Cytotoxic t cell response modifiers

Abstract

The present invention relates to a compound selectively binding to a sensory receptor or selectively altering the expression of a sensory receptor for use in a method for treating or preventing a disease associated with a pathologic cellular cytotoxic T cell (CTL) response. Further, the invention relates to means for detecting a sensory receptor for use in a method for diagnosing cellular resistance against CTL response in a patient. The invention further embraces a method for determining the resistance of a cell against a CTL response in vitro and to a method for identifying agents that influence the response of cells to CTLs.

1 .- 15 . (canceled) 
     
     
         16 . A method for treating or preventing a disease associated with a pathologic cellular cytotoxic T cell (CTL) response in a subject, preferably a disease selected from the group consisting of neoplasia, in particular cancer, an autoimmune disease, an infection and graft-versus-host disease, the method comprising the step of administering to the subject at least one compound selectively binding to at least one sensory receptor or selectively altering the expression or activity of a sensory receptor or selectively altering the expression or activity of a polypeptide involved in cell signaling of a sensory receptor. 
     
     
         17 . The method according to  claim 16 , wherein said sensory receptor: (i) regulates or mediates CTL response; and/or (ii) functions as an immune-checkpoint molecule. 
     
     
         18 . The method according to  claim 16 , wherein the sensory receptor is a receptor localized at the cellular surface, preferably wherein said sensory receptor is selected from the group consisting of
 (a) an olfactory receptor, preferably selected from the group consisting of:
 (i) a taste receptor, in particular a taste receptor type 2 (TAS2R), in particular taste receptor type 2 member 3 (TAS2R3), 
 (ii) an olfactory receptor of family 1, 2 or 51, in particular an olfactory receptor of family 1 subfamily F (OR1F), in particular an olfactory receptor of family 1 subfamily F member 1 (OR1F1), an olfactory receptor of family 2 subfamily J (OR2J), in particular an olfactory receptor of family 2 subfamily J member 2 (OR2J2), or an olfactory receptor of family 51 subfamily E (OR51E), in particular an olfactory receptor of family 51 subfamily E member 2 (OR51E2), and 
 (iii) a vomeronasal receptor, preferably vomeronasal 1, in particular vomeronasal 1 receptor 4 (VN1R4); 
 (iv) a pheromone receptor; and 
   (b) an opsin, in particular opsin 3 (OPN3).   
     
     
         19 . The method according to  claim 16 , wherein the compound enhances an immune response to an activated CTL, in particular in a patient suffering from a neoplasia, in particular cancer, preferably
 wherein the local surface concentration and/or T cell-binding activity of the surface receptor that prevents CTL response is reduced; and/or   wherein said compound is selected from the group consisting of:
 (i) a compound reducing the expression of a sensory receptor and/or of a polypeptide involved in cell signaling of a sensory receptor, preferably an interfering RNA, in particular a small interfering ribonucleic acid (siRNA) or a small hairpin RNA (shRNA), complementary to mRNA encoding for said sensory receptor, and 
 (ii) a compound preventing the T cell-binding activity to said sensory receptor, in particular wherein said compound is an antibody or a sensory receptor-binding fragment. 
   
     
     
         20 . The method according to  claim 16 , wherein the compound suppresses an immune response to an activated CTL, in particular in a patient suffering from an autoimmune diseases and/or graft-versus-host disease, preferably
 wherein the amount and/or T cell-binding activity of the surface receptor that prevents CTL response is enhanced; and/or   wherein the compound is selected from the group consisting of:
 (i) genetic material encoding for a sensory receptor, 
 (ii) a compound enhancing the expression of said sensory receptor, and 
 (iii) a compound cross-linking a surface polypeptide preventing CTL with a T cell, in particular a cross-linking antibody or antibody fragment. 
   
     
     
         21 . A method for determining the resistance of a cell against a CTL response, said method comprising the step of detecting at least one sensory receptor of said cell and/or at least one polypeptide involved in cell signaling of a sensory receptor of said cell. 
     
     
         22 . The method according to  claim 21 , wherein the step of detecting comprises the use of a detector for detecting the at least one sensory receptor of said cell and/or the at least one polypeptide involved in cell signaling of a sensory receptor of said cell. 
     
     
         23 . The method according to  claim 22 , wherein said detector comprises at least one compound selectively binding to the sensory receptor and/or comprises at least one compound selectively altering the expression of the sensory receptor and/or of the polypeptide involved in cell signaling of a sensory receptor, preferably wherein said compound is labeled, in particular fluorescently labeled, labeled by a contrast agent, spin-labeled and/or radioactively labeled, in particular wherein said compound:
 (i) is an interfering RNA, in particular small interfering ribonucleic acid (siRNA) or small hairpin RNA (shRNA), complementary to the messenger ribonucleic acid (mRNA) encoding for the sensory receptor,   (ii) is or comprises one or more amino acid moieties; and/or a polypeptide strand, such as a peptide or protein, preferably an antibody or a fragment thereof, selectively binding to the sensory receptor,   (iii) is or comprises a small molecule having a molecular weight of not more than 500 Da, selectively binding to the sensory receptor, or   (iv) is or comprises one or more types(s) of high molecular weight compounds having a molecular weight higher that 500 Da.   
     
     
         24 . The method according to  claim 21 , wherein said sensory receptor: (i) regulates or mediates CTL response; and/or (ii) functions as an immune-checkpoint molecule 
     
     
         25 . The method according to  claim 21 , wherein the sensory receptor is a receptor localized at the cellular surface, preferably wherein said sensory receptor is selected from the group consisting of
 (a) an olfactory receptor, preferably selected from the group consisting of:
 (i) a taste receptor, in particular a taste receptor type 2 (TAS2R), in particular taste receptor type 2 member 3 (TAS2R3), 
 (ii) an olfactory receptor of family 1, 2 or 51, in particular an olfactory receptor of family 1 subfamily F (OR1F), in particular an olfactory receptor of family 1 subfamily F member 1 (OR1F1), an olfactory receptor of family 2 subfamily J (OR2J), in particular an olfactory receptor of family 2 subfamily J member 2 (OR2J2), or an olfactory receptor of family 51 subfamily E (OR51E), in particular an olfactory receptor of family 51 subfamily E member 2 (OR51E2), and 
 (iii) a vomeronasal receptor, preferably vomeronasal 1, in particular vomeronasal 1 receptor 4 (VN1R4); 
 (iv) a pheromone receptor; and 
   (b) an opsin, in particular opsin 3 (OPN3).   
     
     
         26 . The method of  claim 21 , further comprising the steps:
 (i) providing the cell to be tested for resistance against a CTL response,   (ii) determining the amount of at least one sensory receptor at the surface of said cell, and   (iii) comparing said amount of the sensory receptor with a standard of a comparable cell lacking the sensory receptor at its surface and/or a comparable cell having such amounts at its surface that an extensive resistance to a CTL response occurs.   
     
     
         27 . The method according to  claim 26 , wherein step (ii) comprises contacting the cell with
 (a) at least one compound selectively binding to the at least one sensory receptor, preferably wherein said compound is labeled, in particular fluorescently labeled, labeled by a contrast agent, spin-labeled and/or radioactively labeled, in particular wherein said compound is an antibody or an sensory receptor-binding fragment thereof; and/or   (b) at least one antagonist of the at least one sensory receptor and/or of the at least polypeptide involved in cell signaling of a sensory receptor, in particular wherein said antagonist is interfering RNA, in particular small interfering ribonucleic acid (siRNA) or small hairpin RNA (shRNA), complementary to said sensory receptor, co-incubating the cell contacted with the at least one antagonist with at least one CTL, and subsequently determining the CTL activity, preferably determining the CTL activity by one of the following:
 (i) by determining the viability of the cell co-incubated with the at least one CTL and comparing the viability with that of a sample comprising a comparable cell and CTLs not treated by said antagonist, or 
 (ii) by determining the excretion of polypeptides typical for CTL activity, such as one or more interferon(s) (IFN), one or more interleukin(s) (IL) and/or one or more tumor necrosis factor(s) (TNF), and comparing the results with that of a sample comprising a comparable cell and CTLs not treated by said antagonist. 
   
     
     
         28 . The method according to  claim 26 , wherein step (ii) comprises contacting the cell with an antibody selectively binding to the at least one sensory receptor, or with a fragment thereof binding the at least one sensory receptor. 
     
     
         29 . The method according to  claim 27 , wherein step (ii) of (b) comprises contacting the cell with at least one antagonist of the at least one sensory receptor and/or of the at least one polypeptide involved in cell signaling of a sensory receptor, in particular wherein said antagonist is (a) an antibody selectively binding to the at least one sensory receptor, or with a fragment thereof binding the at least one sensory receptor; or (b) interfering RNA, and determining the excretion of one or more interferon(s) (IFN), one or more interleukin(s) (IL) and/or one or more tumor necrosis factor(s) (TNF). 
     
     
         30 . The method according to  claim 26 , wherein step (ii) comprises detection at the gene expression level, such as by using qPCR primer probes. 
     
     
         31 . The method of  claim 16 , further comprising the steps of:
 (i) providing a cell of the subject to be tested for a pathologic CTL response,   (ii) detecting at least one sensory receptor of said cell and/or at least one polypeptide involved in cell signaling of a sensory receptor, and   (iii) diagnosing the subject with the disease associated with a pathologic CTL response, such as cellular resistance against a CTL response, when the at least one sensory receptor and/or the at least one polypeptide involved in cell signaling of a sensory receptor is detected with the cell.   
     
     
         32 . A composition or kit comprising a cell expressing: (x) at least one surface polypeptide preventing CTL response, in particular a sensory receptor, such as a sensory receptor selected from the group consisting of
 (a) an olfactory receptor, preferably selected from the group consisting of:
 (i) a taste receptor, in particular a taste receptor type 2 (TAS2R), in particular taste receptor type 2 member 3 (TAS2R3), 
 (ii) an olfactory receptor of family 1, 2 or 51, in particular an olfactory receptor of family 1 subfamily F (OR1F), in particular an olfactory receptor of family 1 subfamily F member 1 (OR1F1), an olfactory receptor of family 2 subfamily J (OR2J), in particular an olfactory receptor of family 2 subfamily J member 2 (OR2J2), or an olfactory receptor of family 51 subfamily E (OR51E), in particular an olfactory receptor of family 51 subfamily E member 2 (OR51E2), and 
 (iii) a vomeronasal receptor, preferably vomeronasal 1, in particular vomeronasal 1 receptor 4 (VN1R4); 
 (iv) a pheromone receptor; and 
   (b) an opsin, in particular opsin 3 (OPN3); and/or (y) at least one polypeptide involved in cell signaling preventing CTL response, in particular a polypeptide involved in cell signaling of at least one surface polypeptide preventing CTL response, such as the sensory receptor, and/or a cell obtained from a patient, preferably a human patient that has developed a pathologic CTL response and/or is at risk of developing a pathologic CTL response, such as cellular resistance against a CTL response,   a CTL, such as, pre-activated CD8+ T-cells, in particular CD8+ T cells pre-activated by T cell receptor (TCR) stimulation through a CD3 specific antibody and/or a CD28 specific antibody, and
 an agent of interest being a candidate agent influencing the response of cells to CTLs such as an agent that: 
 (i) is an interfering RNA, in particular small interfering ribonucleic acid (siRNA) or small hairpin RNA (shRNA), complementary to the messenger ribonucleic acid (mRNA) encoding for a polypeptide typically localized at the cell surface and/or a polypeptide involved in cell signaling, 
 (ii) is or comprises one or more amino acid moieties; and/or a polypeptide strand, such a peptide or protein, preferably an antibody or a fragment thereof, 
 (iii) is or comprises a small molecule having a molecular weight of not more than 500 Da, or 
 (iv) is or comprises one or more types(s) of high molecular weight compounds having a molecular weight higher than 500 Da; and/or an agent that is a compound selectively binding to the at least one sensory receptor or selectively altering the expression or activity of the at least one sensory receptor and/or of the at least one polypeptide involved in cell signaling, 
   wherein the kit additionally comprises
 optionally a buffer and/or medium for washing away unviable cells, 
 optionally a bioluminescence precursor compound and cofactors, and 
 optionally a user manual. 
   
     
     
         33 . The composition or kit of  claim 32 , wherein the cell further expresses a polypeptide enabling bioluminescence, in particular a bioluminescent polypeptide such as luciferase. 
     
     
         34 . The composition or kit of  claim 32 , wherein the surface polypeptide preventing CTL response is a sensory receptor. 
     
     
         35 . The composition or kit of  claim 32 , wherein the sensory receptor is selected from the group consisting of
 (a) an olfactory receptor, preferably selected from the group consisting of:
 (i) a taste receptor, in particular a taste receptor type 2 (TAS2R), in particular taste receptor type 2 member 3 (TAS2R3), 
 (ii) an olfactory receptor of family 1, 2 or 51, in particular an olfactory receptor of family 1 subfamily F (OR1F), in particular an olfactory receptor of family 1 subfamily F member 1 (OR1F1), an olfactory receptor of family 2 subfamily J (OR2J), in particular an olfactory receptor of family 2 subfamily J member 2 (OR2J2), or an olfactory receptor of family 51 subfamily E (OR51E), in particular an olfactory receptor of family 51 subfamily E member 2 (OR51E2), and 
 (iii) a vomeronasal receptor, preferably vomeronasal 1, in particular vomeronasal 1 receptor 4 (VN1R4); 
 (iv) a pheromone receptor; and 
   (b) an opsin, in particular opsin 3 (OPN3).