WO2011160845A2 - Phospholipides et lipoprotéines oxydés, et anticorps dirigés contre eux, à titre de biomarqueurs des états inflammatoires et méthodes de traitement - Google Patents

Phospholipides et lipoprotéines oxydés, et anticorps dirigés contre eux, à titre de biomarqueurs des états inflammatoires et méthodes de traitement Download PDF

Info

Publication number
WO2011160845A2
WO2011160845A2 PCT/EP2011/003110 EP2011003110W WO2011160845A2 WO 2011160845 A2 WO2011160845 A2 WO 2011160845A2 EP 2011003110 W EP2011003110 W EP 2011003110W WO 2011160845 A2 WO2011160845 A2 WO 2011160845A2
Authority
WO
WIPO (PCT)
Prior art keywords
oxps
oxcl
inflammatory
oxldl
ldl
Prior art date
Application number
PCT/EP2011/003110
Other languages
English (en)
Other versions
WO2011160845A3 (fr
Inventor
Johan FROSTEGÅRD
Original Assignee
Medirista Biotechnologies Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Medirista Biotechnologies Ab filed Critical Medirista Biotechnologies Ab
Publication of WO2011160845A2 publication Critical patent/WO2011160845A2/fr
Publication of WO2011160845A3 publication Critical patent/WO2011160845A3/fr

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0012—Lipids; Lipoproteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • 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
    • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/50—Determining the risk of developing a disease

Definitions

  • This invention relates to a method for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, by measuring the levels of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti- MDA-LDL and anti-PC, as well as a kit for implementing the method.
  • It also relates to a method for treating or decreasing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, by administering medicaments comprising one or more compounds selected from the list consisting of annexin A5, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, as well as bioactive components and/or parts/fragments thereof.
  • compositions comprising one or more inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL and inhibitors of PC, as well as bioactive components and/or parts/fragments thereof, and also to compositions comprising one or more conjugates selected from the list consisting of oxCL conjugates, oxPS conjugates, oxLDL conjugates, MDA-LDL conjugates and PC conjugates, as well as bioactive components and/or parts/fragments thereof.
  • Levels of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC may be used to indicate if and to which extent treatment according to the present invention, i.e.
  • inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL activity, inhibitors of oxPS activity, inhibitors of oxLDL activity, inhibitors of MDA-LDL activity and inhibitors of PC activity, as well
  • such levels may be used by physicians to evaluate if, and to which extent, immunization according to this invention, i.e. treatment to elevate natural levels of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, and/or to decrease oxCL levels (or activity) and/or oxPS levels (or activity) and/or oxLDL levels (or activity) and/or MDA-LDL levels (or activity), should be the treatment of choice.
  • immunization i.e. treatment to elevate natural levels of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, and/or to decrease oxCL levels (or activity) and/or oxPS levels (or activity) and/or oxLDL levels (or activity) and/or MDA-LDL levels (or activity)
  • rheumatic diseases and inflammatory conditions are in general major health problems in the Western World and increasing in developing countries. These diseases include, but are not limited to, rheumatic conditions like rheumatoid arthritis and systemic lupus erythematosus (examples of autoimmune diseases), the latter often described as a prototypic autoimmune disease 61 , and in text books more than 80 autoimmune rheumatic diseases are described, cf. e.g. Harrison's Principles of Internal Medicine, 17th Edition.
  • CVDs cardiovascular diseases
  • diseases include, but are not limited to, atherosclerosis, stroke, myocardial infarction, and heart failure (including, but not limited to, acute and chronic heart failure); these diseases are often correlated with or preceded by the metabolic syndrome.
  • the metabolic syndrome has moreover been correlated to Type 2 diabetes mellitus, CVD and coronary heart disease, which are also categorized as inflammatory disorders, cf. e.g. Peter W.F. et al., Metabolic Syndrome as a Precursor of Cardiovascular Disease and Type 2 Diabetes Mellitus, Epidemiology, Circulation. 2005;112:3066-3072.
  • Atherosclerosis is an inflammatory disease and the dominating underlying cause of cardiovascular disease, including myocardial infarction (Ml) and heart failure, stroke and claudication.
  • Leukocyte extravasation through the endothelial barrier is important in the pathogenesis of inflammatory disorders. Endothelial cells line the lumina of vessels, thus separating and also connecting the blood and the synovial tissue. It has become clear that, in inflammation, endothelial cells are not only passive bystanders but are actively responding to various stimuli, interacting with the immune system. Thus, endothelium is a target for inflammatory leukocytes and their mediators.
  • endothelial cells themselves produce a number of inflammatory mediators, express cellular adhesion molecules (CAMs) and therefore directly influence the immune system, for example by recruiting and/or activating white blood cells modulating the outcome of the inflammatory response.
  • CAMs cellular adhesion molecules
  • Inflammatory conditions include not only conditions where an undesirable immune response is responsible for a pathological state or parts thereof including autoimmune diseases, infectious conditions, rheumatic conditions, and/or secondary complications, but also those conditions caused by side-effects of medicaments, surgery or other clinical therapy.
  • condition relates to systemic conditions, disease and inflammatory activity or those defined by a tissue and/or organ.
  • One or more infectious conditions becoming inflammatory conditions, as well as the mortality related thereto, are known problems in clinical care of a broad range of patients with very different symptoms, conditions and diseases, including uremia and autoimmune conditions, such as severe Sjogren's syndrome and SLE, but also tuberculosis and HIV, as well as different status of morbidity and risk related to mortality, whether treated in their homes and/or in the hospital.
  • uremia and autoimmune conditions such as severe Sjogren's syndrome and SLE
  • tuberculosis and HIV as well as different status of morbidity and risk related to mortality, whether treated in their homes and/or in the hospital.
  • the risk of being infected with bacteria or virus may be limited by isolation (e.g. avoiding or eliminating contact to people and animals and/or their surroundings) and a high level of hygiene.
  • eliminating or even limiting the risk of viral and/or bacterial infections developing undesirably and/or uncontrollably is a greater challenge.
  • a biological marker pointing out patients at special risk of developing one or more infectious conditions and/or determining the mortality related to such conditions, as well as a prophylactic, palliative and/or curative treatment for those patients may be of great use in the clinical care of patients at all levels of illness.
  • Patients of interest include, but are not limited to, patients with frequent bacterial and/or viral infections, patients with depressed immune systems, patients in poor physical conditions due to operations and/or patients in dialysis.
  • One group of patients of great risk of developing inflammatory conditions are patients admitted into intensive care units (ICUs), which are at great risk of acquiring nosocomial infections. They are susceptible to infection because of the underlying diseases or conditions associated with impaired immunity and are at increased risk of infections during invasive monitoring as well as secondary infections after exposure to broad-spectrum antimicrobials (Eggimann, P. and Pittet, D., 2001 , and Cotran, R.S., 1993) 38,39 .
  • COPD chronic obstructive pulmonary disease
  • Pro-inflammatory cytokines in particular TNF (Tumor Necrosis Factor) may be the driving force behind the disease process (Sevenoaks, M. J. and Stockley, R.A., 2006) 41 .
  • TNF Tumor Necrosis Factor
  • the above reviewed groups of patients with different inflammatory conditions represent only a small section of patients which daily develop inflammatory conditions due to underlying systemic diseases or dysfunctions and/or diseases or dysfunctions defined by tissue and/or organs, or due to side-effects from surgery or clinical treatment, and do not limit the scope of the invention.
  • the above- mentioned conditions do exemplify that inflammatory conditions are a great issue in health care.
  • risk prediction, prophylactic treatment as well as monitoring options and palliative and/or curative treatment related to development of inflammations are of great interest in order to decrease the morbidity as well as mortality of such patients in need.
  • novel biomarkers such as oxCL and anti-oxCL
  • treatment with such agents in immunotherapy has also been revealed to be beneficial in prophylactic, palliative and/or curative treatment of developing inflammatory activity and/or autoimmune conditions and/or secondary inflammatory conditions in disease.
  • novel risk factors and risk markers include anti-citrullin antibodies.
  • Rheumatoid factor (RF) is also used, but is less effective.
  • RF rheumatoid factor
  • inflammatory conditions Palliative treatment of inflammatory conditions varies from condition to condition as well as among patients and their overall physical state.
  • Common choices of treatment in inflammatory conditions are immunosuppressant drugs, non-steroidal- inflammatory drugs (NSAIDs), histamines, anti-rheumatic drugs, such as the disease-modifying anti-rheumatoid drugs (DMARDs) and different chemotherapies, as well as antibacterial, antifungal and antiviral drugs.
  • NSAIDs non-steroidal- inflammatory drugs
  • DMARDs disease-modifying anti-rheumatoid drugs
  • chemotherapies antibacterial, antifungal and antiviral drugs.
  • a specific combination of drugs has to be selected for each patient due to side-effects, disease, physical state and several other circumstances influencing the patient's course of disease. For example, most antibiotics are effective only against certain bacteria. In selecting an antibiotic to treat a person with an infection, doctors estimate which bacteria are likely to be the cause.
  • antibiotics that are effective in the laboratory do not necessarily work in an infected person.
  • the effectiveness of the treatment depends on how well the drug is absorbed into the bloodstream, how much of the drug reaches the sites of infection in the body, and how quickly the body eliminates the drug. These factors may vary from person to person, depending on other drugs being taken, other disorders present, and the person's age.
  • doctors also consider the nature and seriousness of the infection, the drug's possible side effects, the possibility of allergies or other serious reactions to the drug, and the cost of the drug (Merck Manuals, online medical library) 57 .
  • Prediction and modulation of inflammation must take into account a complex biology not only including cytokines and other inflammatory factors and/or cells, but also several other biochemical cascades acting in parallel with the immune system. Such cascades include, but are not limited to, the complement system and coagulation.
  • biomarkers predicting one or more potential developing inflammatory condition may thus be as favorable as biomarkers indicating the patient's status in regard to risking inflammatory conditions.
  • ESRD end-stage renal disease
  • Acute renal failure is a frequent problem in the intensive care unit and is associated with a high mortality in critically ill patients.
  • Patients on maintenance dialysis hemodialysis and peritoneal dialysis 1
  • secondary infections bacterial and/or virus. This risk is among others connected to the exposure of bodily fluids during dialysis procedure as well as the patient's general poor physical condition.
  • Peritoneal dialysis is a treatment for patients with severe chronic kidney failure.
  • the process uses the patient's peritoneum in the abdomen as a membrane across which fluids and dissolved substances (electrolytes, urea, glucose, albumin and other small molecules) are exchanged from the blood.
  • Acute renal failure is defined as decreased glomerular filtration, detected by increased serum creatinine.
  • ARF can be a consequence of three etiological scenarios; prerenal causes, intrarenal causes and postrenal causes, and is characterized in that the failure is preceded by an acute injury, which may be healed and regenerated in some degree, thus not resulting in failure of the kidney.
  • Prerenal causes include, but are not limited to, renal failure due to the ingestion of non-steroidal anti-inflammatory drugs (NSAIDs), angiotensin converting enzyme (ACE) inhibitor initiated acute renal failure in patients suffering from hypoperfusion of the kidneys due to, amongst others, renal artery stenosis, congestive heart failure or intrarenal small vessel disease, dependent on angiotensin ll-mediated vasoconstriction of the efferent arteriole to maintain renal perfusion.
  • NSAIDs non-steroidal anti-inflammatory drugs
  • ACE angiotensin converting enzyme
  • Intrarenal causes for acute renal failure may be further divided into inflammatory diseases and acute tubular necrosis.
  • Inflammatory diseases are included in the non- exhaustive list of conditions; vasculitis, glomerulonephritis and drug-induced injury.
  • Acute tubular necrosis may be caused by a plurality of conditions, including, but not limited to, ischemia, poisons and hemolysis.
  • Notable intrarenal diseases are among others the toxic effects of aminoglycoside antibiotics or rhabdomyolysis, after crush injury of muscle results in precipitation of myoglobin into the renal tubules. The former may be prevented by monitoring especially renal compromised patients and/or elderly patients during treatment with antibiotics.
  • sepsis is a common cause of acute renal failure, including prerenal conditions in combination with intrarenal causes.
  • Postrenal causes for acute renal failure may be conditions, where the urinary tract is obstructed.
  • Chronic renal failure may be caused by a non-exhaustive list of conditions, such as diabetes mellitus, hypertension and glomerulonephritis, as well as polycystic kidney disease, obstructions and/or infections.
  • the pathogenesis of acute renal disease is very different from that of chronic renal disease.
  • an acute injury of the kidney results in death and sloughing of the tubular epithelial cells, often followed by regeneration with establishment of normal architecture in acute renal disease
  • chronic injuries in chronic renal disease result in irreversible loss of nephrotic mass.
  • an increased functional burden is borne by fewer nephrons, leading to increased glomerular filtration pressure and hyperfiltration, subsequently leading to fibrosis and scarring (glomerular sclerosis), speeding up the progression to uremia, which indicates the stadium of renal disease when residual renal function is inadequate and renal failure is in progress.
  • nephrons can be lost without any short term evidence of functional impairment.
  • maintenance dialysis may be required for patients with loss of nephrotic mass over 50%, in periods between regeneration of renal tissue (e.g. during chronic renal disease/failure) or whenever glomerular filtration is severely impaired.
  • CVD cardiovascular disease
  • ESRD end-stage renal disease
  • CVD cardiovascular disease
  • the high risk for cardiovascular disease (CVD) results from the additive effect of multiple factors, including but not limited to hemodynamic overload and several metabolic and endocrine abnormalities more or less specific to uremia.
  • CVD includes disorders of the heart, including, but not limited to, left ventricular hypertrophy [LVH] and cardiomyopathy, as well as disorders of the vascular system, including, but not limited to, atherosclerosis and arteriosclerosis, these two disorders being usually associated and interrelated (Semin Dial. 2003 Mar-Apr; 16(2):85-94).
  • biomarkers related to one or more infectious conditions, inflammatory activity and and/or secondary inflammatory conditions related to renal disease, renal failure and/or renal tubular injury, damage and/or dysfunction at an early stage before a decline in glomerular filtration rate is noted by an increased serum creatinine are of great interest.
  • a method for evaluation of the mortality related to such conditions, activity or diseases may be a powerful tool for physicians in determining treatment and the status of the patient's overall physical condition.
  • biomarkers related to renal failure include tubular enzymes (alpha- and pi- glutathione S-transferase, N-acetyl-glucosaminidase, alkaline phosphatase, gamma-glutamyl transpeptidase, Ala-(Leu-Gly)-aminopeptidase, and fructose-1 ,6- biphosphatase), low-molecular weight urinary proteins (alphal- and beta2- microglobulin, retinol-binding protein, adenosine deaminase-binding protein, and cystatin C), Na+/H+ exchanger, neutrophil gelatinase-associated lipocalin, cysteine- rich protein 61 , kidney injury molecule 1 , urinary interleukins/adhesion molecules, and markers of glomerular filtration, such as proatrial natriuretic peptide (1-98) and cystatin C (Trof RJ et al
  • This inflammatory activity indicates that ARF may be an inflammatory condition itself and that this inflammatory state affects the mortality of the condition.
  • Chronic kidney injuries such as seen in, for example, renal disease and/or failure, are commonly considered a chronic inflammation, supported by the elevated levels of TNF-alpha, IL-1beta and IL-6 in bodily fluids of patients.
  • Modulation of inflammation must therefore take into account the complex cytokine biology in patients with established renal failure.
  • the recognition and use of biomarkers predicting a potential development into this inflammatory state of ARF and/or CRF may thus be as favorable as biomarkers indicating the patient's status in regard to risking secondary infectious diseases, CVD and the development of ARF.
  • Low levels of anti-oxCL have previously been shown to be correlated with higher risk of developing inflammatory conditions.
  • OxCL has been found to have proinflammatory properties, and it has also been shown that a low concentration of anti- oxCL is an effective indicator of inflammatory conditions.
  • anti-oxCL is a suitable biomarker for predicting risk, early pathogenesis, progression of as well as mortality related to one or more infectious conditions, renal disease and/or conditions including renal damage, injury and/or dysfunction.
  • the risk of secondary inflammatory conditions in renal disease, such as CVD and secondary infectious conditions and/or inflammatory activity in renal disease and/or renal failure can be predicted when determining the anti-oxCL level in bodily fluids of patients.
  • Cardiolipin is a dimeric phospholipid which is known to be present in eukaryotic cells, bacteria and Archaebacteria but its functional role is only partly known, (Schlame, M., 2008) 1 . It is more prevalent in cells with high metabolic activity, like heart and skeletal muscle, and especially in mitochondrial membranes. The presence of CL in mitochondria and bacteria is interesting from an evolutionary point of view since mitochondria are likely to have a bacterial origin, (Martin, W. et al., 2001 ) 2 . Also lipoproteins, including low density lipoprotein (LDL), contain CL, in contrast to what has previously has been thought, and two thirds of CL are present in low density lipoprotein (LDL), (Deguchi, H. et al., 2000) 3 .
  • LDL low density lipoprotein
  • CL has a unique dimeric structure, highly enriched in linoleic acid groups susceptible to oxidation (Schlame, M. et al., 2000, and Chicco, AJ., and Sparagna, GC, 2007) 4,5 . It has been suggested to play a role in generation of an electrochemical potential for substrate transport and ATP synthesis both in bacteria and mitochondria, (Belikova, NA. et al., 2007, and Basova, LV. et al., 2007) 6 ⁇ 7 CL that has undergone oxidation (oxCL) promotes delocalization and release of cytochrome c, predisposing to its release from mitochondria and the activation of the cell death programmes, (Chicco, AJ. and Sparagna, GC, 2007, Gonzalvez, F. et al., 2007, and Nakagawa, Y., 2004) 5 ⁇ 8 ⁇ 9
  • Antibodies against CL are known to be thrombogenic both in arteries and veins (when present in exceedingly high levels typically above 2 standard deviations as compared to a normal healthy control group). This is especially the case in autoimmune diseases including SLE.
  • PS also named phosphatidylserine, 1 ,2-diacyl-sn-glycero-3-phospho-L-serine and Ptd-L-Ser
  • PS usually represents less than 10% of the total phospholipids, the greatest concentration being in myelin from brain tissue. However, it may comprise 10 to 20 mol% of the total phospholipid in the plasma membrane and endoplasmic reticulum of the cell.
  • the fatty acid composition of PS varies from tissue to tissue, but does not appear to resemble the precursor phospholipids.
  • 1- stearoyl-2-oleoyl and 1-stearoyl-2-arachidonoyl species predominate, but in brain (especially grey matter), retina and many other tissues 1 -stearoyl-2- docosahexaenoyl species are very abundant. Indeed, the ratio of n-3 to n-6 fatty acids in brain PS is very much higher than in most other lipids. (Wood, R. and Harlow, R.D., 1969, and Yabuuchi, H. and O'Brien, J.S., 1968) 42 ⁇ 43
  • Apoptosis (also known as "controlled cell death”) is a cell suicide program under the influence of hormones, growth factors and cytokines, which depending upon the receptors present on the target cells, may activate a genetically controlled cell elimination process.
  • the cell membrane remains intact and the cell breaks into apoptotic bodies, which are engulfed by macrophages and other phagocytic cells.
  • Apoptosis in contrast to necrosis, is not harmful to the host and does not induce any inflammatory reaction.
  • the principal events that lead to inflammatory conditions are necrosis and/or cell damage induced by chemical/physical injury, anoxia or starvation.
  • Cell damage means leakage of cell contents into the adjacent tissues, resulting in the capillary transmigration of granulocytes to the injured tissue.
  • the accumulation of neutrophils and release of enzymes and oxygen radicals enhances the inflammatory reaction and form part of the factors controlling the accumulation and the tissue load of granulocytes and their histotoxic products in inflammatory processes.
  • oxidized phospholipids In chronic inflammation, oxidized phospholipids have been suggested to be part of the oxidative stress, triggering the chronic conditions; however, further evaluation or elaboration of which oxidized phospholipids may be involved have not been made (Kadi, A. et al., 2004) 44 .
  • PS is known to have an essential role in the chain of events of apoptosis after a response to particular calcium-dependent stimuli. Also, PS is exposed on the surface of platelets upon their activation (Sun, J. et al., 1993) 45 .
  • the normal distribution of PS on the inner leaflet of the membrane bilayer is affected when the enzyme scramblase is stimulated and/or aminophospholipid translocases are inhibited.
  • the enzyme scramblase is able to move PS in both directions across the membrane, whereas aminophospholipid translocases return the lipid to the inner side of the membrane.
  • a receptor on the surface of macrophages and related scavenger cells recognizes the PS and facilitates the removal of the apoptotic cells and their potentially toxic or immunogenic contents in a non-inflammatory manner. Binding of PS to specific proteins, such as apolipoprotein H ⁇ -glycoprotein 1), enhances the recognition and clearance. This process is essential for the development of lung and brain, and it is also relevant to clinical situations where apoptosis plays an important part, such as cancer, chronic autoimmunity, and infections.
  • Annexin A5 has been disclosed in the prevention and treatment of artherosclerosis and plaque rupture (International patent application WO 20 ⁇ 5099744) 49 and treatment of inflammatory disorders, SLE as well as sepsis (International patent application WO 2010043045 50 and Cederholm, A., and Frostegard, J., 2007 51 ) and restenosis (International patent application WO 2009103977) 52 .
  • Phospholipids e.g. PS
  • oxidized phospholipids e.g. oxPS
  • oxPS oxidized phospholipids
  • Especially PS and oxPS have been shown to be present on the surface of apoptotic cells (Kadi, A. et al., 2004, and Letter to editor, 2004) 44 ⁇ 53 .
  • CD36 class B cluster of differentiation 36
  • scavenger receptor CD36
  • macrophage recognition of apoptotic cells via CD36 has been shown to occur via interactions with membrane-associated oxidized PS (oxPS) and, to a lesser extent, oxidized phosphatidylcholine (oxPC), but not nonoxidized PS molecular species (Greenberg, M.E. et al., 2006) 54 .
  • oxPS membrane-associated oxidized PS
  • oxPC oxidized phosphatidylcholine
  • lipoproteins including low density lipoproteins (LDLs) contain PS in contrast to what previously has been thought.
  • antioxidants that are capable of inhibiting PS oxidation have been suggested to interfere with PS externalization and/or its recognition by macrophages and a mechanistic link between the oxidative stress in a cell during apoptosis, and the oxidation of PS stimulating a PS-dependent signaling pathway culminating in the disposal of cells by macrophages has previously been suggested (Kagan, V.E. et al., 2003) 56 .
  • milk fat globule epidermal growth factor 8 (MFG-E8) serves as an opsonin for apoptotic cells and recognizes phosphatidylcholine (PC) and PS and preferably interacts with oxPS (Letter to the editor 2004) 53 .
  • Oxidized low density lipoprotein (oxLDL) is taken up through specific scavenger receptors on endothelial cells, which are not down-regulated when exposed to increasing amounts of oxLDL (as opposed to the uptake of LDL), (Hansson, G.K., 2005) 14 .
  • Inhibition of the scavenger function is generally believed to be atheroprotective, preventing foam cell formation in the vascular wall which is a key process in development of atherosclerosis.
  • mice defective in scavenger receptor function develop less atherosclerosis as compared to control mice, (Febraio, M. et al. 2000) 23 . It should be noted, however, that recent findings indicate that different scavenger receptors may play different roles and the role of scavenger receptors may vary depending on disease stage and type, (Moore, K.J. et al., 2006) 24 .
  • oxPS is predominantly exposed on oxLDL, binding and uptake of oxLDL could through oxPS promote atherogenesis.
  • oxPS is present mainly on other compounds, e.g. apoptotic and/or necrotic cells, other proteins, or even bacteria, it is not clear how this would influence foam cell development, which in principle could be decreased.
  • oxPS has been found to play an important role in regulation of apoptosis as well as blood coagulation, which are both bio- molecular systems interacting with the immune response as reviewed above. Further research is needed to clarify which parts of oxLDL play the major role in foam cell formation, as well as if and to what extent phospholipids and/or their oxidized variants influence such processes.
  • prior art associates oxPS with apoptosis and clearance of apoptotic cells preventing inflammation by acting as an "eat me” marker to phagocytic cells and/or opsonins attracting such engulfing cells, when presented on the membrane surface of apoptotic cell.
  • oxPS and not PS
  • anti-oxPS antibodies against oxPS
  • anti-oxPS antibodies against oxPS
  • the increased levels of oxPS (or increased activity thereof) detected in an animal is a potentially predictive value when assessing these individuals' risk of developing acute and/or chronic inflammatory conditions, when infected by microbes or other infectious agents, or experiencing other deficiencies related to systemic and/or organ and/or tissue related disease, injury and/or damage, which can be interpreted as latent stages of (or prior to) developing inflammatory conditions.
  • the measurement of oxPS and/or anti-oxPS levels in animals prior to, or during any kind of clinical treatment may give confirmational information to a presumption of increased risk of inflammation upon infections at an early stage and most importantly and preferably before onset of such an inflammation.
  • prophylactic and/or palliative treatment at early stages, maybe even before symptoms are detected
  • anti-oxPS could be beneficial, reducing the possibilities of inflammations due to infections for animals at such risk by increasing serum levels of anti-oxPS, thereby decreasing oxPS activity and/or levels of oxPS, under or after the onset of treatment related to the primary disease. In this way, the risk of developing inflammatory conditions or consequences of such inflammations may be constrained dramatically.
  • a prediction marker as well as one general prophylactic, palliative and/or curative treatment of inflammatory disease given by the present invention has a great potential of reducing the need and/or amount of additional and disease specific treatments and has the potential of saving time and decreasing mortality and morbidity related to such conditions.
  • Anti-PC has anti-inflammatory properties, inhibiting inflammatory phospholipids like PAF, and anti-PC may be atheroprotective in humans, since these antibodies are negatively associated with atherosclerosis development. Further, as previously published, low anti-PC levels are independently associated with increased risk of cardiovascular diseases (CVD). Moreover, it has surprisingly been found that oxidized phosphatidylserine (oxPS), and not phosphatidylserine (PS), increase the expression of vascular cell adhesion molecule-1 (VCAM-1) (also known as CD106) on the surface of endothelial cells and increase tumor necrosis factor (TNF) levels, suggesting a stimulatory role of oxPS in inflammatory events.
  • VCAM-1 also known as CD106
  • Increased anti-oxPS levels have been revealed to have a strong negative correlation with both TNF serum levels and VCAM-1 expression on endothelial cells. It has further been shown that decreased levels of anti-oxCL are correlated with infectious conditions, and thus therapy involving an elevation of the patient's serum anti-oxCL levels could potentially decrease the risk of such patients developing infectious conditions into chronic inflammatory conditions.
  • the antibodies could either be endogenously produced as a result of active immunization (vaccination), or exogenously administered (passive immunization).
  • active immunization vaccination
  • passive immunization Likewise, our finding that a combination of low IgM anti-PC with low anti-oxCL or low anti-oxPS increases the excess risk of CVD would suggest that the combination of these markers is useful in assessing disease risk and that a combination of these antibodies could also improve therapy, if raised through active immunization or passive immunization.
  • Immunization in regard to stimulating the patient's own production of natural antibodies against oxCL, oxPS, oxLDL, MDA-LDL and/or PC may reduce the risk of patients with inflammatory conditions and provide a tool for prophylactic, palliative and/or curative treatment of developing inflammatory conditions, and/or inflammatory activity and/or secondary inflammatory conditions in disease and/or primary or secondary infectious conditions.
  • treatment with such combinations may reduce the mortality risk of patients with low natural levels of the above antibodies and/or high levels of oxCL, oxPS, oxLDL, MDA-LDL and/or PC, especially during other medical treatment with increased risk of secondary infections and/or suppressive effect on the overall immune system, such as transfusion medicine, immunosuppressant medicine and/or chemotherapy.
  • Atherosclerosis is an inflammatory disease where oxidized low-density lipoprotein may play an important role through oxCL.
  • CVD cardiovascular disease
  • ESRD end-stage renal disease
  • oxCL The surprising characteristics of oxCL are confirmed by the fact that low concentrations of antibodies to mammal oxCL are an effective indicator of development of and/or mortality related to developing infectious disease, in one embodiment especially related to renal disease and/or conditions, renal damage, injury and/or dysfunction. Moreover, the risk of secondary inflammatory conditions in renal disease, such as, but not limited to, CVD and/or development of ARF, as well as inflammatory activity in renal disease and/or related to renal damage and/or injury and/or dysfunction can be predicted when determining the anti-oxCL level in bodily fluids of patients.
  • oxCL this particular antigen
  • oxCL a particular antigen
  • replacement-therapy with anti-oxCL in patients with low natural anti- oxCL levels, or immunization in regard to stimulating the patient's own production of natural antibodies against anti-oxCL may reduce the risk of developing infectious disease and/or reduce the risk of patients with renal disease and/or conditions, renal damage, injury and/or dysfunction, to develop inflammatory activity and/or secondary inflammatory disease and/or to develop ARF and/or CRF.
  • treatment with anti-oxCL may reduce the mortality risk when administered during renal disease, especially during dialysis.
  • the anti-oxCL levels in the patients' bodily fluids may indicate whether and to which extent an immunization and/or a replacement therapy with anti-oxCL may reduce the morbidity of the disease as well as the morbidity related to developing one or more infectious conditions, ARF and/or CRF.
  • oxidized PS in contrast to native PS, has pro-inflammatory properties and serves as a predictive risk marker for developing inflammatory conditions.
  • VCAM-1 vascular cell adhesion molecule-1
  • CD106 cluster of differentiation 106 receptor
  • oxPS The surprising characteristics of oxPS are confirmed by the fact that low concentrations of anti-oxPS and high levels of oxPS are an effective indicator of developing inflammatory conditions.
  • Anti-oxPS, and not anti-PS has now been found to have inflammation protective abilities and, when present in low levels relative to predetermined cutoff values, anti-oxPS predict an increased risk of developing inflammatory conditions.
  • prediction of increased risk of developing inflammatory conditions can accordingly be based on using oxPS, anti-oxPS or both oxPS and anti-oxPS as markers.
  • Immunization by stimulation of an animal's own production of natural antibodies against anti-oxPS reduces the risk of developing infectious and/or inflammatory conditions, inflammatory activity and/or secondary inflammatory conditions related to damage of tissue and/or organs.
  • Replacement therapy may be conducted by administering inhibitors of oxPS and/or anti-oxPS (or bioactive components and/or parts thereof) by injection and/or administered via infusion products enriched with inhibitors of oxPS and/or anti-oxPS (bioactive components and/or parts thereof) such as, but not limited to, plasma products with anti-oxPS and/or any other suitable means of supplying the patient's bodily fluid with anti-oxPS.
  • Vaccines according to this invention that elevate the animals' natural synthesis of anti-oxPS may also be administered by injection and/or infusion.
  • Several methods of administration may be used, including, but not limited to, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, and intraperitoneal injection or infusion. Long-acting forms of subcutaneous/intramuscular injections such as depot injections are available for various drugs, and may also be a choice of administration in connection with the present invention.
  • Administration to mucosal tissues and lungs may be used, including, but not limited to, liquid and solid preparations for nebulisation and rectal application. Dermal application in the form of emulsions, creams, ointments, lotions, gels or other standard formulations for skin preparations may be used.
  • OxPS conjugates, oxPS as such or bioactive components and/or parts thereof, optionally in combination with any suitable adjuvants, may be administered orally, nasally, rectally or through other mucosal tissues in order to achieve an alternative immune-specific reaction than achieved by injection.
  • Transfusion medicine is the branch of medicine that is concerned with the transfusion of blood and blood components.
  • a blood product is any component of the blood which is collected from a donor for use in a blood transfusion.
  • Whole blood is uncommonly used in transfusion medicine at present; most blood products consist of specific processed components such as red blood cells, blood plasma, or platelets.
  • other blood substitutes such as, but not limited to, cryoprecipitate, plasma frozen within 24 Hours after phlebotomy (PF24), fresh frozen plasma (FFP) or cryosupernatant may also be used.
  • oxPS or anti-oxPS or both oxPS and anti- oxPS are used as biologic risk markers for inflammatory conditions of different underlying conditions, diseases and/or dysfunctions.
  • the invention also relates to use of inhibitors of oxPS activity and/or antibodies against oxPS in a method of replacement therapy, i.e. elevating the patients' serum levels of anti-oxPS in patients with low levels of functional anti-oxPS and/or high levels of oxPS and therefore experiencing an increased risk of developing inflammatory conditions.
  • Means for replacement therapy may be anti-oxPS as such, administered by injection and/or administered through enrichment of infusion products, such as, but not limited to, plasma products with anti-oxPS and/or any other suitable means of supplying the patient's bodily fluid with anti-oxPS.
  • the invention also relates to a vaccine (agent that is suitable for increasing the anti- oxPS immune response in an animal, in particular oxPS conjugates, oxPS as such or bioactive components and/or parts thereof optionally in combination with any suitable adjuvants).
  • a vaccine with oxPS and/or fragments thereof may be a choice of prophylactic treatment prior to surgery, in order to elevate anti-oxPS levels and to decrease the risk of developing post-operative inflammatory conditions due to infections, or prevent inflammation due to treatment with immunosuppressant drugs, in patients with unstable immune systems or other clinical treatments known to increase risk of infection and/or inflammation.
  • the present invention further relates to a kit needed to perform the immunoassay comprising oxPS markers.
  • anti-oxPS levels measured in a sample of animal origin may indicate whether and to which extent an immunization using a vaccine against oxPS and/or a replacement therapy with anti-oxPS or bioactive components and/or parts thereof may reduce the risk and/or the morbidity related to such conditions.
  • oxPS levels measured in a sample of animal origin may indicate whether and to which extent an immunization using a vaccine against oxPS and/or a replacement therapy with anti-oxPS or bioactive components and/or parts thereof may reduce the risk and/or the morbidity related to such conditions.
  • anti-oxPS and oxPS levels measured in a sample of animal origin may indicate whether and to which extent an immunization using a vaccine against oxPS and/or a replacement therapy with anti-oxPS or bioactive components and/or parts thereof may reduce the risk and/or the morbidity related to such conditions.
  • infectious condition is understood in the general meaning of the term, comprising microbial infections, provoking a normal immune response which will be cured in an untreated state (e.g. not develop into a prolonged inflammatory condition).
  • the term "developing infectious condition” means a condition caused by an infectious agent of bacterial, protozoal, viral or parasitic type, developing into an inflammatory condition due to an abnormal increased or decreased immune response to an exogenous stimulus (e.g. virus and/or bacteria and/or toxins originating from infectious agents).
  • an exogenous stimulus e.g. virus and/or bacteria and/or toxins originating from infectious agents.
  • condition means both disease and/or condition, systemic and/or defined by tissue and/or organs.
  • renal disease means both renal disease and/or renal conditions where glomerular filtration is acutely and/or chronically decreased and/or failing due to prerenal, intrarenal and/or postrenal causes.
  • Such renal diseases include, but are not limited to, renal conditions and/or failure including ARF and/or CRF where prerenal causes include, but are not limited to, the ingestion of non-steroidal anti-inflammatory drugs (NSAIDs), angiotensin converting enzyme (ACE) inhibitor initiated acute renal failure in patients suffering from hypoperfusion of the kidneys due to, amongst others, renal artery stenosis, congestive heart failure or intrarenal small vessel disease, dependent on angiotensin ll-mediated vasoconstriction of the efferent arteriole to maintain renal perfusion.
  • NSAIDs non-steroidal anti-inflammatory drugs
  • ACE angiotensin converting enzyme
  • Intrarenal causes may be further divided into inflammatory diseases and acute tubular necrosis.
  • Inflammatory diseases are included in the non-exhaustive list of conditions such as; vasculitis, glomerulonephritis and drug-induced injury.
  • Acute tubular necrosis may be caused by a plurality of conditions, such as, but not limited to, ischemia, poisons and hemolysis.
  • Notable intrarenal causes are among others the toxic effects of aminoglycoside antibiotics or rhabdomyolysis, after crush injury of muscle results in precipitation of myoglobin into the renal tubules.
  • Postrenal causes for renal disease including acute renal failure may not be limited to conditions where the urinary tract is obstructed.
  • CRF may also be caused by a non- exhaustive list of conditions, such as diabetes mellitus, hypertension and glomerulonephritis, as well as polycystic kidney disease, obstructions and/or infections.
  • inflammatory activity means inflammatory activity in tissue originated in a diseased, damaged and/or injured organ not related to a systemic inflammatory condition.
  • Inflammatory activity includes, but is not limited to, acute and chronic tissue and/or organ damages or injuries showing elevated levels of inflammatory biomarkers that include, but are not limited to, TNF, IL-1-beta, IL-6 and IL-8 in bodily fluids of patients.
  • secondary inflammatory condition in disease means inflammatory activity, conditions and/or disease with an onset following the onset of a primary disease, condition and/or tissue or organ damage, injury and/or dysfunction.
  • secondary inflammatory conditions in this invention include both systemic inflammatory conditions and inflammatory conditions defined by tissue and/or organ, and include, but are not limited to, any of the following diseases, conditions and/or activities; auto-immune diseases, type II diabetes, Alzheimer's disease, dementia in general, rheumatic diseases, atherosclerosis, acute and/or chronic inflammatory conditions, type I diabetes, rheumatoid arthritis, psoriasis, psoriatic arthritis, acne, ankylosing spondylitis, Reiter's Syndrome, systemic lupus erythematosus (SLE), dermatomyositis, Sjogren's syndrome, multiple sclerosis, asthma, encephalitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), arthritis, including osteoarthritis, idiopathic inflammatory myopathies (MM), dermatomyositis (DM), polymyositis (PM), inclusion body myositis, and/or an
  • animal means mammals and other vertebrates and invertebrates, in particular mammals such as mice, rats, rabbits, dogs, cats, cattle, horses and humans.
  • samples of animal origin means any natural sample from an animal, including but not limited to tissue and/or fluid, such as, but not limited to, plasma, serum, blood, urine, saliva or samples collected via bronchioalveolar lavage (BAL) or aspiration.
  • tissue and/or fluid such as, but not limited to, plasma, serum, blood, urine, saliva or samples collected via bronchioalveolar lavage (BAL) or aspiration.
  • BAL bronchioalveolar lavage
  • monoclonal or polyclonal antibodies of isotype IgA, IgD, IgE, IgG, IgM, raised against oxCL, oxPS, oxLDL, MDA-LDL, PC, apoBIOO or bioactive components and/or parts/fragments/derivatives thereof refer to any monoclonal or polyclonal antibody produced by immunisation of a suitable mammal, including, but not limited to, mouse, rabbit, goat, sheep, or horse.
  • antibodies against oxCL (anti-oxCL), antibodies against oxPS (anti-oxPS), antibodies against oxLDL (anti-oxLDL), antibodies against MDA-LDL (anti-MDA-LDL), antibodies against apoBIOO (anti- apoBIOO), antibodies against PC (anti-PC) and/or or antibodies against bioactive components and/or parts/fragments/derivatives thereof may be determined using any of the methods and techniques conventional in the art for such determination.
  • a method may comprise an immunoassay e.g.
  • a medicament may be intended for oral, rectal, parenteral and mucosal administration and may be formulated with excipients normally employed for such formulations.
  • the medicament may be administered in a way so as to be compatible with the dosage formulation and in such amount as will be therapeutically effective and immunogenic.
  • vaccine means any agent that is suitable for increasing the animal's natural anti-oxCL response, in particular oxCL conjugates, oxCL as such or bioactive components and/or parts thereof, and/or the animal's natural anti-oxPS response, in particular oxPS conjugates, oxPS as such or bioactive components and/or parts thereof, and/or the animal's natural anti-oxLDL response, in particular oxLDL conjugates, oxLDL as such or bioactive components and/or parts thereof and/or the animal's natural anti-MDA-LDL response, in particular MDA-LDL conjugates, MDA-LDL as such or bioactive components and/or parts thereof and/or the animal's natural anti-apoB100 response, in particular apoBIOO conjugates, apoBIOO as such or bioactive components and/or parts thereof and/or the animal's natural anti-PC response, in particular PC-containing bacteria, such as pneumococcae or parts of it, PC conjugates, PC
  • OxCL in contrast to native CL, has pro-inflammatory properties.
  • OxCL, but not CL induced endothelial cells to express ICAM-1 , intracellular adhesion molecule 1 , and VCAM-1 , vascular cell adhesion molecule 1.
  • Adhesion molecules play an important role in recruiting monocytes into the intimal compartment of arteries, which is an early step in inflammatory processes.
  • oxCL, but not CL has the capacity to induce interleukin 6, IL-6, production.
  • CRP C-reactive protein
  • Fb fibrinogen
  • IL-6 has been identified as an independent risk factor for coronary artery disease (CAD).
  • oxCL could induce leukotriene B4 (LTB4) production in both neutrophils and macrophages. It is also found that oxCL, but not CL, can provoke intracellular calcium mobilization which is an initiation signal for LTB4 production and also in general a sign of cell activation. Leukotrienes are short-lived lipid mediators that have potent pro-inflammatory biological activities.
  • Leukotriene B4 is one of the most potent chemotactic agents for other inflammatory cells and is biosynthesized from arachidonic acid by the sequential action of 5-lipoxygenase (5-LO) and leukotriene A4, LTA4, hydrolase, mainly in cells of myeloid lineage, such as neutrophil and macrophage, (Funk, CD., 2005) 16 .
  • 5-lipoxygenase (5-LO) and leukotriene A4, LTA4 hydrolase mainly in cells of myeloid lineage, such as neutrophil and macrophage, (Funk, CD., 2005) 16 .
  • Two G-protein coupled LTB4 receptors have been identified, BLT1 and BLT2, with high and low affinity for LTB4, respectively, (Yokomizo, T. et al., 1997, and Yokomizo, T. et al., 2000) 17,18 .
  • LTB4 is known to exert broad pro-inflammatory effects, and evidence is accumulating regarding the antimicrobial functions of LTB4, (Serezani, CH. et al., 2005, and Wan, M. et al., 2007) 19 ⁇ 20 Furthermore, the LTB4- BLT1 pathway was found to be important for linking early immune responses and the multiple classes of effector cells associated with acquired immunity, (Goodarzi, K. et al., 2003) 21 . LTB4 may play an important role in atherosclerosis and CVD since mRNA levels for the three key proteins are significantly increased in human atherosclerotic plaque, and more pronounced in patients with ongoing CVD (Qui, H. et al., 2006) 22 .
  • oxCL could play a role in initiating plaque rupture and CVD.
  • Another finding was that oxCL but not CL (or reduced CL), inhibits uptake of oxLDL in macrophages.
  • oxCL is predominantly exposed on oxLDL, binding and uptake of oxLDL through oxCL could promote atherogenesis.
  • oxCL is present mainly on other compounds, e.g. apoptotic cells, other proteins, or even bacteria, it is not clear how this would influence foam cell development, which in principle could be decreased.
  • apoptotic cells are known not to have any pronounced pro-inflammatory effects, this would suggest at least that oxCL is not an important factor exposed during apoptosis. Further research is needed to clarify which parts of oxLDL play the major role in foam cell formation.
  • annexin A5 is abundant in atherosclerotic lesions and that anti-CL can interfere with its binding to endothelial cells, promoting CVD in SLE, (Cederholm, A. et al., 2005) 12 . Rand et al. have demonstrated that annexin A5 can form a crystalline shield over cell surfaces, which could have a protective function. However, this can be disrupted by anti-CL, causing the antiphospholipid antibody syndrome, characterized by arterial and venous thrombosis and also miscarriage, (Rand, JH., and Wu, XX., 1999) 25 .
  • Annexin A5 has recently been implicated in CVD in general also as indicated by its function as a potent anti-atherothrombotic agent in a rabbit model of arterial thrombosis, by interfering with tissue factor expression and by recovery of hypercoagulability, (Cederholm, A. et al., 2005, Thiagarajan, P., and Benedict, CR., 1997, and Ishii, H. et al., 2007) 26"28 .
  • Annexin A5 inhibited the pro-inflammatory effects of oxCL, including induction of adhesion molecules, IL-6.
  • the mechanism could be that annexin A5 binds to phosphatidylserine (PS) of endothelial cells, PS being a pro-thrombotic factor.
  • PS phosphatidylserine
  • annexin A5 can bind to oxCL but not CL ( Figure 5), though the exact mechanisms are not clear. Annexin A5 can thus in principle prevent these oxCL-induced effects by interacting with oxCL, though the exact mechanisms remain unknown.
  • annexin A5 enabling it to form crystalline layers, which by themselves could be antiinflammatory, inhibiting pro-inflammatory effects of oxCL by a mechanistic barrier.
  • CL is synthesized in cells de novo through the action of cardiolipin synthase, which is most active in high-metabolic tissue (where CL itself is most abundant). In mitochondria, CL modification and remodelling occurs, including substantial changes in the acyl composition 1 . Accumulating evidence now also suggests that remodeling defects of CL could play a role in physiology and also pathology such as in diabetes, heart failure and Barth syndrome, (Han, X. et al., 2007, Sparagna, GC.
  • oxCL also could contribute to human chronic inflammatory disease in general, by its pro-inflammatory effects.
  • CL binds easily to proteins, and it is possible that such complexes can become proinflammatory e.g. if exposed to the hypoxic and/or pro-oxidant environment in atherosclerotic plaques, but in principle also in other chronic inflammatory conditions like rheumatoid arthritis (RA) and SLE.
  • RA rheumatoid arthritis
  • oxLDL and foam cells are present in rheumatoid arthritis, RA-synovia, (Winyard, PG. et al., 1993) 33 , and oxLDL is raised and associated with disease activity in RA (unpublished observation).
  • SLE systemic lupus erythematosus
  • the risk of CVD is very high due to a combination of traditional and non-traditional risk factors, (Frostegard, J., 2005) 10 .
  • Both anti-CL and oxLDL are important examples of non-traditional risk factors.
  • oxLDL is raised in CVD in SLE, (Frostegard, J., 2005, and Frostegard, J.
  • antibodies to this particular antigen i.e. oxCL
  • oxCL this particular antigen
  • infectious conditions e.g. developing infectious conditions
  • inflammatory activity in renal disease related to renal damage, injury and/or dysfunction and/or secondary inflammatory conditions in renal disease e.g. oxCL
  • an insufficiently developed natural immunity against oxCL represents an underlying mechanism for development of such diseases and the mortality related thereto.
  • OxCL belongs to a novel class of pathogen-associated molecular patterns (PAMPs) and natural antibodies against oxCL bind oxLDL, (Tuominen, A. et al., 2006) 36 .
  • PAMPs pathogen-associated molecular patterns
  • oxCL can be a novel pro-inflammatory factor, causing development and progression of infectious diseases, also playing a role in mortality related to such one or more infectious conditions and/or secondary inflammatory diseases in renal diseases, activity and/or conditions including CVD, RA and SLE. Further, based on its capacity to inhibit oxCL-effects, it can be hypothesized that antibodies against oxCL could also be developed into a therapeutic agent for prophylactic, palliative and/or curative treatment in diseases where mortality and/or morbidity are increased due to oxCL activity.
  • OxPS contributes to human chronic inflammatory disease in general, by its pro-inflammatory effects. OxPS may easily bind proteins, and it is possible that such complexes can become pro-inflammatory e.g. if exposed to the hypoxic and/or pro-oxidant environment in atherosclerotic plaques, but in principle also in other chronic inflammatory conditions like rheumatoid arthritis (RA) and SLE.
  • RA rheumatoid arthritis
  • oxPS has great potential as a novel proinflammatory factor causing development and progression of infectious and/or inflammatory conditions, including auto-immune as well as inflammatory conditions, such as, but not limited to, RA and SLE.
  • annexin A5 and/or anti-oxPS and/or bioactive components and/or parts thereof could be developed into a therapeutic agent for prophylactic, palliative and/or curative treatment in diseases where morbidity is related to increased oxPS activity and/or decreased anti-oxPS activity.
  • oxPS such as oxPS conjugates, oxPS as such and/or bioactive components and/or parts thereof
  • administration of vaccines against oxPS may be used in immune therapy for protection against developing inflammatory conditions and/or activity in animals at risk thereof, including scenarios that are not limited to preparation of a patient for surgery and/or chemotherapy.
  • Detection of antibodies is a common form of medical diagnostics. For example, in different biochemical assays for disease diagnosis, a titre of antibodies indicative for a particular disease is estimated from the blood and if those antibodies are not present, the person does not have the disease in question. If however antibodies are found, a diagnosis is normally accepted when symptoms are correlating such findings.
  • Several immunodiagnostic methods based on detection of antigen-antibody complex are used to diagnose infectious diseases, for example ELISA, immunofluorescence, Western blot, immunodiffusion, Immunoelectrophoresis, and magnetic immunoassay.
  • Targeted monoclonal antibody therapy has been used to treat diseases such as rheumatoid arthritis, multiple sclerosis, psoriasis, and many forms of cancer including non-Hodgkin's lymphoma, colorectal cancer, head and neck cancer and breast cancer.
  • An immunoassay is a test that measures the concentration of a substance in a sample of animal origin (such as, but not limited to, bodily fluid and/or tissue), using the reaction of an antibody or antibodies to its antigen. Both polyclonal and monoclonal antibodies can be used. Monoclonal antibodies usually bind only to one site of a particular molecule, and therefore provide a more specific and accurate test. Both the presence of antigen or antibodies can be measured.
  • the response (often a fluorescence signal or dye intensity; however also other labelling options may be used) measured is compared to standards of a known concentration. This can be done through the plotting of a standard curve on a graph, the position of the curve at response of the unknown is then examined, and so the quantity of the unknown can be determined.
  • the most common method used for detecting the quantity of antibody or antigen is to label either the antigen or antibody.
  • the label may consist of an enzyme (enzyme immunoassay (EIA, also called enzyme-linked immunosorbent assay or ELISA), colloidal gold (lateral flow assays), radioisotopes such as 1-125 radioimmunoassay (RIA), magnetic labels (magnetic immunoassay - MIA) or fluorescence.
  • enzyme immunoassay also called enzyme-linked immunosorbent assay or ELISA
  • colloidal gold lateral flow assays
  • radioisotopes such as 1-125 radioimmunoassay (RIA)
  • RIA radioimmunoassay
  • MIA magnetic labels
  • fluorescence Other techniques include agglutination, nephelometry, turbidimetry and Western Blot.
  • Immunoassays are normally divided into those that involve labelled reagents and those which involve non-labelled reagents. Those which involve labelled reagents can be subdivided into homogenous and heterogeneous immunoassays. Heterogeneous immunoassays can furthermore be competitive or non-competitive.
  • ELISA or EIA a primary antigen or antibody is affixed to a surface in a dish, well or any other comparable laboratory means and then a sample of interest suspected to contain the corresponding antibody or antigen is added and incubated to let the corresponding antigen/antibody complex react. An additional (i.e.
  • This antibody linked to an enzyme or other form of labelling means is added to the well or dish after an intermediate wash.
  • This antibody is targeted to bind the antigen/antibody complex and/or parts thereof establishing a possibility to detect a signal upon an activation reaction with suitable instruments measuring intensity of the signal chosen.
  • any form of ELISA (it being direct ELISA, indirect ELISA, sandwich ELISA, competitive ELISA or reverse ELISA) can be performed to evaluate either the presence of antigen or the presence of antibody in a sample, and is consequently a useful tool for determining serum antibody concentrations.
  • a sample of bodily fluid e.g. a person's serum
  • a plate to which one of the relevant compounds (for example oxCL, oxPS, oxLDL, or MDA-LDL) is attached.
  • one of the relevant compounds for example oxCL, oxPS, oxLDL, or MDA-LDL
  • antibodies to the compound are present in the serum, they may bind to the compound.
  • the plate can then be washed to remove all other components of the serum.
  • the plate will contain enzyme in proportion to the amount of secondary antibody bound to the plate.
  • a substrate for the enzyme can then be applied, and catalysis by the enzyme leads to a change in colour or fluorescence.
  • ELISA results can then be reported as a number, which can subsequently be compared to the relevant "cutoff point between a positive and negative result.
  • a cutoff value may be determined by comparing it with a known standard. This can be determined by applying a sample of known concentration to a surface and fixing it to the surface to render it immobile. Samples of known concentrations can then be used to generate a standard curve.
  • Immunotherapy is normally defined within medicine as "Treatment of disease by inducing, enhancing, or suppressing an immune response”.
  • Immunotherapies designed to reduce, suppress or more appropriately direct an existing immune response, as in cases of autoimmunity or allergy, are normally termed “suppression immunotherapies”.
  • the active agents of immunotherapy are collectively called “immunomodulators”.
  • the average level of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS anti- oxLDL, anti-MDA-LDL and/or anti-PC in animals depends on the type of bodily fluid sample, the specific species, and may vary between the different population groups.
  • a cutoff value may be chosen so that concentrations of oxCL, oxPS, oxLDL or MDA- LDL higher than said cutoff value are associated with an increased risk of developing a cardiovascular disease, an auto-immune disease or inflammatory condition.
  • a cutoff value may be chosen so that concentrations of oxCL, oxPS, oxLDL or MDA- LDL lower than said cutoff value are not associated with an increased risk of developing a cardiovascular disease, an auto-immune disease or inflammatory condition.
  • a cutoff value may be chosen so that concentrations of anti-oxCL, anti-oxPS, anti- oxLDL, anti-MDA-LDL and/or anti-PC lower than said cutoff value are associated with an increased risk of developing a cardiovascular disease, an auto-immune disease or inflammatory condition.
  • a cutoff value may be chosen so that concentrations of anti-oxCL, anti-oxPS, anti- oxLDL, anti-MDA-LDL and/or anti-PC higher than said cutoff value are not associated with an increased risk of developing a cardiovascular disease, an autoimmune disease or inflammatory condition.
  • the cutoff value may also be determined from the ratio between the oxCL level and the anti-oxCL level, the ratio between the oxPS level and the anti-oxPS level, the ratio between the oxLDL level and the anti-oxLDL level, and the ratio between the MDA-LDL level and the anti-MDA-LDL level.
  • One embodiment of this invention relates to a method for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising monitoring and/or determining the levels of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA- LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, and comparing said levels with predetermined cutoff values.
  • Another embodiment of this invention relates to a method for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising monitoring and/or determining the levels of oxPS or anti-oxPS, and comparing said levels with predetermined cutoff values.
  • Another embodiment of this invention relates to a method for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising monitoring and/or determining the levels of two or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA- LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, and comparing said levels with predetermined cutoff values.
  • the levels of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL and MDA-LDL are compared with predetermined cutoff values for a given population of individuals leading to a diagnosis.
  • the levels of two or more markers selected from the list consisting of oxCL, oxPS, oxLDL and MDA-LDL are compared with predetermined cutoff values for a given population of individuals leading to a diagnosis.
  • One way of testing the levels of oxCL, oxPS, oxLDL and MDA-LDL in such samples may be immunoassays.
  • the level of one or more markers selected from the list consisting of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC is compared with a predetermined cutoff value for a given population of individuals leading to a diagnosis.
  • the level of two or more markers selected from the list consisting of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC is compared with a predetermined cutoff value for a given population of individuals leading to a diagnosis.
  • One way of testing the levels of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and/or anti-PC in such samples may be immunoassays.
  • One preferred embodiment of the method relates to a predetermined cutoff value for a given population of individuals being chosen so that concentrations of oxCL, oxPS, oxLDL and/or MDA-LDL higher than said cutoff value is associated with an increased risk of developing inflammatory conditions.
  • Another preferred embodiment of the method relates to a predetermined cutoff value for a given population of individuals being chosen so that concentrations of oxCL, oxPS, oxLDL and/or MDA-LDL lower than said cutoff value is not associated with an increased risk of developing inflammatory conditions.
  • Another preferred embodiment of the method relates to a predetermined cutoff value for a given population of individuals being chosen so that concentrations of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and/or anti-PC lower than said cutoff value is associated with an increased risk of developing inflammatory conditions.
  • Another preferred embodiment of such method relates to a predetermined cutoff value for a given population of individuals being chosen so that concentrations of anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and/or anti-PC higher than said cutoff value is not associated with an increased risk of developing inflammatory conditions.
  • the inflammatory condition may be caused by infectious agents directly or indirectly (via e.g. toxins).
  • inflammatory conditions comprise one or more primary and/or secondary inflammatory diseases, inflammatory conditions and/or inflammatory activity.
  • Inflammatory condition may be selected from the non-exhaustive group comprising infections, auto-immune diseases, type II diabetes, Alzheimer's disease, dementia in general, rheumatic diseases, cardiovascular disease (CVD), such as atherosclerosis, atheromatous plaque rupture, myocardial infarction, acute coronary syndrome, stroke, transient ischemic attack (TIA), claudication, angina pectoris, high blood pressure, acute and/or chronic inflammatory conditions, type I diabetes, rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Reiter's syndrome, systemic lupus erythematosus, dermatomyositis, Sjogren's syndrome, multiple sclerosis, asthma, acne, encephalitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), arthritis, including osteoarthritis, idiopathic inflammatory myopathies (MM), dermatomyositis (
  • inflammatory conditions include conditions where an undesirable immune response is responsible for a pathological state or parts thereof and include autoimmune diseases, infectious conditions, rheumatic conditions, or secondary complications, also those caused by side-effects of medicaments.
  • condition relates to conditions, disease and activity defined by a tissue and/or organ.
  • said inflammatory conditions are systemic or defined by the tissue and/or organ of origin, such as a renal condition, a renal disease and/or renal failure.
  • said renal condition, renal disease and/or renal failure is an autoimmune disease, a metabolic disease or a disease caused by toxic compounds.
  • said levels of oxCL, oxPS, anti-oxCL, anti-oxPS, anti- oxLDL, anti-MDA-LDL and/or anti-PC are monitored and/or determined in samples of animal origin, wherein said animal is selected from the group comprising mammals, vertebrates or invertebrates and wherein said mammal is selected from the group comprising human, dog, horse, cat mouse, rat, pig or cattle.
  • said samples of animal origin comprise tissue, plasma, serum, blood, urine, saliva or samples collected via bronchioalveolar lavage (BAL) or aspiration.
  • BAL bronchioalveolar lavage
  • said method may be an immunoassay, making use of oxidized cardiolipin (oxCL) or fragments thereof, oxidized phosphatidylserine (oxPS) or fragments thereof, oxidized LDL (oxLDL) or fragments thereof, and a marker of antibody origin for binding to oxCL, oxPS, anti-oxCL, anti-oxPS, anti-oxLDL, anti- MDA-LDL, anti-PC, anti-oxCL/oxCL-antibody-antigen complex, anti-oxPS/oxPS- antibody-antigen complex, anti-oxLDL oxLDL-antibody-antigen complex, anti-MDA- LDUMDA-LDL-antibody-antigen complex and/or anti-PC/PC-antibody-antigen complex selected from ELISA, radioimmunoassay (RIA), flow cytometry or EIA.
  • oxCL oxidized cardiolipin
  • oxPS oxidized phosphati
  • Another embodiment of this invention relates to a method for treating or decreasing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising administering to an animal in need thereof a therapeutically effective amount of one or more compounds selected from the list consisting of annexin A5, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA- LDL and anti-PC, as well as antibodies against components, fragments or derivatives of MDA-LDL or OxLDL (such as apoBIOO or fragments thereof), and also bioactive components and/or parts/fragments of these compounds.
  • Another embodiment of this invention relates to a method for treating or decreasing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising administering to an animal in need thereof a therapeutically effective amount of two or more compounds selected from the list consisting of annexin A5, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA- LDL and anti-PC, as well as antibodies against components, fragments or derivatives of MDA-LDL or OxLDL (such as apoBIOO or fragments thereof), and also bioactive components and/or parts/fragments of these compounds.
  • antibodies can be monoclonal or polyclonal of isotype IgA, IgD, IgE, IgG, IgM against oxCL, oxPS or PC, or bioactive components and/or parts/fragments thereof.
  • the animal is suffering from a renal condition, a renal disease and/or renal failure.
  • Another embodiment of this invention relates to a method for assessing and decreasing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising monitoring and/or determining the level of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, comparing said levels with predetermined cutoff values, and administering to an animal in need thereof a therapeutically effective amount of one or more compounds selected from the list consisting of annexin A5, anti-oxCL, anti- oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, as well as antibodies against components, fragments or derivatives of MDA-LDL or OxLDL (such as apoBIOO or fragments thereof), and also bioactive
  • Another embodiment of this invention relates to a method for assessing and decreasing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto, comprising monitoring and/or determining the level of two or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, comparing said levels with predetermined cutoff values, and administering to an animal in need thereof a therapeutically effective amount of two or more compounds selected from the list consisting of annexin A5, anti-oxCL, anti- oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC, as well as antibodies against components, fragments or derivatives of MDA-LDL or OxLDL (such as apoBIOO or fragments thereof), and also bioactive components and/or parts/fragments of these compounds.
  • compositions comprising one or more inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL and inhibitors of PC, including antibodies, and/or bioactive components and/or fragments thereof, may, according to this invention, be used as a medicament, preferably used for treatment of inflammatory conditions, wherein said treatment comprises prophylactic, palliative and/or curative treatment.
  • compositions comprising two or more inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL and inhibitors of PC, including antibodies, and/or bioactive components and/or fragments thereof, may, according to this invention, be used as a medicament, preferably used for treatment of inflammatory conditions, wherein said treatment comprises prophylactic, palliative and/or curative treatment.
  • this medicament is in the form of a vaccine.
  • compositions comprising one or more inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL and inhibitors of PC, including antibodies, and/or bioactive components and/or fragments thereof are administered by injection and/or infusion, such as, but not limited to, transfusion.
  • Another route of administration could be nasal, rectal or oral, in order to modify and raise anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC levels.
  • compositions comprising two or more inhibitors selected from the list consisting of annexin A5, inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL and inhibitors of PC, including antibodies, and/or bioactive components and/or fragments thereof are administered by injection and/or infusion, such as, but not limited to, transfusion.
  • Another route of administration could be nasal, rectal or oral, in order to modify and raise anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and anti-PC levels.
  • products used in transfusion medicine where the inhibitors of oxCL and/or inhibitors of oxPS and/or inhibitors of oxLDL and/or inhibitors of MDA-LDL and/or inhibitors of PC and/or bioactive components and/or fragments thereof is/are added comprise blood products selected from the group comprising whole blood products, platelet products, cryosupernatant or plasma products.
  • Another embodiment of this invention is the use of one or more conjugates selected from the list consisting of oxCL conjugates, oxPS conjugates, oxLDL conjugates, MDA-LDL conjugates and PC conjugates and/or bioactive components and/or fragments thereof for activation immunotherapy for treatment of inflammatory conditions, optionally adjuvants may be added.
  • Another embodiment relates to the use of one or more inhibitors selected from the list consisting of inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and/or anti-PC and/or bioactive components and/or fragments thereof as a part of infusion products selected from the group comprising whole blood products, platelet products, cryosupernatant or plasma products.
  • Another embodiment of this invention is the use of two or more conjugates selected from the list consisting of oxCL conjugates, oxPS conjugates, oxLDL conjugates, MDA-LDL conjugates and PC conjugates and/or bioactive components and/or fragments thereof for activation immunotherapy for treatment of inflammatory conditions, optionally adjuvants may be added.
  • Another embodiment relates to the use of two or more inhibitors selected from the list consisting of inhibitors of oxCL, inhibitors of oxPS, inhibitors of oxLDL, inhibitors of MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL and/or anti-PC and/or bioactive components and/or fragments thereof as a part of infusion products selected from group comprising whole blood products, platelet products, cryosupernatant or plasma products.
  • Another embodiment of this invention relates to a kit for monitoring and/or determining the level of one or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL, and anti-PC for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto.
  • Another embodiment of this invention relates to a kit for monitoring and/or determining the level of two or more markers selected from the list consisting of oxCL, oxPS, oxLDL, MDA-LDL, anti-oxCL, anti-oxPS, anti-oxLDL, anti-MDA-LDL, and anti-PC for assessing the risk of developing inflammatory conditions, including infectious conditions, and/or the risk of mortality related thereto.
  • Another embodiment of this invention relates to a method of preventing or treating inflammation induced by radiation, for example in radiotherapy of cancer, comprising administering to an animal in need thereof a therapeutically effective amount of a) anti-oxCL and anti-oxPS, or
  • Another embodiment of this invention relates to a topical skin care composition
  • a topical skin care composition comprising
  • Another embodiment of this invention relates to a method of preventing or treating inflammation induced by UVA and/or UVB rays comprising administering to an animal in need thereof a therapeutically effective amount of a topical skin care composition comprising
  • Cardiolipin from bovine heart was purchased as ethanol solution from Sigma (Sigma product C 1649) and was stored at -20°C.
  • Hydro heart cardiolipin (reduced CL) was purchased from Avanti Polar Lipids, Inc.
  • cardiolipin was oxidized in aqueous solutions containing 1 .5 mmol/L tert- butylhydroperoxide and 20 ⁇ /L CuSO4. Both cardiolipin and copper treated cardiolipin were measured with MS-spectrophotometer, and it was confirmed to have been oxidized by copper and tert-butylhydroperoxide.
  • HUVECs human umbilical vascular endothelial cells
  • Cascade Biologies, Inc Portland, Ore
  • Cultures were maintained in EGMTM phenol red-free medium (Clonetics, San Diego, CA), containing 2% of fetal bovine serum and supplements, at 37°C under humidified 5% CO 2 conditions. All experiments were performed at passage 3 to 5.
  • HUVECs were seeded at 6x10 4 cells/2mL density on 6-well plates (NUNC Inc, Naperville, III). After allowing endothelial cells to attach overnight, cells were used for stimulation.
  • oxCL was incubated with annexin A5 (10pg/ml) for a half hour before adding to cells. After 24 hours incubation, detached floating cells were washed away and cells were harvested into Falcon FACS tubes. After centrifuging at 1200 rpm for 5 minutes, cells were resuspended in 300 ⁇ FACS buffer (1 % FBS-PBS), incubated with 10 ⁇ PE- conjugated anti-CD54 (eBioscience) and 10 ⁇ FITC-conjugated anti-Human CD106 (Becton, Dickinson) for 30 minutes on ice.
  • the intercellular adhesion molecule (ICAM-1 ) CD54 and the vascular cell adhesion molecule (VCAM-1 ) CD106 were studied with flow cytometry analysis equipped with CellQuest software. For each sample, 10,000 events were analyzed.
  • the macrophages which transformed from human monocyte-derived THP-1 cells were plated in a 6-well plate at a density of 1 x 10 6 cells/well in DMEM (Invitrogen, USA) containing 10% FBS overnight. Then the cells were washed three times with serum free medium before being incubated with oxCL (2 ig/m ⁇ ). Thereafter, the cells were washed 4 times with ice-cold 0.2% BSA/PBS and once with PBS. The cells were harvested in PBS containing 0.1% BSA and 0.01% NaN3.
  • HUVEC cells as above were seeded at density 10 6 /2ml into 6 well plates. Allowing cell attachment for 24 hours, oxCL (20pg/ml) with or without annexin A5 (20pg/ml) and reduced CL were added and incubated for 24 hours. Cell supernatants were collected. Production of IL-6 was measured with protein multiplex immunoassay kits (from Bioscource, USA) and Bio-PleiTM system (BioRad, USA).
  • Enzyme-Linked Immunosorbent Assay for annexin A5 Binding
  • F96 microtiter polysorb plates (Roskilde, Denmark) were coated with oxCL, CL or
  • Hydro Heart cardiolipin (Biosearch Technologies, Inc, Ca, USA) 10pg/ml incubated overnight at 4°C. After five washings with PBS, the plates were blocked with 2% PBS-BSA for 2h at room temperature. Annexin A5 was added and incubated for 1 hour. After washing, bound annexin A5 was detected by incubating subsequently with rabbit anti-human annexin A5 polyclonal antibodies (Hyphen Biomed, Andresy, France) 1 :2000 and polyclonal goat anti-rabbit Immunoglobulins/AP (DakoCytomation) 1 :3000.
  • the reaction was developed with alkaline phosphatase substrate (Sigma), and optical density (OD) was read at 405nm with an ELISA Multiscan Plus spectrophotometer (Molecular Devices Emax, San Francisco). All samples were measured in duplicates and the coefficient of variation was below 15%.
  • OxLDL was incubated with Dil (Molecular Probes Eugene, Oregon, USA) in lipoprotein-deficient serum (Sigma) at 37°C for 15 hours, and then dialyzed against saline-EDTA buffer for 6 hours.
  • Uptake of Dil-labelled oxLDL was studied either by flow cytometry or fluorescence/confocal microscopy.
  • the macrophages (1 x 10 6 ) were grown overnight on culture slides (Nunc, Naperville, New York), for flow cytometry, the macrophages were plated in a 6-well plate at density of 1 x 10 6 cells/well in DMEM (Invitrogen, USA) containing 10% FBS overnight.
  • the cells were incubated with Dil-oxLDL (5pg/ml), with oxCL (40, 80pg/ml), with CL control (40, 80pg/ml), with unlabelled oxLDL (40 g/ml) or with unlabelled LDL (40pg/ml) for 4 hours.
  • Dil-oxLDL 5pg/ml
  • oxCL 40, 80pg/ml
  • CL control 40, 80pg/ml
  • unlabelled oxLDL 40 g/ml
  • LDL 40pg/ml
  • macrophages were incubated with Dil-ox-LDL (5ug/ml), with annexin A5 (0.01 , 0.04, 0.16, 0.64, 1 , 10, 20, 40 Mg/ml). Thereafter, the cells from above were washed 4 times with 0.2% BSA/PBS and once with PBS.
  • the cells were harvested in PBS containing 0.1% BSA and 0.01 % NaN3. Mean fluorescence intensity was measured by flow cytometry (BD Biosciences, San Jose, CA, USA). For each sample, fluorescence emission above 550nm was measured and a minimum of 10,000 cells were analyzed.
  • Human mononuclear cells were isolated from freshly prepared buffy coats (Karolinska Hospital blood bank, Sweden) by gradient centrifugation on Ficoll-Paque (Amersham Biosciences, Uppsala, Sweden). The mononuclear cells were cultured at a density of 5 ⁇ 10 6 /ml in RPMI-1640 medium with 25mM Hepes, 1% L-glutamine, 1% penicillin-streptomycin and 10% FBS. After 7 days, there were approximately 2 * 10 6 macrophages per well.
  • PMNs Human PMNs were isolated from freshly prepared buffy coats (Karolinska Hospital blood bank, Sweden) by dextran sedimentation, hypotonic lysis of erythrocytes and gradient centrifugation on Lymphoprep (Axis-Shield PoC AS, Oslo, Norway). PMNs were suspended at a density of 10 * 10 6 /ml in Dulbecco's PBS (Gibco (Invitrogen), Paisley, UK). PMN purity (> 95%) and viability (> 98%) was determined using Hemacolor (J.T. Baker, Utrecht, Holland) and Trypan Blue (Sigma Chemical Co.) staining, respectively.
  • Neutrophils were added into black, 96-well plates with transparent bottom (Corning Costar; 5 ⁇ 10 4 cells/well), and the plate was spun down at 120 ⁇ g for 3 min, afterwards changing the medium containing 4 ⁇ FURA-2AM (Fura-2 acetoxymethyl ester), or buffer as appropriate, and the cells were incubated for 30 min at 37°C and 5% CO 2 . Cells were washed four times with 50 ⁇ of a buffer solution (135 mM NaCI, 4.6 mM KCI, 1.2 mM MgCI 2 , 1.5 mM CaCI 2 , 11 mM glucose, 11 mM Hepes, pH 7.4) before a final 45 ⁇ volume of buffer was added to each well.
  • a buffer solution (135 mM NaCI, 4.6 mM KCI, 1.2 mM MgCI 2 , 1.5 mM CaCI 2 , 11 mM glucose, 11 mM Hepes, pH 7.4
  • the plates were transferred to a fluorometer (FluostarTM, BMG Technologies), and 50 ⁇ of different agonists according to experimental designs or buffer solution as control were injected into individual wells and the cells were monitored for the next 120 seconds. Control wells containing cells that had not been exposed to FURA- 2AM were used to subtract background auto-fluorescence. The results are given as the ratio of mean fluorescence intensity (MFI) between 340 and 380 nm, and normalized by control.
  • MFI mean fluorescence intensity
  • the transducer was held in the same position throughout the study by a mechanical arm.
  • a resting B-mode scan was recorded, flow velocity was measured with a 5 MHz pulsed Doppler with a 1.5 mm gate width in the centre of the vessel at a 70 degree angle.
  • Flow-increase was induced by the post ischemic response to deflation of a pneumatic tourniquet placed around the forearm of the patient to 250 mmHg for 4.5 min.
  • a second scan was recorded from 30 s before to 90 s after cuff deflation.
  • Flow velocity was recorded with a pulsed wave Doppler for 15 s before and 15 s after cuff deflation. This was followed by 10 minutes rest for vessel recovery.
  • a third scan at rest was taken and 0.40 mg sublingual nitro-glycerine spray was administered. Three to four minutes after nitro-glycerine, the last scan was recorded. The vessel was measured at a fixed point in all scans. Measures were made on digitalised cine- loops from the anterior wall leading edge of the intima-media echo to the leading edge of the far wall intima-media vessel interface incidentally with the R-wave on the ECG for three consecutive cardiac cycles and the average measurements were used. After reactive hyperemia, measurements were made 50-70 s after cuff deflation. The increase in vessel diameter during hyperemia and after nitro-glycerine administration is expressed as percentages of the first control scan.
  • the Immulon 1 B plate was coated with 50 ⁇ /well of oxCL 10 ⁇ g/ml, and allowed to dry overnight at 4°C. After washing with PBS, the plate was blocked with 2% BSA at room temperature for 2 hours. 1 :50 diluted sera were added in duplicates. The plate was incubated overnight at 4°C. The secondary antibody (anti-lg) was added 100 ⁇ /well, then left overnight at 4°C. After washing five times with PBS, substrate was added (100 ⁇ /well). The ELISA Multiscan Plus Spectrophotometer was used to determine optical density.
  • the PBMC were cultured in 96-well culture plates at density of 2 * 10 5 cells/1 ⁇ /well with stimulation of 1 pg/ml PHA in medium presence with different sera for 48 hours, and the supernatants were measured for TNF-a concentration by commercially available ELISA kits.
  • OxCL-MBSA and MBSA were coupled to a HiTrap NHS column (Amersham Biosciences) separately according to the manufacturer's instructions.
  • Human pooled immunoglobulin (IVIG; Gammaguard, S/D) was diluted in binding buffer (20mM Na 2 HPO 4 ) at 50mg/ml and filtered through 0.45 pm filter before passing through pre- coupled oxCL-MBSA and MBSA Sepharose gel column.
  • Anti-oxCL IgG was eluted by 0.1 M Glycine-HCI buffer. The purified fractions were desalted using PD-10 columns (Amersham Pharmacia Biotech AB). Binding to oxCL (as described for determination of anti-oxCL antibodies) was confirmed.
  • HUVECs human umbilical vascular endothelial cells
  • Cascade Biologies, Inc Portland, Ore
  • Cultures were maintained in EGMTM phenol red-free medium (Clonetics, San Diego, CA), containing 2% of fetal bovine serum and supplements, at 37°C under humidified 5% CO 2 conditions. All experiments were performed at passage 3 to 5.
  • HUVECs were seeded at 6x10 4 cells/2mL density on 6-well plates (NUNC Inc, Naperville, III).
  • oxidized cardiolipin 10pg/ml either with or without anti-oxCL-lgG 0.22 mg/ml. After 24 hours incubation, detached floating cells were washed away and cells were harvested into Falcon FACS tubes. After centrifuging at 1400 rpm for 5 minutes, cells were resuspended in 300 ⁇ FACS buffer (1% FBS-PBS), incubated with 10 ⁇ FITC-conjugated anti-Human CD106 (Becton, Dickinson) for 30 minutes on ice.
  • the vascular cell adhesion molecule (VCAM-1) CD106 was studied with flow cytometry analysis equipped with CellQuest software. For each sample, 10,000 events were analyzed. Brief description of the drawings
  • FIG. 1 Electrospray ionization mass MS spectrometer (Micromass, Beverly, MA) was used to demonstrate that bovine heart cardiolipin was oxidized.
  • Figure 2 Induction of adhesion molecules. Oxidized CL but not CL induces adhesion molecules in endothelial cells, flow cytometry.
  • Endothelial cells were incubated with oxidized CL (20pg/ml) or CL (20pg/ml) for 24 hours, oxidized CL induced ICAM-1 and VCAM-1 production but CL did not show the same effect.
  • Figure 3 Inhibition of oxCL-induced adhesion molecule expression by annexin A5, detected by flow cytometry.
  • Oxidized CL (20pg/ml) was preincubated with annexin A5 (10pg/ml) before stimulating cells for 24 hours.
  • Annexin A5 inhibited oxCL induced endothelial cell expression of ICAM-1 (CD54) and VCAM-1 (CD106).
  • Figure 4 Induction of IL-6 secretion by endothelial cells by oxCL and inhibition by annexin A5.
  • Endothelial cells were incubated with oxCL (20pg/ml) with or without annexin A5 (20 g/ml) and CL (20pg/ml) for 24 hours.
  • Oxidized CL can significantly induce IL-6 production and this effect can be inhibited by annexin A5.
  • CL had no such effect.
  • Figure 5 Binding of oxCL by annexin A5, ELISA. OxCL, CL and reduced CL (5pg/ml) were coated on ELISA plates overnight. Annexin A5 (0.32pg/ml, 0.64pg/ml, 1.28pg/ml, 2.56pg/ml, 5.12pg/ml and 10.24pg/ml) can bind to oxidized cardiolipin and air exposed CL but not reduced cardiolipin.
  • Figure 6 Effect of oxCL on uptake of oxLDL in macrophages, detected by flow cytometry. OxCL competed macrophage uptake of oxLDL, but CL did not have such an effect.
  • THP-1 cells (differentiated MQ), a model for monocyte/macrophage function, were studied for uptake of Dil-oxLDL, the uptake could be competed by oxCL but not non-oxidized CL.
  • Figure 7 OxCL-induced calcium mobilization, fluorometer.
  • Results show that only oxidized cardiolipin can activate the neutrophils and induce the intracellular calcium mobilization. Furthermore, annexin A5 can inhibit the oxCL induced calcium mobilization in neutrophils.
  • Oxidized cardiolipin promotes human neutrophils and macrophages to release leukotriene B 4 , but cardiolipin did not show the similar reaction.
  • Figure 9 Effect of annexin A5 on oxCL induced LTB 4 production from oxCL- stimulated human neutrophils and macrophages, which may give new insights into clinical novel targets for medical treatments of the associated inflammatory conditions, EIA.
  • FIG 10 Antibodies against oxCL were extracted by conjugating bovine serum albumin (BSA) with oxCL, whereupon the oxCL-BSA was loaded on a sepharose containing column and Ig was added. Anti-oxCL could then be extracted (IgM), and used in experiments, where 5 pg/ml of anti-oxCL was added 30 minutes before addition of oxCL. As indicated in Figure 10, anti-oxCL had the capacity to inhibit the proinflammatory effects of oxCL on expression of the adhesion molecule VCAM (CD106) as determined by flow cytometry, as seen by a shift to the left of the histogram.
  • Figure 11 Effects of oxCL on activation of T-cells, flow cytometry.
  • PBMC Human PBMC (Peripheral blood mononuclear leukocytes) were incubated overnight with 5 pg/ml of oxCL or CL. Both CD4 and CD8 positive T cells were studied. Quadrant Q2 represent (%) T-cells positive for CD69 expression and thus activated T-cells. Thus, it is demonstrated that oxCL but not CL can activate CD8-positive and CD4-positive T cells as determined by an increase in Q2 which is highly significant.
  • Figure 12 Protective effect of anti-oxCL on amyloid peptide 1-42 induced cell death. A Neuroblastoma cell line was treated with amyloid peptide1-42 which induced cell death (as determined by propidium iodide; PI).
  • OxPS is positively correlated with endothelial cell expression of VCAM-1.
  • HUVEC cells were stimulated with oxPS or oxPS that was preincubated with annexin A5 for 30 minutes.
  • VCAM- 1 (CD106) expression was measured by flow cytometry (BD Biosciences, San Jose, CA, USA). Relative to the control (curve marked with "control”), the curve shifts to the right (curve marked with "oxPS”), confirming that oxPS does induce VCAM-1 expression.
  • Figure 14 Effect of antibodies against phosphorylcholine (anti-PC) IgM antibodies on lysophosphatidylcholine (LPC) induced necrosis, expressed as fluorescent measure of leakage of lactate dehydrogenase from cells with a damaged membrane in PBMC. (300,000 cells/well, 96 well plates) were incubated with LPC for 2 h and LDH activity in supernatant was measured. Anti-PC or control antibodies which did not bind PC were preincubated with LPC 90 minutes before starting the treatment. The results were expressed as units per liter of LDH.
  • anti-PC phosphorylcholine
  • LPC lysophosphatidylcholine
  • Figure 15 Effect of human IgM antibodies on lysophosphatidylcholine (LPC) induced cytotoxicity, measured as decrease in mitochondrial dehydrogenase activity in PBMC. (3,000,000 cells/well) were incubated with LPC for 18 h and the mitochondrial dependent reduction of MTT to formazin was measured. Anti-PC, total IgM and IgM which did not bind PC (flow through) were preincubated with LPC 90 minutes before starting the treatment. The results were expressed as percentage of viable cells compared with non stimulated cells (control).
  • LPC lysophosphatidylcholine
  • Figure 16 Antibodies against oxidized phosphatidylserine (anti-oxPS) negatively associated with disease activity in SLE.
  • anti-oxPS oxidized phosphatidylserine
  • SLE activity was determined with the Systemic Lupus Activity Measure (SLAM) and also with Systemic Lupus Erythematosus diseases activity index (SLEDAI). Organ damage was determined with Systemic Lupus International Collaborating Clinics (SLICC) damage index.
  • Example 1 Chemical treatments of cardiolipin
  • Native cardiolipin and oxidation product were further analyzed by mass spectrometry, figure 1.
  • Native cardiolipin from bovine heart yielded two major signals, corresponding to the double charged anion (m/z 724.0), the single charged anion (m/z 1447).
  • the oxidized derivative peaks from both double charged and single charged ions in the oxidized fraction were 8 m/z units apart, suggesting progressively oxidized cardiolipins.
  • oxCL can significantly increase both ICAM-1 and VCAM-1 expression, but native CL did not show the same effect (Fig. 2). Both the increased expression of ICAM-1 and VCAM-1 (CD54 and CD106) induced by oxCL were significantly inhibited by annexin A5 (Fig. 3).
  • OxCL could stimulate endothelial cells to produce 564.3 ⁇ 142.02 (pg/ml) IL-6 from the supernatant.
  • Annexin A5 significantly decreased II-6 level to 276.4 ⁇ 28.62 (pg/ml).
  • both native CL and annexin A5 themselves could not significantly increase endothelial cell II-6 levels (Fig. 4). This supports the hypothesis that oxCL stimulates IL-6, which is a known inflammatory marker, e.g. oxCL might be involved in inducing/mediating inflammatory responses.
  • OxLDL could inhibit Dil-oxLDL uptake up to more than 60%, but LDL had almost no effect on the uptake of Dil- oxLDL, which confirmed the specificity of uptake results. Further results suggested oxCL could, in a concentration dependent manner, inhibit Dil-oxLDL uptake while the native CL did not show the competition effects (Fig. 6).
  • Example 7 Negative association between major parameters in systemic lupus erythematosus (SLE), inflammation and anti-oxCL
  • Both CD4 and CD8 positive T-cells from the cell line human PBMC were studied in an incubation setup illustrated in figure 11. Both T-cell types were incubated with buffer (control), oxCL or CL to determine the effect on activation, interpreted by the expression of CD69 surface molecule after stimulation.
  • This quadrant represents the percentage of T-cells in the population tested to be CD69 positive, e.g. activated by the stimuli prior to the data collection.
  • the beta amyloid (Abeta) and especially Abeta peptide (1-42) is an important component of senile plaques in Alzheimer's disease, and is known to be directly responsible for the production of free radicals toxic to brain tissue.
  • Abeta (1-42)- induced free radical oxidative stress in the neurodegeneration observed in AD brain may be one mechanism for neurotoxicity.
  • Human SH-SY5Y neuroblastoma cells were used in cell culture systems. The cells were treated with 5 ⁇ of the peptide for 24 hours. Cell death was determined by the addition of 1 mg/ml_ propidium iodide (PI), which labels the nucleus in dying cells which lack an intact plasma membrane.
  • PI propidium iodide
  • the amount of dead cells was measured as 23.99%, whereas the amyloid peptide 1-42 induced cell death increased that number to 45.54%.
  • amyloid peptide1-42 was incubated together with oxCL-lgG, 30pg/ml, (anti-oxCL), the cell death was reduced to 37.10%.
  • Example 10 Association between IgM anti-oxCL and anti-OxPS and risk of death in patients undergoing haemodialysis
  • IgM anti-oxCL IgM anti-oxCL
  • oxPS IgM anti-oxPS
  • RESULTS The patients with an anti-oxCL value below the median had a higher mortality rate (ROC curve value 60%, p ⁇ 0.05). The patients with anti-OxPS levels under median also had increased mortality rate (p ⁇ 0.05). These patients remained at higher risk of death even after adjustment for age, sex, smoking habits, cardiovascular (CVD) risk factors, albumin and inflammation.
  • CVD cardiovascular
  • OxPS and/or anti-oxPS in inflammatory conditions Materials and methods Chemical treatments of phosphatidylsehne
  • PS is stored at -20°C.
  • PS is oxidized in aqueous solutions containing tert-butylhydroperoxide and CuSO 4 .
  • Both PS and copper treated PS (oxidized product) are then measured with mass spectrometry (MS)-spectrophotometer to confirm that the phosphatidylserine has been oxidized by the copper and tert-butylhydroperoxide.
  • MS mass spectrometry
  • HUVECs human umbilical vascular endothelial cells
  • Cascade Biologies, Inc Portland, Ore
  • Cultures are maintained in EGMTM phenol red-free medium (Clonetics, San Diego, CA), containing 2% of fetal bovine serum and supplements, at 37°C under humidified 5% CO 2 conditions. All experiments are performed at passage 3 to 5.
  • HUVECs are seeded at 6x10 4 cells/2ml_ density on 6-well plates (NUNC Inc, Naperville, III). After allowing endothelial cells to settle and attach overnight, cells are ready for stimulation.
  • oxPS After washing with phosphate-buffered saline (PBS), oxPS is added.
  • annexin A5 from Bender MedSystems GmbH, Austria
  • oxPS is incubated with annexin A5 (10pg/ml) for half an hour before adding to cells.
  • anti-oxPS inhibition study oxPS is incubated with anti-oxPS (10, 15, 20, 25 or 30 pg/ml) for half an hour before adding to cells, or the cells are incubated with oxPS, PS or reduced PS for 24 hours, followed by stimulation with anti-oxPS (10, 15, 20, 25 or 30 pg/ml) for additional 12 hours.
  • intercellular adhesion molecule (ICAM-1) CD54 and the vascular cell adhesion molecule (VCAM-1) CD106 are studied with flow cytometry analysis equipped with CellQuest software. For each sample, 10,000 events are analyzed. Controls for anti-oxPS should be incubated additional 12 hours after the first 24 hours of incubation for proper comparison.
  • the macrophages which may be transformed from Human monocyte-derived THP- 1 cells (American Type Culture Collection, Manassas, VA, U.S.A.) are seeded in a 6-well plate at a density of 1 x 10 6 cells/well in DMEM (Invitrogen, USA) containing 10% fetal bovine serum (FBS) overnight. Then the cells are washed three times with serum free medium before being incubated with PS, reduced PS or oxPS (2pg/ml). Thereafter, the cells are washed 4 times with ice-cold 0.2% bovine serum albumin (BSA)/PBS and once with PBS. The cells are harvested in PBS containing 0.1% BSA and 0.01% NaN3.
  • BSA bovine serum albumin
  • human umbilical vein endothelial cells from the above setup are seeded at density 10 6 /2ml into 6 well plates. Cells are allowed to attach for 24 hours. OxPS (20pg/ml), with or without annexin A5 (20pg/ml) and reduced PS, are then added in combination or to separate wells and incubated for 24 hours. A combination comprising anti-oxPS may also be added. Cell supernatants are then collected and IL-6 production can be measured with protein multiplex immunoassay kits (from Bioscource, USA) and Bio- PleiTM system (BioRad, USA).
  • Induction of IL-6 by oxPS and inhibition by annexin A5 are determined by Luminex and detected by intensity of fluorescence.
  • Human mononuclear cells are isolated from freshly prepared buffy coats (Karolinska Hospital blood bank, Sweden) by gradient centrifugation on Ficoll-Paque (Amersham Biosciences, Uppsala, Sweden). The mononuclear cells are cultured at a density of 5 * 10 6 /ml in RPMI-1640 medium with 25mM Hepes, 1% L-glutamine, 1% penicillin-streptomycin and 10% FBS. After 7 days, approximately 2 ⁇ 10 6 macrophages per well may be expected.
  • PMNs Human PMNs are isolated from freshly prepared buffy coats (Karolinska Hospital blood bank, Sweden) by dextran sedimentation, hypotonic lysis of erythrocytes and gradient centrifugation on Lymphoprep (Axis-Shield PoC AS, Oslo, Norway). The PMNs are then suspended at a density of 10 x 10 6 /ml in Dulbecco's PBS (Gibco (Invitrogen), Paisley, UK). PMN purity and viability is determined using Hemacolor (J.T. Baker, Utrecht, Holland) and Trypan Blue (Sigma Chemical Co.) staining, respectively. Intracellular calcium mobilization
  • neutrophils are added into black, 96-well plates with transparent bottom (Corning Costar; 5 * 10 4 cells/well), and the plate is spun down at 120 * g for 3 min, afterwards changing the medium to 4 ⁇ Fura-2- acetoxymethyl ester (FURA-2AM) or buffer as appropriate.
  • the cells are then incubated for 30 min at 37°C and 5% CO 2 .
  • Cells are washed four times with 50 ⁇ of a buffer solution (135 mM NaCI, 4.6 mM KCI, 1.2 mM MgCI 2 , 1.5 mM CaCI 2 , 11 mM glucose, 11 mM Hepes, pH 7.4) before a final 45 pi volume of buffer is added to each well.
  • the plates are transferred to a fluorometer (FluostarTM, BMG Technologies), and 50 ⁇ of different agonists according to experimental designs or buffer solution as control are injected into individual wells and the cells are monitored for the next 120 seconds.
  • Control wells containing cells that have not been exposed to FURA-2AM are used to subtract background auto-fluorescence.
  • the results are measured as the ratio of mean fluorescence intensity (MFI) between 340 and 380 nm, and normalized by control. Determination of oxPS levels
  • oxPS may be conjugated with BSA, which acts as carrier protein.
  • Anti-oxPS is extracted by use of columns loaded with oxPS-BSA. Such antibodies against oxPS are generated and another monoclonal antibody against low density lipoprotein (LDL) is coated on ELISA plates.
  • monoclonal antibodies e.g. anti-oxPS
  • serum is added and monoclonal anti-oxPS is added again; thus, all oxPS containing particles are measured.
  • a fluid or sample subject for the oxPS determination such as serum, is added. After incubation and washing of the plate, bound oxPS is detected with an antibody against oxPS conjugated with a color detection marker.
  • Immulon 1 B plate is coated with 50 ⁇ /well of oxPS 10 ⁇ g/ml, and allowed to dry overnight at 4°C. After washing with PBS, the plate is blocked with 2% BSA at room temperature for 2 hours. 1 :50 diluted sera is added in duplicates. The plate is incubated overnight at 4°C. The secondary antibody (anti-lg) is added 100pl/well and incubated overnight at 4°C, followed by 5 rounds of washing with PBS before substrate is added ( ⁇ /well). ELISA Multiscan Plus Spectrophotometer can be used to determine optical density.
  • PBMCs Peripheral blood mononuclear cells
  • PHA polyhydroxyalkanoates
  • the cells are washed after the first 48 hours and stimulated an additional 12 hours with pure medium, anti-oxPS, annexin A5 or a combination thereof, before harvesting and evaluation by commercially available ELISA kits.
  • OxPS-mannosylated BSA (mBSA) or PS-mBSA and mBSA are coupled to a HiTrap N-hydroxysuccinimide (NHS) column (Amersham Biosciences) separately according to the manufacturer's instructions.
  • Human pooled immunoglobulin (IVIG; Gammaguard, S/D) is diluted in binding buffer (20mM Na 2 HP0 4 ) at 50mg/ml and filtered through 0.45pm filter before passing through pre-coupled oxPS-mBSA and mBSA Sepharose gel column.
  • Anti-oxPS IgG is eluted by 0.1 M Glycine-HCI buffer. The purified fractions are desalted using PD-10 columns (Amersham Pharmacia Biotech AB).
  • HUVECs human umbilical vascular endothelial cells
  • Cascade Biologies, Inc Portland, Ore
  • Cultures are maintained in EGMTM phenol red-free medium (Clonetics, San Diego, CA), containing 2% of fetal bovine serum and supplements, at 37°C under humidified 5% CO 2 conditions. All experiments are performed at passage 3 to 5.
  • HUVECs are seeded at 6x10 4 cells/2mL density on 6-well plates (NUNC Inc, Naperville, III).
  • the cells are incubated with oxidized phosphatidylserine (oxPS) 10 g/ml, either with or without anti-oxPS-lgG 0.22 mg/ml. After 24 hours incubation, detached floating cells are washed away and cells are harvested into Falcon FACS tubes. After centrifuging at 1400 rpm for 5 minutes, cells are resuspended in 300 ⁇ FACS buffer (1% FBS-PBS), incubated with 10 ⁇ FITC-conjugated anti-Human CD106 (Becton, Dickinson) for 30 minutes on ice. The vascular cell adhesion molecules VCAM-1 (CD106) and ICAM-1 (CD54) are then studied with flow cytometry analysis equipped with CellQuest software. For each sample, 10,000 cells should be analyzed.
  • oxPS oxidized phosphatidylserine
  • OxPS or conjugates containing epitopes of oxPS are used to immunize mouse models of atherosclerosis/chronic inflammation, such as, but not limited to, apoE knock out models, or other relevant models for chronic inflammatory diseases. Increased anti-oxPS levels after such immunization are then monitored.
  • Another method involving immunization of mouse models is, when monoclonal antibodies against oxPS are produced, and used in the abovementioned mouse models of atherosclerosis/chronic inflammation, such as, but not limited to, apoE knock out models, or other relevant models for chronic inflammatory diseases, to evaluate effects on inflammation.
  • Different oxPS dependent changes may then be determined, including vascular inflammation or inflammation in tissues such as kidney, or inflammation-related changes in brain, such as amyloid measurements or other dementia markers.
  • Example 11 Chemical treatments of PS; Electrospray ionization mass MS spectrometer (Micromass, Beverly, MA) is used to demonstrate that commercially available phosphatidylserine is oxidized.
  • Example 12 Anti-oxPS binds oxPS (and not PS)
  • Binding to oxPS (as described for determination of anti-oxPS antibodies) can be confirmed when performing an extraction of anti-oxPS IgG from intravenous immunoglobulin (IVIG) as described above. It is expected that the results show that anti-oxPS does not bind PS.
  • IVIG intravenous immunoglobulin
  • Example 13 Endothelial cells and adhesion molecules
  • Example 14 IL-6 production Endothelial cells can be incubated with oxPS (20pg/ml) with or without annexin A5, anti-oxPS (20pg/ml) and PS (20pg/ml) for 24 hours, oxidized PS is expected to significantly induce IL-6 production and this effect is moreover expected to be inhibited by annexin A5 and anti-oxPS. PS is not expected to show such effect.
  • both native PS and annexin A5 themselves should not be expected to significantly increase endothelial cell IL-6 levels.
  • OxLDL is expected to inhibit Dil- oxLDL uptake, but LDL should not have such an effect on the uptake of Dil-oxLDL then confirming the specificity of uptake results.
  • OxPS could in a concentration dependent manner inhibit Dil-oxLDL uptake while the native PS is expected not to show the competition effects.
  • OxPS is expected to compete with the macrophage uptake of oxLDL, but PS should not have such effect.
  • Example 16 Intracellular calcium mobilization
  • MFI mean fluorescence intensities
  • Example 17 Anti-oxPS is negatively correlated with endothelial cell expression of VCAM-1
  • Example 19 Inhibitors of oxPS and their effect on oxPS induced VCAM-1 expression on endothelial cells
  • HUVECs are seeded, incubated for settlement and attachment and stimulated with oxPS or PS for 24 hours, followed by 12 hours of incubation before harvest and evaluation by flow cytometry, with pure medium, anti-oxPS, annexin A5 or a combination thereof. Comparative data is expected to reveal that the inhibitors of oxPS decrease oxPS induced VCAM-1 expression on the HUVECs.
  • Inhibitors that bind oxPS, but not PS, are obviously expected to be part of the invention. Such inhibitors should have an inhibitory effect on oxPS activity similar to anti-oxPS and annexin A5, however, should not (as annexin A5) inhibit PS.
  • Example 21 Inhibitors of oxPS and their effect on oxPS induced TNF levels
  • PBMCs peripheral blood mononuclear cells
  • PHA Polyhydroxyalkanoates
  • TNF-a concentrations are measured by commercially available ELISA kits. Comparative data is expected to reveal that the inhibitors of oxPS decrease oxPS induced TNF levels in PBMCs.
  • OxPS or conjugates containing epitopes of oxPS are used to immunize mouse models of atherosclerosis/chronic inflammation, such as, but not limited to, apoE knock out models, or other relevant models for chronic inflammatory diseases. Increased anti-oxPS levels after such immunization are monitored.
  • Example 23 Inhibition of lysophosphatidylcholine induced cell death by anti- PC IgM
  • PBMC Peripheral blood mononuclear cells
  • LPC lysophosphatidylcholine
  • LDC lactate dehydrogenase
  • Antibodies against phosphorylcholine (anti-PC IgM) or control antibodies which did not bind PC were preincubated with LPC 90 minutes before starting the treatment. The results were expressed as units per liter of LDH. As can be seen from Figure 14, anti-PC IgM reduced cell death more than total IgM and flowthrough IgM.
  • Serum samples were obtained from 226 subjects with established hypertension (diastolic pressure >95 mm Hg) prior to their entry into the Swedish component of the European Lacidipine Study on Atherosclerosis (ELSA) 58, 59 Samples were collected following a 4-week washout period without medication to minimize the effects of treatment on the measured parameters. Blood pressure, cholesterol and triglyceride levels were determined as described previously 58, 59 . One hundred and fifteen of the subjects were subsequently assigned to treatment with the ⁇ -blocker atenolol, and 111 of the subjects were assigned to treatment with the calcium antagonist lacidipine. Carotid ultrasound determinations were performed and analyzed as detailed elsewhere 58"60 .
  • IgM antibodies to PC-BSA were determined by enzyme-linked immunosorbent assay (ELISA). Pooled serum from the same donors was used as an internal standard and tested on every plate. The plateau of antibody binding was reached with the antigen concentration of 10pg/ml. F96 microtiter polysorb plate was therefore coated with PC-BSA(10pg/ml) 50pl/well in PBS. Coated plates were incubated overnight at 4°C. After five washings with PBS, the plates were blocked with 2% BSA-PBS for 2h at room temperature and washed as described above.
  • ELISA enzyme-linked immunosorbent assay
  • Serum samples were diluted (1 :30) in 0.2% BSA-PBS and added at 50pl/well.
  • IgM antibodies to anti-oxCL and anti-oxPS were determined by ELISA as described.
  • Cardiolipin and phosphatidylserine were oxidized in aqueous solutions containing 1.5 mmol/L tert-butylhydroperoxide and 20 pmol/L CuSO4.
  • LDL was isolated from plasma of healthy donors by sequential preparative ultra- centrifugation and oxidized by use of copper ions (OxLDL) or derivatized with MDA (MDA-LDL) as described 61 .
  • OxLDL and MDA-LDL were determined by ELISA essentially as described 61 .
  • OxLDL or MDA-LDL was diluted to 2 pg/ml in coating buffer (carbonate-bicarbonate buffer 50 mM pH 9.7), and 100 ⁇ /well was used to coat ELISA plates (Costar 2581). The plates were kept at 4°C overnight, washed 4 times with PBS, and then blocked with 20% adult bovine serum in PBS (20% ABS- PBS) for 2 hours in room temperature. They were then incubated with 100 ⁇ serum, diluted 1 :30 in 20% ABS-PBS at 4°C overnight.
  • Model 1 indicates exposure to high levels of any of the antibodies
  • Model 2 exposure to having high levels of both antibodies.
  • PC phosphorylcholine
  • MDA-LDL MDA-LDL
  • Anti-MDA-LDL .67 .36 1.2 .18 (highest 25 th
  • Anti-PC (highest 10 th .36 .15 0.87 .024 percentile)
  • Table 8 Prediction of changes in IMT with baseline levels of IgM autoantibodies to phosphorylcholine (PC) or Ox-LDL.
  • Example 25 Increased risk of CVD from combination of low IgM anti-PC with low anti-oxCL or low anti-oxPS
  • Example 26 Association between low levels of IgM anti-oxPS and risk for myocardial infarction and stroke (CVD) among 60-year olds in Sweden county, followed for 5 years. Study of 60-year olds
  • IgM anti-oxPS is a novel protection marker for CVD. Active immunization or passive immunization to raise anti-oxPS levels could be a novel therapy against chronic inflammation including CVD and atherosclerosis.
  • Table 9 Association between low levels of IgM anti-oxPS and risk for myocardial infarction and stroke (CVD) among 60-year olds in Sweden county, followed for 5 years.
  • Serum samples were obtained from 226 subjects with established hypertension (diastolic pressure >95 mm Hg) prior to their entry into the Swedish component of the European Lacidipine Study on Atherosclerosis (ELSA) 58, 59 Samples were collected following a 4-week washout period without medication to minimize the effects of treatment on the measured parameters. Blood pressure, cholesterol and triglyceride levels were determined as described previously 58, 59 One hundred and fifteen of the subjects were subsequently assigned to treatment with the ⁇ -blocker atenolol, and 111 of the subjects were assigned to treatment with the calcium antagonist lacidipine. Carotid ultrasound determinations were performed and analysed as detailed elsewhere 58"60 .
  • IgM antibodies to anti-oxCL and anti-oxPS were determined by ELISA as described.
  • Cardiolipin and phosphatidylserine were oxidized in aqueous solutions containing 1.5 mmol/L tert-butylhydroperoxide and CuS0 4 in 20 ⁇ /L LDL was isolated from plasma of healthy donors by sequential preparative ultra-centrifugation and derivatized with MDA (MDA-LDL) as described 61 .
  • MDA-LDL was diluted to 2 g/ml in coating buffer (carbonate-bicarbonate buffer 50 mM pH 9.7), and 100 ⁇ /well was used to coat ELISA plates (Costar 2581). The plates were kept at 4°C overnight, washed 4 times with PBS, and then blocked with 20% adult bovine serum in PBS (20% ABS-PBS) for 2 hours in room temperature. They were then incubated with 100 ⁇ serum, diluted 1 :30 in 20% ABS-PBS at 4°C overnight.
  • coating buffer carbonate-bicarbonate buffer 50 mM pH 9.7
  • Anti-oxPS is a protection marker for atherosclerosis development, where high levels, above 75 th , 80 th and 90 th percentile, were associated with a favourable atherosclerosis development (p ⁇ 0.05).
  • Anti-oxPS is a protection factor against chronic inflammatory diseases such as atherosclerosis, suggesting that anti-oxPS can be raised through active immunization (vaccination) or passive immunization (where antibodies are administered), as a novel therapy.
  • Table 12 Basic characteristics of the study group at enrolment. Results are presented as means (SD) or percentage (%) and mg/dL for lipids.
  • Table 13 Prediction of increase in atherosclerosis measurements during 5 years. High anti-oxPS levels predict decreased risk.
  • Example 28 Antibodies against oxidized phosphatidylserine (anti-oxPS) negatively associated with vulnerable plaque in SLE Introduction
  • CVD cardiovascular disease
  • SLE systemic lupus erythematosus
  • anti-PC phosphorylcholine
  • Potential mechanisms include anti-inflammatory properties and inhibition of uptake of oxidized low density lipoprotein in macrophages.
  • the objective herein was to study atherosclerosis in SLE in detail and in relation to traditional and non-traditional risk factors including novel factors antibodies against oxidized phosphatidylserine (anti-oxPS) and antibodies against oxidized cardiolipin (anti-oxCL).
  • IMT carotid intima- media thickness
  • cIMa calculated intima-media area
  • plaque occurrence were determined by B-mode ultrasound as a surrogate measure of atherosclerosis.
  • Plaques were graded according to echogenicity and grouped as 1-4, with 1 being echolucent, and considered most vulnerable.
  • Antibodies were studied by ELISA.
  • Plaque occurrence in the carotid arteries is increased in SLE and independently associated with age, LDL and low anti-PC levels. Vulnerable plaques were more common in SLE and associated independent of age, negatively with anti-oxPS and anti-oxCL. Anti-oxPS could be a novel risk marker also with a therapeutic potential in SLE, by itself or in combination with anti-oxCL and anti-PC.
  • Example 29 Antibodies against oxidized phosphatidylserine (anti-oxPS) negatively associated with disease activity in SLE
  • CVD cardiovascular disease
  • SLE systemic lupus erythematosus
  • the objective herein was to study disease manifestations SLE in detail and in relation to traditional and non-traditional risk factors including novel factors antibodies against oxidized phosphatidylserine (anti-oxPS) and antibodies against oxidized cardiolipin (anti-oxCL) and also antibodies against phosphorylcholine (anti-PC).
  • SLE activity was determined with the Systemic Lupus Activity Measure (SLAM) and also with Systemic Lupus Erythematosus diseases activity index (SLEDAI).
  • SLEDAI Systemic Lupus Erythematosus diseases activity index
  • Organ damage was determined with Systemic Lupus International Collaborating Clinics (SLICC) damage index.
  • Antibodies were determined by ELISA.
  • Anti-oxPS was negatively associated with SLAM >6; a sign of high disease activity.
  • Anti-oxPS is associated with disease activity in SLE, negatively. Anti-oxPS could be a novel risk marker also with a therapeutic potential in SLE.
  • Example 30 Anti-oxPS negatively associated with TNF in serum in SLE
  • TNF oxidized phosphatidylserine
  • Anti-oxPS is negatively associated with the important proinflammatory cytokine TNF, implying it is anti-inflammatory.
  • Example 31 Treatment with anti-oxCL and anti-oxPS to prevent radiation damage
  • a major problem is radiation side effects, causing serious damage to tissues like lungs, Gl-tract and endothelium.
  • oxidized cardiolipin and oxidized phosphatidylserine oxCL and oxPS
  • oxCL and oxPS oxidized cardiolipin and oxidized phosphatidylserine
  • oxCL and oxPS oxidized cardiolipin and oxidized phosphatidylserine
  • Our previous results indicate anti-inflammatory properties in antibodies against oxCL and oxPS (anti-oxCL and anti-oxPS) and also that annexin A5 inhibits the inflammatory effects of oxCL and oxPS.
  • the following experiment can illustrate treatment with anti-oxCL and anti-oxPS to prevent radiation damage.
  • mice Three groups of mice are subjected to total-body irradiation (5-15 Gy (or other relevant doses)) and euthanized after 24 h.
  • One group is pre-treated with anti-oxCL and anti-oxPS, one group is pre-treated with anti-oxCL and anti-oxPS and annexin A5, and one group is untreated control.
  • Mouse lung endothelial cells and GIT tissue can be analyzed 48 h after gamma radiation and apoptosis and inflammatory changes detected.
  • Example 32 Topical skin care composition
  • Leukotrienes enhance the bactericidal activity of alveolar macrophages against Klebsiella pneumoniae through the activation of NADPH oxidase. Blood. 2005; 106: 1067-75.
  • Antiphospholipid antibodies are directed against epitopes of oxidized phospholipids. Recognition of cardiolipin by monoclonal antibodies to epitopes of oxidized low density lipoprotein. J Clin Invest. 1996;98:815-25.
  • Annexin A5 as a novel player in prevention of atherothrombosis in SLE and in the general population, Cederholm A. and Frostegard J., 01.06.2007 Annals of the New York Academy of Science, 1108, 96-103
  • Milk fat globule epidermal growth factor 8 (MFG-E8) binds to oxidized phosphatidylserine: implications for macrophage clearance of apoptotic cells, August 2004 Cell Death and differentiation, 11 , 8, 943-945).
  • Zanchetti A Bond MG, Hennig M, Neiss A, Mancia G, Dal Palu C, Hansson L, Magnani B, Rahn KH, Reid J, Rodicio J, Safar M, Eckes L, Ravinetto R.
  • Zanchetti A Bond MG, Hennig M, Neiss A, Mancia G, Dal Palu C, Hansson L, Magnani B, Rahn KH, Reid JL, Rodicio J, Safar M, Eckes L, Rizzini P.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biophysics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Rheumatology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Cardiology (AREA)
  • Genetics & Genomics (AREA)
  • Pain & Pain Management (AREA)
  • Zoology (AREA)

Abstract

Cette invention concerne un procédé permettant d'évaluer le risque de développer des états inflammatoires, et/ou le risque de mortalité associé à ceux-ci, comprenant la surveillance et/ou la détermination des taux d'un ou de plusieurs phospholipides et lipoprotéines oxydés, et des anticorps dirigés contre eux, et la comparaison desdits taux à des valeurs de seuil prédéterminées, ainsi qu'un kit pour la mise en œuvre de ladite méthode. Une méthode permettant de traiter ou de réduire le risque de développer des états inflammatoires, et/ou le risque de mortalité associé à ceux-ci, par administration de médicaments comprenant une annexine A5 et/ou des anticorps dirigés contre un ou plusieurs phospholipides et lipoprotéines oxydés, ainsi que des composants bioactifs et/ou des parties/fragments de ceux-ci est également décrite. Cette invention concerne également des compositions comprenant un ou plusieurs inhibiteurs de phospholipides et de lipoprotéines oxydés, ainsi que des composants bioactifs et/ou des parties/fragments de ceux-ci ; et des compositions comprenant un ou plusieurs conjugués de phospholipides et de lipoprotéines oxydés, ainsi que des composants bioactifs et/ou des parties/fragments de ceux-ci.
PCT/EP2011/003110 2010-06-24 2011-06-24 Phospholipides et lipoprotéines oxydés, et anticorps dirigés contre eux, à titre de biomarqueurs des états inflammatoires et méthodes de traitement WO2011160845A2 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US35810210P 2010-06-24 2010-06-24
US61/358,102 2010-06-24
US37344110P 2010-08-13 2010-08-13
US61/373,441 2010-08-13
US201161449276P 2011-03-04 2011-03-04
US61/449,276 2011-03-04

Publications (2)

Publication Number Publication Date
WO2011160845A2 true WO2011160845A2 (fr) 2011-12-29
WO2011160845A3 WO2011160845A3 (fr) 2012-03-15

Family

ID=44358157

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2011/003110 WO2011160845A2 (fr) 2010-06-24 2011-06-24 Phospholipides et lipoprotéines oxydés, et anticorps dirigés contre eux, à titre de biomarqueurs des états inflammatoires et méthodes de traitement

Country Status (1)

Country Link
WO (1) WO2011160845A2 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10060908B2 (en) 2008-12-19 2018-08-28 Medirista Biotechnologies Ab Oxidized cardiolipin as a pro-inflammatory factor
WO2018220224A1 (fr) * 2017-06-02 2018-12-06 Medirista Biotechnologies Ab Antigènes associés aux lipides et anticorps contre eux
JP2020506206A (ja) * 2017-02-09 2020-02-27 アネキシン ファーマシューティカルズ アーベー 治療用組成物
CN112041682A (zh) * 2018-04-19 2020-12-04 国立大学法人东京大学 用于辅助对象中的疾病的诊断的方法及套件
WO2021097146A1 (fr) * 2019-11-12 2021-05-20 Abcentra, Llc Méthodes et compositions pour le traitement du cancer
EP3746478A4 (fr) * 2018-01-29 2022-01-19 The Regents of The University of California Thérapies et méthodes pour traiter des maladies et des troubles médiés par tlr2
WO2023057588A1 (fr) * 2021-10-06 2023-04-13 Iltoo Pharma Constructions chimériques d'interleukine 2 ont une spécificité de ciblage vis-à-vis des tissus enflammés

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050222030A1 (en) 2001-02-21 2005-10-06 Anthony Allison Modified annexin proteins and methods for preventing thrombosis
WO2005099744A1 (fr) 2004-04-15 2005-10-27 Athera Biotechnologies Ab Utilisation de l'annexine v pour la prevention de l'atherothrombose et de la rupture de la plaque
WO2009007764A2 (fr) 2007-07-10 2009-01-15 Egis Gyógyszergyár Nyilvánosan Müködö Préparations pharmaceutiques contenant des silicones hautement volatiles
WO2009103977A1 (fr) 2008-02-22 2009-08-27 Athera Biotechnologies Ab Composés et procédés pour prévention ou le traitement de la resténose
WO2010004550A2 (fr) 2008-07-06 2010-01-14 Lamos Inc. Structure thermoélectrique à deux éléments ainsi que dispositifs et systèmes utilisant ladite structure
WO2010043045A1 (fr) 2008-10-17 2010-04-22 London Health Sciences Centre Research Inc. Annexine et son utilisation pour le traitement de troubles inflammatoires

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2009328507A1 (en) * 2008-12-19 2011-07-14 Medirista Biotechnologies Ab Oxidized cardiolipin as a novel pro-inflammatory factor

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050222030A1 (en) 2001-02-21 2005-10-06 Anthony Allison Modified annexin proteins and methods for preventing thrombosis
WO2005099744A1 (fr) 2004-04-15 2005-10-27 Athera Biotechnologies Ab Utilisation de l'annexine v pour la prevention de l'atherothrombose et de la rupture de la plaque
WO2009007764A2 (fr) 2007-07-10 2009-01-15 Egis Gyógyszergyár Nyilvánosan Müködö Préparations pharmaceutiques contenant des silicones hautement volatiles
WO2009103977A1 (fr) 2008-02-22 2009-08-27 Athera Biotechnologies Ab Composés et procédés pour prévention ou le traitement de la resténose
WO2010004550A2 (fr) 2008-07-06 2010-01-14 Lamos Inc. Structure thermoélectrique à deux éléments ainsi que dispositifs et systèmes utilisant ladite structure
WO2010043045A1 (fr) 2008-10-17 2010-04-22 London Health Sciences Centre Research Inc. Annexine et son utilisation pour le traitement de troubles inflammatoires

Non-Patent Citations (64)

* Cited by examiner, † Cited by third party
Title
"Milk fat globule epidermal growth factor 8 (MFG-E8) binds to oxidized phosphatidylserine: implications for macrophage clearance of apoptotic cell", CELL DEATH AND DIFFERENTIATION, vol. 11, no. 8, August 2004 (2004-08-01), pages 943 - 945
ALBANO E.: "Oxidative mechanism in the pathogenesis of alcoholic liver disease", MOLECULAR ASPECTS OF MEDICINE, vol. 29, 1 February 2008 (2008-02-01), pages 1 - 9,9
ALLON M., RADEVA M., BAILEY J, BEDDHU S., BUTTERLY D., COYNE D.W., DEPNER7, JENNIFER T.A., GASSMAN J., KAUFMAN A. M.: "for the HEMO Study Group, The spectrum of infection-related morbidity in hospitalized haemodialysis patients", NEPHROL. DIAL. TRANSPLANT., vol. 20, 2005, pages 1180 - 1186
BASOVA LV, KURNIKOV IV, WANG L, RITOV VB, BELIKOVA NA, VLASOVA, , PACHECO AA, WINNICA DE, PETERSON J, BAYIR H, WALDECK DH: "Cardiolipin switch in mitochondria: shutting off the reduction of cytochrome c and turning on the peroxidase activity", BIOCHEMISTRY, vol. 46, 2007, pages 3423 - 34
BELIKOVA NA, JIANG J, TYURINA YY, ZHAO Q, EPPERLY MW, GREENBERGER J, KAGAN VE: "Cardiolipin-specific peroxidase reactions of cytochrome C in mitochondria during irradiation-induced apoptosis", INT J RADIAT ONCOL BIOL PHYS., vol. 69, 2007, pages 176 - 86, XP022201156, DOI: doi:10.1016/j.ijrobp.2007.03.043
CARLSSON AC, WANDELL PE, DE FAIRE U, HELLENIUS ML: "Risk factors associated with newly diagnosed high blood pressure in men and women", AM J HYPERTENS, vol. 21, no. 7, 2008, pages 771 - 777
CEDERHOLM A, FROSTEGARD J.: "Annexin A5 in cardiovascular disease and systemic lupus erythematosus", IMMUNOBIOLOGY, vol. 210, 2005, pages 761 - 8
CEDERHOLM A, SVENUNGSSON E, JENSEN-URSTAD K, TROLLMO C, ULFGREN AK, SWEDENBORG J, FEI GZ, FROSTEGARD J: "Decreased binding of annexin v to endothelial cells: a potential mechanism in atherothrombosis of patients with systemic lupus erythematosus", ARTERIOSCLER THROMB VASC BIOL., vol. 25, 2005, pages 198 - 203, XP009051391
CEDERHOLM A., FROSTEGARD J.: "Annexin A5 as a novel player in prevention of atherothrombosis in SLE and in the general population", ANNALS OF THE NEW YORK ACADEMY OF SCIENCE, vol. 1108, 1 June 2007 (2007-06-01), pages 96 - 103
CHICCO AJ, SPARAGNA GC.: "Role of cardiolipin alterations in mitochondrial dysfunction and disease", AM J PHYSIOL CELL PHYSIOL., vol. 292, 2007, pages C33 - 44
COTRAN R.S.: "Textbook of Rheumatology", 1993, SAUNDERS, article "Endothelial cells", pages: 327 - 336
DE FAIRE U, SU J, HUA X, FROSTEGARD A, HALLDIN M, HELLENIUS ML, WIKSTROM M, DAHLBOM I, GRONLUND H, FROSTEGARD J: "Low levels of IgM antibodies to phosphorylcholine predict cardiovascular disease in 60-year old men: effects on uptake of oxidized LDL in macrophages as a potential mechanism", J AUTOIMMUN., vol. 34, no. 2, 2010, pages 73 - 79, XP026886085, DOI: doi:10.1016/j.jaut.2009.05.003
DEGUCHI H, FERNANDEZ JA, HACKENG TM, BANKA CL, GRIFFIN JH: "Cardiolipin is a normal component of human plasma lipoproteins", PROC NATL ACAD SCI U S A., vol. 97, 2000, pages 1743 - 8
EGGIMAN P., PITTET D., INFECTION IN THE ICU CHEST., vol. 120, 2001, pages 2059 - 2059
FEBBRAIO M, PODREZ EA, SMITH JD, HAJJAR DP, HAZEN SL, HOFF HF, SHARMA K, SILVERSTEIN RL: "Targeted disruption of the class B scavenger receptor CD36 protects against atherosclerotic lesion development in mice", J CLIN INVEST., vol. 105, 2000, pages 1049 - 56, XP002487573, DOI: doi:10.1172/JCI9259
FROSTEGARD J, SVENUNGSSON E, WU R, GUNNARSSON, LUNDBERG IE, KLARESKOG L, HORKKO S, WITZTUM JL: "Lipid peroxidation is enhanced in patients with systemic lupus erythematosus and is associated with arterial and renal disease manifestations", ARTHRITIS RHEUM., vol. 52, 2005, pages 192 - 200, XP002573122, DOI: doi:10.1002/art.20780
FROSTEGARD J, ULFGREN AK, NYBERG P, HEDIN U, SWEDENBORG J, ANDERSSON U, HANSSON GK: "Cytokine expression in advanced human atherosclerotic plaques: dominance of pro-inflammatory (Th1) and macrophage-stimulating cytokines", ATHEROSCLEROSIS, vol. 145, 1999, pages 33 - 43, XP000994998, DOI: doi:10.1016/S0021-9150(99)00011-8
FROSTEGARD J.: "Atherosclerosis in patients with autoimmune disorders", ARTERIOSCLER THROMB VASC BIOL., vol. 25, 2005, pages 1776 - 85, XP055025588, DOI: doi:10.1161/01.ATV.0000174800.78362.ec
FROSTEGARD. J.: "Atherosclerosis in Patients with Autoimmune Disorders", ARTHEROSCLER. THROMB. BIOL., 23 June 2005 (2005-06-23)
FUNK CD: "Leukotriene modifiers as potential therapeutics for cardiovascular disease", NAT REV DRUG DISCOV., vol. 4, 2005, pages 664 - 72
GONZALVEZ F, GOTTLIEB E.: "Cardiolipin: setting the beat of apoptosis", APOPTOSIS, vol. 12, 2007, pages 877 - 85, XP019497513, DOI: doi:10.1007/s10495-007-0718-8
GOODARZI K, GOODARZI M, TAGER AM, LUSTER AD, VON ANDRIAN UH: "Leukotriene B4 and BLT1 control cytotoxic effector T cell recruitment to inflamed tissues", NAT IMMUNOL., vol. 4, 2003, pages 965 - 73
GREENBERG M.E, SUN M., ZHANG R., FEBBRAIO M., SILVERSTEIN R., HAZEN S.L.: "Oxidized phosphatidylserine-CD36 interactions play an essential role in macrophage-dependent phagocytosis of apoptotic cells", JEM HOME, vol. 203, no. 12, November 2006 (2006-11-01), pages 2613
HALLDIN M, ROSELL M, DE FAIRE U, HELLENIUS ML: "The metabolic syndrome: prevalence and association to leisure-time and work-related physical activity in 60-year-old men and women", NUTR METAB CARDIOVASC DIS., vol. 17, no. 5, 2007, pages 349 - 357, XP022127003, DOI: doi:10.1016/j.numecd.2006.01.002
HAMSTEN A, NORBERG R, BJORKHOLM M, DE FAIRE U, HOLM G.: "Antibodies to cardiolipin in young survivors of myocardial infarction: an association with recurrent cardiovascular events", LANCET, vol. 1, 1986, pages 113 - 6
HAN X, YANG J, YANG K, ZHAO Z, ABENDSCHEIN DR, GROSS RW: "Alterations in myocardial cardiolipin content and composition occur at the very earliest stages of diabetes: a shotgun lipidomics study", BIOCHEMISTRY, vol. 46, 2007, pages 6417 - 28
HANSSON GK.: "Inflammation, atherosclerosis, and coronary artery disease", N ENGL J MED., vol. 352, 2005, pages 1685 - 95, XP003014526, DOI: doi:10.1056/NEJMra043430
HORKKO S, MILLER E, DUDL E, REAVEN P, CURTISS LK, ZVAIFLER NJ, TERKELTAUB R, PIERANGELI SS, BRANCH DW, PALINSKI W: "Antiphospholipid antibodies are directed against epitopes of oxidized phospholipids. Recognition of cardiolipin by monoclonal antibodies to epitopes of oxidized low density lipoprotein", J CLIN INVEST., vol. 98, 1996, pages 815 - 25
ISHII H, HIRAOKA M, TANAKA A, SHIMOKADO K, YOSHIDA M: "Recombinant Annexin-2 inhibits the progress of diabetic nephropathy in a diabetic mouse model via recovery of hypercoagulability", THROMB HAEMOST., vol. 97, 2007, pages 124 - 8
KADL A., BOCHKOV VALERY N., HUBER J., LEITINGER N.: "Apoptotic cells as sources for biologically active oxidized phospholipids", ANTIOXIDANTS & REDOX SIGNALING, vol. 6, no. 2, April 2004 (2004-04-01), pages 311 - 320
KAGAN V.E., BORISENKO G.G., TYURINA Y.Y., TYURIN V.A., JIANG J., LIU S.X., SHVEDOVA A.A., FABIASK J.P, UTHAISANG W., FADEEL B.: "Appetizing rancidity of apoptotic cells for macrophages: oxidation, externalization, and recognition of phosphatidylserine", AM. J. PHYSIOL. LUNG CELL PHYSIOL., vol. 285, 2003, pages L1 - L17
MARTIN W, HOFFMEISTER M, ROTTE C, HENZE K: "An overview of endosymbiotic models for the origins of eukaryotes, their ATP-producing organelles (mitochondria and hydrogenosomes), and their heterotrophic lifestyle", BIOL CHEM., vol. 382, 2001, pages 1521 - 39
MOORE KJ, FREEMAN MW: "Scavenger receptors in atherosclerosis: beyond lipid uptake", ARTERIOSCLER THROMB VASC BIOL., vol. 26, 2006, pages 1702 - 11, XP002487576, DOI: doi:10.1161/01.ATV.0000229218.97976.43
NAKAGAWA Y.: "Initiation of apoptotic signal by the peroxidation of cardiolipin of mitochondria", ANN N Y ACAD SCI., vol. 1011, 2004, pages 177 - 84
PETER W.F. ET AL.: "Metabolic Syndrome as a Precursor of Cardiovascular Disease and Type 2 Diabetes Mellitus, Epidemiology", CIRCULATION, vol. 112, 2005, pages 3066 - 3072
POBER J.S., COTRAN R.S.: "Cytokines and endothelial cell biology", PHYSIOL. REV., vol. 70, 1990, pages 427 - 434
QIU H, GABRIELSEN A, AGARDH HE, WAN M, WETTERHOLM A, WONG CH, HEDIN U, SWEDENBORG J, HANSSON GK, SAMUELSSON B: "Expression of 5-lipoxygenase and leukotriene A4 hydrolase in human atherosclerotic lesions correlates with symptoms of plaque instability", PROC NATL ACAD SCI U S A., vol. 103, 2006, pages 8161 - 6
RAND JH, WU XX: "Antibody-mediated disruption of the annexin-V antithrombotic shield: a new mechanism for thrombosis in the antiphospholipid syndrome", THROMB HAEMOST., vol. 82, 1999, pages 649 - 55
SCHLAME M, RUA D, GREENBERG ML: "The biosynthesis and functional role of cardiolipin", PROG LIPID RES., vol. 39, 2000, pages 257 - 88
SCHLAME M, TOWBIN JA, HEERDT PM, JEHLE R, DIMAURO S, BLANCK TJ: "Deficiency of tetralinoleoyl-cardiolipin in Barth syndrome", ANN NEUROL., vol. 51, 2002, pages 634 - 7
SCHLAME M.: "Cardiolipin synthesis for the assembly of bacterial and mitochondrial membranes", J LIPID RES., vol. 49, 2008, pages 1607 - 20
SEMIN DIAL., vol. 16, no. 2, March 2003 (2003-03-01), pages 85 - 94
SEREZANI CH, ARONOFF DM, JANCAR S, MANCUSO P, PETERS-GOLDEN M: "Leukotrienes enhance the bactericidal activity of alveolar macrophages against Klebsiella pneumoniae through the activation of NADPH oxidase", BLOOD, vol. 106, 2005, pages 1067 - 75
SEVENOAKS M.J, STOCKLEY R.A.: "Chronic obstructive Pulmonary Disease, inflammation and co-morbidity- a common inflammation phenotype?", RESPIRATORY RESEARCH, vol. 7, 2006, pages 70, XP021013349, DOI: doi:10.1186/1465-9921-7-70
SIMMONS ET AL., KIDNEY INT., vol. 65, no. 4, April 2004 (2004-04-01), pages 1357 - 65
SJOBERG BG., ATHEROSCLEROSIS., 2008
SPARAGNA GC, CHICCO AJ, MURPHY RC, BRISTOW MR, JOHNSON CA, REES ML, MAXEY ML, MCCUNE SA, MOORE RL: "Loss of cardiac tetralinoleoyl cardiolipin in human and experimental heart failure", J LIPID RES., vol. 48, 2007, pages 1559 - 70
SU J, GEORGIADES A, WU R, THULIN T, DE FAIRE U, FROSTEGARD J: "Antibodies of IgM subclass to phosphorylcholine and oxidized LDL are protective factors for atherosclerosis in patients with hypertension", ATHEROSCLEROSIS, vol. 188, no. 1, 2006, pages 160 - 166, XP028071487, DOI: doi:10.1016/j.atherosclerosis.2005.10.017
SUN J., BIRD P., SALEM H.H.: "Interaction of Annexin Vand platelets: Effects on platelet function and protein S binding", THROMBOSIS RESEARCH, vol. 69, 1993, pages 289 - 296
THIAGARAJAN P, BENEDICT CR: "Inhibition of arterial thrombosis by recombinant annexin V in a rabbit carotid artery injury model", CIRCULATION, vol. 96, 1997, pages 2339 - 47, XP002338645
TROF RJ ET AL., SCHOCK, vol. 26, no. 3, September 2006 (2006-09-01), pages 245 - 53
TUOMINEN A, MILLER YI, HANSEN LF, KESANIEMI YA, WITZTUM JL, HORKKO S: "A natural antibody to oxidized cardiolipin binds to oxidized low-density lipoprotein, apoptotic cells, and atherosclerotic lesions", ARTERIOSCLER THROMB VASC BIOL., vol. 26, 2006, pages 2096 - 102, XP002573119, DOI: doi:10.1161/01.ATV.0000233333.07991.4a
TYURINA YY, TYURIN VA, KAPRALOVA VI, WASSERLOOS K, MOSHER M, EPPERLY MW, GREENBERGER JS, PITT BR, KAGAN VE: "Oxidative Lipidomics of gamma-Radiation-Induced Lung Injury: Mass Spectrometric Characterization of Cardiolipin and Phosphatidylserine Peroxidation", RADIAT RES
VAARALA 0, ALFTHAN G, JAUHIAINEN M, LEIRISALO-REPO M, AHO K, PALOSUO T: "Crossreaction between antibodies to oxidised low-density lipoprotein and to cardiolipin in systemic lupus erythematosus", LANCET, vol. 341, 1993, pages 923 - 5
VAN HEERDE W.L., SAKARIASSEN K.S., HEMKER H.C., SIXMA J.J., REUTELINGSPERGER C.P., DE GROOT P.G.: "Annexin V inhibits the procoagulant activity of matrices of TNF- stimulated endothelium under blood flow conditions", ARTERIOSCLER. THROMB. VASC. BIOL., vol. 14, 1994, pages 824 - 830
WAN M, SABIRSH A, WETTERHOLM A, AGERBERTH B, HAEGGSTROM JZ: "Leukotriene B4 triggers release of the cathelicidin LL-37 from human neutrophils: novel lipid-peptide interactions in innate immune responses", FASEB J., vol. 21, 2007, pages 2897 - 905, XP002603099
WINYARD PG, TATZBER F, ESTERBAUER H, KUS ML, BLAKE DR, MORRIS CJ: "Presence of foam cells containing oxidised low density lipoprotein in the synovial membrane from patients with rheumatoid arthritis", ANN RHEUM DIS., vol. 52, 1993, pages 677 - 80
WOOD, R., HARLOW, R.D.: "Structural analyses of rat liver phosphoglycerides", ARCH. BIOCHEM. BIOPHYS., vol. 135, 1969, pages 272 - 281, XP024804998, DOI: doi:10.1016/0003-9861(69)90540-2
WU R, DE FAIRE U, LEMNE C, WITZTUM JL, FROSTEGARD J: "Autoantibodies to OxLDL are decreased in individuals with borderline hypertension", HYPERTENSION, vol. 33, no. 1, 1999, pages 53 - 59
YABUUCHI H., O'BRIEN J.S.: "Positional distribution of fatty acids in glycerophosphatides of bovine gray matter", J. LIPID RES., vol. 9, 1968, pages 65 - 67
YOKOMIZO T, IZUMI T, CHANG K, TAKUWA Y, SHIMIZU T: "A G-protein-coupled receptor for leukotriene B4 that mediates chemotaxis", NATURE, vol. 387, 1997, pages 620 - 4
YOKOMIZO T, KATO K, TERAWAKI K, IZUMI T, SHIMIZU T: "A second leukotriene B(4) receptor, BLT2. A new therapeutic target in inflammation and immunological disorders", J EXP MED., vol. 192, 2000, pages 421 - 32
ZANCHETTI A, BOND MG, HENNIG M, NEISS A, MANCIA G, DAL PALU C, HANSSON L, MAGNANI B, RAHN KH, REID J: "Risk factors associated with alterations in carotid intima-media thickness in hypertension: baseline data from the European Lacidipine Study on Atherosclerosis", J HYPERTENS., vol. 16, no. 7, 1998, pages 949 - 961
ZANCHETTI A, BOND MG, HENNIG M, NEISS A, MANCIA G, DAL PALU C, HANSSON L, MAGNANI B, RAHN KH, REID JL: "Calcium antagonist lacidipine slows down progression of asymptomatic carotid atherosclerosis: principal results of the European Lacidipine Study on Atherosclerosis (ELSA), a randomized, double-blind, long-term trial", CIRCULATION, vol. 106, no. 19, 2002, pages 2422 - 2427

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10060908B2 (en) 2008-12-19 2018-08-28 Medirista Biotechnologies Ab Oxidized cardiolipin as a pro-inflammatory factor
JP2020506206A (ja) * 2017-02-09 2020-02-27 アネキシン ファーマシューティカルズ アーベー 治療用組成物
JP7365478B2 (ja) 2017-02-09 2023-10-19 アネキシン ファーマシューティカルズ アーベー 治療用組成物
US11801283B2 (en) 2017-02-09 2023-10-31 Annexin Pharmaceuticals Ab Therapeutic compositions
WO2018220224A1 (fr) * 2017-06-02 2018-12-06 Medirista Biotechnologies Ab Antigènes associés aux lipides et anticorps contre eux
EP3746478A4 (fr) * 2018-01-29 2022-01-19 The Regents of The University of California Thérapies et méthodes pour traiter des maladies et des troubles médiés par tlr2
US11530259B2 (en) 2018-01-29 2022-12-20 The Regents Of The University Of California Therapies and methods to treat TLR2-mediated diseases and disorders
CN112041682A (zh) * 2018-04-19 2020-12-04 国立大学法人东京大学 用于辅助对象中的疾病的诊断的方法及套件
CN112041682B (zh) * 2018-04-19 2024-04-02 国立大学法人东京大学 用于辅助对象中的疾病的诊断的方法及套件
WO2021097146A1 (fr) * 2019-11-12 2021-05-20 Abcentra, Llc Méthodes et compositions pour le traitement du cancer
WO2023057588A1 (fr) * 2021-10-06 2023-04-13 Iltoo Pharma Constructions chimériques d'interleukine 2 ont une spécificité de ciblage vis-à-vis des tissus enflammés

Also Published As

Publication number Publication date
WO2011160845A3 (fr) 2012-03-15

Similar Documents

Publication Publication Date Title
US20210389310A1 (en) Oxidized cardiolipin as a pro-inflammatory factor
Jensen et al. Novel metabolic biomarkers of cardiovascular disease
Upadhyay Emerging risk biomarkers in cardiovascular diseases and disorders
Verhoye et al. Circulating oxidized low-density lipoprotein: a biomarker of atherosclerosis and cardiovascular risk?
Onat et al. Complement C3 and cleavage products in cardiometabolic risk
WO2011160845A2 (fr) Phospholipides et lipoprotéines oxydés, et anticorps dirigés contre eux, à titre de biomarqueurs des états inflammatoires et méthodes de traitement
Papac-Milicevic et al. Malondialdehyde epitopes as targets of immunity and the implications for atherosclerosis
Nanayakkara et al. Plasma asymmetric dimethylarginine (ADMA) concentration is independently associated with carotid intima-media thickness and plasma soluble vascular cell adhesion molecule-1 (sVCAM-1) concentration in patients with mild-to-moderate renal failure
Fredrikson et al. Identification of immune responses against aldehyde-modified peptide sequences in apoB associated with cardiovascular disease
Kontush et al. Functionally defective high-density lipoprotein: a new therapeutic target at the crossroads of dyslipidemia, inflammation, and atherosclerosis
Rahman et al. IgM antibodies against malondialdehyde and phosphorylcholine are together strong protection markers for atherosclerosis in systemic lupus erythematosus: Regulation and underlying mechanisms
Lin et al. HSP60 in heart failure: abnormal distribution and role in cardiac myocyte apoptosis
EP1735349B1 (fr) Méthode pour déterminer le risque de maladies cardiovasculaires ischémiques en utilisant des conjugués de la phosphorylcholine
Haq et al. Consumption of β-casomorphins-7/5 induce inflammatory immune response in mice gut through Th2 pathway
Ling et al. NETosis as a pathogenic factor for heart failure
Pawlak et al. Oxidized LDL to autoantibodies against oxLDL ratio–the new biomarker associated with carotid atherosclerosis and cardiovascular complications in dialyzed patients
Goetze et al. Reproducible determination of high-density lipoprotein proteotypes
Carter et al. Complement C3 and C-reactive protein in male survivors of myocardial infarction
Cho High-density lipoproteins as biomarkers and therapeutic tools: Volume 1. Impacts of lifestyle, diseases, and environmental stressors on HDL
Allen et al. Role of oxidized phospholipids in cardiovascular pathology
Pawlak et al. Inflammation but not oxidative stress is associated with β-chemokine levels and prevalence of cardiovascular disease in uraemic patients
Vik et al. Serum osteoprotegerin in young survivors of myocardial infarction
Ralapanawa Biomarkers of Inflammation and Oxidative Stress in the Prediction and Management of Acute Coronary Syndrome
Le Oxidized lipids and lipoproteins: indices of risk or targets for management
Wu et al. Proteomics Analysis of Lipid Metabolism and Inflammatory Response in the Liver of Rabbits fed on a High Cholesterol Diet

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11736274

Country of ref document: EP

Kind code of ref document: A2

DPE1 Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101)
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11736274

Country of ref document: EP

Kind code of ref document: A2