# US 7,608,392 B2 · Diagnostic assay for measuring a cell mediated immune response > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,608,392 B2 - Document ID: US7608392B2 - Status: Granted - Kind: B2 - Application number: 10/477,571 - Filed: 2003-11-06 - Published: 2005-01-20 - Granted: 2009-10-27 - Earliest claimed: 2002-11-08 - Assignees: Cellestis Limited - Inventors: James Stuart Rothel; Steven Paul Wild; Angela Cosgriff - Web page: https://patentgrep.com/patent/US7608392B2 ## CPC - G01N 33/5091: {for testing the pathological state of an organism} - G01N 33/5047: {Cells of the immune system} - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present invention relates generally to a diagnostic assay and, more particularly, an assay for measuring cell-mediated immune reactivity. Even more particularly, the present invention provides an assay and a kit for measuring a cell-mediated response to an antigen using whole blood or other suitable biological sample. The assay may be conducted using ligands to immune effector molecules or at the nucleic acid level, screening for expression of genes encoding the immune effector molecules. The assay is useful in therapeutic and diagnostic protocols for human, livestock and veterinary and wild life applications. ## Claims ### Claim 1 A method for measuring a CMI response in a subject, said method comprising; collecting a whole blood sample from said subject wherein said whole blood sample comprises cells of the immune system which are capable of producing immune effector molecules following stimulation by an antigen, incubating a mixture comprising the whole blood sample, the antigen and an amount of an isolated simple sugar effective to enhance the stimulation by the antigen, and optionally heparin, and measuring the presence of, or elevation in, the level of an immune effector molecule wherein the presence or level of said immune effector molecule is indicative of the capacity of said subject to mount a cell-mediated immune response. ### Claim 2 (depends on 1) The method of claim 1 wherein the subject is a human. ### Claim 3 (depends on 1) The method of claim 1 wherein the whole blood is collected in a tube comprising antigen. ### Claim 4 (depends on 1) The method of claim 1 wherein the whole blood is collected in a tube comprising heparin. ### Claim 5 (depends on 3) The method of claim 3 wherein the tube comprises heparin. ### Claim 6 (depends on 1) The method of claim 1 wherein the whole blood sample is incubated with the antigen for from about 5 to about 50 hours. ### Claim 7 (depends on 1) The method of claim 1 wherein the immune effector molecule is a cytokine. ### Claim 8 (depends on 7) The method of claim 7 wherein the cytokine is IFN-γ. ### Claim 9 (depends on 7) The method of claim 7 wherein the cytokine is GM-CSF. ### Claim 10 (depends on 7) The method of claim 7 wherein the cytokine is an interleukin. ### Claim 11 (depends on 7) The method of claim 7 wherein the cytokine is a TNF-α. ### Claim 12 (depends on 1, 2) The method of claim 1 or 2 wherein the subject is infected by a pathogenic agent. ### Claim 13 (depends on 12) The method of claim 12 wherein the pathogenic agent is HBV. ### Claim 14 (depends on 12) The method of claim 12 wherein the pathogenic agent is HIV. ### Claim 15 (depends on 1) The method of claim 1 wherein the immune cells are selected from NK cells, T-cells, B-cells, dendritic cells, macrophages or monocytes. ### Claim 16 (depends on 15) The method of claim 15 wherein the immune cells are T-cells. ### Claim 17 (depends on 1) The method of claim 1 wherein the simple sugar is dextrose. ### Claim 18 (depends on 1) The method of claim 1 wherein the antigen is a peptide. ### Claim 19 (depends on 1) The method of claim 1 wherein the antigen is a polypeptide. ### Claim 20 (depends on 1) The method of claim 1 wherein the antigen is a protein. ### Claim 21 (depends on 1) The method of claim 1 wherein the antigen is a glycoprotein. ### Claim 22 (depends on 1) The method of claim 1 wherein the antigen is a carbohydrate. ### Claim 23 (depends on 1) The method of claim 1 wherein the antigen is selected from a phospholipid, phosphoprotein and a phospholipoprotein. ### Claim 24 (depends on 1) The method of claim 1 wherein the antigen is a TB-specific antigen. ### Claim 25 (depends on 24) The method of claim 24 wherein the TB-specific antigen is ESAT-6. ### Claim 26 (depends on 24) The method of claim 24 wherein the TB-specific antigen is CFP-10. ### Claim 27 (depends on 24) The method of claim 24 wherein the TB-specific antigen is TB7. ### Claim 28 (depends on 1) The method of claim 1 wherein the antigen is PPD from Mycobacterium tuberculosis. ### Claim 29 (depends on 1) The method of claim 1 wherein the antigen is PPD from Mycobacteriun avium. ### Claim 30 (depends on 1) The method of claim 1 wherein the antigen is tetanus toxoid. ### Claim 31 (depends on 1) The method of claim 1 wherein the immune effectors are detected with antibodies specific for same. ### Claim 32 (depends on 31) The method of claim 31 wherein the immune effectors are detected using ELISA. ### Claim 33 (depends on 31) The method of claim 31 wherein the immune effectors are detected using ELISpot. ### Claim 34 A method of treatment of a subject having a pathogenic infection, an autoimmune disorder or cancer or a propensity for developing such a disorder, said method comprising assessing the ability for said subject to mount a cell mediated immune response by the method of measuring a CMI response in a subject, said method of measuring comprising, collecting a whole blood sample from said subject wherein said whole blood sample comprises cells of the immune system which are capable of producing immune effector molecules following stimulation by an antigen, incubating a mixture comprising the whole blood sample, antigen and an amount of an isolated simple sugar effective to enhance the stimulation by the antigen, and optionally heparin, and measuring the presence of or elevation in the level of an immune effector molecule wherein the presence or level of said immune effector molecule is indicative of the capacity of said subject to mount a cell-mediated immune response and selecting a suitable therapeutic protocol. ### Claim 35 (depends on 34) The method of claim 34 wherein the subject is a human. ### Claim 36 (depends on 34) The method of claim 34 wherein the whole blood is collected in a tube comprising antigen. ### Claim 37 (depends on 34) The method of claim 34 wherein the whole blood is collected in a tube comprising heparin. ### Claim 38 (depends on 36) The method of claim 36 wherein the tube comprises heparin. ### Claim 39 (depends on 34) The method of claim 34 wherein the whole blood sample is incubated with the antigen for from about 5 to about 50 hours. ### Claim 40 (depends on 34) The method of claim 34 wherein the immune effector molecule is a cytokine. ### Claim 41 (depends on 40) The method of claim 40 wherein the cytokine is IFN-γ. ### Claim 42 (depends on 40) The method of claim 40 wherein the cytokine is GM-CSF. ### Claim 43 (depends on 40) The method of claim 40 wherein the cytokine is an interleukin. ### Claim 44 (depends on 40) The method of claim 40 wherein the cytokine is a TNF-α. ### Claim 45 (depends on 34) The method of claim 34 wherein the immune cells are selected from NK cells, T-cells, B-cells, dendritic cells, macrophages or monocytes. ### Claim 46 (depends on 45) The method of claim 45 wherein the immune cells are T-cells. ### Claim 47 (depends on 34) The method of claim 34 wherein the simple sugar is dextrose. ### Claim 48 (depends on 34) The method of claim 34 wherein the antigen is selected from a peptide. ### Claim 49 (depends on 34) The method of claim 34 wherein the antigen is selected from a polypeptide. ### Claim 50 (depends on 34) The method of claim 34 wherein the antigen is selected from a protein. ### Claim 51 (depends on 34) The method of claim 34 wherein the antigen is selected from a glycoprotein. ### Claim 52 (depends on 34) The method of claim 34 wherein the antigen is selected from a carbohydrate. ### Claim 53 (depends on 34) The method of claim 34 wherein the antigen is selected from a phospholipid, phosphoprotein and phospholipoprotein. ### Claim 54 (depends on 34) The method of claim 34 wherein the antigen is a TB-specific antigen. ### Claim 55 (depends on 54) The method of claim 54 wherein the TB-specific antigen is ESAT-6. ### Claim 56 (depends on 54) The method of claim 54 wherein the TB-specific antigen is CFP-10. ### Claim 57 (depends on 54) The method of claim 54 wherein the TB-specific antigen is TB-7. ### Claim 58 (depends on 34) The method of claim 34 wherein the antigen is PPD from Mycobacterium tuberculosis. ### Claim 59 (depends on 34) The method of claim 34 wherein the antigen is PPD for Mycobacterium avium. ### Claim 60 (depends on 34) The method of claim 34 wherein the antigen is phytohemagglutinin. ### Claim 61 (depends on 34) The method of claim 34 wherein the antigen is tetanus toxoid. ### Claim 62 (depends on 34) The method of claim 34 wherein the immune effectors are detected with antibodies specific for same. ### Claim 63 (depends on 62) The method of claim 62 wherein the immune effectors are detected using ELISA. ### Claim 64 (depends on 63) The method of claim 63 wherein the immune effectors are detected using ELISpot.