WO2006097388A2 - Variants d'acides nucleiques dans le gene c1qr1 associe a une immunite innee modifiee - Google Patents

Variants d'acides nucleiques dans le gene c1qr1 associe a une immunite innee modifiee Download PDF

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WO2006097388A2
WO2006097388A2 PCT/EP2006/050650 EP2006050650W WO2006097388A2 WO 2006097388 A2 WO2006097388 A2 WO 2006097388A2 EP 2006050650 W EP2006050650 W EP 2006050650W WO 2006097388 A2 WO2006097388 A2 WO 2006097388A2
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clqrl
nucleic acid
disease
subject
risk
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PCT/EP2006/050650
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WO2006097388A3 (fr
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Lieve Nuytinck
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Innogenetics N.V.
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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
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • 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
    • C12Q2600/156—Polymorphic or mutational markers
    • 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
    • C12Q2600/16—Primer sets for multiplex assays

Definitions

  • the present invention relates to a method and kit for identifying a subject at risk of, or having, an indication associated with altered innate immunity.
  • the present invention is based on the determination of the ClQRl genotype and/or serum level.
  • Immunity to infection is mediated by two systems, the acquired (or adaptive) immune system and the innate (or natural) immune system.
  • the innate immunity system is an evolutionary ancient form of immunity and offers the main resistance to microbial pathogens within the first minutes, hours or days of an infection (Fujita et al., 2002).
  • PAMPs pathogen-associated molecular patterns
  • PRR pattern-recognition receptors
  • PRRs As soon as the PRRs identify the corresponding predetermined carbohydrate pattern on a pathogen, they immediately trigger effector cells to destroy the invading microorganism, rather than after having to undergo a proliferative cycle, as is the case for the time-delayed adaptive immune response.
  • PRRs can be divided into three classes: signaling, endocytic, and secreted (Medzhitov R. et al., 2004).
  • Innate immunity refers to antigen-nonspecific defense mechanisms that a host uses immediately or within several hours after exposure to an antigen. This is the immunity one is born with and is the initial response by the body to eliminate microbes and prevent infection. . cascade can be activated through three distinct pathways, i.e. the classical, the alternative and the lectin-pathway.
  • the lectin pathway involves carbohydrate recognition by PRR, such as collectins, and the subsequent activation of associated enzymes that are known as MBL-associated proteins (mannose binding protein serine protease or MASP), MASP-I, MASP-2 and MASP-3, and its truncated form, small MBL-associated protein (sMAP, also called Mapl9) (Matsushita et al., 2000; Fujita T., 2002; Lu et al., 2002).
  • MBL-associated proteins mannose binding protein serine protease or MASP
  • MASP-I mannose binding protein serine protease
  • MASP-2 MASP-2
  • MASP-3 truncated form, small MBL-associated protein
  • Mapl9 small MBL-associated protein
  • Collectins belong to the super family of mammalian C-type lectins. The following eight collectins have been identified so far: mannose-binding lectin (MBL), surfactant protein A (SP-A), surfactant protein D (SP-D), collectin liver 1 (CL-Ll), collectin placenta 1 (CL-Pl), conglutinin, collectin of 43 kDa (CL-43) and collectin of 46kDa (CL-46) (van de Wetering JK et al. 2004).
  • MBL mannose-binding lectin
  • SP-A surfactant protein A
  • SP-D surfactant protein D
  • collectin liver 1 CL-Ll
  • CL-Pl collectin placenta 1
  • conglutinin collectin of 43 kDa
  • CL-43 collectin of 46kDa
  • CL-46 van de Wetering JK et al. 2004.
  • CIq complement 1, q subcomponent
  • CIqRp complement receptor
  • CRT calreticulin
  • binding protein for the globular head of CIq gClqbp
  • MCL mannose binding lectin
  • SP-A surfactant protein A
  • CIqRp also known as the AA4 antigen in rodents
  • mNI-11 pro-adhesive monoclonal antibody
  • mNI-11 pro-adhesive monoclonal antibody
  • CRl CD35
  • C4b C3b
  • iC3b C3b
  • MBL MBL
  • CRT collagenous domains of CIq
  • CRT belongs to the family of heat-shock proteins, the most abundant and ubiquitous soluble intracellular proteins. Though CRT does not have a transmembrane domain, it seems to mediate phagocytosis of the apoptotic cells through association . , , has been termed gClqbp. This protein is located in mitochondria, suggesting that gClqbp is not a cell surface receptor itself.
  • Binding of MBL to microorganisms can result in inactivation of the organism by activation of the complement cascade.
  • SP-A can prevent the formation of active Cl complex.
  • CIqRp was a type I transmembrane protein encoded by a 1959 bp cDNA. Sequence comparison revealed a 21 amino acid N-terminal signal peptide and a 156 amino acid C-type lectin-like domain also termed a carbohydrate recognition domain. The protein also contained five epidermal growth factor-like domains, three of which were predicted to have Ca2+ binding activity as well as a single transmembrane domain and a short cytoplasmic tail containing a tyrosine kinase recognition motif.
  • CIqRp The cellular distribution of CIqRp is specific and the molecule was found abundantly on endothelial cells and platelets (neutrophils, microglial central nervous system glial cells and monocytes, but not macrophages, also express CIqRp) (McGreal et al. 2002; Fonseca et al. 2001). Notably, CIqRp was not detected on lymphocytes, fibroblasts or smooth muscle cells, which have been shown to respond to CIq stimulation. This indicates that CIqRp is not a component of all CIq receptors and may only be involved in particular CIq- mediated functions. Its expression by endothelial cells and platelets suggests functions for this protein beyond mediating phagocytic enhancement and, if CIqRp is merely a subcomponent of a larger CIq receptor complex, it may have functions which are completely independent
  • CIqRp may mediate the enhancement of phagocytosis in monocytes and macrophages upon interaction with soluble defense collagens and may play a role in intercellular adhesion.
  • the present invention provides a method and kit for identifying a subject at risk of, or having, an indication associated with altered innate immunity, based on the ClQRl genotype, concentration or functionality.
  • the present invention provides a method and kit for identifying a subject at risk of, or having, an indication associated with altered innate immunity, variant in the ClQRl gene.
  • the present invention provides a method and kit for identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising detecting in a sample the presence or absence of at least one nucleic acid variant in the ClQRl gene, or part thereof, whereby the presence of at least one nucleic acid variant identifies whether a subject is at risk of or has an indication associated with an altered innate immunity.
  • Specific regions of interest in the ClQRl genes are exon 1 and/or exon 2.
  • the methods and kits of the present invention can also be carried out in combination with other methods for identifying a subject at risk of, or having, an indication associated with altered innate immunity.
  • the methods and kits are carried out in combination with a method for the detection of the presence or absence of a nucleic acid variant, or other markers, in any other gene.
  • Any detection method is part of the present invention.
  • Preferred methods and means for the detection of the presence or absence of the nucleic acid variants of the present invention are hybridization, sequencing, PCR, MLPA, OLA, primer extension and restriction site analysis.
  • the method and kits of the present invention identify a subject at risk of, or having, an indication associated with altered innate immunity, and comprises measuring the concentration or functionality of the ClQRl protein in a biological sample, wherein an increased or decreased ClQRl concentration or altered functionality compared to a reference value representing a known health status indicates that said subject is at risk or suffers from a disorder associated with altered innate immunity.
  • a further embodiment of the present invention relates to a method for selecting an appropriate treatment or therapeutic agent for a subject at risk of, or having, an indication associated with altered innate immunity, comprising determining the status or activity of the innate immunity by the methods of the present invention and selecting an appropriate treatment or therapeutic agent.
  • Figure 1 human ClQRl wt gDNA sequence (SEQ ID NO:1): exon sequences and possible polymorphism positions are indicated in respectively grey and bold/boxed. Nucleotide +1 is the A of the ATG-translation initiation codon which is bold and underlined.
  • LOCUS NT Ol 1387 9001 bp DNA linear CON 20-AUG-2004 DEFINITION Homo sapiens chromosome 20 genomic contig.
  • /db_xref GeneID:22918
  • /db_xref MIM: 120577” mRNA join(1024..3105,3390.-8006)
  • the determination of the nucleic acid sequence and/or the ClQRl concentration or functionality makes it possible to estimate or identify whether a subject is at risk of, or has, an indication associated with altered innate immunity.
  • the method of the present invention determines the presence of both variant and normal nucleic acids of the ClQRl gene in a sample.
  • the term "ClQRl gene” refers to the gene ClQRl and also to analogous or derivatives thereof. "Part thereof refers to the region of interest, i.e. the region of the ClQRl gene comprising a nucleic acid variant. More particular, a part thereof refers to the 5'UTR, the promotor region, exon 1, intron 1 and/or exon 2.
  • the current invention relates to a method of identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising detecting the presence or absence of at least one nucleic acid variant in the ClQRl gene, whereby the presence of at least one nucleic acid variant identifies whether a subject is at risk of, or has, an indication associated with an altered innate immunity.
  • sequence and may consist of deoxyribonucleotides or ribonucleotides, nucleotide analogues or modified nucleotides, or may have been adapted for therapeutic purposes.
  • a nucleic acid that is up to about 100 nucleotides in length is often also referred to as an oligonucleotide.
  • the reference nucleic acid and protein sequences indicated in the current invention are derived from GeneBank (NCBI) and indicated by their respective accession number, as is well known to the person skilled in the art.
  • NCBI GeneBank
  • the nomenclature for the ClQRl nucleotide and amino acid changes as used herein is generally accepted and recommended by den Dunnen and Antonarakis (2000). Frequent updates of the nomenclature for the description of sequence variations are provided on the web-site of the Human Genome Variation Society. Accordingly, the nucleotide numbering of the coding DNA and RNA reference sequence is as follows:
  • nucleotide +1 is the A of the ATG-translation initiation codon
  • the ClQRl gene (complement component 1, q subcomponent, receptor 1) is located at chromosome 20 and comprises 2 exons. Synonyms are: Complement component CIq receptor precursor (CIqRp), C lq/MBL/SP A receptor, CD93 antigen, Cell surface antigen AA4 and Lymphocyte antigen 68.
  • the reference nucleic acid sequence for ClQRl is NT Ol 1387.8 (gDNA; Version: NT Ol 1387.8 GL27501067) and NM_012072.2 (mRNA).
  • the reference protein sequence is NP 036204.1.
  • nucleic acid variant or “polymorphism” or “variant” as used in the present invention, means that the nucleic acid sequence at a certain position in the ClQRl gene differs relative to one or more reference nucleic acid sequences.
  • nucleic acid polymorphism The most simple nucleic acid polymorphism is a polymorphism affecting a single nucleotide, i.e. a single nucleotide polymorphism or SNP. Nucleic acid polymorphisms further include any number of contiguous and/or non-contiguous relative to the primary nucleotide sequence of one or more reference nucleic acids.
  • the term "polymorphic position" or "position” refers to the nucleic acid position at which a nucleic acid polymorphism arises. Nucleic acid sequences comprising at least one such polymorphism are referred to as "polymorphic nucleic acid sequences", “polymorphic polynucleotides", “polymorphic sequences” or the like.
  • the polymorhism or nucleic acid variant can be an insertion, deletion, substitution, tandem repeat or similar.
  • haplotype means a particular pattern of sequential polymorphisms found on a single chromosome.
  • allele is one of several alternative forms of a gene or DNA sequence at a specific chromosomal location (locus). At each autosomal locus an individual possesses two alleles, one inherited from the father and one from the mother.
  • locus a specific chromosomal location
  • the present invention relates to a method according to the present invention, wherein the ClQRl genotype has at least one variant allele of the ClQRl gene (heterozygous).
  • the method of the invention relates to a method according to the present invention, wherein the ClQRl genotype has two variant alleles of the ClQRl gene (homozygous).
  • the ClQRl gene has 2 exons.
  • the invention relates in particular to any polymorphism located within the ClQRl gene as identified in SEQ ID NO:1, or in the corresponding cDNA or mRNA sequence.
  • the polymorphism is located in exon 1 and/or exon 2 of the
  • Exon 1 may contain a nucleic acid variant at position 1621 of the gDNA sequence.
  • nucleic acid variant is 1621C>T, resulting in the amino acid change P541 S and is identified as rs3746731 (dbSNP NCBI).
  • the exon 2 region of the ClQRl gene is also polymorphic.
  • a nucleic acid variant may be present at nucleic acid position 6573 of the gDNA sequence. More specific, the nucleic acid variant is 6573T>C and is identified as rs7492. , - .
  • the nucleic acid sequence of the wild type human ClQRl gene is identified by SEQ ID NO:1.
  • the allele may be normal as in the reference sequence, or it may be a variant, such as a structural or a non-structural variant.
  • analogue is a compound (or molecule) that is a (chemical) structural derivative of ClQRl. It is also used to describe a molecule which may be structurally similar (but not identical) to another, and which exhibits many or some of the same biological functions of ClQRl .
  • An analogue is to be understood as being any peptide sequence capable of the same biological functions as wild-type ClQRl, including recombinant ClQRl.
  • innate immunity refers to the natural ability of an organism to defend itself against invasions by pathogens.
  • Pathogens as used herein, may include, but are not limited to bacteria, fungi, parasites, viruses and algae.
  • innate immunity includes immune responses that affect other diseases, such as cancer, inflammatory diseases, neurological diseases, autoimmune diseases and various infections.
  • an "indication or condition associated with aberrant, modified or altered innate immunity” refers to any indication or disease resulting from a decreased or increased defense mechanism.
  • a decreased defense can increase the susceptibility for infection or inflammation or can increase risk for acquiring of a particular disease.
  • An increased defense might result in neurological disease, autoimmune disease or inflammatory diseases.
  • ClQRl deficiencies are associated with an increased risk for infections, inflammation, neurological disease and autoimmune conditions, and influence the severity and/or course of several diseases. Accordingly, ClQRl deficiencies can be linked with increased susceptibility for disease and/or prognosis for more severe or more frequent disease, or worse outcomes due to complication. In general, ClQRl deficiencies can be . , considered and include eventual ClQRl replacement therapy.
  • the risk for developing a disorder associated with an altered activity of innate immunity can be determined.
  • the present invention relates to a method of identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising detecting the presence or absence of at least one nucleic acid variant in the ClQRl gene. More specific, the present invention relates to a method of identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising detecting the presence or absence of at least one nucleic acid variant in the ClQRl gene, whereby the presence of at least one nucleic acid variant identifies a subject at risk of, or having an increased susceptibility for disease. In a further embodiment, the presence of at least one ClQRl nucleic acid variant identifies a subject at risk of, or having an increased severity of disease.
  • the presence of at least one ClQRl nucleic acid variant identifies a subject at risk of, or having a modified response to therapy for a disease. Furthermore, the presence of at least one ClQRl nucleic acid variant identifies a subject at risk of, or having increased risk of transplant rejection.
  • the altered innate immunity is associated with an increased susceptibility for or severity of infection, an autoimmune disease, cystic fibrosis, a cardiovasular disease, a neurological disease or cancer.
  • infection encompasses bacterial, viral, fungal, parasitic or algae infection. Examples are sepsis/severe sepsis/septic shock and otitis media. Sepsis is defined as presence of infection and several of other parameters of general clinical nature, inflammatory, hemodynamic and tissue perfusion parameters. Severe sepsis is the presence of sepsis complicated by organ dysfunction. Septic shock is defined as the presence of severe sepsis accompanied by acute circulatory failure. Otitis media is an infection of the middle ear.
  • the altered innate immunity is associated with one or more of the following autoimmune diseases: rheumatoid arthritis (RA), spondyloarthropathy, systemic lupus erythematosus (SLE), Sjogren's disease, , , , , . disease, asthma, atopic dermatitis, dermatomyositis or Behcet's disease.
  • RA rheumatoid arthritis
  • SLE systemic lupus erythematosus
  • Sjogren's disease , , , , . disease, asthma, atopic dermatitis, dermatomyositis or Behcet's disease.
  • the altered innate immunity is associated with one or more of the following cancers: (1) solid tumors such as colon cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, liver cancer, kidney cancer or brain cancer, and (2) haematological malignancies such as a) Leukemias: acute myeloid leukemia (AML), acute lymphoid leukemia (ALL), chronic myeloid leukemia (CML) and chronic lymphoid leukemia (CLL), b) Lymphomas: Hodgkins and non-Hodgkin's lymphomas and c) Myelomas.
  • AML acute myeloid leukemia
  • ALL acute lymphoid leukemia
  • CML chronic myeloid leukemia
  • CLL chronic lymphoid leukemia
  • Lymphomas Hodgkins and non-Hodgkin's lymphomas
  • Myelomas Myelomas.
  • the altered innate immunity is associated with one or more of the following cardiovascular diseases: bypass failure, atherosclerosis, myocardial reperfusion injury, coronary artery disease or heart disease.
  • the altered innate immunity is associated with one or more of the following neurological diseases: Alzheimer's disease, myasthenia gravis, multiple sclerosis, microbial infections, head trauma and stroke, Pick's disease, Parkinson's disease, dementia with Lewy bodies, Huntington disease, chromosome 13 dementias, Down's syndrome, cerebrovascular disease, Rasmussen's encephalitis, viral meningitis, NPSLE, amyotrophic lateral sclerosis, Creutzfeldt-Jacob disease, Gerstmann-Straussler-Scheinker disease, transmissible spongiform encephalopathies or ischemic reperfusion damage.
  • the altered innate immunity is associated with an increased severity of disease influencing the course of a disease.
  • the disease status can be aggravated leading to a higher mortality.
  • autoimmune disease for example RA
  • the severity or damage to the joints can be more pronounced, as measured by radiology.
  • the altered innate immunity is associated with a modified response to therapy for a specific disease resulting in adverse effects. This can for example by seen in vaccinations or in NSAID therapy.
  • the present invention relates to a method of identifying a subject at risk of, or having, an increased or decreased susceptibility for (recurrent) otitis media comprising detecting the presence or absence of at least one nucleic acid variant in the ClQRl gene.
  • the presence of the nucleic acid variant can be associated .
  • nucleic acid variant has a protective effect. More specific, the nucleic acid variant is located in exon 1 of the ClQRl gene. Even more specific, the nucleic acid variant is located at position 1621 of the ClQRl gene.
  • the present invention relates to a method of identifying a subject at risk of, or having, an increased or decreased susceptibility for Rheumatoid Arthritis (RA) comprising detecting the presence or absence of at least one nucleic acid variant in the ClQRl gene.
  • the presence of the nucleic acid variant can be associated with an increased or decreased (reduced) risk for RA.
  • the nucleic acid variant has a protective effect. More specific, the nucleic acid variant is located in exon 2 of the ClQRl gene. Even more specific, the nucleic acid variant is located at position 6573 of the ClQRl gene.
  • defect refers to an alteration in the genomic DNA sequence when compared to the wild type sequence. This may result in altered (enhanced or decreased) expression levels or may result in changes (increase or decrease) in the functionality of the encoded protein, or even may result in a change of function of the protein ("gain of function").
  • the "subject" on which the method of the present invention is carried out can be any subject of which the risk of an altered innate immunity needs to be determined.
  • the subject may be a non-human subject such as (but not limited to) a cow, a pig, a sheep, a goat, a horse, a monkey, a rabbit, a dog, a cat, a mouse, a rat, a hamster, a zebrafish, a pufferfish (Fugu), a fly, a worm or C. elegans. More preferably, the subject is a primate. Even more preferably, the subject is a human.
  • nucleic acid from any nucleated cell can be used as the starting point for such assay techniques and may be isolated according to standard nucleic acid preparation procedures well known to those of skill in the art.
  • Many current methods for the detection of allelic variation are reviewed by Nollau et al. (1997), and in standard textbooks, for example "Laboratory Protocols for Mutation Detection", Ed. by U. Landegren, Oxford University Press, 1996 and "PCR", ⁇ ind , , (incorporated herein by reference).
  • the method of the present invention can be carried out in vivo or in vitro. Preferred, however, is in vitro detection of nucleic acid variants in the ClQRl gene in a biological sample obtained from the subject.
  • biological sample means a tissue sample or a body fluid sample.
  • a tissue sample includes (but is not limited to) buccal cells, a brain sample, a skin sample or organ sample (e.g. liver).
  • body fluid refers to all fluids that are present in the body including but not limited to blood, plasma, serum, lymph, urine, saliva or cerebrospinal fluid.
  • the biological sample may also be obtained by subjecting it to a pretreatment if necessary, for example, by homogenizing or extracting. Such a pretreatment may be selected appropriately by those skilled in the art depending on the biological sample to be subjected.
  • a nucleic acid comprising an intended sequence prepared from a biological sample may be prepared from DNA (e.g. gDNA or cDNA) or RNA (e.g. mRNA). Release, concentration and isolation of the nucleic acids from the sample can be done by any method known in the art. Currently, various commercial kits are available such as the QIAamp Blood Kit from Qiagen (Hilden, Germany) for the isolation of nucleic acids from blood samples, or the 'High pure PCR Template Preparation Kit' (Roche Diagnostics, Basel, Switzerland) or the DNA purification kits (PureGene, Gentra, Minneapolis, US). Other, well-known procedures for the isolation of DNA or RNA from a biological sample are also available (Sambrook et al., 1989; Ausubel et al., 2003).
  • the nucleic acid may be amplified.
  • amplification procedures can be accomplished by those methods known in the art, including, for example, the polymerase chain reaction (PCR), ligase chain reaction (LCR), nucleic acid sequence-based amplification (NASBA), strand displacement amplification, rolling circle amplification, T7- polymerase amplification, and reverse transcription polymerase reaction (RT-PCR).
  • PCR polymerase chain reaction
  • LCR ligase chain reaction
  • NASBA nucleic acid sequence-based amplification
  • strand displacement amplification strand displacement amplification
  • rolling circle amplification rolling circle amplification
  • T7- polymerase amplification T7- polymerase amplification
  • RT-PCR reverse transcription polymerase reaction
  • sequences are well-known in the art.
  • the present invention is not limited by any particular method used to detect the target sequences disclosed herein. Examples of such methods are described by Gut (2001) and Syvanen (2001), and include, but are not limited to, hybridization methods such as reverse dot blot, LiPA, geneChip microarrays, DASH, PNA and LNA probes, TaqMan (5 'nuclease assay) and molecular beacons; allele-specific PCR methods such as intercalating dye, FRET primers and Alphascreen; primer extension methods such as ARMS, kinetic PCR, SNPstream, GBA, multiplex minisequencing, SNaPshot, pyrosequencing, MassExtend, Mass Array, Goodassay, microarray miniseq, APEX, sequence specific priming (SSP), microarray primer extension, Tag arrays, code
  • hybridization methods such as reverse dot blot, LiPA, geneChip microarrays, DASH, P
  • the detection of the presence or absence of a nucleic acid variant is determined by hybridization, sequencing, PCR, MLPA, OLA, primer extension or restriction site analysis.
  • the present invention also provides oligonucleotides, i.e. primers and probes, in order to amplify and/or detect nucleic acid variants and/or the wild type sequence of the ClQRl gene.
  • the wild type sequence of the ClQRl gene is identified by SEQ ID NO:1.
  • Such primers or probes, specifically hybridizing to the target nucleic acid are of any convenient length such as to comprise at least 8, 9, 10, 11, 12, 13, 14 or 15 nucleotides and up to 40 nucleotides, up to 30 nucleotides or more conveniently up to 25 nucleotides in length.
  • a preferred length of the primers or probes is thus 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides.
  • primers or probes will comprise nucleotide sequences entirely complementary to the corresponding wild type or variant locus in the ClQRl gene. However, if required one or more nucleotides may be added or one or more mismatches may be introduced, provided that the discriminatory power of the oligonucleotide primer or probe is not unduly affected. Specific length and sequence of the probes and primers will depend on the complexity of the required nucleic acid target, as well as on the reaction conditions such as temperature and ionic strength. In general, the hybridization . under which a nucleotide sequence can bind to related or non-specific sequences. For example, high temperature and lower salt increases stringency such that non-specific binding or binding with low melting temperature will dissolve.
  • the primers or probes of the invention may carry one or more labels to facilitate detection.
  • the primer or probe consists of 10 to 30 nucleotides, preferably 15 to 30 nucleotides, and is capable of specifically forming a hybrid with a part of the ClQRl gene and is at least one or more selected from the group consisting of: 1) an oligonucleotide capable of hybridizing under a stringent condition with the sequence as represented by SEQ ID NO:1, or the complement thereof;
  • an oligonucleotide capable of hybridizing under a stringent condition with the sequence as represented by SEQ ID NO:1 wherein one or more nucleotides was subjected to a variation such as a substitution, deletion, insertion or addition, or the complement thereof.
  • the present invention relates to an isolated oligonucleotide consisting of 10 to 30 nucleotides, optionally 15 to 30 nucleotides, for detecting the presence of one or more nucleic acid variants in SEQ ID NO: 1 , or the complementary strand. More specific, the nucleic acid variants are located in exon 1 and/or exon 2, and in particular at position 1621 or 6573 of SEQ ID NO:1.
  • the polymorphism located in the ClQRl gene may also be detected in vitro by determining in the isolated ClQRl protein, as identified in the present invention, the presence or absence of an amino acid change by sequencing said protein.
  • the amino acid change may also be detected by any conventional method known in the art, for example by mass-spectroscopy, gel electrophoresis, MALDI-TOF mass spectroscopy,
  • ficolin genes i.e. FCNl, FCN2, FCN3
  • BPI Bacterial/permeability-increasing protein
  • CD 14 CD 14 antigen precursor
  • beta-catenin CNNBI, Cadherin Associated Protein beta I
  • ILlO Interleukin 10
  • LBP Lipopolysaccharide binding protein
  • RP 105 LY64, lymphocyte antigen 64 homolog radioprotective
  • MBL2 Mannose Binding Protein
  • MASP-2 MD-I (RP105-associated), MD-2 (MD2 Protein, Lymphocyte antigen 96), MYD88 (Myeloid differentiation primary response gene 88), NODl (Caspase recruitment domain 4, CARD4), NOD2 (Caspase recruitment domain family, member 15, CARDl 5)
  • NODl Caspase recruitment domain 4, CARD4
  • NOD2 Caspase recruitment domain family, member 15, CARDl 5
  • Toll Like Receptor genes i.
  • the present invention also relates to a method of identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising the step of detecting the presence or absence of a nucleic acid variant in the ClQRl gene and detecting the presence or absence of one or more nucleic acid variants in any other gene.
  • the "other" gene is selected from the group consisting of: the ficolin genes (i.e.
  • FCNl FCN2, FCN3
  • BPI Bacterial/permeability- increasing protein
  • CD 14 CD 14 antigen precursor
  • beta-catenin CNNBI, Cadherin Associated Protein beta
  • ILlO Interleukin 10
  • LBP Lipopolysaccharide binding protein
  • RP 105 LY64, lymphocyte antigen 64 homolog radioprotective
  • MBL2 Mannose Binding Protein
  • MASP-2 MD-I (RP105-associated)
  • MD-2 MD2 Protein, Lymphocyte antigen 96
  • MYD88 Myeloid differentiation primary response gene 88
  • NODl Caspase recruitment domain 4, CARD4
  • NOD2 Caspase recruitment domain family, member 15, CARDl 5
  • TLRl TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 or TLRlO.
  • TLRl TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 or TLRl
  • the present invention also encompasses a method for determining whether a subject has a risk of developing a disease wherein the nucleic acid variants in the ClQRl gene are detected by their protein phenotype.
  • the invention discloses how decreased or increased levels of ClQRl and lack of ⁇ , embodiment, the method encompasses the measurement of one or more proteins.
  • the present invention relates to a method for identifying a subject at risk of, or having, an indication associated with altered innate immunity comprising the steps of: a) determining the concentration or functionality of the ClQRl protein in a sample, b) identifying if said subject is at risk of, or has, an indication associated with altered innate immunity.
  • the current invention provides a method of identifying a subject at risk of, or having, an indication associated with altered innate immunity, comprising measuring the concentration or functionality of the ClQRl protein in a biological sample, wherein an increased or decreased ClQRl concentration or altered functionality compared to a reference value representing a known health status indicates that said subject is at risk of or suffers from a disorder associated with altered innate immunity.
  • a "ClQRl protein” is a protein encoded by the ClQRl gene as described in the present invention, or a variant thereof.
  • concentration refers to the presence or absence and/or amount of a certain protein.
  • a change in the concentration of a protein refers to a measurable increase or decrease, including total absence or presence, in the protein concentration when compared to a control subject.
  • a known health status or "control subject”, as defined in the present invention is a subject of the same species as the subject under examination which is free from, or not at direct risk of developing a disease associated with altered innate immunity.
  • the healthy subject can be of the same weight, age, and gender as the subject who is being diagnosed or prognosed for an altered innate immunity. In some cases, it might be preferred to use a reference value from the subject which is diagnosed.
  • the concentration obtained upon analyzing the subject under examination relative to the concentration obtained upon analyzing a control subject will depend on the particular analytical protocol and detection technique that is used. Accordingly, those skilled in the art will understand that, based on the present description, any laboratory can establish, for the ClQRl protein, a suitable "reference range”, “reference level range”, “concentration range” or “range of levels” (those terms are used interchangeable) characteristic for control subjects according to the . subject under examination can then be compared with this reference range and based on this comparison, a conclusion can be drawn as to whether the subject has a risk of developing a disease as described herein.
  • the reference value can be that of a level or concentration of the ClQRl protein in a sample, preferably a body fluid, from a subject not suffering from a disease associated with altered innate immunity.
  • the healthy subject can be of the same weight, age, and gender as the subject who is being diagnosed or prognosed for an altered innate immunity. In some cases, it might be preferred to use a reference value from the subject which is diagnosed.
  • the ClQRl protein that is detected in the method of the present invention may be detected by any method known to those skilled in the art. They can be identified by their structure, by partial amino acid sequence determination, by functional assay, by enzyme assay, by various immunological methods, or by biochemical methods.
  • the lectin pathway function is assessed using plates coated with mannan, followed by incubation of the serum in buffer containing Ca 2+ , Mg 2+ and an inhibitory antibody directed against CIq. The formation of the membrane attack complex is subsequently detected by use of a specific monoclonal antibody directed against C5b- 9.
  • Biochemical methods include (but are not limited to) capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC), hyperdiffusion chromatography, two-dimensional liquid phase electrophoresis (2 -D- LPE; Davidsson et al. 1999) or detection of the migration pattern in gel electrophoreses.
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis SDS- PAGE
  • SDS- PAGE is a widely used approach for separating proteins from complex mixtures (Patterson and Aebersold, 1995). It can be performed in one- or two-dimensional (2- D) configuration. For less complicated protein preparation, one-dimensional SDS- PAGE is preferred over 2-D gels, because it is simpler.
  • 2-D gel electrophoresis incorporates isoelectric focusing (IEF) in the first dimension and SDS-PAGE in the second dimension, leading to a separation by charge and size (O'Farrell, 1975).
  • IEF isoelectric focusing
  • 2-D PAGE is a powerful technique for separating very complex protein preparations, resolving up to 10 000 proteins from mammalian tissues and other complex proteins (Klose and Kobalz, 1995; Celis et al., 1996; Yan et al., 1997).
  • the ClQRl proteins of the present invention can be identified by their isoelectric focusing point (pi) and their molecular weight (MW) in kilodaltons (kD).
  • the level of ClQRl protein can also be detected by an immunoassay.
  • an "immunoassay” is an assay that utilizes an antibody to specifically bind to the antigen (i.e. the ClQRl protein). The immunoassay is thus characterized by detection of specific binding of the ClQRl protein to an antibody.
  • Immunoassays for detecting ClQRl proteins may be either competitive or noncompetitive. Noncompetitive immunoassays are assays in which the amount of captured analyte (i.e. the ClQRl protein) is directly measured. In competitive assays, the amount of analyte (i.e.
  • the ClQRl protein) present in the sample is measured indirectly by measuring the amount of an added (exogenous) analyte displaced (or competed away) from a capture agent (i.e. the antibody) by the analyte (i.e. the ClQRl protein) present in the sample.
  • a capture agent i.e. the antibody
  • analyte i.e. the ClQRl protein
  • a known amount of the (exogenous) ClQRl protein is added to the sample and the sample is then contacted with the antibody.
  • the amount of added (exogenous) ClQRl protein bound to the antibody is inversely proportional to the concentration of the ClQRl protein in the sample before the exogenous ClQRl protein is added.
  • the antibodies can be bound directly to a solid substrate where they are immobilized. These immobilized antibodies then capture the ClQRl protein of interest present in the test sample.
  • Other immunological methods include but are not limited to fluid or gel precipitation reactions, immunodiffusion (single or double), agglutination assays, Immunoelectrophoresis, radioimmunoassays (RIA), enzyme-linked immunosorbent assays (ELISA), TRIFMA (Christiansen et al., 1999), Western blots, liposome immunoassays (Monroe et al., 1986), complement- fixation assays, immunoradiometric assays, fluorescent immunoassays, protein A . Wild (2001), Ghindilis et al. (2002) and Kilpatrick (2002).
  • the method of the present invention may also be used in determining whether and which therapeutic agent might be suitable for a patient being at risk of, or having an indication associated with altered innate immunity.
  • the therapeutic agent may be used to prevent or treat the indication or disease.
  • preventing a disease means inhibiting or reversing the onset of the disease, inhibiting or reversing the initial signs of the disease, inhibiting the appearance of clinical symptoms of the disease.
  • treating a disease includes substantially inhibiting the disease, substantially slowing or reversing the progression of the disease, substantially ameliorating clinical symptoms of the disease or substantially preventing the appearance of clinical symptoms of the disease.
  • kits for identifying a subject at risk of, or having, an indication associated with altered innate immunity can be based on the detection of nucleic acid variants in the ClQRl gene of said subject or it can be based on the detection of ClQRl proteins. Accordingly, the kit of the present invention comprises reagents that selectively detect a nucleic acid variant in the ClQRl gene or that selectively detect a ClQRl protein.
  • a kit based on the detection of nucleic acid variants in the ClQRl gene may comprise: (a) a means or reagent for detecting the presence or absence of one or more nucleic acid variants in the ClQRl gene of said subject; and/or (b) optionally, a means for determining, from the nucleic acid variants detected with the means of step (a), whether the subject is at risk of, or has, an indication associated with altered innate immunity. More preferred, the kit comprises a means for detecting the presence or absence of one or more nucleic acid variants in exon 1 and/or exon 2 of the ClQRl gene. In a preferred embodiment of the present invention, the kit comprises: nucleic acid variants at the positions 1621 and/or 6573 of the ClQRl gene of said subject; and
  • step (c) optionally, a means for determining, from the nucleic acid variants detected with the means of step (a), whether the subject is at risk of, or has, an indication associated with altered innate immunity.
  • the means or reagents in step (a) of said kit may comprise:
  • the means or reagent in step (a) of said kit comprise at least one oligonucleotide probe suitable for detection of a target ClQRl nucleic acid.
  • the target ClQRl nucleic acid is located in exon 1 and/or exon 2. Even more specific, the target ClQRl nucleic acid is located at position 1621 and/or 6573 of the ClQRl gene.
  • the oligonucleotide suitable for detection of a target ClQRl nucleic acid is a SSP (sequence specific primer). More specifically, the oligonucleotide suitable for detection of a target _ . acid to form an invase cleavage structure.
  • the cleavage structure can be detected by a cleavage agent or enzyme such as a structure-specific nuclease, a 5 'nuclease, a FEN-I endonuclease or a polymerase.
  • the means or reagent in step (a) also includes at least one pair of primers suitable for amplification of a target ClQRl poiynucieic acid.
  • the target poiynucieic acid is exon 1 and/or exon 2 of the ClQRl gene, or part thereof.
  • the target ClQRl poiynucieic acid comprises or consists of position 1621 and/or 6573 of the ClQRl gene.
  • hybridization buffer means a buffer allowing a hybridization reaction between the oligonucleotides and the poiynucieic acids present in the sample, or the amplified products, under the appropriate stringency conditions.
  • wash solution means a solution enabling washing of the hybrids formed under the appropriate stringency conditions.
  • the means for detecting the presence or absence of nucleic acid variants in the ClQRl gene is an ESfVADER assay (see e.g. WO97/27214, incorporated herein by reference)
  • the means for detecting the presence or absence of nucleic acid variants in the ClQRl gene is a line probe assay (LiPA; Stuyver et al., 1996; Stuyver et al., 1997; Van Geyt et al., 1998).
  • the selected set of probes is immobilized to a membrane strip in a line fashion.
  • oligonucleotides may be coupled to microspheres or chips.
  • an assay that provides for simultaneous detection includes (but is not limited to) the xMAPTM technology (Luminex®, Austin, Texas, USA) and the PamGene technology (PamGene, 's-Hertogenbosch, The Netherlands).
  • ClQRl gene detected with the means of step (a), whether the subject is at risk of, or has, an indication associated with altered innate immunity include a table, a chart, or similar, generally referred to as "a predisposition risk algorithm", indicating the presence of a predisposition risk algorithm.
  • ClQRl nucleic acid variants or haplotypes that confer a risk for or the existence of an indication associated with altered innate immunity.
  • the term "chart” refers to graphical presentation, visual aid, diagram, plan, graph, sheet, map or the like including the relevant information. The determination of the risk can be performed manually or with the use of a computer.
  • the kit of the present invention may include, in additions to the means of steps (a), a means for detection other risk factors, e.g. nucleic acid variants in a gene, for an indication associated with an altered innate immunity.
  • the kit additionally includes a means for detecting the genotype of or a nucleic acid variant in at least one of the genes selected from the group consisting of: the ficolin genes (FCNl, FCN2, FCN3, BPI (Bacterial/permeability-increasing protein), CD14 (CD 14 antigen precursor), beta-catenin (CTNNBI, Cadherin Associated Protein beta), ILlO (Interleukin 10), LBP (Lipopolysaccharide binding protein), RP 105 (LY64, lymphocyte antigen 64 homolog radioprotective), MBL2 (Mannose Binding Protein), MASP-2, MD-I (RP105-associated), MD-2 (MD2 Protein, Lymphocyte antigen 96), MYD88 (Myeloid differentiation primary response
  • kits based on the detection of ClQRl proteins may comprise an antibody that specifically recognizes the ClQRl protein that is detected.
  • a preferred kit for carrying out the method of the invention comprises: - an antibody (primary antibody) which forms an immunological complex with the ClQRl protein to be detected;
  • - a monoclonal antibody (secondary antibody) which specifically recognizes the ClQRl protein to be detected; - antibody;
  • Table 1 identifies the ClQRl nucleic acid and protein variants studied in the present examples.
  • the statistical analysis of the data is based on the determination of odds ratios (OR) using standard procedures.
  • An odds ratio is calculated by dividing the odds in the treated or exposed (case) group by the odds in the control group.
  • the odds of an event are calculated as the number of events divided by the number of non-events. If the odds of an event are greater than one the event is more likely to happen than not (the odds of an event that is certain to happen are infinite); if the odds are less than one the chances are that the event won't happen (the odds of an impossible event are zero).
  • the strength of association was reported as odds ratios (OR) (with 95% lower (LCL) and upper
  • (UCL) confidence limit indicating the factor by which the risk of developing a disorder or disease is increased (OR>1), or indicating the factor by which the risk of developing a disorder or disease is decreased (OR ⁇ 1) (protective effect). o can be confident (to a computed probability, such as 90 or 95%) that the population value being estimated will be found. Confidence intervals indicate the strength of evidence; where confidence intervals are wide, they indicate less precise estimates of effect. The larger the trial's sample size, the larger the number of outcome events and the greater becomes the confidence that the true relative risk reduction is close to the value stated. Thus the confidence intervals narrow and "precision" is increased.
  • Example 1 Detection of nucleic acid variants in the ClQRl gene from patients with Recurrent Otitis Media (OM) and from control subjects
  • the control group (C) consisted of 172 healthy individuals. From each patient, informed consent to participate in the study is available.
  • the relevant coding sequences (see Table 1) of the genes under study were amplified using biotinylated oligonucleotides.
  • the polymorphisms were detected by use of a reverse hybridization method (Dot Probe Assay) with probes designed to recognize the polymorphisms as given in Table 1. After stringent washing at 56°C, hybridized probes were incubated with a streptavidine-alkaline phosphatase conjugate. The presence of a hybridized probe was revealed using NBIT/BCIP color development. Details on the reverse hybridization are described in Stuyver et al. (1996), Stuyver et al. (1997) and Van Geyt et al. (1998). 17 patients with recurrent OM and 172 C-diagnosed subjects were genotyped for 2 SNPs in the ClQRl gene.
  • the 95% confidence interval (95% CI) is the interval computed from the sample data which, were the study repeated multiple times, would contain the true effect 95% of the time.
  • Example 2 Detection of nucleic acid variants in the ClQRl gene from patients with Rheumatoid arthritis and from control subjects
  • the control group (C) consisted of 172 healthy individuals. From each patient, informed consent to participate in the study is available.
  • the relevant coding sequences (see Table 1) of the genes under study were amplified using biotinylated oligonucleotides.
  • the polymorphisms were detected by use of a reverse hybridization method (Dot Probe Assay) with probes designed to recognize the polymorphisms as given in Table 1. After stringent washing at 56°C, hybridized probes were incubated with a streptavidine-alkaline phosphatase conjugate. The presence of a hybridized probe was revealed using NBIT/BCIP color development. . , al. (1997) and Van Geyt et al. (1998).
  • the 95% confidence interval (95% CI) is the interval computed from the sample data which, were the study repeated multiple times, would contain the true effect 95% of the time.
  • M major allele (wt)
  • m minor allele (variant)

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Abstract

L'invention concerne un procédé et un kit permettant d'identifier un sujet présentant un risque de développer une indication, ou possédant un telle indication associée à une immunité innée modifiée. Les procédé et kit selon l'invention sont fondés sur la détermination de génotypes ClQRl et/ou les taux de sérum.
PCT/EP2006/050650 2005-02-08 2006-02-03 Variants d'acides nucleiques dans le gene c1qr1 associe a une immunite innee modifiee WO2006097388A2 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5965439A (en) * 1996-11-18 1999-10-12 The Regents Of The University Of California Host defense enhancement
US6268142B1 (en) * 1997-05-29 2001-07-31 Interleukin Genetics, Inc. Diagnostics and therapeutics for diseases associated with an IL-1 inflammatory haplotype
US20040001803A1 (en) * 2001-12-03 2004-01-01 Hancock Robert E.W. Effectors of innate immunity determination
WO2004048933A2 (fr) * 2002-11-21 2004-06-10 Wyeth Methodes de diagnostic de rcc et autres tumeurs solides
WO2004072265A2 (fr) * 2003-02-11 2004-08-26 Wyeth Procedes pour controler in vivo des activites de medicaments

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5965439A (en) * 1996-11-18 1999-10-12 The Regents Of The University Of California Host defense enhancement
US6268142B1 (en) * 1997-05-29 2001-07-31 Interleukin Genetics, Inc. Diagnostics and therapeutics for diseases associated with an IL-1 inflammatory haplotype
US20040001803A1 (en) * 2001-12-03 2004-01-01 Hancock Robert E.W. Effectors of innate immunity determination
WO2004048933A2 (fr) * 2002-11-21 2004-06-10 Wyeth Methodes de diagnostic de rcc et autres tumeurs solides
WO2004072265A2 (fr) * 2003-02-11 2004-08-26 Wyeth Procedes pour controler in vivo des activites de medicaments

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
CHANOCK S J ET AL: "SNPing away at innate immunity." THE JOURNAL OF CLINICAL INVESTIGATION. AUG 1999, vol. 104, no. 4, August 1999 (1999-08), pages 369-370, XP002343688 ISSN: 0021-9738 *
CHOI E H ET AL: "Genetic polymorphisms in molecules of innate immunity and susceptibility to infection with Wuchereria bancrofti in South India." GENES AND IMMUNITY. AUG 2001, vol. 2, no. 5, August 2001 (2001-08), pages 248-253, XP008052161 ISSN: 1466-4879 *
ELWARD K ET AL: ""Eat me" and "don't eat me" signals govern the innate immune response and tissue repair in the CNS: Emphasis on the critical role of the complement system" MOLECULAR IMMUNOLOGY 2003 UNITED KINGDOM, vol. 40, no. 2, 2003, pages 85-94, XP002343690 ISSN: 0161-5890 *
FOSTER C B ET AL: "Mining variations in genes of innate and phagocytic immunity: current status and future prospects." CURRENT OPINION IN HEMATOLOGY. JAN 2000, vol. 7, no. 1, January 2000 (2000-01), pages 9-15, XP008052162 ISSN: 1065-6251 *
KISHORE U ET AL: "CLQ: STRUCTURE, FUNCTION, AND RECEPTORS" IMMUNOPHARMACOLOGY, ELSEVIER SCIENCE PUBLISHERS BV, vol. 49, no. 1/2, August 2000 (2000-08), pages 159-170, XP001078864 ISSN: 0162-3109 *
MCGREAL E ET AL: "Structure-function studies of the receptors for complement C1q." BIOCHEMICAL SOCIETY TRANSACTIONS, vol. 30, no. 6, November 2002 (2002-11), pages 1010-1014, XP002343745 ISSN: 0300-5127 *
NEPOMUCENO R R ET AL: "CDNA CLONING AND PRIMARY STRUCTURE ANALYSIS OF C1QRP, THE HUMAN C1Q/MBL/SPA RECEPTOR THAT MEDIATES ENHANCED PHAGOCYTONIS IN VITRO" IMMUNITY, CELL PRESS, US, vol. 6, no. 2, February 1997 (1997-02), pages 119-129, XP002058777 ISSN: 1074-7613 *
NORSWORTHY PETER J ET AL: "Murine CD93 (C1qRp) contributes to the removal of apoptotic cells in vivo but is not required for C1q-mediated enhancement of phagocytosis." JOURNAL OF IMMUNOLOGY, vol. 172, no. 6, 15 March 2004 (2004-03-15), pages 3406-3414, XP002343686 ISSN: 0022-1767 cited in the application *
SONTHEIMER RICHARD D ET AL: "C1q: its functions within the innate and adaptive immune responses and its role in lupus autoimmunity." THE JOURNAL OF INVESTIGATIVE DERMATOLOGY. JUL 2005, vol. 125, no. 1, July 2005 (2005-07), pages 14-23, XP002343691 ISSN: 0022-202X *

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