Methods of characterizing host responsiveness to interferon by ex vivo induction of interferon-responsive markers

1. A method for treating a subject having hepatitis with an interferon-based therapy, the method comprising:

(A) having at least a first and second sample of whole blood from said subject sent to a laboratory prior to interferon administration, for the laboratory to perform an assay comprising the following steps (1)-(4):

(1) exposing said first sample of whole blood from said subject to interferon alpha 2b in a solvent for an amount of time sufficient for said interferon alpha 2b to alter the expression of one or more markers whose expression is potentially altered in response to exposure to IFN alpha 2b selected from the group consisting of TNFSF10, CXCL10, GBP, and BST2;

(2) exposing said second sample of whole blood from said subject to the solvent without said interferon alpha 2b for said amount of time;

(3) quantifying the effect of said interferon alpha 2b as a change in expression of said one or more markers by a method comprising:

(i) isolating mRNA from said first whole blood sample,

(ii) contacting said mRNA from said first whole blood sample with a reverse transcriptase to generate a stimulated complementary DNA (cDNA), and

(iii) contacting said stimulated cDNA with sense and antisense primers that are specific for said one or more markers and a DNA polymerase to generate stimulated amplified DNA; and

(iv) isolating mRNA from said second whole blood sample,

(v) contacting said mRNA from said second whole blood sample with a reverse transcriptase to generate control complementary DNA (cDNA), and

(vi) contacting said control cDNA with sense and antisense primers that are specific for said one or more markers and a DNA polymerase to generate control amplified DNA;

(4) calculating a change in the amount of marker mRNA in said first whole blood sample compared to the amount of marker mRNA in said second whole blood sample;

(B) treating the subject with an interferon-based therapy if the results of said assay indicate there is a difference in the amount of marker mRNA quantified from said first blood sample as compared to said second blood sample.

2. The method of claim 1 , wherein said one or more markers comprise a marker that increases in response to interferon alpha 2b.

3. The method of claim 1 , wherein said one or more markers comprise a marker that decreases in response to interferon alpha 2b.

4. The method of claim 1 , wherein said interferon alpha 2b is present in a concentration from about 1 to about 100,000 units per mL.

5. The method of claim 1 , wherein said exposing is for a time between one hour and seven hours and wherein said exposing occurs at about thirty-seven (37) degrees Celsius.

6. The method of claim 1 , wherein said whole blood comprises human blood that is optionally heparinzed.

7. The method of claim 1 , wherein said hepatitis is caused by the hepatitis B virus or the hepatitis C virus.

8. The method of claim 1 , wherein a medical professional administers the recommended interferon-based therapy.

9. A method for treating a subject having hepatitis with interferon therapy based on the ongoing efficacy of the interferon therapy, the method comprising:

having a first sample of whole blood collected from said subject prior to interferon alpha 2b administration sent to a laboratory;

having a second sample of whole blood collected from said subject after interferon alpha 2b administration sent to a laboratory, for the laboratory to perform an assay comprising the following steps (1)-(2):

(1) quantifying the effect of one or more markers whose expression is potentially altered in response to exposure to IFN2 alpha b selected from the group consisting of TNFSF10, CXCL10, GBP, and BST2 by a method comprising:

(i) isolating mRNA from said first whole blood sample,

(ii) contacting said mRNA from said first whole blood sample with a reverse transcriptase to generate a first complementary DNA (cDNA), and

(iii) contacting said first cDNA with sense and antisense primers that are specific for said one or more markers and a DNA polymerase to generate a first amplified DNA; and

(iv) isolating mRNA from said second whole blood sample,

(v) contacting said mRNA from said second whole blood sample with a reverse transcriptase to generate a second complementary DNA (cDNA), and

(vi) contacting said second cDNA with sense and antisense primers that are specific for said one or more markers and a DNA polymerase to generate a second amplified DNA;

(2) determining the efficacy of interferon alpha 2b therapy by comparing the expression of said one or more markers in said first whole blood sample and in said second whole blood sample by comparing the amount of said first and said second amplified DNA; and

(I) continuing said interferon alpha 2b therapy based on the efficacy of the therapy, said efficacy being indicated by a difference in the amount of marker mRNA quantified from said second blood sample as compared to said first blood sample in results from said assay; or

(II) discontinuing said interferon therapy based on a lack of efficacy of the therapy, said lack of efficacy being indicated by a lack of difference in the amount of marker mRNA quantified from said second blood sample as compared to said first blood sample in results from said assay.

10. The method of claim 9 , wherein said hepatitis is hepatitis B virus or hepatitis C virus.

11. The method of claim 1 , further comprising comparing said difference by comparing to a normal effect of interferon expression of said one or more markers, wherein said normal effect is calculated as an average change in expression levels of said one or more markers in a panel of control individuals.

12. The method of claim 1 , wherein said difference is a statistically significant difference.