. expression of the RNA), or to the translation level (i.e. expression of the protein) of the marker. In one embodiment of the invention, the expression of the VKC markers is assessed at the protein level. Methods for determining a protein level in a sample are well-known in the art.
  • VKC markers are assessed at the RNA level. Methods for assessing the transcription level of a marker are well known in the prior art.
  • RT-PCR examples include, but are not limited to, RT-PCR, RT-qPCR, Northern Blot, hybridization techniques such as, for example, use of microarrays, and combination thereof including but not limited to, hybridization of amplicons obtained by RT-PCR, sequencing such as, for example, next-generation DNA sequencing (NGS) or RNA-seq (also known as “Whole Transcriptome Shotgun Sequencing”) and the like.
  • NGS next-generation DNA sequencing
  • RNA-seq also known as “Whole Transcriptome Shotgun Sequencing”
  • the method comprises the steps of:
  • total RNA from the sample are not retro-transcribing to obtain cDNA.
  • the method comprises the steps of:
  • the expression profile of markers of the signature of the invention is measured using a polynucleotide microarray, so that the expression profiles of each of the markers of the signature of the invention are simultaneously measured.
  • the method comprises the steps of:
  • the method comprises the steps of:
  • RNA preferably the total mRNA
  • sequencing the total RNA preferably the total mRNA from the sample from the subject
  • sequencing the total RNA preferably the total mRNA from the reference sample
  • sequencing the total RNA preferably the total mRNA from the reference sample
  • a marker of the invention is considered as differentially expressed in the sample from the subject as compared to a reference sample if both expression levels differ by a factor of at least 2, preferably at least 2.5, 3, 4 or 5, more preferably at least 10, 15, 20 or 25 and even more preferably at least 50, 100, 150, 200, 250, 300, 400 or 500.
  • the method of the invention further comprises a step of comparing the results of markers expression with signs and symptoms commonly tested in VKC. Accordingly, in one embodiment, the method of the invention comprises a step of comparing the results of markers expression with results of ocular itching, watery eye, photophobia, blurred vision, stringy white mucous exudate, palpebral, limbal and/or corneal manifestations.
  • the present invention also relates to a kit for measuring the expression profile of markers of the signature of the invention, and/or for implementing the method of the invention.
  • the kit comprises means for determining the expression of the markers of the signature of the invention.
  • the expression profile is measured at the protein level, and the kit of the invention comprises means for total protein extraction, as well as antibodies for detecting the markers of the invention.
  • the expression profile is measured at the RNA level, and the kit of the invention comprises means for total RNA extraction, means for reverse transcription of total RNA, and means for quantifying the expression of RNA corresponding to the markers of the invention.
  • the expression profile is measured at the RNA level
  • the kit of the invention comprises means for total RNA extraction and means for quantifying the expression of RNA corresponding to the markers of the invention, without reverse transcription of total RNA.
  • the means for determining the expression of the markers are PCR primers, for example qPCR primers, specific for said markers.
  • said means for determining the expression of the markers are probes to detect qPCR amplicons obtained with qPCR primers as hereinabove described.
  • said means for quantifying the expression of RNA corresponding to the markers of the invention is PCR, for example qPCR.
  • the kit of the invention also comprises primers for amplifying reference genes.
  • kits of the invention comprises means for total RNA extraction, means for reverse transcription of total RNA, and/or reagents for carrying out a quantitative PCR as hereinabove described (such as, for example, primers, buffers, enzyme, and the like).
  • kit of the invention comprises means for total RNA extraction, and means for quantification of the expression of RNA corresponding to the markers of the invention.
  • the kit of the invention also comprises a reference sample.
  • the means for determining the expression of the markers of the signature comprise probes specific for said markers.
  • probes are reporter and capture probes used generate a target-probes complex with RNA, and/or plates for counting of the reporter probes.
  • the means for determining the expression of the markers of the signature is a microarray comprising probes specific for said markers.
  • said means for quantifying the expression of RNA corresponding to the markers of the invention is a microarray.
  • the present invention thus also relates to microarrays for measuring the RNA expression profile of markers of the signature of the invention, and/or for implementing the non-invasive method of the invention.
  • the microarray of the invention comprises DNA probes, which may be hybridized to the retro-transcribed RNA corresponding to the markers of the invention.
  • VKC vernal keratoconjunctivitis
  • healthy individuals corresponding to“normal” patients
  • VKC patients were identified according to usual signs and symptoms for VKC, such as family and medical history, ocular itching, watery eye, photophobia, blurred vision, stringy white mucous exudate, palpebral, limbal and/or corneal manifestations.
  • Total RNAs were extracted according to standard procedures with commercially available extraction and purification kits.
  • RNA integrity number RIN
  • UV spectroscopy was used to quantitate mRNA.
  • NanoString nCounter technology with immunology human Code Set was used to analyze expressed transcripts (nCounter GX Human Immunology Kit). Briefly, total mRNA (100 ng) is hybridized with the reporter and capture probes to generate target-probes complex. The probes in excess are washed away and the target-probes complexes are bind onto the plate for counting of the reporter (colored) probes. The number of colored probes is directly linked to the number of target sequences and allows for a direct quantitation of the expressed target sequences.

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Abstract

The present invention relates to a signature comprising at least 2 or 3 markers. The present invention also relates to a method for the detection of vernal keratoconjunctivitis (VKC) in a subject, wherein said method comprises assessing the expression of markers of a signature of the invention in a sample from said subject; and to a kit for implementing this method.

Description

GENE SIGNATURE FOR THE DETECTION OF VERNAL
KERATOCONJUNCTIVITIS
FIELD OF INVENTION The present invention relates to the field of vernal keratoconjunctivitis detection and prognosis. More specifically, the present invention relates to a signature based on differential gene expression in different conditions of vernal keratoconjunctivitis, for the detection and/or prognosis of the disease in a subject.
BACKGROUND OF INVENTION The main types of ocular allergy are seasonal allergic conjunctivitis (SAC), perennial allergic conjunctivitis (PAC), vernal keratoconjunctivitis (VKC), atopic keratoconjunctivitis (AKC), and giant papillary conjunctivitis (GPC). Diagnosis of allergic conjunctivitis is generally made by thorough history and careful clinical observation. Type I hypersensitivity reactions are important in these diseases, although they are not the only pathophysiologic mechanism. In particular, VKC is a chronic, bilateral, and severe form of allergic inflammation affecting the ocular surface and can cause severe damage to the ocular surface, leading to corneal scarring and vision loss if not treated properly. Main clinical symptoms of VKC are ocular itching, watery eye, photophobia, and blurred vision, while signs are essentially stringy white mucous exudate, palpebral, limbal and corneal manifestations. Contrary to SAC, VKC involves various different types of immune reactions, with raised IgE levels in tears and serum, and mast cells and eosinophils in the conjunctival epithelium. It is therefore important to diagnose properly the allergic background responsible for VKC. For that purpose it is important to assess the biomarker profile of VKC patients. Indeed, markers may help in the diagnosis of VKC and in the prescription of the most appropriate treatment option as well as in a better follow-up of patient’s response to the treatment. Recently, the international patent application WO2012038574 discloses a method or diagnosing ocular allergy in a child by analyzing histamine, ECF, MBP, ECP, EDN and/or E-selectin markers. However, this set of markers is used to diagnose several types of allergic conjunctivitis, i.e. seasonal conjunctivitis (SAC), perennial conjunctivitis (PAC) and vernal keratoconjunctivitis (VKC). VKC have to be distinguished from other allergic conjunctivitis, in particular from AKC with which it shares the main symptoms and signs,^while the underlying mechanism might be different. Moreover, complications may arise without therapy to control symptoms. For example, in some patients symptoms may persist beyond childhood, which in some cases may represent a conversion to an adult form of AKC. Consequently, there exists a need to correctly identify VKC in patients. Therefore, the present invention relates to a signature of VKC of particular clinical relevance for the detection of this disease in a subject, as well as for the graduation of its severity for providing the appropriate treatment to patients.
SUMMARY The present invention relates to a method for the detection of vernal keratoconjunctivitis (VKC) in a subject, wherein the method comprises assessing the expression of markers of a signature comprising at least two VKC markers in a sample from said subject, wherein the at least two VKC markers are markers whose expression is different between a VKC patient and a healthy person. In one embodiment, the signature comprises at least 2 markers, preferably at least 3, preferably at least 4, more preferably at least 5 markers. In one embodiment, the VKC markers are selected from the list of markers of Table 1, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 2, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 3, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 4, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 5, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 6, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 7, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 8, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 9, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 10, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 11, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 12, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 13, fragments, variants and equivalents thereof. In another embodiment, the VKC markers are selected from the list of markers of Table 14, fragments, variants and equivalents thereof. In one embodiment, the sample is conjunctival superficial cells of said subject. According to an embodiment, the said subject is a human, preferably a child. In one embodiment, the method of the invention further comprises a step of comparing the expression of VKC markers with a reference expression profile. In one embodiment, the said method comprises the steps of:
- extracting total RNA from the sample from the subject,
- determining the expression profile of the markers of the signature, and - comparing said expression profile with a reference expression profile determined in a reference sample. The present invention also relates to a genechip specific for vernal keratoconjunctivitis (VKC), comprising at least 3 of the genes selected from the group comprising markers of the lists of Table 1, preferably of Table 2, more preferably of Table 3, even more preferably of Table 4, Table 5, Table 6, Table 7 Table 8, Table 9, Table 10, Table 11, Table 12, Table 13 or Table 14, fragments, variants and equivalents thereof. The present invention further relates to a kit for implementing the method according to the invention, wherein the kit comprises means for determining the expression of the markers of the signature.
DEFINITIONS
In the present invention, the following terms have the following meanings:
- “Prognosis” refers to the likelihood of vernal keratoconjunctivitis progression during the natural history of the disease, or to the likelihood of a beneficial response to a specific treatment, wherein a beneficial response means an improvement in any measure of patient status including, but not limited to, ocular itching, watery eye, photophobia, and blurred vision. Accordingly, a“prognostic signature” refers to a signature that may be used for the prognosis of a subject. In one embodiment, the term “prognostic signature” also includes“predictive signature”, wherein said term refers to a signature that may be used for anticipating the response of a subject to a specific treatment.
- “Signature” refers to a group of markers (i.e. at least 2, preferably at least 3, more preferably at least 5, and even more preferably at least 10 markers) whose combined expression profile is indicative of a biological condition, or of a particular prognosis or of a particular response of a subject to a treatment. In one embodiment, the signature of the invention comprises or consists of one marker.
- A“marker” corresponds to a nucleotide sequence isolated from the genome, preferably to a gene in the genome, i.e. each marker is identifiable as all or a portion of a gene. A marker may thus correspond to an entire gene, or to an EST (wherein EST stands for Expressed Sequence Tag) derived from this gene.
- “Expression” refers interchangeably to expression of a marker, including the encoded polypeptide or protein. Expression of a marker may be determined, for example, by immunoassay using one or more antibody(ies) that bind(s) with the polypeptide. Alternatively, expression of a marker may be determined by measurement of mRNA levels, for example, by RT-PCR, RT-qPCR (wherein qPCR stands for quantitative PCR), or using a microarray, or using sequencing methods. In one embodiment, the term“expression” of a marker may also refer to modification of a protein or peptide, preferably to post-translational modification of a protein or peptide.
- “Subject” or“patient” refers to an animal, preferably a mammal, more preferably a human. In one embodiment, the subject is a patient, i.e. a recipient of health care services. Preferably, the subject is a VKC patient or the subject is susceptible or suspected of having VKC. In one embodiment, the subject is a child, i.e. a person under the age of 18, preferably under the age of 14.
- “About”^preceding a figure means plus or less 10% of the value of said figure.
DETAILED DESCRIPTION The identification of biomarkers of inflammation and immunology involved in VKC will help in the characterization of the pathways implicated in disease development and severity and in the diagnosis of VKC. A better characterization of the disease background and a better understanding of the mechanism and pathways implicated in disease development will lead to a better management of its complication and of its treatment. Using microarray analysis, the expression of 579 mRNA was analyzed in patients suffering from VKC or in healthy people. Of the 579 genes analyzed, 335 were induced or down-regulated greater than 2-fold in VKC patients as compared to healthy control. The results of this analysis include a set of differentially expressed genes that can be used as a“signature” for the identification of VKC and for the severity of VKC. The present invention relates to a signature for VKC disease, wherein said signature comprises markers whose expression is different between a VKC patient and a“normal” patient, i.e. a patient who does not have VKC. Throughout the invention, the term“normal patient” may be replaced by the term“healthy patient” or“healthy person”. As used herein, a VKC patient is a patient suffering from VKC. In one embodiment, the signature of the invention comprises at least 2 markers, preferably at least 3 markers, more preferably at least 5 markers, and even more preferably at least 8 markers. Methods for determining markers are well-known from the skilled artisan, and include, without limitation, comparing the transcriptome (in an embodiment wherein expression relates to transcription of a marker) or proteome (in an embodiment wherein expression relates to translation of a marker) in a VKC patient and a“normal” patient. An example of such a method, based on the comparison of transcriptomes, is presented in the Examples. In one embodiment, the genes are identified as differentially expressed in VKC, when there is at least about a 2 fold difference in expression from normal, at least about 2.5 fold, 3 fold, 5 fold and 10 fold. In a further embodiment, the genes are identified as differentially expressed in VKC, when there is at least about a 20 fold difference in expression from normal, including at least about 25 fold, 30 fold, 35 fold, 40 fold, 45 fold, 50 fold, 60 fold, 70 fold, 80 fold and 90 fold. In a further embodiment, the genes are identified as differentially expressed in VKC, when there is at least about a 100 fold difference in expression from normal, including at least about 125 fold, 150 fold, 200 fold, 300 fold, 400 fold, and 500 fold. However, some genes can show a higher difference in expression than others. These genes can be more involved or alternatively, equally involved in the manifestation of disease as a gene that is less differentially expressed. In one embodiment, the difference in expression is positive or negative. In other words, in one embodiment, genes may be induced or down-regulated, respectively. Markers whose expression is different between a VKC patient and a“normal” patient are hereinafter referred as“VKC markers”. The present invention thus also relates to“VKC markers”. In one embodiment, VKC markers are selected from the list of the 335 VKC markers of Table 1 below, as well as their variants, fragments or equivalents. Table 1 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 2.
Figure imgf000008_0001
Figure imgf000009_0001
Figure imgf000010_0001
Figure imgf000011_0001
Figure imgf000012_0001
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Figure imgf000014_0001
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Figure imgf000021_0001
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Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
Figure imgf000027_0001
Figure imgf000028_0001
In one embodiment, a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence comprising at least 25 contiguous nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 contiguous nucleotides of said nucleotide sequence SEQ ID NO: X. In another embodiment, a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence comprising the nucleotide sequence SEQ ID NO: X and additional nucleic acids in 3’ and/or 5’ of SEQ ID NO: X, wherein the number of additional nucleic acids ranges from 1 to 500, preferably from 1 to 200, more preferably from 1 to 100 nucleotides. In another embodiment, a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence that typically differs from said nucleotide sequence SEQ ID NO: X in one or more substitutions, deletions, additions and/or insertions. In one embodiment, said substitutions, deletions, additions and/or insertions may affect 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleic acids. In another embodiment, a variant of a nucleotide sequence SEQ ID NO: X is a nucleotide sequence of at least 25, preferably of at least 50, 100, 150, 200, 300, 400, 500, 1000, 1500, 2000 or 3000 nucleotides having at least 75%, 80%, 90%, 95%, or at least 96%, 97%, 98%, 99% identity with the nucleotide sequence SEQ ID NO: X. The term“identity” or“identical”, when used in a relationship between the sequences of two or more polypeptides, refers to the degree of sequence relatedness between polypeptides, as determined by the number of matches between strings of two or more amino acid residues.“Identity” measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (i.e.,“algorithms”). Identity of related polypeptides can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM J. Applied Math. 48, 1073 (1988). Preferred methods for determining identity are designed to give the largest match between the sequences tested. Methods of determining identity are described in publicly available computer programs. Preferred computer program methods for determining identity between two sequences include the GCG program package, including GAP (Devereux et al., Nucl. Acid. Res. \2, 387 (1984); Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul et al., J. MoI. Biol.215, 403- 410 (1990)). The BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB/NLM/NIH Bethesda, Md. 20894; Altschul et al., supra). The well-known Smith Waterman algorithm may also be used to determine identity. In one embodiment of the invention, a fragment is a nucleotide sequence of at least 25 nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 nucleotides. In one embodiment of the invention, a fragment of a sequence SEQ ID NO: X is a sequence of at least 25 contiguous nucleotides, preferably of at least 50, 100, 150, 200 or at least 500 contiguous nucleotides of SEQ ID NO: X. In one embodiment, an equivalent of a nucleotide sequence SEQ ID NO: X, preferably of a gene having the sequence SEQ ID NO: X, is a nucleotide sequence, preferably a gene involved in the same pathway than the nucleotide sequence SEQ ID NO: X. In another embodiment, VKC markers are selected from the list of the 315 VKC markers of Table 2 below, as well as their variants, fragments or equivalents. Table 2 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 2.
Figure imgf000030_0001
Figure imgf000031_0001
Figure imgf000032_0001
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Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
In another embodiment, VKC markers are selected from the list of the 206 VKC markers of Table 3 below, as well as their variants, fragments or equivalents. Table 3 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 5.
Figure imgf000049_0001
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Figure imgf000060_0001
Figure imgf000061_0001
In another embodiment, VKC markers are selected from the list of the 193 VKC markers of Table 4 below, as well as their variants, fragments or equivalents. Table 4 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 5.
Figure imgf000061_0002
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Figure imgf000071_0001
Figure imgf000072_0001
In another embodiment, VKC markers are selected from the list of the 149 VKC markers of Table 5 below, as well as their variants, fragments or equivalents. Table 5 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 10.
Figure imgf000073_0001
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Figure imgf000081_0001
Figure imgf000082_0001
In another embodiment, VKC markers are selected from the list of the 138 VKC markers of Table 6 below, as well as their variants, fragments or equivalents. Table 6 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 10.
Figure imgf000082_0002
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Figure imgf000089_0001
Figure imgf000090_0002
In another embodiment, VKC markers are selected from the list of the 95 VKC markers of Table 7 below, as well as their variants, fragments or equivalents. Table 7 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 25.
Figure imgf000090_0001
Figure imgf000091_0001
Figure imgf000092_0001
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Figure imgf000095_0001
Figure imgf000096_0001
In another embodiment, VKC markers are selected from the list of the 86 VKC markers of Table 8 below, as well as their variants, fragments or equivalents. Table 8 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 25.
Figure imgf000096_0002
Figure imgf000097_0001
Figure imgf000098_0001
Figure imgf000099_0001
Figure imgf000100_0001
Figure imgf000101_0001
In another embodiment, VKC markers are selected from the list of the 63 VKC markers of Table 9 below, as well as their variants, fragments or equivalents. Table 9 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 50.
Figure imgf000102_0001
Figure imgf000103_0001
Figure imgf000104_0001
Figure imgf000105_0001
In another embodiment, VKC markers are selected from the list of the 57 VKC markers of Table 10 below, as well as their variants, fragments or equivalents. Table 10 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 50.
Figure imgf000106_0001
Figure imgf000107_0001
Figure imgf000108_0001
Figure imgf000109_0001
In another embodiment, VKC markers are selected from the list of the 36 VKC markers of Table 11 below, as well as their variants, fragments or equivalents. Table 11 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 100.
Figure imgf000109_0002
Figure imgf000110_0001
Figure imgf000111_0001
In another embodiment, VKC markers are selected from the list of the 32 VKC markers of Table 12 below, as well as their variants, fragments or equivalents. Table 12 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 100.
Figure imgf000112_0001
Figure imgf000113_0001
Figure imgf000114_0001
In another embodiment, VKC markers are selected from the list of the 14 VKC markers of Table 13 below, as well as their variants, fragments or equivalents. Table 13 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 500.
Figure imgf000114_0002
Figure imgf000115_0001
Table 13 In another embodiment, VKC markers are selected from the list of the 12 VKC markers of Table 14 below, as well as their variants, fragments or equivalents. Table 14 comprises VKC markers identified in the conditions of the Example and presenting a fold-change, positive or negative, superior or equal to 500.
Figure imgf000115_0002
Figure imgf000116_0001
In one embodiment of the invention, the signature of the invention comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 VKC markers. In one embodiment of the invention, the signature of the invention comprises at least 1 marker, preferably at least 2 or 3 markers, selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, even more preferably from the list of Table 5, even more preferably from the list of Table 6, even more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 1 marker, preferably at least 2 or 3 markers, selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7, even more preferably from the list of Table 8, even more preferably from the list of Table 9, even more preferably from the list of Table 10, even more preferably from the list of Table 11, even more preferably from the list of Table 12, even more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 2 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 3 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 3 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 4 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 4 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of 5 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises at least 5 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 5, preferably from the list of Table 6, more preferably from the list of Table 7. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 8, preferably from the list of Table 9, more preferably from the list of Table 10. In one embodiment of the invention, the signature of the invention comprises or consists of at least 6 markers selected from the list of Table 11, preferably from the list of Table 12, more preferably from the list of Table 13, even more preferably from the list of Table 14. In one embodiment, the signature of the invention comprises or consists of 1, 2, 3 markers or more selected from the list of Table 7 and 1, 2, 3 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6. In one embodiment, the signature of the invention comprises or consists of 1 marker selected from the list of Table 7, and 1, 2, 3, 4, 5 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6. In another embodiment, the signature of the invention comprises or consists of 2 markers selected from the list of Table 7, and 1, 2, 3, 4 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6. In another embodiment, the signature of the invention comprises or consists of 3 markers selected from the list of Table 7, and 1, 2, 3 markers or more selected from the list of Table 1, preferably from the list of Table 2, more preferably from the list of Table 3, even more preferably from the list of Table 4, 5 or 6. In one embodiment, the signature of the invention comprises or consists of 1, 2, 3 markers or more selected from the list of Table 14 and 1, 2, 3 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13. In one embodiment, the signature of the invention comprises or consists of 1 marker selected from the list of Table 14, and 1, 2, 3, 4, 5 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13. In another embodiment, the signature of the invention comprises or consists of 2 markers selected from the list of Table 14, and 1, 2, 3, 4 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13. In another embodiment, the signature of the invention comprises or consists of 3 markers selected from the list of Table 14, and 1, 2, 3 or more other markers selected from the list of Table 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13. In one embodiment, the VKC markers of the invention are selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the VKC markers of the invention are selected from the group comprising CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the VKC markers of the invention are selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises at least 2 markers selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of 2 markers selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises at least 3 markers selected from the group comprising or consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of 3 markers selected from the group consisting of CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises the markers CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention consists of the markers CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18 and CCL2. In another embodiment, the signature of the invention comprises or consists of CCL18 and CCL24. In another embodiment, the signature of the invention comprises or consists of CCL18 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL18 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2 and CCL24. In another embodiment, the signature of the invention comprises or consists of CCL2 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL2 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL2 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL24 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL24. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL24 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL18, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL18, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL18, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CCL3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL24 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and CXCR1. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and IL1RL1. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRA5. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRB2. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and LILRB3. In one embodiment, the signature of the invention comprises or consists of CCL2, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL2, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL2, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and CCL4. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL24, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL24, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and CSF3R. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, CCL4 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL3, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL3, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and CXCR1. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, CSF3R and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CCL4, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CCL4, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and IL1RL1. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, CXCR1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CSF3R, LILRA5 and LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CSF3R, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRA5. In another embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, IL1RL1 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of CXCR1, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of CXCR1, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and LILRB2. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRA5 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of IL1RL1, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of IL1RL1, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5, LILRB2 and LILRB3. In another embodiment, the signature of the invention comprises or consists of LILRA5, LILRB2 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRA5, LILRB3 and NLRP3. In one embodiment, the signature of the invention comprises or consists of LILRB2, LILRB3 and NLRP3. In a particular embodiment of the invention, the signature of the invention comprises or consists of one, two or three of CCL4, CCL3 and CSF3R. In a particular embodiment of the invention, the signature of the invention comprises at least the three markers CCL4, CCL3 and CSF3R. In a particular embodiment of the invention, the signature of the invention consists of CCL4, CCL3 and CSF3R. In a particular embodiment of the invention, the signature of the invention comprises or consists of two, three or four of CCL4, CCL3, CSF3R and NLRP3. In a particular embodiment of the invention, the signature of the invention comprises at least the four markers CCL4, CCL3, CSF3R and NLRP3. In a particular embodiment of the invention, the signature of the invention consists of CCL4, CCL3, CSF3R and NLRP3. In a particular embodiment of the invention, the signature of the invention comprises or consists of two, three, four or five of CCL4, CCL3, CSF3R, NLRP3 and LILRB3. In a particular embodiment of the invention, the signature of the invention comprises at least the five markers CCL4, CCL3, CSF3R, NLRP3 and LILRB3. In a particular embodiment of the invention, the signature of the invention consists of CCL4, CCL3, CSF3R, NLRP3 and LILRB3.In one embodiment, the VKC marker of the invention is not TGFB. In one embodiment, the VKC marker of the invention is not PDGF. In one embodiment, the VKC marker of the invention is not TNF. In one embodiment, the VKC marker of the invention is not IL2R. In one embodiment, the VKC marker of the invention is not IL4. In one embodiment, the VKC marker of the invention is not IL6. In one embodiment, the VKC marker of the invention is not IL6R or IL6sR. In one embodiment, the VKC marker of the invention is not IL7. In one embodiment, the VKC marker of the invention is not IL8. In one embodiment, the VKC marker of the invention is not IFNG. In one embodiment, the VKC marker of the invention is not IL1B. In one embodiment, the VKC marker of the invention is not IL13. In one embodiment, the VKC marker of the invention is not IL5. In one embodiment, the VKC marker of the invention is not CD1A. In one embodiment, the VKC marker of the invention is not CD4. In one embodiment, the VKC marker of the invention is not CD45R0. In one embodiment, the VKC marker of the invention is not CD45RA. In one embodiment, the VKC marker of the invention is not HLA-DR. In one embodiment, the VKC marker of the invention is not MMP9. In one embodiment, the VKC marker of the invention is not MMP1. In one embodiment, the VKC marker of the invention is not TIMP-1. In one embodiment, the VKC marker of the invention is not IL10. In one embodiment, the signature of the invention is an array or a genechip that includes the genes that are identified as differentially expressed in VKC, which can be referred to as a“human VKC genechip”. In one embodiment, the array or the genechip of the invention is specific for VKC. A variety of genechips can be produced that are specific to different aspects of the disease. In one embodiment, a genechip can be produced that includes only those genes that are expressed in mild disease or in severe disease. In a further embodiment, a genechip can be produced with only those genes that are identified as possessing key roles in each aspect of the disease. In one embodiment, the array or the genechip of the invention comprises at least 2 of the genes selected from the group comprising markers of Table 1, preferably of Table 2, more preferably of Table 3, even more preferably of Table 4, Table 5, Table 6, Table 7, Table 8, Table 9, Table 10, Table 11, Table 12, Table 13 or Table 14, fragments, variants and equivalents thereof. In one embodiment, the array or the genechip of the invention comprises at least 2 of the genes according to the invention, preferably at least 3, 4, 5, 6, 7, 8, 9 or 10 of the genes according to the invention. The present invention also relates to a signature as hereinabove described, for the identification of VKC in a subject. As used herein, the term“identification of VKC in a subject” means identification of a subject suffering from VKC. In one embodiment, the term“identification” may be replaced by“determination”,“detection” or“diagnostic”. The present invention further relates to a signature as hereinabove described, for the prognosis of VKC in a subject. The present invention further relates to a method for the detection of VKC in a subject, wherein said method comprises assessing the expression of markers in a sample of said subject, whose expressions are different between a VKC patient or a“normal” patient. In one embodiment, the method of the invention is a method to identify a subject suffering from VKC. The present invention also relates to a method for the prognosis of the VKC severity in a subject. In one embodiment, the more the VKC marker of the invention is differentially expressed (upregulated or down-regulated) compared to a healthy person, the more severe is the disease in the subject. In one embodiment, the method of the invention is a non-invasive method. In one embodiment of the invention, the method of the invention is for determining a personalized course of treatment of the subject. Indeed, according to the prognosis obtained, a personalized treatment may be administered to the subject. In one embodiment of the invention, the expression of at least 2, preferably of at least 3, more preferably of at least 4, even more preferably 5 markers is assessed. In one embodiment of the invention, the expression of at least 6 markers is assessed. In one embodiment, the subject is susceptible or suspected of having VKC. In one embodiment, the subject suffer from at least one of the following symptoms: ocular itching, watery eye, photophobia, and blurred vision. In a further embodiment, the subject suffer from at least one of the following signs: stringy white mucous exudate, palpebral, limbal and corneal manifestations. In one embodiment, the subject is at risk of developing VKC. In one embodiment, the risks of developing VKC are family or medical history of allergies or atopic diseases, such as for example asthma, rhinitis, and eczema. In one embodiment, the subject is a VKC patient. According to this embodiment, the severity of the disease is not yet determined. Still according to this embodiment, the signature or the method may be for identifying the severity of the disease and thereby identifying the appropriate treatment, such as, for example, lubricating eye drops, mast cell stabilizers, corticosteroids, steroids, theophylline or antihistamines. In one embodiment, the signature or the method may be for assessing the likelihood of a beneficial response of the patient to a specific anti-VKC treatment. In one embodiment, the subject of the invention is young. As used herein, the term “young” means that the subject is at most 20 years old, at most 15 or 10 years old. In one embodiment, the subject is a child. As used herein, the term“child” refers to a human being (person) during the period between birth and puberty. By“puberty” it means the time in which sexual and physical characteristics mature person because of hormonal changes. In a particular embodiment, the present invention child is considered a person of up to 14 years (inclusive). In one embodiment, the subject is a man. In another embodiment, the subject is a woman. In one embodiment of the invention, the method of the invention for the detection of VKC or the prognosis of VKC severity in a subject comprises determining the expression profile of markers of a signature of the invention in a sample of said subject. According to a preferred embodiment, the sample was previously taken from the subject, i.e. the method of the invention does not comprise a step of recovering a sample from the subject. Consequently, according to this embodiment, the method of the invention is a non-invasive method. The term“sample”, as used herein, includes fluid, cells or tissue can be obtained from the eye of a subject, such as, for example, conjunctiva, cornea, tears, aqueous humor, and the like. Such samples may be obtained by conventional methods known to those skilled in the art. In a particular embodiment, said biological sample is a sample of tears. The term “tear” refers to aqueous fluid secreted by the lacrimal glands to wet and clean the conjunctiva of the eye through the eyelid movement. The tear sample can be obtained by conventional methods known to those skilled in the art, for example by an absorbent, a capillary, one absorbing sponge, paper or similar device. The sample can be obtained from subjects previously diagnosed or undiagnosed, as well as subjects undergoing treatment, or who have been previously treated for this disease. In one embodiment, the sample of the invention is a biological sample. In one embodiment of the invention, the sample is conjunctival superficial cells or tears. In a preferred embodiment, the sample is conjunctival superficial cells. As used herein, the term“conjunctival superficial cells” means cells from superficial layers of the ocular surface epithelium. In one embodiment, the ocular surface epithelium englobes two, three or more layers of cells. In one embodiment, conjunctival superficial cells include conjunctival epithelial cells and inflammatory cells integrated in the tissue, i.e. the ocular surface epithelium. In one embodiment of the invention, the method of the invention comprises a step of comparing the expression profile of the markers of the signature of the invention measured in the sample of the subject with a reference expression profile, measured in a reference sample. A reference expression profile can be relative to an expression profile derived from population studies, including without limitation, such subjects having similar age range, subjects in the same or similar ethnic group, similar VKC history and the like. In one embodiment, the reference expression profile is constructed using algorithms and other methods of statistical and structural classification. In one embodiment of the invention, the reference expression profile is derived from the measurement of the expression profile of markers of a signature of the invention in a control sample derived from one or more substantially healthy subjects, also called “normal” patients. As used herein, a“substantially healthy subject” has not been previously diagnosed or identified as having or suffering from VKC. In one embodiment of the invention, the reference expression profile is derived from the previous measurement of the expression profile of markers of a signature of the invention in a reference sample derived from the same subject, such as, for example, the expression profile measured one month before, preferably six months before, more preferably one year before or more. In another embodiment of the invention, the reference expression profile is derived from the measurement of the expression profile of markers of a signature of the invention in a reference population. In one embodiment, the reference sample is thus derived from a reference population. In one embodiment, the reference population comprises substantially healthy subjects, preferably at least 5, more preferably at least 10, more preferably at least 20 and even more preferably at least 50 substantially healthy subjects (or“normal” patients). In one embodiment, the reference expression profile corresponds to the mean expression profile of the markers of the signature of the invention measured in the reference population. In one embodiment of the invention, the reference expression profile corresponds to the median expression profile of the markers of the genetic signature of the invention measured in the reference population. In one embodiment of the invention, the expression of the VKC markers of the invention corresponds to the transcription level (i.e. expression of the RNA), or to the translation level (i.e. expression of the protein) of the marker. In one embodiment of the invention, the expression of the VKC markers is assessed at the protein level. Methods for determining a protein level in a sample are well-known in the art. Examples of such methods include, but are not limited to, immunohistochemistry, Multiplex methods (Luminex), western blot, enzyme-linked immunosorbent assay (ELISA), sandwich ELISA, fluorescent-linked immunosorbent assay (FLISA), enzyme immunoassay (EIA), radioimmunoassay (RIA) and the like. In another embodiment of the invention, the expression of the VKC markers is assessed at the RNA level. Methods for assessing the transcription level of a marker are well known in the prior art. Examples of such methods include, but are not limited to, RT- PCR, RT-qPCR, Northern Blot, hybridization techniques such as, for example, use of microarrays, and combination thereof including but not limited to, hybridization of amplicons obtained by RT-PCR, sequencing such as, for example, next-generation DNA sequencing (NGS) or RNA-seq (also known as “Whole Transcriptome Shotgun Sequencing”) and the like. In one embodiment, the method comprises the steps of:
- extracting total RNA from the sample from the subject, - determining the expression profile of the markers of the signature, and - comparing said expression profile with a reference expression profile determined in a reference sample. According to this embodiment, total RNA from the sample are not retro-transcribing to obtain cDNA. In another embodiment, the method comprises the steps of:
- extracting total RNA from the sample from the subject,
- retro-transcribing these total RNA, thereby obtaining total cDNA,
- specifically amplifying by PCR, preferably by qPCR, the cDNA corresponding to the VKC markers of the signature of the invention, thereby determining the expression profile of the markers of the signature, and - comparing said expression profile with a reference expression profile determined in a reference sample. In one embodiment, the expression profile of markers of the signature of the invention is measured using a polynucleotide microarray, so that the expression profiles of each of the markers of the signature of the invention are simultaneously measured. In one embodiment, the method comprises the steps of:
- extracting total RNA from the sample from the subject, retro-transcribing these total RNA, thereby obtaining total cDNA from the sample, and sequencing the total cDNA from the sample from the subject,
- extracting total RNA from the reference sample, retro-transcribing these total RNA, thereby obtaining total cDNA from the reference sample, and sequencing the total cDNA from the reference sample, and
- comparing the results of the cDNA sequencing and identifying markers which are differentially expressed between the sample from the subject and the reference sample. In another embodiment, the method comprises the steps of:
- extracting total RNA from the sample from the subject, and sequencing the total RNA, preferably the total mRNA, from the sample from the subject, - extracting total RNA from the reference sample, and sequencing the total RNA, preferably the total mRNA from the reference sample, and - comparing the results of the RNA, preferably mRNA, sequencing and identifying markers which are differentially expressed between the sample from the subject and the reference sample. In one embodiment of the invention, a marker of the invention is considered as differentially expressed in the sample from the subject as compared to a reference sample if both expression levels differ by a factor of at least 2, preferably at least 2.5, 3, 4 or 5, more preferably at least 10, 15, 20 or 25 and even more preferably at least 50, 100, 150, 200, 250, 300, 400 or 500. In one embodiment, the method of the invention further comprises a step of comparing the results of markers expression with signs and symptoms commonly tested in VKC. Accordingly, in one embodiment, the method of the invention comprises a step of comparing the results of markers expression with results of ocular itching, watery eye, photophobia, blurred vision, stringy white mucous exudate, palpebral, limbal and/or corneal manifestations. The present invention also relates to a kit for measuring the expression profile of markers of the signature of the invention, and/or for implementing the method of the invention. In one embodiment, the kit comprises means for determining the expression of the markers of the signature of the invention. In one embodiment of the invention, the expression profile is measured at the protein level, and the kit of the invention comprises means for total protein extraction, as well as antibodies for detecting the markers of the invention. In another embodiment, the expression profile is measured at the RNA level, and the kit of the invention comprises means for total RNA extraction, means for reverse transcription of total RNA, and means for quantifying the expression of RNA corresponding to the markers of the invention. In another embodiment, the expression profile is measured at the RNA level, and the kit of the invention comprises means for total RNA extraction and means for quantifying the expression of RNA corresponding to the markers of the invention, without reverse transcription of total RNA. In one embodiment, the means for determining the expression of the markers are PCR primers, for example qPCR primers, specific for said markers. In one embodiment, said means for determining the expression of the markers are probes to detect qPCR amplicons obtained with qPCR primers as hereinabove described. In one embodiment, said means for quantifying the expression of RNA corresponding to the markers of the invention is PCR, for example qPCR. In one embodiment of the invention, the kit of the invention also comprises primers for amplifying reference genes. Reference genes are genes expressed at a constant level among different tissues and/or conditions. Examples of reference genes include, but are not limited to, E-actin, genes encoding ribosomal proteins and the like. In one embodiment of the invention, the kit of the invention comprises means for total RNA extraction, means for reverse transcription of total RNA, and/or reagents for carrying out a quantitative PCR as hereinabove described (such as, for example, primers, buffers, enzyme, and the like). In one embodiment of the invention, the kit of the invention comprises means for total RNA extraction, and means for quantification of the expression of RNA corresponding to the markers of the invention. In one embodiment, the kit of the invention also comprises a reference sample. In one embodiment, the means for determining the expression of the markers of the signature comprise probes specific for said markers. In one embodiment, probes are reporter and capture probes used generate a target-probes complex with RNA, and/or plates for counting of the reporter probes. In another embodiment, the means for determining the expression of the markers of the signature is a microarray comprising probes specific for said markers. In one embodiment, said means for quantifying the expression of RNA corresponding to the markers of the invention is a microarray. The present invention thus also relates to microarrays for measuring the RNA expression profile of markers of the signature of the invention, and/or for implementing the non-invasive method of the invention. In one embodiment of the invention, the microarray of the invention comprises DNA probes, which may be hybridized to the retro-transcribed RNA corresponding to the markers of the invention.
EXAMPLES The present invention is further illustrated by the following example. Materials and Methods Conjunctival superficial cells were harvested by impression cytology from patients with vernal keratoconjunctivitis (VKC) of various etiologies and from healthy individuals (corresponding to“normal” patients). VKC patients were identified according to usual signs and symptoms for VKC, such as family and medical history, ocular itching, watery eye, photophobia, blurred vision, stringy white mucous exudate, palpebral, limbal and/or corneal manifestations. Total RNAs were extracted according to standard procedures with commercially available extraction and purification kits. The quality and quantity of mRNA was confirmed via the determination of the RNA integrity number (RIN) with an Agilent 2100 bioanalyzer, and UV spectroscopy was used to quantitate mRNA. NanoString nCounter technology with immunology human Code Set was used to analyze expressed transcripts (nCounter GX Human Immunology Kit). Briefly, total mRNA (100 ng) is hybridized with the reporter and capture probes to generate target-probes complex. The probes in excess are washed away and the target-probes complexes are bind onto the plate for counting of the reporter (colored) probes. The number of colored probes is directly linked to the number of target sequences and allows for a direct quantitation of the expressed target sequences. Expression levels are tabulated in Excel files and analyzed relatively to the controls to give“fold-change” values. Results Lists of markers differentially expressed between VKC patients and“normal” patients with a fold-change superior or equal to 2 are presented in Table 15. The average“fold- change” is also indicated. Table 15: Lists of markers differentially expressed between VKC patients and“normal” patients.
Figure imgf000146_0001
Figure imgf000147_0001
Figure imgf000148_0001
Figure imgf000149_0001
Figure imgf000150_0001

Claims

CLAIMS 1. A method for the detection of vernal keratoconjunctivitis (VKC) in a subject, wherein said method comprises assessing the expression of markers of a signature comprising at least two VKC markers in a sample from said subject, wherein said at least two VKC markers are selected from the list of markers of Table 2, fragments, variants and equivalents thereof.
2. The method according to claim 1, wherein said signature comprises at least 3 markers, preferably at least 4 markers, more preferably at least 5 markers.
3. The method according to claim 1 or 2, wherein said VKC markers are selected from the list of markers of Table 3, fragments, variants and equivalents thereof.
4. The method according to anyone of claims 1 to 3, wherein VKC markers are selected from the list of markers of Table 4, fragments, variants and equivalents thereof.
5. The method according to anyone of claims 1 to 4, wherein VKC markers are selected from the list of markers of Table 5, fragments, variants and equivalents thereof.
6. The method according to anyone of claims 1 to 5, wherein VKC markers are selected from the list of markers of Table 6, fragments, variants and equivalents thereof.
7. The method according to anyone of claims 1 to 6, wherein VKC markers are selected from the list of markers of Table 7, fragments, variants and equivalents thereof.
8. The method according to anyone of claims 1 to 7, wherein VKC markers are selected from the list of markers of Table 8, fragments, variants and equivalents thereof.
9. The method according to anyone of claims 1 to 8, wherein VKC markers are selected from the list of markers of Table 9, fragments, variants and equivalents thereof.
10. The method according to anyone of claims 1 to 9, wherein VKC markers are selected from the list of markers of Table 10, fragments, variants and equivalents thereof.
11. The method according to anyone of claims 1 to 10, wherein VKC markers are selected from the list of markers of Table 11, fragments, variants and equivalents thereof.
12. The method according to anyone of claims 1 to 11, wherein VKC markers are selected from the list of markers of Table 12, fragments, variants and equivalents thereof.
13. The method according to anyone of claims 1 to 12, wherein VKC markers are selected from the list of markers of Table 13, fragments, variants and equivalents thereof.
14. The method according to anyone of claims 1 to 13, wherein VKC markers are selected from the list comprising CCL18, CCL2, CCL24, CCL3, CCL4, CSF3R, CXCR1, IL1RL1, LILRA5, LILRB2, LILRB3 and NLRP3.
15. The method according to anyone of claims 1 to 14, wherein said sample is conjunctival superficial cells or tears of said subject.
16. The method according to anyone of claims 1 to 15, wherein said subject is a human, preferably a child.
17. The method according to anyone of claims 1 to 16, further comprising comparing said expression with a reference expression profile.
18. The method according to anyone of claims 1 to 17, wherein said method comprises the steps of: - extracting total RNA from the sample from the subject,
- determining the expression profile of the markers of the signature, and^ - comparing said expression profile with a reference expression profile determined in a reference sample.
19. A genechip specific for vernal keratoconjunctivitis (VKC), comprising at least 3 of the genes selected from the group comprising markers of the list of Table 2, more preferably of Table 3, even more preferably of Table 4, Table 5, Table 6 or Table 7, fragments, variants and equivalents thereof.
20. A kit for implementing the method according to anyone of claims 1 to 18, wherein said kit comprises means for determining the expression of the markers of the signature.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012038574A1 (en) 2010-09-20 2012-03-29 Bioftalmik S.L. Method for diagnosing an eye allergy in a child
KR20150145986A (en) * 2014-06-20 2015-12-31 연세대학교 산학협력단 Diagnosis kit for ocular disease

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012038574A1 (en) 2010-09-20 2012-03-29 Bioftalmik S.L. Method for diagnosing an eye allergy in a child
KR20150145986A (en) * 2014-06-20 2015-12-31 연세대학교 산학협력단 Diagnosis kit for ocular disease

Non-Patent Citations (20)

* Cited by examiner, † Cited by third party
Title
"Affymetrix GeneChip Human Genome U133 Array Set HG-U133A", pages 1 - 3,19,289, XP002751195, Retrieved from the Internet <URL:Affymetrix GeneChip Human Genome U133 Array Set HG-U133A> [retrieved on 20020311] *
"Biocomputing: Informatics and Genome Projects", 1993, ACADEMIC PRESS
"Computational Molecular Biology", 1988, OXFORD UNIVERSITY PRESS
"Computer Analysis of Sequence Data", 1994, HUMANA PRESS
"Sequence Analysis Primer", 1991, M. STOCKTON PRESS
A M A. EL-ASRAR ET AL: "Expression of T lymphocyte chemoattractants and activation markers in vernal keratoconjunctivitis", BRITISH JOURNAL OF OPHTHALMOLOGY, vol. 86, no. 10, 1 October 2002 (2002-10-01), GB, pages 1175 - 1180, XP055367385, ISSN: 0007-1161, DOI: 10.1136/bjo.86.10.1175 *
A. LEONARDI ET AL: "Identification of human tear fluid biomarkers in vernal keratoconjunctivitis using iTRAQ quantitative proteomics", ALLERGY, vol. 69, no. 2, 13 December 2013 (2013-12-13), United Kingdom, pages 254 - 260, XP055275215, ISSN: 0105-4538, DOI: 10.1111/all.12331 *
ALTSCHUL ET AL., J. MOL. BIOL., vol. 215, 1990, pages 403 - 410
ALTSCHUL ET AL.: "BLAST Manual", NCB/NLM/NIH
ANDREA LEONARDI ET AL: "Th1 and Th2-type cytokines in chronic ocular allergy", GRAEFE'S ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY ; INCORPORATING GERMAN JOURNAL OF OPHTHALMOLOGY, SPRINGER, BERLIN, DE, vol. 244, no. 10, 15 March 2006 (2006-03-15), pages 1240 - 1245, XP019443121, ISSN: 1435-702X, DOI: 10.1007/S00417-006-0285-7 *
ANDREA LEONARDI: "Vernal keratoconjunctivitis: pathogenesis and treatment", PROGRESS IN RETINAL AND EYE RESEARCH, vol. 21, no. 3, 1 May 2002 (2002-05-01), GB, pages 319 - 339, XP055275217, ISSN: 1350-9462, DOI: 10.1016/S1350-9462(02)00006-X *
CARILLO ET AL., SIAM J. APPLIED MATH., vol. 48, 1988, pages 1073
DEVEREUX ET AL., NUCL. ACID. RES., vol. 2, 1984, pages 387
EL-ASRAR A M ABU ET AL: "Immunopathogenesis of conjunctival remodelling in vernal keratoconjunctivitis", EYE, NATURE PUBLISHING GROUP, GB, vol. 20, no. 1, 1 January 2006 (2006-01-01), pages 71 - 79, XP009099730, ISSN: 0950-222X, DOI: 10.1038/SJ.EYE.6701811 *
JUN SHOJI ET AL: "Antibody Array-Generated Cytokine Profiles of Tears of Patients with Vernal Keratoconjunctivitis or Giant Papillary Conjunctivitis", JAPANESE JOURNAL OF OPHTHALMOLOGY ; THE OFFICIAL ENGLISH-LANGUAGE JOURNAL OF THE JAPANESE OPHTHALMOLOGICAL SOCIETY, SPRINGER-VERLAG, TO, vol. 50, no. 3, 1 May 2006 (2006-05-01), pages 195 - 204, XP019389329, ISSN: 1613-2246, DOI: 10.1007/S10384-005-0319-4 *
LEONARDI A ET AL: "Tear levels and activity of matrix metalloproteinase (MMP)-1 and MMP-9 in vernal keratoconjunctivitis", INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE - IOVS, ASSOCIATION FOR RESEARCH IN VISION AND OPHTHALMOLOGY, US, vol. 44, no. 7, 1 July 2003 (2003-07-01), pages 3052 - 3058, XP008066620, ISSN: 0146-0404, DOI: 10.1167/IOVS.02-0766 *
LEONARDI ANDREA ET AL: "Vernal Keratoconjunctivitis-like Disease in Adults", AMERICAN JOURNAL OF OPHTHALMOLOGY, ELSEVIER, AMSTERDAM, NL, vol. 155, no. 5, 23 January 2013 (2013-01-23), pages 796 - 803, XP028547247, ISSN: 0002-9394, DOI: 10.1016/J.AJO.2012.11.018 *
METZ ET AL: "Phenotypic characterization of T cells infiltrating the conjunctiva in chronic allergic eye disease", JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, ELSEVIER, AMSTERDAM, NL, vol. 98, no. 3, 1 September 1996 (1996-09-01), pages 686 - 696, XP005154240, ISSN: 0091-6749, DOI: 10.1016/S0091-6749(96)70103-3 *
UCHIO ET AL: "Tear levels of interferon-gamma, interleukin (IL) -2, IL-4 and IL-5 in patients with vernal keratoconjunctivitis, atopic keratoconjunctivitis and allergic conjunctivitis", CLINICAL & EXPERIMENTAL ALLERGY : JOURNAL OF THE BRITISH SOCIETY FOR ALLERGY AND CLINICAL IMMUNOLOGY, vol. 30, no. 1, 1 January 2000 (2000-01-01), UK, pages 103 - 109, XP055275212, ISSN: 0954-7894, DOI: 10.1046/j.1365-2222.2000.00699.x *
VON HEINJE, G.: "Sequence Analysis in Molecular Biology", 1987, ACADEMIC PRESS

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