Diagnosis and treatment of preeclampsia

Abstract

The present invention relates to biomarkers for preeclampsia as well as treatment of this disease. In particular, the invention relates to methods for diagnosis or aiding in the diagnosis of preeclampsia of a pregnant female mammal to detect elevated levels of free haemoglobin, particularly free fetal haemoglobin. This facilitates and makes possible early diagnosis and clinical intervention when a preeclamptic condition is found. In addition, the invention relates to a method to treat female mammals with preeclampsia with the purpose to reverse the pathological conditions associated with this disease.

1 - 8 . (canceled) 
     
     
         9 . Fetal haemoglobin as a marker for preeclampsia. 
     
     
         10 . A method for diagnosing or aiding in diagnosing preeclampsia comprising the following steps: i) obtaining a non-foetal biological sample from a pregnant female mammal, ii) measuring the level of free fetal haemoglobin or measuring the level of free fetal haemoglobin and the level of total free haemoglobin in said biological sample. 
     
     
         11 . A method according to  claim 10  further comprising the step of iii) comparing the level of free fetal haemoglobin in the sample with a reference value or comparing the ratio between the level of free fetal haemoglobin and the level of total free haemoglobin in the sample with a reference value. 
     
     
         12 . A method for monitoring the progression or regression of preeclampsia comprising the steps of i) measuring the level of free fetal haemoglobin or measuring the level of free fetal haemoglobin and the level of total free haemoglobin, in a first biological sample isolated from a pregnant female mammal, ii) measuring the level of free fetal haemoglobin or measuring the level of free fetal haemoglobin and the level of total free haemoglobin, in a second biological sample isolated from said pregnant female mammal at a later time. 
     
     
         13 . A method according to  claim 12  further comprising the step of iii) comparing the values measured in step i) and ii). 
     
     
         14 . A method for assessing the efficacy of a treatment of preeclampsia comprising the steps of i) measuring the level of free fetal haemoglobin or measuring the level of free fetal haemoglobin and the level of total free haemoglobin, in a first biological sample obtained from a pregnant female mammal before treatment or at time ti, ii) measuring the level of free fetal haemoglobin or measuring the level of free fetal haemoglobin and the level of total free haemoglobin, in a second biological sample obtained from the same pregnant female mammal at a later time t 2 . 
     
     
         15 . A method according to  claim 14  further comprising the steps of iii) comparing the values measured in step i) and ii). 
     
     
         16 . A method according to  claim 10 , wherein the biological sample is a blood sample, a plasma sample, a urine sample or a sample of placental tissue. 
     
     
         17 - 24 . (canceled) 
     
     
         25 . An assay kit for use in a method defined in  claim 10 , the kit comprising i) means for detecting, in a biological sample of a pregnant female mammal, levels of free fetal and, if relevant, total haemoglobin, and ii) instructions for using said detecting means. 
     
     
         26 . A kit according to  claim 25  comprising i) a solid surface precoated with fetal haemoglobin, ii) one or more antibodies or an immune reactive fragment therefore that binds specifically to isolated free fetal haemoglobin from a pregnant female mammal or an immunogenic peptide, fragment or epitope thereof, iii) means for detecting the formation of an antigen-antibody complex, and iv) optionally, instructions for use. 
     
     
         27 . A kit according to  claim 26 , further comprising components to measure the total haemoglobin concentration in a sample. 
     
     
         28 . A kit according to  claim 27 , wherein the components needed for measuring the total haemoglobin concentration in a sample comprises, i) a solid surface precoated with haemoglobin ii) one or more antibodies or an immune reactive fragment therefore that binds specifically to isolated free haemoglobin from a pregnant female mammal or an immunogenic peptide, fragment or epitope thereof, iii) means for detecting the formation of an antigen-antibody complex, and iv) optionally, instructions for use. 
     
     
         29 . A kit according to  claim 26 , wherein said solid surface i) is one or more well(s) in a microtiter plate. 
     
     
         30 . A kit according to  claim 26 , wherein the antibody or antibody fragment i) is specific for the haemoglobin alpha chain, the haemoglobin beta chain, the haemoglobin delta chain, the haemoglobin gamma chain or an immunogenic peptide, fragment or epitope thereof. 
     
     
         31 . A kit according to  claim 26 , wherein said antibody i) is monoclonal or polyclonal antibody. 
     
     
         32 . A kit according to  claim 31 , wherein said antibody is obtained from rabbits. 
     
     
         33 . A kit according to  claim 26 , wherein the means for detection of an antigen-antibody complex comprises a labeled mono- or polyclonal antibody. 
     
     
         34 . A kit according to  claim 33 , wherein the labeled antibody binds specifically to an antibody of ii) or an immunogenic peptide, fragment or epitope thereof. 
     
     
         35 . A kit according to  claim 34 , wherein the labeled antibody is an enzyme-labeled antibody such as e.g. an alkaline phosphatase labeled antibody. 
     
     
         36 . A kit according to  claim 26 , wherein the means for detecting the formation of an antigen-antibody complex comprises a swine-anti-rabbit IgG-alkaline phosphatase antibody. 
     
     
         37 . A kit according to  claim 25 , further comprising standard dilution series of haemoglobin in bottles ready for use. 
     
     
         38 - 42 . (canceled) 
     
     
         43 . A method for preventing or treating preeclampsia comprising administering to a pregnant female mammal in need thereof an effective amount of one or more substances selected from haemoglobin binding agents, heme-binding agents, heme-degrading agents and/or iron-binding agents. 
     
     
         44 . A method of prognosis for preeclampsia comprising the following steps
 i) obtaining a biological sample from a pregnant female mammal;   ii) measuring the level of human leukocyte antigen DPA1 (HLA-DPA1), in said biological sample; and   iii) comparing the level of HLA-DPA1 in the sample with a reference value.   
     
     
         45 . A method according to  claim 44 , wherein steps (a) to (c) are performed to determine if said pregnant female is or is not at increased risk of developing preeclampsia, or is or is not at increased risk of developing a severe form of preeclampsia. 
     
     
         46 . A method according to  claim 44 , wherein an expression or a high expression of HLA-DPA1 indicates a better prognosis than no expression of HLA-DPA1. 
     
     
         47 . An assay kit for the prognosis or aiding in the prognosis of preeclampsia, according to the method of  claim 44 , comprising means for detecting, in a biological sample of a pregnant female mammal, levels of HLA-DPA1 and instructions for using said detecting means.