Method for detecting antigen specific or mitogen-activated T cells
The present invention comprises a method for the quantitative or qualitative detection of antigen-specific CD4+ T cells and/or CD8+ T cells in a subject, said method comprising quantitatively or qualitatively detecting the expression of cell surface marker CD25 and one or more of cell surface markers CD134 and CD137 in a suitable lymphocyte-containing sample from said subject in response to exposure to an antigen. A method for determining the immunocompetence of a subject and a method for isolating antigen-specific CD4+ and/or CD8+ T cells is also disclosed.
1. A method for the quantitative or qualitative detection of antigen-specific CD4+ T cells in a subject, said method comprising quantitatively or qualitatively detecting co-expression of cell surface markers CD4, CD25 and CD134 by a cell in a whole blood sample from said subject following in vitro exposure of the whole blood sample to an antigen.
2. The method of claim 1 comprising the following steps:
(i) culturing the whole blood sample from the subject in the presence of the antigen; and
(ii) quantitatively or qualitatively detecting co-expression of CD4, CD25 and CD134 by a cell in the cultured whole blood sample.
3. The method of claim 1 wherein the antigen is selected from the group consisting of tuberculin, Hepatitis C Virus (HCV) core antigen, HCV nonstructural protein 3 (NS3), cytomegalovirus (CMV) phosphoprotein 65 (pp65), CMV lysate, Herpes Simplex Virus (HSV)-1 lysate, HSV-2 lysate, vaccinia lysate, tetanus toxoid (TT), purified protein derivative (PPD) from Mycobacterium tuberculosis, Streptococcus antigen streptokinase, Human Immunodeficiency virus (HIV)-1 p24, and pools of overlapping peptides from HIV-1 Gag, Env, Pol or other HIV-1 accessory proteins.
4. A method for measuring CD 4 + T-cell mediated immunocompetence of a subject, said method comprising quantitatively or qualitatively detecting the co-expression of cell surface markers CD4, CD25 and CD134 by a cell in a whole blood sample from said subject following in vitro exposure of the whole blood sample to an antigen, wherein the detection of cells co-expressing CD4, CD25 and CD134 is indicative of the subject having CD 4 + T cell-mediated immunocompetence.
5. The method of claim 4 comprising the following steps:
(i) culturing the whole blood sample from the subject in the presence of the antigen; and
(ii) quantitatively or qualitatively detecting co-expression of CD4, CD25 and CD134 by a cell in the cultured whole blood sample.
6. The method of claim 4 wherein said step of culturing consists of culturing the whole blood sample in the presence of the antigen.
7. The method of claim 4 wherein the antigen is selected from the group consisting of tuberculin, Hepatitis C Virus (HCV) core antigen, HCV nonstructural protein 3 (NS3), cytomegalovirus (CMV) lysate, CMV phosphoprotein 65 (pp65), Herpes Simplex Virus (HSV)-1 lysate, HSV-2 lysate, vaccinia lysate, tetanus toxoid (TT), purified protein derivative (PPD) from Mycobacterium tuberculosis, Streptococcus antigen streptokinase, Human Immunodeficiency virus (HIV)-1 p24, and pools of overlapping peptides from HIV-1 Gag, Env, Pol or other HIV-1 accessory proteins.
8. The method of claim 1 wherein the detecting step is performed within about 24 to 48 hours of commencement of the culturing step.
9. The method of claim 1 wherein the detecting step is performed within about 40 to 44 hours of commencement of the culturing step.
10. The method of claim 1 wherein the whole blood sample is a heparinised whole blood sample.
11. The method of claim 1 wherein the detecting step comprises the use of at least one monoclonal antibody selected from the group of monoclonal antibodies which specifically bind to one of CD25 and CD134.
12. The method of claim 1 wherein the detecting step comprises the use of flow cytometry.
13. The method of claim 1 wherein the method further comprises a step of isolating cells co-expressing CD4, CD25 and CD134.
14. The method of claim 13 wherein the isolating step comprises the use of at least one monoclonal antibody selected from the group of monoclonal antibodies which specifically bind to one of CD25 and CD134.
15. The method of claim 13 wherein the isolating step comprises the use of flow cytometry.
16. The method of claim 1 wherein the subject is human.