WO2005071408A1 - Method of diagnosing infectious disease by measuring the level of soluble trem-1 in a sample - Google Patents
Method of diagnosing infectious disease by measuring the level of soluble trem-1 in a sample Download PDFInfo
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- WO2005071408A1 WO2005071408A1 PCT/GB2005/000273 GB2005000273W WO2005071408A1 WO 2005071408 A1 WO2005071408 A1 WO 2005071408A1 GB 2005000273 W GB2005000273 W GB 2005000273W WO 2005071408 A1 WO2005071408 A1 WO 2005071408A1
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/12—Pulmonary diseases
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/26—Infectious diseases, e.g. generalised sepsis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/56—Staging of a disease; Further complications associated with the disease
Definitions
- This invention relates generally to the field of immunology. More particularly, the present invention relates to inflammation and the use of markers that allow the prompt diagnosis of infectious disease (for example of bacterial or fungal) origin and the follow up of infected patients during pharmacological treatment. These markers have particular applications in the diagnosis of pneumonia and sepsis.
- infectious disease for example of bacterial or fungal
- the Inventors describe a rapid detection test of the soluble form of the human TREM-1 receptor (sTREM-1) in bronchoalveolar fluid of mechanically ventilated patients to accurately diagnose bacterial or fungal pneumonia. Many non-infectious processes lead to fever and new pulmonary infiltrates in the mechanically ventilated patient, rendering the diagnosis of pneumonia (and especially ventilator-associated pneumonia) very challenging.
- the systemic signs of infection are non-specific findings and can be caused by any condition that releases cytokines.
- Pugin etal. Am Rev Respir Dis 1991;143:1121-9 combined body temperature, white biood cells count, volume and appearance of tracheal secretions, ratio of the partial pressure of arterial oxygen to the fraction of inspired oxygen (PaO2/FiO2), chest-X-ray, and tracheal aspirate cultures into a clinical pulmonary infection score (CPIS) and reported that a score >6 was associated with a high likelihood of pneumonia.
- CPIS clinical pulmonary infection score
- TNF ⁇ tumor necrosis factor- ⁇
- IL-1 ⁇ interleukin-1 ⁇
- other cytokines have been demonstrated to be increased in various types of pulmonary infectious challenges with potential prognostic implications.
- the Inventors were unable to determine accurate discriminating cut-off level of such mediators for the diagnosis of pneumonia.
- the Inventors using an easy-to-perform immunoblot technique, demonstrate herein that a soluble form of TREM-1 (sTREM-1) is released locally in the bronchoalveolar lavage fluid from patients suffering from pneumonia with a sensitivity of at least 98 percent.
- sTREM-1 was detected in only 6 out of 64 patients without pneumonia. Bronchoalveolar lavage fluid levels of sTREM-1 were not correlated to any of the clinical or biological parameters tested and stood as an independent parameter of high specificity. In a multiple logistic regression analysis, presence of sTREM-1 in bronchoalveolar lavage fluid was shown to be the best predictor of pneumonia with an odds ratio as high as 41.52. Presence of sTREM-1 by itself was more accurate that any clinical findings or laboratory values in identifying the existence of bacterial or fungal pneumonia. Thus rapid detection of sTREM-1 in bronchoalveolar lavage fluid is useful in establishing or excluding the diagnosis of bacterial or fungal pneumonia.
- PCT procalcitonin
- CRP C-reactive protein
- SIRS systemic inflammatory response syndrome
- Sepsis is further stratified into severe sepsis when there is evidence of organ hypoperfusion, made evident by ' signs of organ dysfunction such as hypoxaemia, oliguria, lactic acidosis or altered cerebral function.
- Septic shock is severe sepsis complicated by hypotension defined as systolic blood pressure less than 90 mmHg despite adequate fluid resuscitation.
- Sepsis and SIRS may be complicated by the failure of two or more organs, termed multiple organ failure (MOF), due to disordered organ perfusion and oxygenation.
- MOF multiple organ failure
- a systemic inflammatory response may occur in severe inflammatory conditions such as pancreatitis and burns. The appearance of signs of an inflammatory response is less well defined following traumatic insults.
- gram-negative bacteria are implicated in 50 to 60% of sepsis with gram-positive bacteria accounting for a further 35 to 40% of cases. The remainder of cases are due to the less common causes of fungi, viruses and protozoa.
- SIRS Systemic Inflammatory Response Syndrome
- Early recognition of sepsis and Systemic Inflammatory Response Syndrome (SIRS) in the critically ill patient may avoid the increased morbidity, mortality and length of stay associated with multiple organ failure.
- SIRS Systemic Inflammatory Response Syndrome
- the present invention is directed towards circumventing the existing problems associated with diagnosing sepsis to provide an accurate and consistent method of detection.
- the Inventors describe the value of assaying the soluble form of TREM-1 (sTREM-1) in plasma samples of newly admitted critically ill patients with suspected sepsis as a new approach to accurately diagnose infectious processes.
- sTREM-1 soluble form of TREM-1
- Early identification of infection has a major impact on the clinical course, management and outcome of critical patients.
- Critical care physicians have at their disposal a variety of indicators to serve as a guide in discriminating infectious from non-infectious conditions in newly admitted patients.
- the diagnosis of sepsis becomes clear after completing the medical history and physical examination of a newly admitted patient (Bates DW, et al. Ann Intern Med. 1990;113:495-500).
- plasmatic sTREM-1 level assay offers a higher degree of certainty than other currently available candidates. This accuracy can usefully guide physicians in their clinical decision-making and stepwise approach to the complex management of critically ill patients.
- the immunoblot technique used here can be performed within 3 to 4 hours and may provide valuable information long before blood culture results are back. Moreover, it is of low cost and can be applied to small series or even individual samples.
- the results reported here demonstrate that rapid measurement of the plasmatic sTREM-1 levels may improve the ability of clinicians to differentiate patients with sepsis from those with systemic inflammation of non-infectious origin. This should be especially useful among patients in whom the diagnosis is not clinically straightforward.
- the immunoblot technique described is rapid, accurate, of low cost and can be applied to small series or even individual samples. Use of this test to assess plasmatic sTREM-1 levels should lead to a more accurate diagnosis of sepsis in patients admitted in ICUs with a clinical suspicion of infection.
- the triggering receptor expressed on myeloid cells-1 is a member of the Ig-superfamily, the expression of which is up-regulated on phagocytic cells in the presence of bacteria or fungi (Bouchon A et al. Nature 2001;230:1103-7).
- the inventors have determined that TREM-1 is shed or secreted from the membrane of activated phagocytes and can be found in a soluble form in body fluids and is therefore a useful diagnostic marker.
- the presence of a soluble form of TREM-1 (sTREM-1) in bronchoalveolar lavage (BAL) fluid from mechanically ventilated patients is shown herein to be a good indicator of infectious pneumonia.
- BAL bronchoalveolar lavage
- the present invention provides methods and compositions for the clinical screening and diagnosis of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- the present invention provides methods and compositions for monitoring the effectiveness of the treatment of disease of bacterial or fungal origin, for example, pneumonia or sepsis, for selecting participants in clinical trials relating disease of bacterial or fungal origin, for identifying subjects most likely to respond to a particular therapeutic treatment for disease of bacterial or fungal origin and for screening and development of drugs for treatment of disease of bacterial or fungal origin.
- the invention provides a method of diagnosis of disease of bacterial or fungal origin in a subject, which method comprises the step of measuring the level of sTREM-1 in a biological sample obtained from said subject.
- the disease is an inflammatory state
- said method is capable of identifying a microbial origin for said inflammatory state.
- inflammatory states pneumonia and sepsis of bacterial or fungal origin.
- the invention provides a method of diagnosis of pneumonia in a subject, which method comprises the step of measuring the level of sTREM-1 in a biological sample obtained from said subject.
- the invention provides a method of diagnosing sepsis of bacterial or fungal origin in a subject, which method comprises the step of measuring the level of sTREM-1 in a biological sample obtained from said subject.
- the invention provides methods of diagnosing or monitoring disease of bacterial or fungal origin, for example, pneumonia or sepsis in a patient, comprising: measuring the level of sTREM-1 in a sample from the patient, wherein the level is an indicator of presence or extent of disease of bacterial or fungal origin in the patient.
- sTREM-1 is a soluble form of the TREM-1 Receptor which can be detected in certain body fluid samples by an antibody raised against the TREM-1 Receptor.
- sTREM-1 means, an inflammation of the lung caused by infection by extracellular pathogens such as bacterial infection, and non-bacterial infections (for example, infection by Blastomyces dermatitidis, Histoplasma capsulatum, Coccidioides, Sporothrix schenckii, Pneumocystis carinii, Cryptococcus, Aspergillus, or Mucorsp.), protozoal infections or parasitic infections (for example, those caused by Toxoplasma gondii, Strongyloides stercoralfs, Ascaris, hookworm, Dirofilaria, Paragonimus, or Entamoeba histolytica) where increased expression of sTREM-1 can be detected.
- non-bacterial infections for example, infection by Blastomyces dermatitidis, Histoplasma capsulatum, Coccidioides, Sporothrix schenckii, Pneumocystis carinii, Cryptococcus, Aspergill
- Pneumonia includes "Lobar Pneumonia” (which occurs in one lobe of the lung) and Bronchopneumonia (tends to be irregularly located in the lung). Furthermore, pneumonia is often classified into two categories that may help predict the organisms that are the most likely culprits. "Community- acquired (pneumonia contracted outside the hospital). Pneumonia” in this setting often follows a viral respiratory infection. It affects nearly 4 million adults each year. It is likely to be caused by Streptococcus pneumoniae, the most common pneumonia-causing bacteria. Other organisms, such as atypical bacteria called Chlarnydia or Mycoplasma pneumonia are also common causes of community-acquired pneumonia.
- SIRS Systemic Inflammatory Response Syndrome
- extracellular pathogens such as bacterial infection, for example bacteremia (the presence of bacteria in the blood) with or without organ failure, and non- bacterial infections, such as fungemia (for example, yeast infection by Candida albicans), protozoal infections or parasitemia (such as in filariasis, malaria, and trypanosomiasis) where increased expression of sTREM-1 can be detected.
- bacterial infection for example bacteremia (the presence of bacteria in the blood) with or without organ failure
- non- bacterial infections such as fungemia (for example, yeast infection by Candida albicans), protozoal infections or parasitemia (such as in filariasis, malaria, and trypanosomiasis) where increased expression of sTREM-1 can be detected.
- fungemia for example, yeast infection by Candida albicans
- protozoal infections or parasitemia such as in filariasis, malaria, and trypanosomiasis
- the measurement of the level of sTREM-1 comprises the steps of (a) contacting said biological sample with a compound capable of binding sTREM-1 ; and (b) detecting the level of sTREM-1 present in the sample by observing the level of binding between said compound and sTREM- 1.
- the assay or measurement of the sample for the levels of sTREM-1 present in the sample may be carried out using standard protocols known in the art. For example, where the observation of binding between sTREM-1 and the compound capable of binding sTREM-1 takes place, this observation may be carried out using known methodologies.
- the binding may be detected through use of a competitive immunoassay, a non-competitive assay system using techniques such as western blots, a radioimmunoassay, an ELISA (enzyme linked immunosorbent assay), a "sandwich” immunoassay, an immunoprecipitation assay, a precipitin reaction, a gel diffusion precipitin reaction, an immunodiffusion assay, an agglutination assay, a complementfixation assay, an immunoradiometric assay, a fluorescent immunoassay, a protein A immunoassay, an immunoprecipitation assay, an immunohistochemical assay, a competition or sandwich ELISA, a radioimmunoassay, a Western blot assay, an immunohistological assay, an immunocytochemical assay, a dot blot assay, a fluorescence polarization assay, a scintillation proximity assay, a homogeneous time resolved fluorescence
- the determination of the incidence of of bacterial or fungal origin may comprise deriving the likelihood ratio using a multivariate analysis based on distribution parameters from a set of reference data derived from analysis of the levels of sTREM-1 in patients in which disease of bacterial or fungal origin is absent, present or in remission.
- the invention therefore also provides diagnostic means capable of measuring levels of sTREM-1 and/or comparing said levels to known levels that are indicative of the disease state of the patient.
- diagnostic means can take the form of a stick test, for example carrying the necessary reagents to perform the method of the invention and to produce, for example, a colorimetric result which can be compared against a colour chart.
- the method according to the first aspect of the invention can comprise the further step of c) correlating the detected level of sTREM-1 with the presence or absence of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- a correlation can be made by comparing the measured level of sTREM-1 in the sample with a mean level in samples obtained from a control population of individuals not having disease of bacterial or fungal origin, to indicate the presence or extent of disease of bacterial or fungal origin in the patient.
- the method according to the first aspect of the invention can be used in monitoring the progression or remission of disease of bacterial or fungal origin, in other words, to indicate the progression or remission of the disease.
- Such methods can be used to monitor the effectiveness and/or progress of therapy in a subject.
- the method further comprises the steps of measuring the level of sTREM-1 in a second or further sample from the patient, the first and second or further samples being obtained at different times; and comparing the levels in the samples to indicate the progression or remission of the disease of bacterial or fungal origin.
- the diagnostic methods according to the present invention are carried out ex vivo.
- Biological samples for analysis by the methods of the invention can be obtained using methods known in the art from various sources, in particular from body fluids such as whole blood, blood serum, blood plasma, urine and bronchoalveolar lavage fluid.
- the sample should be a sample treated such that any sTREM-1 present is not removed prior to the assay or is rendered undetectable.
- a preferred biological sample is a sample of bronchoalveolar lavage fluid.
- a preferred biological sample is a sample of blood serum.
- the methods of the invention are applicable to mammals, for example humans, non-human primates, sheep, pigs, cows, horses, goats, dogs, cats and rodents, such as mouse and rat.
- the biological sample tested by the methods of the invention is a human sample.
- the biological sample should generally contain protein molecules from the test subject and is handled such that proteins in the sample are not rendered undetectable by the compound chosen to detect them.
- the term "compound capable of binding sTREM-1” means polypeptides, ligands, antibodies or otherwise discriminating entities which predominantly, preferably specifically, bind to sTREM-1.
- Such binding compounds, or "sTREM-1 binding partners” can be a naturally occurring sTREM-1 binding molecule, for example a ligand for the TREM-1 Receptor or sTREM-1 and natural and synthetic variants thereof.
- binding compounds include, a chemically modified or genetically modified derivative of a sTREM-1 binding molecule, an artificially (for example chemically produced) sTREM-1 binding molecule or a recombinant or engineered soluble sTREM-1 binding molecule.
- antibodies which bind predominately, preferably specifically or exclusively to, sTREM-1 including, but not limited to, those antibodies which are: mono-or polyclonal antibodies (for example, raised against sTREM-1), bi-specific, multi-specific, human, humanized, chimeric antibodies, single chain antibodies, antibodies derived from phage display techniques, Fab fragments, F(ab')2 fragments, disulfide- linked Fvs, and fragments containing either a VL or VH domain or even a complementary determining region (CDR) that specifically binds to sTREM-1.
- mono-or polyclonal antibodies for example, raised against sTREM-1
- bi-specific, multi-specific, human, humanized, chimeric antibodies single chain antibodies
- antibodies derived from phage display techniques Fab fragments, F(ab')2 fragments, disulfide- linked Fvs, and fragments containing either a VL or VH domain or even a complementary determining region (CDR) that specifically
- modified immunoglobulins are also included within the scope of the invention, for example a fusion of the TREM-1 -Receptor to one or more immunoglobulin-derived protein domains, for example to confer solubility and/or stability, for example human IgG or IgM Fc fragments.
- substances or products mimicking the tertiary structure of a ligand for the TREM-1 -Receptor can be used as binding partners specific for sTREM-1. It is possible to design such on the basis of computer modelling. The product can be produced synthetically using chemical means. Use of recombinant DNA technology to engineer the required structure is also possible as is chemical modification.
- a compound capable of binding sTREM-1 is an antibody raised against the TREM-1 receptor, a fragment thereof or a variant thereof, provided that it is capable of binding sTREM-1.
- a compound capable of binding sTREM-1 is an antibody raised against the TREM-1 receptor, a fragment thereof or a variant thereof, provided that it is capable of binding sTREM-1.
- such an antibody is one raised against TREM-1 human Fc (TREM-1 -Fc) fusion protein (see Example 1 herein).
- the invention provides a compound capable of binding sTREM-1 for use in the diagnosis, prognosis, treatment or monitoring of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- the invention provides use of a compound capable of binding sTREM-1 in a method of treatment or diagnosis of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- the invention provides use of a compound capable of binding sTREM-1 in the manufacture of a medicament for the diagnosis, prognosis, treatment or monitoring of the treatment disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- the methods described herein can furthermore be used as screening assays to identify a subject with, or at risk of developing, disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- Such assays can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat disease of bacterial or fungal origin.
- an agent e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate
- agents e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate
- agents e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate
- such methods can be used to determine whether a subject
- the present invention provides methods for determining whether a subject can be effectively treated with an agent for disease of bacterial or fungal origin, for example, pneumonia or sepsis in which a test sample is obtained and TREM-1 is detected.
- a further embodiment of the invention provides a pharmaceutical composition comprising a compound capable of binding sTREM-1 together with a pharmaceutically acceptable diluent, carrier orexcipient for use in the diagnosis or treatment of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- a pharmaceutically acceptable diluent, carrier orexcipient for use in the diagnosis or treatment of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- the invention also provides a compound capable of binding sTREM-1 for use in, or used in, a method of diagnosis or treatment of disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- pharmaceutically acceptable diluent, carrier or excipient is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art.
- a third aspect of the invention provides a method of identifying agonists or antagonists of sTREM-1 said method comprising comparing the level of binding in a sample containing said sTREM-1 and a compound capable of binding sTREM-1, in the presence and absence of a compound to be tested. Also provided by are agonists or antagonists of sTREM-1 identified according to the method of this aspect of the invention.
- kits, associated reagents and contacting means comprising at least one compound capable of binding sTREM-1 and reagents for detecting binding of said compound to sTREM-1.
- kits comprising at least one compound capable of binding sTREM-1 and means for contacting said compound with a sample containing sTREM-1.
- the kit can comprise, for example: (1) a binding compound (e.g., attached to a solid support) that binds to sTREM-1; and, optionally, (2) a second, different binding compound e.g. an antibody, which binds to either the sTREM-1 or the first binding compound and is conjugated to a detectable agent.
- kits can also comprise, e.g., a buffering agent, a preservative, or a protein stabilizing agent.
- the kit can also comprise components necessary for detecting the detectable agent (e.g., an enzyme or a substrate).
- the kit can also contain a control sample or a series of control samples which can be assayed and compared to the test sample contained.
- Each component of the kit is usually enclosed within an individual container, and all of the various containers are within a single package, along with instructions for determining whether the subject from which. the sample is derived is suffering from or is at risk of developing disease of bacterial or fungal origin, for example, pneumonia or sepsis.
- an antibody raised against the TREM-1 -Receptor, a fragment thereof or a variant thereof can function as a compound c pable of binding sTREM-1.
- Antibodies are preferably raised against the human TREM-1 -Receptor (triggering receptor expressed on myeloid cells) for which the cDNA sequence is given in [SEQ ID NO:1].
- the TREM-1 -Receptor is expressed on human myeloid cells, is a transmembrane protein of the immunoglobulin superfamily (Ig-SF).
- the TREM-1 -Receptor is a transmembrane glycoprotein having the amino acid sequence of [SEQ I D NO:2] that is selectively expressed on blood neutrophils and a subset of monocytes but not on lymphocytes and other cell types.
- the invention encompasses antibodies raised against isolated or recombinantly prepared TREM proteins or polypeptides or fragments, homologues, derivatives, or variants thereof, as defined herein, as "TREM-1 -Receptor-derived polypeptides"
- TREM-1 -Receptor-derived polypeptides In accordance with the definition of "compound capable of binding sTREM-1", such antibodies raised against “TREM-1 -Receptor-derived polypeptides" predominantly, preferably specifically, bind sTREM-1.
- Such antibodies may be tested for binding with cells expressing the TREM-1 receptor and preferably also a sample from a patient known to have been suffering from pneumonia or sepsis of bacterial or fungal origin.
- the term "homologue,” especially "TREM-1 -Receptor homologue” as used herein refers to any member of a series of peptides to which antibodies capable of binding sTREM-1 can be raised-. TREM-1 -Receptor homologues can be from either the same or different species of animals.
- variant refers either to a naturally occurring allelic variation of a given peptide or a recombinantly prepared variation of a given peptide or protein in which one or more amino acid residues have been modified by amino acid substitution, addition, or deletion.
- the term "derivative" as used herein refers to a variation of given peptide or protein that are otherwise modified, i.e., by covalent attachment of any type of molecule, preferably having bioactivity, to the peptide or protein, including non-naturally occurring amino acids.
- the human TREM-1 -Receptor cDNA is 884-nucleotide long (Fig. 1; [SEQ ID NO:1]) and the open reading frame of TREM-1 -Receptor is nucleotides 48 to 752 of [SEQ ID NO:1], which encodes a transmembrane protein comprising the 234 amino acid sequence shown in Fig. 2 [SEQ 1 D NO:2].
- the human TREM-1 -Receptor cDNA can be found in the GenBank database under accession number AF 196329.
- the putative transmembrane domain starts from amino acid residues 201 to 229 of [SEQ ID NO:2J and contains a charged lysine residue at position 217. Its cytoplasmic tail consists of 5 amino acid residues and appears to contain no signaling motifs.
- antibodies for binding sTREM-1 are raised against a TREM-1 -Receptor-derived polypeptide comprising at least an extracellular domain comprising amino acid residues 17 to 200 of [SEQ ID NO:2].
- antibodies suitable for use in the invention are those antibodies having the ability to bind sTREM- 1 which are raised against homologues of TREM-1-Receptor from either the same or different species of animal, preferably from mammals, more preferably from rodents, such as mouse and rat, and most preferably from human.
- Homologues of the TREM-1 -Receptor nucleic acid molecule i.e., [SEQ ID NO:1]
- aspects of fiie invention can be also applied in the framework of multiple diagnosis of a subject.
- a method of screening a patient for presence or susceptibility to disease comprising performing a plurality of diagnostic tests on a tissue sample from the patient for a plurality of diseases
- the invention provides the improvement wherein one of the diagnostic tests comprises measuring the level of sTREM-1.
- a diagnostic means for detecting disease of bacterial or fungal origin for example, pneumonia or sepsis
- a diagnostic kit comprising such a diagnostic means
- a method of treatment of infection which includes the step of screening an individual for disease of bacterial or fungal origin, for example, pneumonia or sepsis, wherein disease of bacterial or fungal origin is correlated with the levels of sTREM-1 in a sample from said individual, and if disease of bacterial or fungal origin is identified, treating that individual to prevent or reduce the infection
- a bronchoalveolar lavage sample from a patient with lung-related symptoms would be used to diagnose pneumonia based upon elevated levels of sTREM-1.
- a blood serum sample from a patient exhibiting symptoms of SIRS would be used to diagnose sepsis of bacterial or fungal origin based upon elevated levels of sTREM-1.
- the invention also provides a method of diagnosing disease of bacterial or fungal origin in a subject, which method comprises the step of measuring the level of sTREM-1 and the step of measuring the level of TREM- 1 -Ligand in one or more biological samples obtained from said subject.
- the Inventors have developed an immuno-enzymatic (in this case ELISA) based method for the detection of soluble TREM-1.
- the invention provides a method of diagnosing disease of bacterial or fungal origin in a subject, which method comprises the step of measuring the level of sTREM-1 in a biological sample obtained from said subject and wherein the level of sTREM-1 is measured by an immunochemical technique.
- immunochemical techniques are indirect immunofluorescence (I IF), immunoperoxydase (POD), western immunoblotting (WB), radioimmunoprecipitation (RIPA), enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and agglutination assays.
- I IF indirect immunofluorescence
- POD immunoperoxydase
- WB western immunoblotting
- RIPA radioimmunoprecipitation
- ELISA enzyme linked immunosorbent assay
- RIA radioimmunoassay
- agglutination assays agglutination assays.
- an ELISA method using an anti-human Trem-1 antibody is used to measure the level of sTREM-1.
- WO2004081233 describes a method of diagnosing bacterial or fungal sepsis in a subject by measuring the level of TREM-1 -Ligand in a biological sample obtained from the subject and compounds capable of binding TREM- 1 -Ligan
- the level of TREM-1 -Ligand present in the sample is measured by observing the level of binding between these compounds and TREM-1 -Ligand.
- the Inventors have determined that measurement of both soluble TREM-1 (as described herein) and membrane associated TREM-1 Ligand (as described in WO2004081233) in newly admitted critically ill patients allows the rapid identification those with infection.
- Figure 1 shows Human TREM-1 -Receptor cDNA [SEQ ID NO:1].
- Figure 2 shows Human TREM-1-Receptor amino acid sequence [SEQ ID NO:2].
- Figure 3 shows the levels of sTREM-1 in bronchoalveolar lavage fluid from patients according to diagnosis. Individual values are plotted and the bars represent the means of the values. P ⁇ 0.001 between CAP and NP and between VAP and NP.
- Figure 4. shows receiver-operating-characteristic curves for various cut-off levels of bronchoalveolar lavage fluid sTREM-1, Tumor necrosis factor- ⁇ and Interleukin-1 ⁇ in differentiating between presence and absence of pneumonia.
- sTREM-1 0.93 (95% confidence interval, 0.92 to 0.95)
- Tumor necrosis factor- ⁇ 0.64 (95% confidence interval, 0.62 to 0.69)
- lnterleukin-1 ⁇ 0.69 (95% confidence interval, 0.67 to 0.72)
- Figure 5 shows bronchoalveolar lavage fluid (BAL) supernatants examined by Western blot analysis using 21 C7, an anti TREM-1 monoclonal antibody: Lane 1 : positive control (sTREM-1 , 50 pg/mL) Lane 2: BAL supernatant from a patient with pneumonia Lane 3: BAL supernatant from a patient without pneumonia
- Figure 6. shows a flow-chart of the patients admitted to the ICU during the study period.
- Figure 8 shows Receiver-operating-characteristic curves for various cut-off levels of plasmatic C-Reactive Protein, Procalcitonin and sTREM-1 in differentiating between presence and absence of infection. Areas under the ROC. curves for: C-Reactive Protein: 0.77 (95% confidence interval, 0.69 to 0.85) Procalcitonin: 0.85 (95% confidence interval, 0.81 to 0.89) sTREM-1: 0.97 (95% confidence interval, 0.94 to 1.0)
- FIG. 9 shows admission plasmatic levels of C-Reactive Protein
- Figure 10 shows a standard curve for an immuno-enzymatic assay to detect soluble TREM-1 in the sera of patients with suspected sepsis
- Figure 11 shows the kinetics of immuno-enzymatic assay to detect soluble TREM-1 (panel A) and the cytofluorimetric analysis of TREM-1 Ligand (panel B) in a patient having SIRS without infection (HSR34) and in a sepsis patient (HSR37).
- FIG. 16 shows TREM-1 expression pattern on monocytes during septic shock according to outcome. Results are expressed as Mean Fluorescence Intensity. Respective p values are depicted above time points. 'Baseline' corresponds to the first determination and 'Last value' to the last determination of TREM-1 before intensive care unit discharge or death.
- EXAMPLE 1 Production of antibodies against TREM-1 Receptor which are capable of binding sTREM-1
- Antibodies were raised against a fusion protein of the TREM-1 receptor with the human lgG Fc region.
- the cDNA fragment encoding the TREM-1 extracellular region was amplified by PCR and cloned into an expression vector containing the exons for hinge, CH2, and CH3 region of human lgG1 (see Bouchon et al. The Journal of Immunology, 2000, 164: 4991-4995). Briefly, the 760-bp TREM-1 was amplified by RT-PCR, cloned into pCR2.1 (Invitrogen, Carlsbad, CA), and sequenced.
- PCR primers used were: 5'-GCTGGTGCACAGGAAGGATG [SEQ ID NO: 3] 3'-GGCTGGAAGTCAGAGGACATT [SEQ ID NO: 4]
- This chimeric gene was transfected into mouse myeloma cell line J558L, screening of culture supernatants, and purification of TREM-1-Fc can then be performed, as previously described (Traunecker, et al., 1991, Trends Biotechnol. 9:109)).
- Anti-TREM-1 monoclonal antibodies were produced by immunising BALB/c mice with TREM-1-Fc .
- mice received an initial injection of 100 ⁇ g of TREM-1 -Fc fusion protein (TREM-1 -Fc), mixed 1:1 (vol/vol) with Alu-Gel-S (Serva Biochemicals, Paramus, NJ), behind the neck.
- TREM-1 -Fc TREM-1 -Fc fusion protein
- Alu-Gel-S Serva Biochemicals, Paramus, NJ
- mice were sacrificed and draining lymph node cells were isolated and fused with the myeloma fusion partner, Ag8.653, using polyethylene glycol 4000.
- Hybridoma supernatants were screened in two steps. First, an ELISA was performed using TREM-1 -Fc in the coating step and human-adsorbed alkaline phosphatase-labeled goat anti-mouse lgG as secondary antibody. Supernatants from clones that were positive in ELISA were then tested by FACS ® analysis for staining cells by flow cytometry.
- EXAMPLE 2 Rapid detection of the soluble form of TREM-1 (sTREM) in the diagnosis of pneumonia
- SAPS II score SAPS II score
- SOFA score body temperature
- leukocyte count ratio of the partial pressure of arterial oxygen to the fraction of inspired oxygen (Pa0 2 /FiO z ); serum levels of C reactive protein and procalcitonin
- presence of shock defined as systolic arterial pressure lower than 90 mm Hg with signs of peripheral hypoperfusion or need for continuous infusion of vasopressor or inotropic agents
- duration of previous mechanical ventilation and use of previous antimicrobial therapy.
- a clinical pulmonary infection score (CPIS) was calculated as previously described in Pugin J, et al. Am Rev Respir Dis 1991;143:1121-9. The duration of mechanical ventilation, length of ICU stay and ICU mortality were also recorded.
- Mini-bronchoalveolar lavages (BAL) and microbiological specimen processing were performed as described in detail in Papazian L et al. Am J Respir Crit Care Med 1995;152:1982-91 and Duflo F et al. Anesthesiology 2002;1:74-9. Briefly, mini-bronchoalveolar lavage was performed using the Combicath, a single-sheathed, 50-cm, sterile, plugged, telescopic catheter (Plastimed, St Leu La Foret, France).
- the recovered BAL fluid (13+3mL out of 20mL of instilled saline serum) was divided into two samples: one was used for direct microscopic examination and quantitative culture; the other was centrifuged at 10000 revolutions per minute for 30 min and the supernatant was frozen at -80°C until used for sTREM-1 and cytokine measurements.
- concentration of micro-organisms considered significant for the potential diagnosis of pneumonia was >10 3 CFU/mL of BAL fluid.
- Post hoc diagnosis of pneumonia was made from a combination of already mentioned clinical criteria with microbiological evidence of microbial infection. These criteria were similar to those used for ventilator-associated pneumonia described in Pugin J et al. Am Rev Respir Dis 1991;143:1121-9.
- Pneumonia was considered to be absent when an alternative cause for pulmonary infiltrate was established and there was non-significant bacterial growth in culture of BAL in association with full recovery from fever, infiltrate, and leukocytosis without antimicrobial therapy.
- Two intensivists reviewed all medical records pertaining to the patient and independently classified the diagnosis as community-acquired pneumonia, ventilator-associated pneumonia or no pneumonia. A consensus concerning the diagnosis was achieved in all cases. Both intensivists were blinded to the results of sTREM-1 and cytokines levels.
- sTREM-1 and cytokines assays Assessment of sTREM-1 levels in BAL fluid samples was performed using an immunoblot technique with 21C7, a monoclonal murine lgG1 directed against human TREM-1 prepared as described in Example 1 . Briefly, 100 ⁇ L of each BAL fluid supernatant was dotted on a nitrocellulose membrane, dried, and overcoated in phosphate buffer-saline (PBS) supplemented with 3% bovine serum albumin. The nitrocellulose sheet was then incubated for 60 min in the presence of diluted 1:2000 diluted 21 C7.
- PBS phosphate buffer-saline
- the sheet was further incubated for 60 min with diluted 1:1000 diluted goat anti- mouse immunoglobuiins (Dako, Glostrup, Denmark), washed in PBS supplemented with 20% dimethylsulfoxyde and incubated for 30 min with diluted 1: 000 diluted horseradish peroxydase-conjugated streptavidin (Bio- Rad, Cergy, France).
- the enzyme substrate chromogen Opti-4CN Bio-Rad was then added, and colour developed in proportion to the amount of sTREM- 1 bound to the membrane.
- Each sheet also contained calibration samples of a known concentration of sTREM-1 (0 to 200 pg/mL).
- sTREM-1 concentration from each sample was determined by comparing the optical densities of the samples to the standard curve. All measurements were performed in duplicate and results are expressed as the mean concentration in picograms per millilitre of bronchoalveolar lavage fluid.
- the sensitivity of this technique allows the detection of sTREM-1 level as low as 5 pg/mL and the entire procedure takes less than 3 hours. The coefficient of variation of the assay was lower than 5 percent.
- Tumor necrosis factor- ⁇ and interleukin-1 ⁇ were determined in BAL fluid by solid-phase ELISA method according to the recommendations of the manufacturer (BD Biosciences, Le Pont de Claix, France). The sensitivity of the technique allows the detection of levels as low as 2 pg/mL for tumor necrosis factor- ⁇ and 3.9 pg/mL for interleukin-1 ⁇ .
- ROC Receiver-operating- characteristic
- SAPS II scoref mean 52 ⁇ 17 53 ⁇ 20 50 ⁇ 15 53 ⁇ 1 0.76 ( ⁇ SD)
- Acute respiratory failure Neurologic 41 (27.7) 7 (18) 15 (33) 19 (30) 0.45 Shock 37 (25) 6 (16) 16 (35) 15 (23) 0.18 Miscellaneous 28 (19) 2 (5) 11 (24) 15 (23) 0.08
- ARDS Acute respiratory distress syndrome
- ARDS of extra-pulmonary origin
- Clinical characteristics of the three groups did not differ significantly at inclusion (table 1).
- P values are comparisons between CAP, VAP and NP groups
- Monomicrobial pneumonia n(%) 36 (95) 37 (80) Polymicrobial pneumonia, n(%) 2 (5) 9 (20) Total number of pathogens*, n 40 58 Bacilli, n(%) Pse ⁇ domonas aeruginosa 12 (20.7) Haemophilus influenzae 10 (25) 10 (17.2) Acinetobacter baumanii 4 (6.9) Serratia marcescens 6 (10.3) Kiebsiella species 11 ((22..55)) 6 (10.3) Legionnella pneumophilia 3 (7.5) Miscellaneous 2 (5) 2 (3.4)
- Organisms shown are those that were isolated at significant concentrations from quantitative cultures of bronchoalveolar lavage fluid (>10 3 colony-forming units/mL). Legionnella pneumophilia infection was diagnosed by the detection the soluble urinary antigen.
- sTREM-1 Tumor Necrosis Factor- ⁇ and lnterleukin-1 ⁇ levels
- the levels of sTREM-1 were higher in BAL fluid from community- acquired and ventilator-associated pneumonia patients than from no- pneumonia patients (P ⁇ 0.001) but did not differ significantly between community-acquired and ventilator-associated pneumonia patients (figure 3).
- Tumor necrosis factor- ⁇ and interleukin-1 ⁇ levels showed the same trend (P ⁇ 0.001) but with a large overlap of values.
- sTREM-1 was detected in BAL fluid among 36 out of 38 community-acquired pneumonia patients (sensitivity: 95 percent, 2 false negatives), 46 out of 46 ventilator-associated pneumonia patients (sensitivity: 100 percent), and in 6 out of 64 no-pneumonia patients (6 false positives).
- sensitivity 95 percent, 2 false negatives
- 46 out of 46 ventilator-associated pneumonia patients sensitivity: 100 percent
- 6 out of 64 no-pneumonia patients 6 false positives.
- the capacity of sTREM-1 to differentiate pneumonia from no pneumonia was assessed with a ROC curve analysis (figure 4).
- the area under the ROC curve when sTREM-1 was used to differentiate pneumonia from no pneumonia was 0.93 (95 percent Cl 0.92 to 0.95, PO.001).
- a sTREM-1 cut-off value of 5 pg/mL (which represented the technique's threshold of detection) had a sensitivity of 98 percent (95 percent Cl, 95 to 100) a specificity of 90 percent (95 percent Cl, 84 to 96).
- the Inventors determined that the presence of sTREM-1 in BAL fluid was the strongest independent predictor of pneumonia with an odds ratio of 41.52 (table 5).
- the best clinical predictor of pneumonia was a clinical pulmonary infection score >6 (odds ratio: 2.98).
- Table 5 Multiple logistic-regression analysis of factors used for differentiating between patients with and those without pneumonia PREDICTOR P Value ODDS RATIO (95% Confidence Interval) CPIS * >6 0.002 2.98 (1.51 to 5.86) BALTNF ⁇ >150 pg/mL 0.004 2.44 (1.82 to 5.75) BAL lL-1 ⁇ >75 pg/mL 0.003 2.70 (1.97 to 13.18) BAL sTREM-1>5 pg/mL ⁇ 0.001 41.52 (20.90 to 77.62) CPIS: clinical pulmonary infection 1 score « These results demonstrate that rapid detection of the sTREM-1 in bronchoalveolar lavage fluid improves the ability of clinicians to differentiate patients with bacterial or fungal pneumonia from those without pneumonia.
- the immunoblot technique is rapid, accurate, of very low cost and can be applied to small series or even individual samples. Use of this test to detect the presence of sTREM-1 in bronchoalveolar lavage fluid will lead to more accurate diagnoses of pneumonia in mechanically ventilated patients. Microbiological documentation was obtained in all cases of community-acquired and ventilator-associated pneumonia. When pneumonia was considered to be absent, either a non-infectious alternative cause for pulmonary infiltrate was established or patients fully recovered from fever, infiltrate, and leukocytosis without antimicrobial therapy.
- EXAMPLE 3 Diagnostic value of plasmatic levels of the soluble form of triggering receptor expressed on myeloid cells (TREM)-1 in critically ill patients with suspected sepsis
- Clinically suspected infection was defined as an explicit statement by the attending physician indicating the suspicion of an ongoing infection, combined with the initiation of a diagnostic work-up to identify or rule out infection and the prescription of antimicrobial therapy.
- Patients were not enrolled if they were older than 80 years of age or were immunocompromised (treatment with corticosteroids, bone marrow or organ transplant recipients, leukopenia [white blood cells count ⁇ 1 G/L] or neutropenia [polymorphonuclear granulocyte count ⁇ 0.5 G/L], hematologic malignancy or acquired immune deficiency syndrome). Patients who presented with early death or discharge (within 12 hours after admission) or complete absence of antimicrobial treatment were also excluded. Patients originated either from the emergency room, the general wards, or from the operating room. Approval of the institutional review board and informed consent from patients or their relatives were obtained before inclusion.
- Plasmatic PCT concentrations were measured using an immunoassay with a sandwich technique and a chemiluminescent detection system, according to the manufacturer's protocol (LumiTest; Brahms Diagnostica, Berlin, Germany). Assessment of plasmatic sTREM-1 levels was performed as described in Example 2.
- the sheet was further incubated for 60 min with 1:1000 diluted goat anti-mouse immunoglobulins (Dako, Glostrup, Denmark), washed in PBS supplemented with 20% dimethylsulfoxyde and incubated for 30 min with 1 :1000 diluted horseradish peroxydase-conjugated streptavidin (Bio-Rad, Cergy, France).
- the enzyme substrate chromogen Opti-4CN Bio- Rad
- Each sheet also contained calibration samples of a known concentration of sTREM-1 (0 to 5000 ng/mL).
- sTREM-1 concentration from each sample was determined by plotting the optical densities of the samples to the standard curve. All measurements were performed in duplicate and results expressed as mean concentration in nanograms per mL of plasma. The sensitivity of this technique allows the detection of sTREM-1 levels as low as 5 ng/mL and the entire procedure takes less than 3 hours. The coefficient of variation of the assay was lower than 5 percent.
- the predictors included clinical and laboratory findings along with information on plasmatic sTREM-1 level.
- logistic regression analysis which requires binary outcome events, subjects classified as confirmed sepsis, severe sepsis, or septic shock (sepsis syndrome) were compared to patients with SIRS and initial suspicion of infection.
- Receiver-operating-characteristic (ROC) curves were constructed to illustrate various cut-off values of sTREM-1, PCT and CRP. Sensitivity, specificity, and positive and negative predictive values of each parameter were calculated according to standard methods. These values were calculated for the cut-off that represented the best discrimination as derived from the areas under ROC curves. Analysis was completed with Statview software (Abacus Concepts, Berkeley CA) and a two- tailed P ⁇ 0.05 was considered significant.
- Diagnosis was established as SIRS in 29 patients (38 %), sepsis or severe sepsis (grouped as 'Sepsis') in 22 patients (29 %) and septic shock in 25 patients (33 %).
- Clinical characteristics did not differ significantly at inclusion between septic and non- septic patients (Table 6).
- Infections were microbiologically proven in 40 of 49 infected patients (82 %) with 55 % Gram-negative, 42 % Gram-positive bacteria, and 3 % fungal infections. The major sources of infection were the respiratory tract (55 %) and abdomen (22 %). Twenty-four percent of infected patients had a documented bloodstream infection. Neither site of infection nor microbial strains differed between surviving and non-surviving patients (Table 7).
- Baseline Plasmatic Levels of CRP, PCT and sTREM-1 Baseline plasmatic levels of CRP, PCT and sTREM-1 were higher among septic patients than among subjects with SIRS only (Table 6, Figure 7). Plasmatic sTREM-1 levels appeared to be most helpful in differentiating patients with sepsis from those with SIRS. Mean plasmatic sTREM-1 levels on admission were 229 ng/mL for SIRS; 1836 ng/mL for sepsis and 1413 ng/mL for septic shock (PO.001). The accuracy of the candidate parameters to distinguish patients with SIRS from those with septic conditions was highly variable (Table 8).
- plasmatic sTREM-1 levels yielded the highest discriminative value with an area under the ROC curve (AUC) of 0.97 (95 % confidence interval [Cl], 0.94 to 1.0) followed by PCT (AUC, 0.85; Cl, 0.81 to 0.89) and CRP (AUC, 0.77; Cl, 0.69 to 0.85; p ⁇ 0.001).
- AUC area under the ROC curve
- PCT AUC, 0.85; Cl, 0.81 to 0.89
- CRP AUC, 0.77; Cl, 0.69 to 0.85; p ⁇ 0.001
- sTREM-1 yielded a sensitivity of 96 % (95 % Cl, 0.92 to 100 %) and a specificity of 89 % (Cl, 82 to 95 %) to differentiate patients with SIRS from those with sepsis or septic shock.
- CRP area under the ROC curve
- the study population was large and comprised a diverse group of critically ill adult patients admitted to a medical ICU in various phases of infectious and ⁇ on-infectious conditions, which allowed a generalization of the study findings.
- the diagnosis was determined by blinded investigators without knowledge of the plasmatic sTREM-1 levels and the patients were classified as having SIRS of non-infectious origin after incorporation of all other available clinical and laboratory data (Bone RC, et al. Chest. 1992;101:1644-55.).
- the inventors' study was designed as a real-life study, not including control patients without suspected infection but only patients with a high pre-test probability of sepsis, covering the spectrum of patients that is likely to be encountered in the future use of this test.
- Example 4 Use of an immuno-enzymatic assay to detect soluble TREM-1 in the sera of patients with suspected sepsis
- the method is as follows:
- Coating buffer carbonate pH 9,6: 0.015 M Na 2 C0 3 (0.794g in 500 ml H 2 0), 0.035 M NaHC0 3 (1 ,47g in 500 ml H 2 0)
- Wash buffer 0.1% Tween 20 in PBS, pH 7,4
- Substrate solution 30 mM potassium citrate ,pH 4.1, immediately before use, add 1 tablet of 3,3',5,5'-Tetramethylbenzidine (Sigma # T-3405) for 10 ml of buffer and add 2,5 ⁇ l of H 2 0 2 30%.
- Time 0 represents the day of admission into the Intensive Care Unit. Samples were obtained every 48 hours until day 15.
- FIG. 12 shows the time course of median (with interquartile range) plasma levels of sTREM-1 in surviving (squares) and non-surviving (triangles) patients.
- monocyte see Figure 14
- polymorphonuclear see Figure 15
- TREM-1 cell surface expression of TREM-1 was analysed in flow cytometry after labeling with a mouse monoclonal antibody anti-human TREM- 1, PE-labelled (clone 193015, R&D, Abingdon, UK). Results were expressed as Mean Fluorescence Intensity (MFI).
- MFI Mean Fluorescence Intensity
- MFI on monocytes from patients with septic shock are significantly higher than MFI from non-septic patients or healthy controls.
- FIG. 16 shows TREM-1 expression pattern on monocytes during septic shock according to outcome. Results are expressed as Mean Fluorescence Intensity. Respective p values are depicted above time points. 'Baseline' corresponds to the first determination and 'Last value' to the last determination of TREM-1 before intensive care unit discharge or death. These results demonstate that among patients with sepsis, those with lower levels of TREM- 1 expression on monocytes, but not on neutrophils, can be predicted to have a positive outcome.
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JP2006550296A JP4911600B2 (en) | 2004-01-27 | 2005-01-27 | Method for diagnosing infectious diseases by measuring soluble TREM-1 levels in a sample |
EP05702028A EP1709444B1 (en) | 2004-01-27 | 2005-01-27 | Method of diagnosing infectious disease by measuring the level of soluble trem-1 in a sample |
US10/587,356 US8021836B2 (en) | 2004-01-27 | 2005-01-27 | Method of diagnosing infectious disease by measuring the level of soluble TREM-1 in a sample |
AU2005207111A AU2005207111A1 (en) | 2004-01-27 | 2005-01-27 | Method of diagnosing infectious disease by measuring the level of soluble TREM-1 in a sample |
ES05702028T ES2386966T3 (en) | 2004-01-27 | 2005-01-27 | Method to diagnose infectious diseases by measuring the level of soluble TREM-1 in a sample |
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6420526B1 (en) * | 1997-03-07 | 2002-07-16 | Human Genome Sciences, Inc. | 186 human secreted proteins |
US6420326B1 (en) | 1997-08-13 | 2002-07-16 | The Procter & Gamble Company | Glass cleaner compositions having good surface lubricity and alkaline buffer |
WO2002058721A1 (en) | 2000-12-08 | 2002-08-01 | Baylor College Of Medicine | Trem-1 splice variant for use in modifying immune responses |
US20030165875A1 (en) | 2001-03-20 | 2003-09-04 | Marco Colonna | Novel receptor TREM (triggering receptor expressed on myeloid cells) and uses thereof |
WO2004035732A2 (en) * | 2002-08-29 | 2004-04-29 | Five Prime Therapeutics, Inc. | Human polypeptides encoded by polynucleotides and methods of their use |
WO2004081233A1 (en) * | 2003-03-10 | 2004-09-23 | Bioxell S.P.A. | Diagnostic and prognostic compounds and method |
-
2004
- 2004-01-27 GB GBGB0401730.7A patent/GB0401730D0/en not_active Ceased
-
2005
- 2005-01-27 US US10/587,356 patent/US8021836B2/en active Active
- 2005-01-27 JP JP2006550296A patent/JP4911600B2/en active Active
- 2005-01-27 ES ES05702028T patent/ES2386966T3/en active Active
- 2005-01-27 EP EP05702028A patent/EP1709444B1/en active Active
- 2005-01-27 AU AU2005207111A patent/AU2005207111A1/en not_active Abandoned
- 2005-01-27 CA CA002554769A patent/CA2554769A1/en not_active Abandoned
- 2005-01-27 WO PCT/GB2005/000273 patent/WO2005071408A1/en active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6420526B1 (en) * | 1997-03-07 | 2002-07-16 | Human Genome Sciences, Inc. | 186 human secreted proteins |
US6420326B1 (en) | 1997-08-13 | 2002-07-16 | The Procter & Gamble Company | Glass cleaner compositions having good surface lubricity and alkaline buffer |
WO2002058721A1 (en) | 2000-12-08 | 2002-08-01 | Baylor College Of Medicine | Trem-1 splice variant for use in modifying immune responses |
US20030165875A1 (en) | 2001-03-20 | 2003-09-04 | Marco Colonna | Novel receptor TREM (triggering receptor expressed on myeloid cells) and uses thereof |
WO2004035732A2 (en) * | 2002-08-29 | 2004-04-29 | Five Prime Therapeutics, Inc. | Human polypeptides encoded by polynucleotides and methods of their use |
WO2004081233A1 (en) * | 2003-03-10 | 2004-09-23 | Bioxell S.P.A. | Diagnostic and prognostic compounds and method |
Non-Patent Citations (7)
Title |
---|
BOUCHON A ET AL., NATURE, vol. 230, 2001, pages 1103 - 7 |
BOUCHON A ET AL: "CUTTING EDGE: INFLAMMATORY RESPONSES CAN BE TRIGGERED BY TREM-1, A NOVEL RECEPTOR EXPRESSED ON NEUTROPHILS AND MONOCYTES", JOURNAL OF IMMUNOLOGY, THE WILLIAMS AND WILKINS CO. BALTIMORE, US, vol. 164, 2000, pages 4991 - 4995, XP002951620, ISSN: 0022-1767 * |
BOUCHON A ET AL: "TREM-1 amplifies inflammation and is a crucial mediator of septic shock", NATURE, MACMILLAN JOURNALS LTD. LONDON, GB, vol. 410, no. 6832, 26 April 2001 (2001-04-26), pages 1103 - 1107, XP002285055, ISSN: 0028-0836 * |
COLONNA M ET AL: "TREM-1 (TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS): A NEW PLAYER IN ACUTE INFLAMMATORY RESPONSES", JOURNAL OF INFECTIOUS DISEASES, CHICAGO, IL, US, vol. 187, no. SUPPL 2, 15 June 2003 (2003-06-15), pages S397 - S401, XP008033778, ISSN: 0022-1899 * |
GINGRAS MARIE-CLAUDE ET AL: "TREM-1, MDL-1, and DAP12 expression is associated with a mature stage of myeloid development", MOLECULAR IMMUNOLOGY, vol. 38, no. 11, March 2002 (2002-03-01), pages 817 - 824, XP002332587, ISSN: 0161-5890 * |
NATHAN C ET AL: "TREM-1: A new regulator of innate immunity in sepsis syndrome", NATURE MEDICINE, NATURE PUBLISHING, CO, US, vol. 7, no. 5, 2001, pages 530 - 532, XP002291851, ISSN: 1078-8956 * |
NOCHI HITOSHI ET AL: "Modulation of hepatic granulomatous responses by transgene expression of DAP12 or TREM-1-Ig molecules", AMERICAN JOURNAL OF PATHOLOGY, PHILADELPHIA, PA, US, vol. 162, no. 4, April 2003 (2003-04-01), pages 1191 - 1201, XP002285056, ISSN: 0002-9440 * |
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Also Published As
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ES2386966T3 (en) | 2012-09-07 |
US20070281319A1 (en) | 2007-12-06 |
EP1709444A1 (en) | 2006-10-11 |
AU2005207111A1 (en) | 2005-08-04 |
CA2554769A1 (en) | 2005-08-04 |
US8021836B2 (en) | 2011-09-20 |
GB0401730D0 (en) | 2004-03-03 |
EP1709444B1 (en) | 2012-06-06 |
JP4911600B2 (en) | 2012-04-04 |
JP2007519916A (en) | 2007-07-19 |
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