Novel assay for detecting immune responses involving antigen specific cytokine and/or antigen specific cytokine secreting T-cells

An immunoassay method for detecting MIG, a Th−1 cytokine that induces MIG expression, or cells which secrete said cytokine that induces MIG expression, comprising:

• a. obtaining a biological sample from a mammalian host which has been exposed to an immunoreactive substance;
• b. detecting MIG expression in said sample;
• and
• c. correlating said detection to the presence in said sample of said cytokine which induces MIG expression or cells that secrete said cytokine that induces MIG expression.

A method of assessing the effectiveness of a compound or system in inducing an immune response comprising:

• a. obtaining a tissue sample from a mammalian host which has been exposed to an immunoreactive substance;
• b. culturing said sample in the presence of said immunoreactive substance;
• and
• c. detecting the induction of the immune response of said host by up-regulation of MIG expression.

An immunoassay method for detecting IFN-γ or IFN-γ-secreting cells, comprising:

• a. obtaining a tissue sample from a host which has been exposed to an immunoreactive substance;
• b. culturing said sample in the presence of said immunoreactive substance;
• c. detecting expression in said sample of a chemokine whose production is upregulated by IFN-γ;
• and
• c. correlating said detection to the presence of IFN-γ or IFN-γ-secreting cells in said sample.

A method of monitoring the immunoresponsiveness of a mammal, comprising:

• obtaining a tissue sample from a subject that has been exposed to a known immunoreactive substance;
• detecting in said sample expression of a chemokine whose production is upregulated by IFN-γ;
• and
• correlating said detection to the presence of cells which secrete IFN-γ and thus to the immunoresponsiveness of said mammal.

A method of monitoring the immunoresponsiveness of a mammal, comprising:

• obtaining a biological fluid sample from a subject that has been exposed to a known immunoreactive substance;
• detecting MIG in said biological fluid sample;
• and
• correlating said detection to the immunoresponsiveness of said mammal.

An immunoassay kit for detecting in a sample a Th−1 cytokine which upregulates the production of MIG, or cells that secrete said cytokine, comprising:

• a. a biological sample;
• b. an immunoreactive substance;
• c. an indicator for MIG;
• and
• d. a detector which detects said indicator.

An immunoassay kit for detecting IFN-γ or IFN-γ secreting cells in a sample comprising:

• a. a biological sample;
• b. an immunoreactive substance;
• c. an indicator for a chemokine whose production is upregulated by IFN-γ;
• and
• d. a detector which detects said indicator.

An immunoassay for detecting IFN-γ, or cells which secrete IFN-γ, comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human host which has been exposed to an immunoreactive peptide;
• culturing said sample for a period of 4 to 16 hours in the presence of said peptide;
• detecting MIG expression in said sample via flow cytometry;
• and
• correlating said detection to the presence in said sample of IFN-γ or cells that secrete IFN-γ ;
• wherein said expression of MIG is genetically and specifically restricted, was induced by said IFN-γ, and said induction was mediated by CD8+ cells.

A method of assessing the effectiveness of a compound or system in inducing an immune response comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human host which has been exposed to an immunoreactive peptide;
• culturing said sample for 4 to 16 hours in the presence of said peptide.
• detecting, via flow cytometry, the induction of the immune response of said human host by up-regulation of MIG expression;
• wherein said expression of MIG is genetically and specifically restricted, was induced by IFN-γ, and said induction was mediated by CD8+ cells.

An immunoassay for detecting IFN-γ produced in a specifically and genetically restricted response to an antigen or antigenic peptide, comprising:

• exposing a human host to said antigen or antigenic peptide;
• obtaining a sample of plasma or serum from host;
• and
• detecting, via flow cytometry, the presence of MIG in said sample and correlating the presence of MIG in said sample to said host's production of IFN-γ in specific response to said antigenic peptide.

A method of monitoring the immunoresponsiveness of a human, comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human subject that has been exposed to a known antigen or antigenic peptide;
• culturing said sample in the presence of said known antigen or antigenic peptide for 4 to 16 hours;
• detecting, via flow cytometry, MIG expression in said cultured sample;
• and
• correlating said detection to the presence of IFN-γ secreting cells in said sample and thus to the immunoresponsiveness of said human subject;
• wherein said expression of MIG is genetically and specifically restricted, was induced by IFN-γ, and said induction was mediated by CD8+ cells.

A method of monitoring the immunoresponsiveness of a human subject, comprising:

• obtaining a sample of serum or plasma from a human subject that has been exposed to a known antigen or antigenic peptide;
• detecting MIG in said sample;
• and
• correlating said detection to the immunoresponsiveness of said human subject.

An immunoassay kit for detecting IFN-γ or IFN-γ secreting cells in a sample comprising:

• a. a sample including peripheral blood mononuclear cells;
• b. an antigen or antigenic peptide;
• c. a fluorescently labeled antibody to MIG;
• and
• d. a detector which detects said fluorescently labeled antibody via flow cytometry.

An immunoassay for detecting IFN-γ, or cells which secrete IFN-γ, comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human host which has been exposed to an immunoreactive peptide;
• culturing said sample for a period of 4 to 16 hours in the presence of said peptide;
• detecting MIG expression in said sample via flow cytometry;
• and
• correlating said detection to the presence in said sample of IFN-γ or cells that secrete IFN-γ;
• wherein said expression of MIG is genetically and specifically restricted, was induced by said IFN-γ, and said induction was mediated by CD4+ cells.

A method of assessing the effectiveness of a compound or system in inducing an immune response comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human host which has been exposed to an immunoreactive peptide;
• culturing said sample for 4 to 16 hours in the presence of said peptide.
• detecting, via flow cytometry, the induction of the immune response of said human host by up-regulation of MIG expression;
• wherein said expression of MIG is genetically and specifically restricted, was induced by IFN-γ, and said induction was mediated by CD4+ cells.

A method of monitoring the immunoresponsiveness of a human, comprising:

• obtaining a sample including peripheral blood mononuclear cells from a human subject that has been exposed to a known antigen or antigenic peptide;
• culturing said sample in the presence of said known antigen or antigenic peptide for 4 to 16 hours;
• detecting, via flow cytometry, MIG expression in said cultured sample;
• and
• correlating said detection to the presence of IFN-γ secreting cells in said sample and thus to the immunoresponsiveness of said human subject;
• wherein said expression of MIG is genetically and specifically restricted, was induced by IFN-γ, and said induction was mediated by CD4+ cells.

An immunoassay method for detecting a chemokine, a Th−1 cytokine that induces expression of said chemokine, or cells which secrete said Th−1 cytokine that induces expression of said chemokine, comprising:

• obtaining a tissue sample from a mammalian host which has been exposed to an immunoreactive substance;
• culturing said tissue sample in the presence of said immunoreactive substance;
• detecting expression of said chemokine in said sample;
• and
• correlating said detection to the presence of said Th−1 cytokine which induces expression of said chemokines, or cells that secrete said cytokine that induces expression of said chemokine.