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WO2012026885A1 - Diagnostic précoce de la respiration sifflante de l'enfance et de l'eczéma avec des médiateurs de monocytes de sang de cordon ombilical - Google Patents

Diagnostic précoce de la respiration sifflante de l'enfance et de l'eczéma avec des médiateurs de monocytes de sang de cordon ombilical Download PDF

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WO2012026885A1
WO2012026885A1 PCT/SG2011/000292 SG2011000292W WO2012026885A1 WO 2012026885 A1 WO2012026885 A1 WO 2012026885A1 SG 2011000292 W SG2011000292 W SG 2011000292W WO 2012026885 A1 WO2012026885 A1 WO 2012026885A1
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Kaw Yan Chua
Bee Wah Lee
I-Chun Kuo
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National University of Singapore
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National University of Singapore
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/20—Dermatological disorders

Definitions

  • the invention relates generally to methods and kits for determining predisposition or diagnosis of inflammatory disorders such as wheezing / eczema.
  • Asthma and Eczema are both caused by allergens and so are termed an allergic reaction. Other such allergic reactions include hay fever. Allergic / inflammatory disorders are commonly associated with an immature immune response
  • Wheezing in infants is a common symptom and is predominantly caused by viral infections. Some children will outgrow these symptoms but a subset of viral infections may play a role in the inception and exacerbation of asthma in older childhood (1).
  • Several possible predisposing factors that lead to the development of wheeze include prenatal smoking, maternal asthma, premature lung development and the hosts' immune responses.
  • Eczema a chronic or intermittent skin disease characterized by the cutaneous reactivity and pruritus, is commonly diagnosed during infancy and is considered a significant risk factor for wheezing and the subsequent development of asthma (4).
  • cord blood study There is a lack of published reports on the cord blood study in association with infant eczema. The main focus of cord blood studies till now has been in relation to the atopic dermatitis (4-7).
  • the innate immune response is of interest as the immaturity of the infant immune system is capable of altering the outcome of microbial, particularly viral, infections; leading to severe forms of illness due to enhanced immunological responses.
  • these enhanced immunological responses are responsible for the generation of various lineages of T-helper cells such as Thl, Th2, and Thl7 contribute to the inflammatory process in the airways 8 .
  • One objective is to identify the relationships between inflammatory disorders such as wheezing / eczema phenotypes and the intrinsic differences on the production of cytokine, chemokine and soluble mediators from cord blood mononuclear cells (CBMCs). These cytokine, chemokine and mediators secreted by CBMCs are analysed by combined multi- factor analysis approach to corelate with the susceptibility of wheeze or eczema during infancy. This early diagnosis at birth will be useful for early preventive or management measures to control the disease progression in early childhood.
  • CBMCs cord blood mononuclear cells
  • an aspect of the invention includes a method of diagnosing risk of an individual to suffer from an inflammatory disorder, the method comprising:
  • IL-10 Interleukin 10
  • IL-10 Interleukin 10
  • IL-10 Interleukin 10
  • the marker proteins are selected from the group consisting of cluster of differentiation 14 (CD 14) and tumor necrosis factor receptor type II (TNF-RII) or isoforms or fragments of TNF-RII and CD 14; tissue inhibitor of matrix metalloproteinase-1 (TIMP1) and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII; and CD 14, TIMP 1 and TNF-RII or isoforms or fragments of CD 14, TIMP1 and TNF-RII; and
  • the individual is diagnosed with the risk to suffer from an inflammatory disorder, if the measured level of IL-10 is lower than the level measured in the control sample and the level of the at least two substances are higher than the level measured in the control sample.
  • the sample from the individual is derived from one selected from the group consisting of blood, body fluid, whole blood, blood serum, pus, extracellular fluid and cord blood mononuclear cells (CBMCs).
  • CBDCs cord blood mononuclear cells
  • sample is serum free.
  • the substance representative for the quantity of the respective marker protein is the protein itself or a nucleic acid molecule encoding for the protein.
  • the nucleic acid is RNA comprising, mRNA or microRNA or siRNA.
  • the control sample is from an individual not suffering from an inflammatory disorder or not having the risk to suffer from an inflammatory disorder.
  • the individual is of an age of 0 days, or less than 100 years, or less than 90 years, or less than 80 years, or less than 70 years, or less than 60 years, or less than 50 years, or less than 40 years, or less than 30 years, or 20 years, or 15 years, or 10 years, or 9 years, or 8 years, or 7 years, or 6 years, or 5 years, or 4 years, or 3 years.
  • the individual is a new-born baby or an infant, or a toddler (1 to 3 years), or a teenager (10 to 19 years) or an adult (more than 20 years).
  • the inflammatory disorder is inflammatory skin disease.
  • dermatitis or eczema selected from the group consisting of atopic eczema, contact dermatitis, xerotic eczema, seborrhoeic dermatitis, dyshidrosis, discoid eczema, venous eczema, dermatitis herpetiformis, neurodermatitis, autoeczematisation, nummular dermatitis, statis dermatitis, perioral dermatitis and combinations thereof.
  • the dermatitis/eczema is selected from the group consisting of dryness of skin, crusting, flaking, blistering, cracking, oozing, bleeding, recurring skin rashes, skin redness, skin edema (swelling) and combinations thereof.
  • the marker proteins further comprise any of the proteins selected from the group consisting of interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-a (TNF-O!), growth regulated oncogene-a (GRO-o;), urokinase-type plasminogen activator receptor (u-PAR), basic fibroblast growth factor (bFGF), epithelial neutrophil-activating protein-78 (ENA-78), Agouti-related peptide (AgRP), Sialic acid-binding Ig-like lectin 5 (Siglec-5), platelet-derived growth factor AA (PDGF-AA), Activin A, tyrosine kinase with immunoglobulin-like and EGF-like domains (Tie-1), matrix metalloproteinase-1 (MMP-1), or isoforms or fragments thereof.
  • the marker protein CD 14 is a soluble form of CD 14.
  • the marker protein TNF-RII is a soluble form of TNF-RII.
  • the level of at least two substances representative for the quantity of respective marker proteins selected from the group consisting of CD 14, TIMP1, TNF-RII, isoforms and fragments is increased by at least 1 fold, or at least 2 folds, or at least 3 folds, or at least 4 folds, or at least 5 folds, or at least 6 folds, or at least 7 folds or at least 8 folds or at least 9 folds or at least 10 folds or between about 1 to about 10 folds, as compared to the control group.
  • the level of at least two substances representative for the quantity of respective marker proteins selected from the group consisting of CD 14, TIMP1, TNF-RII, isoforms and fragments is increased by 1 fold, or 1.5 folds, or 2 folds, or 2.5 folds, or 3 folds, or 3.5 folds, or 4 folds, or 4.5 folds, or 5 folds, or 5.5 folds, or 6 folds, or 6.5 folds, or 7 folds, or 7.5 folds, or 8 folds, or 8.5 folds, or 9 folds, or 9.5 folds, or 10 folds, as compared to the control group.
  • the method further comprises an increased level of the substance representative for the quantity of IL-6 or an isoform or fragment thereof, compared to the control group.
  • a decreased level of the substances representative for the quantity of respective marker proteins selected from the group consisting of IL-8, TNF- ⁇ , isoforms and fragments thereof, compared to the control sample is indicative of the risk of an individual to suffer from an inflammatory skin disease.
  • the inflammatory disorder is a respiratory tract-related disorder such as asthma.
  • the method may further comprise the steps of: a) measuring in the sample from the individual a level of a further substance selected from the group consisting of interleukin 1-/3 (IL-1/3) and interleukin 23 (IL-23); or isoforms or fragments of IL-1/3 and IL-23; interleukin-6 (IL-6) and IL-23 or isoforms or fragments of IL-6 and IL-23; and IL-1/3, IL-23, IL-6 or isoforms or fragments of IL-1/3, IL-23 and IL-6; and
  • the individual is diagnosed with the risk to suffer from a respiratory tract-related disorder, if the measured level is increased from the level measured in the control sample.
  • the fragment of IL-23 is IL-23p40 or IL23pl9 or IL-23p40 homodimer.
  • the marker proteins may further comprise any of the proteins selected from the group consisting of, IL-8, IL-2, interferon- ⁇ (IFN- ⁇ ), IL-5, tumor necrosis factor-a (TNF-a), CC chemokine 1-309, IL-16, macrophage derived chemokine (MDC), platelet derived growth factor-BB (PDGF-BB), FMS-like tyrosine kinase 3 ligand (FU-3L), IL-la, insulin-like growth factor-binding protein 6 (IGFBP-6), macrophage migration inhibitory factor (MIF), or isoforms or fragments thereof.
  • IGFBP-6 insulin-like growth factor-binding protein 6
  • MIF macrophage migration inhibitory factor
  • an increased level of at least two substances representative for the quantity of respective marker proteins selected from the group consisting of IL-1/3, IL-23, IL- 6, isoforms and fragments thereof compared to the control sample is indicative of the risk of an individual to suffer from a respiratory tract-related disorder.
  • an increased level of the substances representative for the quantity of respective marker proteins selected from the group consisting of IL-8, IL-2, IFN- ⁇ , IL-5, TNF-a, isoforms and fragments thereof compared to the control sample is indicative of the risk of an individual to suffer from a respiratory tract-related disorder.
  • the respiratory tract-related disorder may be caused by a viral infection. [035].
  • the respiratory tract-related disorder may be a wheezing disorder, or asthma or bronchiolitis or combinations thereof.
  • Another aspect of the invention includes a method of identifying a compound which can modulate the inflammatory response caused by an inflammatory disorder in an individual, the method comprising
  • IL-10 Interleukin 10
  • TIMP1 and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII CD 14, TIMP1 and TNF-
  • the compound can inhibit the inflammatory response caused by the inflammatory disorder if the measured marker level deviates from a marker level of the individual prior to being exposed with the compound or the measured marker level is substantially the same as the marker level measured in the control sample.
  • the measured marker level of the individual exposed with the compound may be lower than the marker level of the individual prior to being exposed with the compound.
  • the method may be carried out in an array such as a protein array or a gene array.
  • the protein array may be a multiplex-beads-based assay, or a plate-based ELISA, or a protein array on biochips.
  • the gene array may be a mRNA microarray, or a microRNA array, or a methylated DNA microarray or the gene array is used in a quantitative PCR method.
  • the sample prior to measuring in the sample from the individual the sample is exposed to a stimulator substance such as an adjuvant, lipopolysaccharide (LPS) or phytohaemagglutinin (PHA).
  • a stimulator substance such as an adjuvant, lipopolysaccharide (LPS) or phytohaemagglutinin (PHA).
  • the method may further comprise the steps of:
  • the compound can modulate the inflammatory response caused by a respiratory tract-related disorder if the measured marker level deviates from a marker level of the individual prior to being exposed with the compound or the measured marker level is substantially the same as the marker level measured in the control sample.
  • Another aspect of the invention includes a protein array kit for carrying out the methods of any one of claims 1 to 38, the protein array kit comprising
  • the first and second detection probes are selected from the group consisting of Interleukin 10 (IL-10), CD14 and TNF-RII or isoforms or fragments of CD14 and TNF-RII; TIMPl and TNF-RII or isoforms or fragments of TIMPl and TNF-RII; and CD 14, TIMPl, and TNF-RII or isoforms or fragments of CD 14, TIMPl, and TNF-RII.
  • IL-10 Interleukin 10
  • TIMPl and TNF-RII or isoforms or fragments of TIMPl and TNF-RII and CD 14, TIMPl, and TNF-RII or isoforms or fragments of CD 14, TIMPl, and TNF-RII.
  • the protein array kit may further comprise a third detection probe that is adapted to be immobilized on the array support; and a forth detection probe that is bound to a second detectable marker; wherein the third and forth detection probes are adapted to bind to a substance selected from the group consisting of IL-1
  • CBMCs were stimulated with 1 ug/mL of LPS.
  • CBMCs were stimulated with 5 ug/mL PHA.
  • FIG. 3 CBMCs derived cytokine profiles.
  • A IL-10 production from CBMCs stimulated with 1 ug/mL of LPS
  • B IL- 10 production from CBMCs in response to 5 ug/mL of PHA stimulation.
  • Figure 5 Results from a cytokine antibody array have identified soluble mediators in LPS stimulated culture supernatant associated with eczema, the soluble mediators CD14, sTNFRII and TIMP-1 are expressed at higher levels in the CBMCs of eczema subjects compared to the healthy controls and wheeze subjects. What is interesting about these soluble mediators is that Scdl4 and STFRII have been reported to be downregulators of IL-8, which might explain the attenuated IL-8 cytokine profile we saw in the eczema subjects. [052].
  • Figure 6. Flow chart of study profile.
  • the closed circles ( ⁇ ) represents non- allergen sensitized subjects, while the open circles ( O ) represent the allergen sensitized subjects.
  • the band in the middle of the boxplot represents the median value, and the ends of the whiskers represent the minimum and maximum value of the data.
  • Figure 9 3D Scatter plot of the healthy control and in wheeze group in respect to their correlation to the variables from the factor analysis. Factor analysis of LPS stimulated CBMCs showed that infant wheeze has higher combined responses from cytokines IL-6, IL- 23p40 and IL-1 3 compared to healthy controls. Figure was drawn with Sigma Plot (Systat Software, Inc., Chicago, IL).
  • Figure 10 Intracellular cytokine staining of LPS stimulated CBMCs show CD 14+ cell is main IL-6 producer.
  • the technology allows the early diagnosis of common childhood diseases wheeze and eczema.
  • the assay can be done immediately afterbirth with cord blood mononuclear cells.
  • the readout of these predictive mediators can be done with a mini-protein array in the future.
  • the results generated can be used for preventive managements of these diseases in high risk babies during the first few years of life to reduce the overall medical cost spent in the healthcare system.
  • CBMCs Cord blood mononuclear cells
  • CBMC cord blood mononuclear cell
  • Cytokine, chemokine and soluble mediator responses of CBMC to both the Toll- like- receptor 4 (TLR4) agonist lipopolysaccharide (LPS) and T cell mitogen phytohaemagglutinin (PHA) were assessed to target both the innate antigen presenting cells and naive T cells, respectively.
  • TLR4 Toll- like- receptor 4
  • PHA T cell mitogen phytohaemagglutinin
  • a method of diagnosing risk of an individual to suffer from an inflammatory disorder where measuring in a sample from an individual a level of Interleukin 10 (IL-10) and a level of at least two substances representative for the quantity of respective marker proteins, wherein the marker proteins are selected from the group consisting of cluster of differentiation 14 (CD 14) and tumor necrosis factor receptor type II (TNF-RII) or isoforms or fragments of TNF-RII and CD 14; tissue inhibitor of matrix metalloproteinase-1 (TIMP1) and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII; and CD 14, TIMP1 and TNF-RII or isoforms or fragments of CD 14, TIMP1 and TNF-RII; and comparing the measured level of IL-10 with a level measured in a control sample; comparing the measured level of the at least two substances with a level measured in a control sample; wherein the individual is diagnosed with the risk to suffer from an inflammatory disorder,
  • IL-10 Interleuk
  • a method of diagnosing risk of an individual to suffer from a respiratory tract-related disorder where measuring in a sample from an individual a level of at least two substances representative for the quantity of respective marker proteins, wherein the marker proteins is selected from the group consisting of interleukin 1-/3 (IL-1 3) and interleukin 23 (IL-23); or isoforms or fragments of IL-1/3 and IL- 23; interleukin-6 (IL-6) and IL-23 or isoforms or fragments of IL-6 and IL-23; and IL-1/3, IL- 23, IL-6 or isoforms or fragments of IL-1/3, IL-23 and IL-6; and comparing the measured level of the at least two substances with a level measured in a control sample; wherein the individual is diagnosed with the risk to suffer from a respiratory tract-related disorder, if the measured level deviates from the level measured in the control sample.
  • the marker proteins is selected from the group consisting of interleukin 1-/3 (IL-1 3) and inter
  • the measurement of at least two substances representative for the quantity of respective marker proteins can also be used in a method of identifying a compound which can modulate the inflammatory response caused by an inflammatory disorder such as a respiratory tract-related disorder in an individual.
  • Protein array kits are preferably used for carrying out the methods.
  • Abnormalities associated with inflammation comprise a large, officially unrelated group of disorders which underlie a vast variety of human diseases.
  • the immune system is often involved with inflammatory disorders, demonstrated in allergic reactions resulting in abnormal inflammation.
  • a large variety of proteins are involved in inflammation, and any one of them is open to a genetic mutation which impairs or otherwise dysregulates the normal function and expression of that protein. Examples of disorders associated with inflammation include: Asthma;
  • Allergy is a hypersensitivity disorder of the immune system whereby normally harmless environmental substances known as allergens cause acquired reactions that are predictable, and rapid. Strictly, allergy is one of four forms of hypersensitivity. It is characterized by excessive activation of certain white blood cells and IgE antibody resulting in an extreme inflamatory response. Common allergic reactions include eczema, hives, hay fever, asthma, food allergies and reaction to stinging insect venom.
  • the inflammatory disorder is an inflammatory skin disease such as dermatitis, or eczema.
  • Dermatitis/eczema may be selected from the group consisting of atopic eczema, contact dermatitis, xerotic eczema, seborrhoeic dermatitis, dyshidrosis, discoid eczema, venous eczema, dermatitis ⁇ , neurodermatitis, autoeczematisation, nummular dermatitis, statis dermatitis, perioral dermatitis and combinations thereof.
  • Symptoms may include dryness of skin, crusting, flaking, blistering, cracking, oozing, bleeding, recurring skin rashes, skin redness, skin edema (swelling) and combinations thereof.
  • the inflammatory disorder is a respiratory tract-related disorder such as asthma.
  • the respiratory tract-related disorder may be caused by a viral infection.
  • the respiratory tract-related disorder may be a wheezing disorder, or asthma or bronchiolitis or combinations thereof.
  • Samples can be taken from blood, body fluid, whole blood, blood serum, pus, extracellular fluid and cord blood mononuclear cells (CBMCs). Preferably samples are taken from CBMCs and are examined serum free.
  • the substance representative for the quantity of the respective marker protein is the protein itself or a nucleic acid molecule encoding for the protein.
  • the nucleic acid may be RNA such as mRNA or microRNA or siRNA.
  • the control sample is taken from an individual not suffering from an inflammatory disorder or not having the risk to suffer from an inflammatory disorder.
  • the individual from which the sample or the control sample are taken are of the same age of 0 days (at birth), or less than 100 years, or less than 90 years, or less than 80 years, or less than 70 years, or less than 60 years, or less than 50 years, or less than 40 years, or less than 30 years, or 20 years, or 15 years, or 10 years, or 9 years, or 8 years, or 7 years, or 6 years, or 5 years, or 4 years, or 3 years.
  • the individual is a new-born baby or an infant, or a toddler (1 to 3 years), but the individual may also be or a teenager (10 to 19 years) or an adult (more than 20 years).
  • the marker proteins may further comprise any of the proteins selected from the group consisting of interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-a (TNF-a), growth regulated oncogene-a (GRO-a), urokinase-type plasminogen activator receptor (u-PAR), basic fibroblast growth factor (bFGF), epithelial neutrophil-activating protein-78 (ENA-78), Agouti-related peptide (AgRP), Sialic acid- binding Ig-like lectin 5 (Siglec-5), platelet-derived growth factor AA (PDGF-AA), Activin A, tyrosine kinase with immunoglobulin-like and EGF-like domains (Tie-1), matrix metalloproteinase-1 (MMP-1), or isoforms or fragments thereof.
  • IL-6 interleukin-6
  • IL-8 interleukin-8
  • TNF-a tumor necrosis factor-a
  • the marker protein CD 14 is a soluble form of CD 14 or the marker protein TNF-RII is a soluble form of T F-RII.
  • the method of diagnosing risk of an individual to suffer from an inflammatory disorder is an inflammatory skin disease preferably the level of at least two substances selected from the group consisting of CD 14, TIMP1, TNF-RII, isoforms and fragments is increased by at least 1 fold, or at least 2 folds, or at least 3 folds, or at least 4 folds, or at least 5 folds, or at least 6 folds, or at least 7 folds or at least 8 folds or at least 9 folds or at least 10 folds or between about 1 to about 10 folds, as compared to the control group.
  • the level of at least two substances selected from the group consisting of CD 14, TIMP1, TNF-RII, isoforms and fragments is increased by at 1 fold, or 1.5 folds, or 2 folds, or 2.5 folds, or 3 folds, or 3.5 folds, or 4 folds, or 4.5 folds, or 5 folds, or 5.5 folds, or 6 folds, or 6.5 folds, or 7 folds, or 7.5 folds, or 8 folds, or 8.5 folds, or 9 folds, or 9.5 folds, or 10 folds, as compared to the control group.
  • the method of diagnosing risk of an individual to suffer from an inflammatory disorder is an inflammatory skin disease preferably the level of the substance representative for the quantity of IL-6 or an isoform or fragment thereof, is also increased compared to the control group.
  • the method of diagnosing risk of an individual to suffer from an inflammatory disorder is an inflammatory skin disease
  • the substances representative for the quantity of respective marker proteins selected from the group consisting of IL-8, TNF-a, isoforms and fragments thereof, being decreased compared to the control sample is indicative of the risk of an individual to suffer from an inflammatory skin disease.
  • the sample from the individual is measure for a level of a further substance selected from the group consisting of interleukin 1-/3 (IL-1/3) and interleukin 23 (IL-23); or isoforms or fragments of IL-1/3 and IL-23; interleukin-6 (IL-6) and IL-23 or isoforms or fragments of IL-6 and IL-23; and IL-1/3, IL-23, IL-6 or isoforms or fragments of IL-1/3, IL-23 and IL-6; and compared to the measured level of the further substance with a level measured in a control sample; wherein the individual is diagnosed with the risk to suffer from a respiratory tract-related disorder, if the measured level is increased from the level measured in the control sample.
  • the fragment of IL-23 is IL-23p40 or IL23pl9.
  • the marker proteins may further comprise any of the proteins selected from the group consisting of, IL-8, IL-2, interferon- ⁇ (IFN- ⁇ ), IL-5, tumor necrosis factor-a (TNF-a), CC chemokine 1-309, IL-16, macrophage derived chemokine (MDC), platelet derived growth factor-BB (PDGF-BB), FMS-like tyrosine kinase 3 ligand (Flt-3L), IL-la, insulin-like growth factor-binding protein 6 (IGFBP-6), macrophage migration inhibitory factor (MIF), or isoforms or fragments thereof.
  • IGFBP-6 insulin-like growth factor-binding protein 6
  • MIF macrophage migration inhibitory factor
  • the level of the substances representative for the quantity of respective marker proteins selected from the group consisting of IL-1/3, IL-23, IL-6, isoforms and fragments thereof is increased compared to the control sample.
  • An increase being indicative of the risk of an individual to suffer from a respiratory tract-related disorder.
  • An increased level of the substances representative for the quantity of respective marker proteins selected from the group consisting of IL-8, IL-2, IFN- ⁇ , IL-5, TNF-a, isoforms and fragments thereof compared to the control sample may be further indicative of the risk of an individual to suffer from a respiratory tract-related disorder.
  • a method of identifying a compound which can modulate the inflammatory response caused by an inflammatory disorder in an individual comprising measuring in a sample obtained from the individual a level of at least two substances representative for the quantity of respective marker proteins, wherein the marker proteins are selected from the group consisting of Interleukin 10 (IL-10), CD 14 and TNF-RII or isoforms or fragments of CD 14 and TNF-RII; TIMP1 and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII; and CD 14, TIMP1 and TNF-RII or isoforms or fragments of CD 14, TIMP1 and TNF-RII; wherein the individual is suffering from an inflammatory skin disease or has a risk to suffer from an inflammatory skin disease and has been exposed with the compound prior to obtaining the sample from the individual; and comparing the measured marker level of the at least two substances with a marker level measured in a control sample, wherein the compound can inhibit the inflammatory response caused by the inflammatory disorder
  • the methods of the invention are carried out in an array.
  • the array may be a protein array or a gene array.
  • the protein array may be a multiplex-beads-based assay, or a plate-based ELISA, or a protein array on biochips.
  • the array is a gene array the gene array may be an mRNA microarray, or a microR A array, or a methylated DNA microarray or the gene array is used in a quantitative PCR method.
  • An example of an array system suitable for carrying out the invention is depicted in Figure 13. Other array systems are known to those skilled in the art.
  • the methods of the invention prior to measuring in the sample from the individual, include the step of exposing the sample to a stimulator substance.
  • the stimulator substance is an adjuvant.
  • the stimulator substance is lipopolysaccharide (LPS) or phytohaemagglutinin (PHA).
  • the method of identifying a compound which can modulate the inflammatory response caused by an inflammatory disorder is to modulate a respiratory tract-related disorder
  • the method may further comprising the steps of: measuring in the sample obtained from the individual a level of a further substance selected from the group consisting of IL-1/3 and IL-23 or isoforms or fragments of IL-1/3 and IL-23; IL-6 and IL-23 or isoforms or fragments of IL-6 and IL-23; and IL-1 3, IL-23, IL-6 or isoforms or fragments of IL-1/3, IL-23 and IL-6; and wherein the individual is suffering from a respiratory tract-related disorder or has a risk to suffer from a respiratory tract-related disorder and wherein the individual has been exposed with the compound prior to obtaining the sample from the individual; and comparing the measured marker level of the further substance with a marker level measured in a control sample, wherein the compound can modulate the inflammatory response caused by a respiratory tract-related disorder
  • a first detection probe that is adapted to be immobilized on the array support; and iii) a second detection probe that is bound to a detectable marker;
  • first and second detection probes are selected from the group consisting of Interleukin 10 (IL-10), CD14 and TNF-RII or isoforms or fragments of CD14 and TNF-RII; TIMP1 and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII; and CD 14, TIMP1, and TNF-RII or isoforms or fragments of CD 14, TIMP1, and TNF-RII.
  • IL-10 Interleukin 10
  • TIMP1 and TNF-RII or isoforms or fragments of TIMP1 and TNF-RII and CD 14, TIMP1, and TNF-RII or isoforms or fragments of CD 14, TIMP1, and TNF-RII.
  • the protein array kit may be used in a sandwich ELISA or in other methods known in the art.
  • the protein array kit may further comprise a third detection probe that is adapted to be immobilized on the array support; and a forth detection probe that is bound to a second detectable marker; wherein the third and forth detection probes are adapted to bind to a substance selected from the group consisting of IL-1/3 and IL-23 or isoforms or fragments of IL-1/3 and IL-23; IL-6 and IL-23 or isoforms or fragments of IL-6 and IL-23; and IL-1/3, IL- 23, IL-6 or isoforms or fragments of IL-1/3, IL-23 and IL-6.
  • CBMCs Cord blood mononuclear cells
  • LPS lipopolysaccharide
  • PHA phytohemagglutinin
  • pro-inflammatory cytokines IL-6, IL-8, IFN- ⁇ , IL-2, IL-5 and TNF-a
  • LPS lipopolysaccharide
  • PHA phytohemagglutinin
  • Umbilical cord blood was collected by withdrawing blood from an umbilical vein after delivery and processed within 24 hours.
  • Cord blood mononuclear cells CBMCs
  • CBMCs Cord blood mononuclear cells
  • LPS lipopolysaccharide
  • PHA phytohaemagglutinin
  • cytokine production was performed using the fluorescence activated cell sorter (FACS)-Array (BD Bioscience), which is a new flow cytometry platform for fast and sensitive analysis of cytokine proteins by multiplexed bead assays.
  • FACS fluorescence activated cell sorter
  • BD Bioscience Fluorescence activated cell sorter
  • the advantage is the usage of small sample volumes; fewer sample dilutions and values are obtained in substantially less time as compared to conventional ELISA.
  • Cytokines measured were IL-10, IL-lft IL-6, IL-8, IL-12p70, TNF-a, IFN- ⁇ , IL-2, IL-5, IL-23p40 and IL-13.
  • the detection limit was 10 pg/mL for all cytokines.
  • factor analysis was used. Cytokines in the physiological setting do not act alone but in concert with other cytokines in a network of groups.
  • the factor analysis is a data reduction technique which takes a large set of variables (cytokines) and groups together inter-correlated sets of variables (cytokines), this summary of data are known as factors. Factors with an eigenvalue greater than 1 were formed using only the healthy control subjects to describe cytokine profile of the healthy children. Factor-specific loading was calculated for each original variable which indicates the degree of correlation between the specific factor formed and the variable. To perform additional analysis by using factors as variables, factor scores were generated by the statistical software SPSS.
  • Factor scores are a linear combination of all the variables weighted by the corresponding factor loadings. These factor scores were then compared between groups by Mann-Whitney U test. Logistic regression was used to determine factor scores as possible predictors of wheeze and eczema while adjusting for confounding factors (9).
  • Table 1 Summary of factor analysis including factor loadings from a model of cytokine responses in the LPS stimulated CBMCs of healthy control subjects. Comparisons of factor scores between groups were compared using Mann-Whitney U test.
  • Soluble mediators which were differentially expressed in the eczema and wheeze group were detected in the LPS stimulated CBMCs.
  • Our preliminary studies from a cytokine antibody array shows that LPS stimulated CBMC culture supernatants have higher soluble CD 14, soluble TNF-RII and TIMP-1 in infants who developed eczema compared to infants who developed wheeze and health controls (Figure 5). These targets were selected for further studies to validate the predictive power for early diagnosis of childhood eczema.
  • CD 14 receptor forms a complex with LPS binding protein (LBP) and LPS resulting in the secretion of a variety of cytokines.
  • LBP LPS binding protein
  • Natural monocyte activating agents such as LPS are capable of inducing protease mediated shedding of membrane bound CD 14 receptor from cell surface and sCD14 could be released as an acute phase protein.
  • sCD14 may serve as a modulator of the binding of LPS on mCD14 by competing for LPS binding (10,11). Studies have shown that sCD14 measured in cord blood serum have been shown to be present in differential levels in atopic dermatitis and wheeze infants. It is known to be a regulatory factor capable of interfering with CD40 signalling in B cells and inhibiting IL-6 production (12).
  • sTNFRII Soluble Tumour Necrosis Factor Receptor 2
  • TIMP-1 are endogenous inhibitors that inhibit activity of matrix metalloproteinases (MMPs) which are proteolytic enzymes produced by inflammatory cells. TIMP-1 has been shown to bind to pro and active forms of MMP-9. It would be interesting to investigate TIMP-1 as an imbalance of this mediator via the involvement of different cytokines has been reported in inflammatory disease (14). Preliminary results have shown that cord blood supernatants of infants with eczema are capable of producing more TIMP-1 than wheeze patients. Profile
  • the primary clinical outcome measure was development of eczema and episodes of wheezing which were diagnosed by clinicians 11
  • the secondary outcome measures were allergen sensitization, asthma and allergic rhinitis.
  • Infants were evaluated by a paediatrician at 1, 3, 6, and 12 months of age, which involved a detailed history, recording of anthropometric data and clinical examination.
  • Questionnaires were also administered at these visits to record clinical disease and environmental exposures, including day care, size of sibship, use of antibiotics, and smoking and pets.
  • Biweekly phone calls were performed for the first 6 months, after which monthly phone contacts were carried out to collect data on the health status of the children.
  • Eczema was defined as a pruritic rash over the face and/ or extensors with a chronic relapsing course as described by Hanifin and Rajka and modified by Seymour for infants 12 , and the severity assessed by The Scoring Atopic Dermatitis (SCORAD) index
  • Serum was collected from cord blood and at 12 months, samples were stored at -70°C before being assayed.
  • the total IgE of the serum at 1 year was measured using the fluoroenzymeimmunoassay method (UniCAPs Phadiatop, Pharmacia Diagnostics, Uppsala, Sweden) with a detection limit of 0.35 kU/L.
  • Skin prick test was performed at 12 months of age using a standardized technique with common allergen extracts such as (Alyostal, Stallergenes Laboratoires, Antony Cedex, France), milk, egg yolk, egg white, dust mite allergens - Dermatophagoides pteronyssinus (Greer Laboratories, Lenoir, NC, USA) and Blomia tropicalis manufactured in-house 14 .
  • a histamine dihydrochloride solution (10 mg/mL) was used as a positive control and solvent (50% Cocas 50% Gly) as a negative control.
  • a weal >3mm in diameter above the negative control was considered positive 15 .
  • Umbilical cord blood was collected into blood bags (Terumo, Somerset, USA) prepared with heparin and RPMI medium (Gibco Life Technology, Grand Island, NY, USA). Cord blood is drawn from an umbilical vein after delivery and processed within 24 hours. The heparinized blood was centrifuged at 700 rpm to remove plasma layer before cord blood mononuclear cells (CBMCs) were isolated by Ficoll - Hypaque (GE Healthcare, Uppsala, Sweden) gradient centrifugation.
  • CBMCs cord blood mononuclear cells
  • CBMCs were thawed and washed 3 times with Hanks Balanced Salt Solution (HBSS) (Sigma, St Louis, Mo, USA) before mixing the cells with trypan blue to determine cell viability. The cells normally retained 60% cell viability after thawing.
  • HBSS Hanks Balanced Salt Solution
  • the CBMCs were cultured in AIM-V medium (Gibco Life Technology, Grand Island, NY, USA) and 1 ⁇ 10 "4 M 2-Mercaptoethanol (Gibco Life Technology, Grand Island , NY , USA ) at 5 ⁇ 10 5 viable cells/well in 200 ⁇ medium.
  • cytokine production was performed using the fluorescence activated cell sorter (FACS)-Array (BD Bioscience, Franklin Lakes, NJ , USA). Cytokines measured were IL-10, IL-1 3 , IL-6, IL-8, IL-12p70, TNF-a , IFN- ⁇ , IL-2 , IL-5, IL-23p40 and IL-13 using the BD bead array flexset (BD Bioscience, Franklin Lakes, NJ, USA). The detection limit was lOpg/mL for all cytokines. The responses presented are those evoked by the LPS / PHA stimulus after subtraction by responses from unstimulated cultures.
  • FACS fluorescence activated cell sorter
  • factor analysis was used. Cytokines in the physiological setting do not act alone but in concert with other cytokines in a network of groups.
  • the factor analysis is a data reduction technique which takes a large set of variables (cytokines) and groups together intercorrelated sets of variables (cytokines), this summary of data are known as factors. Factors with an eigenvalue greater than 1 were formed using only the healthy control subjects to describe cytokine profile of the healthy children. Factor-specific loading was calculated for each original variable which indicates the degree of correlation between the specific factor formed and the variable. To perform additional analysis by using factors as variables, factor scores were generated by the statistical software SPSS.
  • Factor scores are a linear combination of all the variables weighted by the corresponding factor loadings. These factor scores were then compared between groups by Mann- Whitney test. Logistic regression was used to determine factor scores as possible predictors of wheeze and eczema while adjusting for confounding factors I6 .
  • Table 2 Demographic characteristics of the children with eczema and wheeze and the healthy control subjects. Values were presented as median with interquartile range and numbers with percentages for categorical data. Mann- Whitney tests; otherwise ⁇ 2 tests were used.
  • Table 4 Allergen sensitization chart of infants at 24 months of age.
  • Table 6 Cytokine responses to lipopolysaccharide (LPS) and phytoheamagglutinin (PHA) in infants with wheeze. P values have been adjust for risk factors birth height, birth weight, birth order, mode of delivery and maternal asthma using logistic regression analysis.
  • LPS lipopolysaccharide
  • PHA phytoheamagglutinin
  • Table 7 Summary of factor analysis, including factor loadings from a model of cytokine responses in the LPS and PHA stimulated CBMCs of healthy control subjects
  • the PHA stimulated CBMC cytokine profiles are shown in Figure 8.
  • cytokine IL-23 a novel member of IL- 12 cytokine family. It is a heterodimeric cytokine produced by antigen presenting cells (monocytes, dendritic cells) which compose of a pi 9 subunit specific for IL-23 and a p40 subunit shared with IL-12 9 .
  • IL-23 is a signature cytokine that is important for the differentiation of CD161 + precursors cells from cord blood and newborn thymus into Thl7 cells, and for polarizing neonatal CD4 + and CD8 + T cells into IL-17 producing cells 19 ' 20 ' 22 ' 24 .
  • CD8+ T cells the dominant pathogenesis role of CD8+ T cells in RSV infection has been demonstrated by two animal studies using experimental respiratory syncytial virus infection mouse model 25 ' 26 '.
  • IL-23 requires an interaction with a heterodimeric complex composed of IL-12R/31 and IL23R to allow signal transduction and this polarizes the differentiation of naive T cells into CD4+ Thl7 cells receptor 22 ' 21 .
  • Thl7 specific transcription factor RORC2 is necessary by pro-inflammatory cytokines IL-6 and IL-10 through signal transducer and activator of transcription 3 (STAT3).
  • STAT3 signal transducer and activator of transcription 3
  • IL-23 or IL-1 3 were enough to drive Thl7 development from naive CD4+ T cells and IL-6 is able to further enhance this development ' ' ' .
  • monocytes activated by lipopolysaccharide (LPS) which produce IL-1/3 and IL-6 were the most efficient antigen presenting cells (APCs) for TH17 differentiation which lead to the hypothesis that changes in the APC function especially in monocytes precedes inappropriate development and differentiation into Thl7 cells 18 .
  • CBMCs have been shown to produce higher levels of IL-6 or IL-8 in response to RSV 29, 30 or LPS 6 stimulation.
  • the IL-8 is primary neutrophil chemokine attractant molecules, secreted in the epithelial cells upon viral infection. The main function of these neutrophils will be to adhere to epithelial cells and augment epithelial damage and detachment of epithelial cells 31 .
  • IL-6 has not been directly associated with wheeze outcome in infants
  • IL-6 as a pro-inflammatory cytokine has also been reported to be produced in higher levels in CBMC of neonates with high risk of allergy and also those who subsequently had allergic disease 35, 36 .
  • IL-6 derived from antigen presenting cells (APCs) is able to polarize CD4+ T cells into Th2 cells and inhibit the development of T regulatory cells which is an important protective arm of the immune response 37 .
  • LPS and PHA stimulated IL-10 did not appear to be the key cytokine that is differentially expressed in disease and healthy groups in this study despite it's role as a potential immunoregulator by inhibiting the progression or expression of inflammatory diseases 7 ⁇ 45> 46 .
  • Attenuated IL-10 was reported to be associated with the wheezing phenotype at 5 years of age, but the effects of this cytokine was strengthened by generating a IL-10/IL-5 response ratio for each subject 7 .
  • This study was also contradictory to another project by COAST on high risk children which also investigated relationships between cord blood cytokine profile responses and susceptibility to viral infections during the first year of life. There were no comparable relationships between the IL-5 and IL-10 responses to subsequent viral infections in that cohort 5 .
  • IL-8 was found to be significantly suppressed in the eczema subjects not only in comparison to the wheeze but also to the healthy subjects.
  • a combination of LPS stimulated TNF- ⁇ , IL-10 and IL-8 cytokines in the eczema group has lower factor scores (lower production) relative to healthy control subjects.
  • the suppression in IL-8 cytokine seen in eczema infants lead us to believe that other soluble factors such as soluble receptors can play a role in regulating this pro-inflammatory cytokine
  • the invention described herein may include one or more range of values (eg size, concentration etc).
  • a range of values will be understood to include all values within the range, including the values defining the range, and values adjacent to the range which lead to the same or substantially the same outcome as the values immediately adjacent to that value which defines the boundary to the range.
  • Copenhaver CC Gern JE, Li Z, Shult PA, Rosenthal LA, Mikus LD, et al. Cytokine response patterns, exposure to viruses, and respiratory infections in the first year of life. Am J Respir Crit Care Med 2004; 170: 175-80.
  • Severity scoring of atopic dermatitis the SCORAD index. Consensus Report of the European Task Force on Atopic Dermatitis. Dermatology 1993; 186:23-31.
  • T helper 17 cells In humans requires T cell receptor ligation in the context of Toll-like receptor- activated monocytes. Proc Natl Acad Sci U S A 2007; 104:17034-9.

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Abstract

L'invention concerne des procédés de diagnostic du risque qu'un individu souffre d'un trouble inflammatoire, un procédé d'identification d'un composé qui peut moduler la réponse inflammatoire causée par un trouble inflammatoire chez un individu, et des kits de puce de protéine pour conduire les procédés. Les procédés comprenant la mesure dans un échantillon d'un individu du taux d'interleukine 10 (IL-10) et d'un taux d'au moins deux substances choisies dans le groupe constitué de l'agrégat de différenciation 14 (CD14) et du récepteur de facteur de nécrose tumoral de type II (TNF-RII) ou des isoformes ou fragments de TNF-RII et CD14 ; l'inhibiteur tissulaire de métalloprotéase de matrice 1 (TIMP1) et TNF-RII ou des isoformes ou fragments de TIMP1 et TNF-RII ; et CD14, TIMP1 et TNF-RII ou des isoformes ou fragments de CD14, TIMP1 et TNF-RII ; et la comparaison du niveau mesuré à un niveau mesuré dans un témoin.
PCT/SG2011/000292 2010-08-24 2011-08-24 Diagnostic précoce de la respiration sifflante de l'enfance et de l'eczéma avec des médiateurs de monocytes de sang de cordon ombilical Ceased WO2012026885A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105389479A (zh) * 2014-08-27 2016-03-09 霍夫曼-拉罗奇有限公司 用于分析核酸扩增反应的分析方法和系统
RU2702236C1 (ru) * 2019-05-08 2019-10-07 федеральное государственное бюджетное образовательное учреждение высшего образования "Башкирский государственный медицинский университет" Министерства здравоохранения Российской Федерации Способ прогнозирования перехода средне-тяжёлого течения нумулярной микробной экземы в тяжёлое течение

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008104953A2 (fr) * 2007-02-28 2008-09-04 The Procter & Gamble Company Procédés et cibles d'identification de composés permettant la régulation d'une infection par rhinovirus
WO2009123737A2 (fr) * 2008-04-03 2009-10-08 Becton, Dickinson And Company Détection avancée d'une sepsie

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008104953A2 (fr) * 2007-02-28 2008-09-04 The Procter & Gamble Company Procédés et cibles d'identification de composés permettant la régulation d'une infection par rhinovirus
WO2009123737A2 (fr) * 2008-04-03 2009-10-08 Becton, Dickinson And Company Détection avancée d'une sepsie

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ASADULLAH, K. ET AL.: "Interleukin-10 therapy - review of a new approach", PHARMACOLOGICAL REVIEWS, vol. 55, no. 2, 2003, pages 241 - 269 *
SHEEHAN, K. C. F. ET AL.: "Monoclonal antibodies specific for murine p55 and p75 tumor necrosis factor receptors: identification of a novel in vivo role for p75", THE JOURNAL OF EXPERIMENTAL MEDICINE, vol. 181, 1995, pages 607 - 617 *
ZIEGLER-HEITBROCK, L.: "The CD14+ CD16+ blood monocytes: their role in infection and inflammation", JOURNAL OF LEUKOCYTE BIOLOGY, vol. 81, 2007, pages 584 - 592 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105389479A (zh) * 2014-08-27 2016-03-09 霍夫曼-拉罗奇有限公司 用于分析核酸扩增反应的分析方法和系统
CN105389479B (zh) * 2014-08-27 2021-08-10 霍夫曼-拉罗奇有限公司 用于分析核酸扩增反应的分析方法和系统
RU2702236C1 (ru) * 2019-05-08 2019-10-07 федеральное государственное бюджетное образовательное учреждение высшего образования "Башкирский государственный медицинский университет" Министерства здравоохранения Российской Федерации Способ прогнозирования перехода средне-тяжёлого течения нумулярной микробной экземы в тяжёлое течение

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