Method for the diagnosis and/or prognosis of inflammatory states

Abstract

The invention relates to a method for the diagnosis and/or prognosis of inflammatory states.

1 . A method for the identification and quantification of the expression of membrane receptors present on the surface of target granulocytes, comprising contacting at least one soluble receptor-binding domain (RBD) with target granulocytes from a mammal, said identification and quantification taking place at a given time or during a given time interval, and allowing the diagnosis and/or prognosis of inflammatory states in a mammal. 
     
     
         2 . The method according to  claim 1 , wherein the method is for the identification and quantification of the expression of membrane receptors present on the surface of target granulocytes (neutrophils, eosinophils, basophils and mast cells), said identification and quantification taking place at a given time or during a given time interval, and allowing the diagnosis and/or prognosis of inflammatory states, provided that when only one RBD is used, said membrane receptor is not GLUT1. 
     
     
         3 . The method according to  claim 1 , wherein said at least one soluble receptor-binding domain is a set of three to twenty soluble receptor-binding domain, preferably a set of three to twelve soluble receptor-binding domain in particular three, four, five, six seven, eight, nine, ten, eleven, or twelve receptor-binding domain. 
     
     
         4 . The method according to  claim 1 , wherein said at least one soluble receptor-binding domain is a set of three to twenty soluble receptor-binding domain, preferably a set of three to twelve soluble receptor-binding domain in particular three, four, five, six seven, eight, nine, ten, eleven, or twelve receptor-binding domain, provided that at least one soluble receptor-binding domain of said set does not interact with GLUT1 membrane receptor. 
     
     
         5 . The method according to  claim 1 , wherein said target granulocytes are selected from the list consisting of neutrophils, eosinophils, basophils and mast cells. 
     
     
         6 . The method according to  claim 1 , wherein said target granulocytes are neutrophils and said inflammatory state is cystic fibrosis. 
     
     
         7 . The method according to  claim 1 , wherein said neutrophils are blood neutrophils or lung neutrophils. 
     
     
         8 . The method according to  claim 1 , wherein said target granulocytes are eosinophils and said inflammatory state is allergy and/or asthma. 
     
     
         9 . The method according to  claim 1 , wherein said target granulocytes are basophils and said inflammatory state is allergy. 
     
     
         10 . The method according to  claim 1 , wherein said RBD is selected from the list consisting of: SEQ ID NO: 1 to 31. 
     
     
         11 . The method according to  claim 1  according to  claim 10 , wherein said RBD is selected from the list consisting of: Amphotropic Murine Leukemia Retrovirus (AMLV, SEQ ID NO:1), Feline endogenous retrovirus (RD114, SEQ ID NO:3), Koala endogeneous Retrovirus (KoRV, SEQ ID NO: 20), Human T Leukaemia Virus-2 (HTLV2, SEQ ID NO:28), Bovine Leukaemia Virus (BLV, SEQ ID NO: 30), or Porcine Endogeneous Retrovirus-A (Pery A, SEQ ID NO:21), 
     
     
         12 . The method according to  claim 1 , wherein said at least one soluble receptor-binding domain is a combination of two soluble receptor-binding domain (RBD). 
     
     
         13 . The method according to  claim 1  according to  claim 12 , wherein said combination is the combination of HTLV-2 RBD (SEQ ID NO: 28) and KoRV RBD (SEQ ID NO: 20) and said membrane receptors are GLUT1 and PiT1 respectively, said membrane receptors being expressed in particular in lung neutrophils and blood neutrophils. 
     
     
         14 . The method according to  claim 1  according to  claim 13 , wherein the expression of said membrane receptors in lung neutrophils is increased compared with the expression of said membrane receptor in blood neutrophils. 
     
     
         15 . The method according to  claim 1  according to  claim 12 , wherein said combination is the combination of PERVA RBD (SEQ ID NO: 21) and BLV RBD (SEQ ID NO: 30) and said membrane receptors are PAR and a membrane receptor interacting with BLV respectively, said membrane receptors being expressed in particular in lung neutrophils and blood neutrophils. 
     
     
         16 . The method according to  claim 1  according to  claim 15 , wherein the expression of said PAR membrane receptor in lung neutrophils is decreased compared with the expression of said membrane receptor in blood neutrophils and said receptor interacting with BLV in lung neutrophils is increased compared with the expression of said membrane receptor in blood neutrophils. 
     
     
         17 . Process of in vitro diagnosis and/or prognosis of an inflammatory state in a mammal, comprising the identification and quantification of the expression of at least one membrane receptors, said identification and quantification being as defined as defined in  claim 1 , present on the surface of target granulocytes. 
     
     
         18 . Process of in vitro diagnosis and/or prognosis of an inflammatory state in a mammal, comprising the identification and quantification of the expression of at least one membrane receptors, said identification and quantification being as defined as defined in  claim 1 , present on the surface of target granulocytes, provided that when only one RBD is used, said membrane receptor is not GLUT1, and when two or more RBD are used, at least one of said soluble receptor-binding domain does not interact with GLUT1 membrane receptor. 
     
     
         19 . Process of in vitro diagnosis and/or prognosis of an inflammatory state according to  claim 16 , comprising the following steps:
 a. contacting at least one soluble receptor-binding domain, optionally marked with a tag, with target granulocytes of a diseased mammal to form at last one complex, said at least one complex being constituted by said at least one soluble receptor-binding domain and at least one membrane receptor of said target granulocytes,   b. identifying said at least complex formed,   c. quantifying the expression of each membrane receptor of said target granulocytes able to form said complex,   d. contacting said at least one soluble receptor-binding domain of step a. with target granulocytes of a control mammal and identifying each complex formed as in step b. and quantifying the expression of each membrane receptor of said target granulocytes able to form said complex as in step c.   e. comparing the level of expression of membrane receptors in step c and d., an overexpression or underexpression of membrane receptors of target granulocytes of said diseased mammal compared with control mammal indicating an inflammatory state   
     
     
         20 . Process according to  claim 19 , wherein said control mammal is the same mammal species as the diseased mammal. 
     
     
         21 . Process according to  claim 20 , wherein said granulocytes are neutrophils, in particular blood neutrophils and lung neutrophils. 
     
     
         22 . Process according to  claim 19 , wherein the inflammatory state is cystic fibrosis. 
     
     
         23 . Process of in vitro diagnosis and/or prognosis of cystic fibrosis according to  claim 21 , comprising the following steps:
 a. contacting HTLV-2 RBD (SEQ ID NO: 28) and/or KoRV RBD (SEQ ID NO: 20) optionally marked with a tag, with lung neutrophils of a mammal to form at least one complex,   b. identifying said at least one complex formed and being constituted by HTLV-2 receptor-binding domain and GLUT1 membrane receptor and/or KoRV receptor-binding domain and Pill membrane receptor of said lung neutrophils,   c. quantifying the expression of said GLUT1 and/or Pill membrane receptor of said lung neutrophils able to form said complex,   d. contacting said HTLV-2 RBD and/or KoRV RBD with blood neutrophils and identifying and quantifying the expression of said GLUT1 and/or Pill membrane receptor of said blood neutrophils able to form said complex,   e. comparing the level of expression of each membrane receptor, an overexpression of GLUT1 and/or Pill in lung neutrophils compared with blood neutrophils indicating a pulmonary inflammatory state during cystic fibrosis.   
     
     
         24 . Process of in vitro diagnosis and/or prognosis of cystic fibrosis according to  claim 21 , comprising the following steps:
 a. contacting PERVA RBD (SEQ ID NO: 21) and and/or BLV RBD (SEQ ID NO: 30) optionally marked with a tag, with lung neutrophils of a mammal to form at least one complex,   b. identifying said at least one complex formed and being constituted by PERVA receptor-binding domain and PAR membrane receptor of said lung neutrophils, and/or BLV receptor-binding domain and a membrane receptor interacting with BLV,   c. quantifying the expression of said PAR and/or a membrane receptor interacting with BLV of said lung neutrophils able to form said complex,   d. contacting said PERVA RBD and/or BLV RBD with blood neutrophils and identifying and quantifying the expression of each said PAR and/or a membrane receptor interacting with BLV of said blood neutrophils able to form said complex,   e. comparing the level of expression of each membrane receptor, an overexpression of said membrane receptor interacting with BLV in blood neutrophils compared with lung neutrophils and/or an underexpression of PAR in blood neutrophils compared with lung neutrophils indicating a pulmonary inflammatory state during cystic fibrosis.   
     
     
         25 . Process according to  claim 20 , wherein said granulocytes are eosinophils. 
     
     
         26 . Process according to  claim 20 , wherein said granulocytes are basophils. 
     
     
         27 . Process according to  claim 20 , wherein said granulocytes are mast cells. 
     
     
         28 . Method for in vitro measuring the therapeutic efficacy of a potential anti-inflammatory drug in a mammal, comprising the following steps:
 a. identifying and quantifying the expression of at least one membrane receptor, said identification and quantification being as defined as defined in  claim 1 , present on the surface of target granulocytes,   b. contacting said granulocytes with a drug liable to treat said inflammatory state to give treated granulocytes,   c. identifying and quantifying the expression of at least one membrane receptor, present on the surface of treated granulocytes,   d. comparing the level of expression of said at least one membrane receptor before and after contacting with said drug, an increase and/or a decrease of the expression of said at least one membrane receptor after contacting indicating a therapeutic efficacy of said drug depending of said inflammatory state.   
     
     
         29 . Method for in vitro measuring the therapeutic efficacy of a potential anti-inflammatory drug in a mammal, according to  claim 28 , wherein step a is carried out provided that when only one RBD is used, said membrane receptor is not GLUT1, and when two or more RBD are used, at least one of said soluble receptor-binding domain does not interact with GLUT1 membrane receptor.