Methods and products related to treatment and prevention of hepatitis c virus infection

Abstract

The invention provides methods for identifying and treating subjects having hepatitis C infections. In some instances, the subjects are those that are non-responsive to non-CpG therapy. Preferably, the subjects are treated with C class CpG immunostimulatory nucleic acids having semi-soft backbone.

1 . A method of treating a subject having an HCV infection that was not successfully treated using a previous non-CpG therapy comprising
 administering to a subject in need of such treatment a CpG immunostimulatory nucleic acid in an amount effective to treat the infection.   
     
     
         2 . The method of  claim 1 , wherein the non-CpG therapy includes interferon-alpha. 
     
     
         3 . The method of  claim 2 , wherein the interferon-alpha is interferon-alpha-2b, interferon-alpha-2a or consensus interferon-alpha. 
     
     
         4 . The method of  claim 2 , wherein the non-CpG therapy includes interferon-alpha and Ribavirin. 
     
     
         5 . The method of  claim 2 , wherein the non-CpG therapy includes pegylated interferon-alpha and Ribavirin. 
     
     
         6 . The method of  claim 1 , wherein the CpG immunostimulatory nucleic acid is an A class CpG immunostimulatory nucleic acid. 
     
     
         7 . The method of  claim 1 , wherein the CpG immunostimulatory nucleic acid is a B class CpG immunostimulatory nucleic acid 
     
     
         8 . The method of  claim 1 , wherein the CpG immunostimulatory nucleic acid is a C class CpG immunostimulatory nucleic acid. 
     
     
         9 . The method of  claim 1 , further comprising the step of administering interferon-alpha to the subject. 
     
     
         10 . The method of  claim 9 , wherein the interferon-alpha is interferon-alpha-2b, interferon-alpha-2a or consensus interferon alpha. 
     
     
         11 . The method of  claim 9 , wherein the interferon-alpha is administered substantially simultaneously with the CpG immunostimulatory nucleic acid. 
     
     
         12 . The method of  claim 1 , wherein the CpG immunostimulatory nucleic acid comprises a backbone modification. 
     
     
         13 . The method of  claim 12 , wherein the backbone modification is a phosphorothioate backbone modification. 
     
     
         14 . The method of  claim 1 , wherein the CpG immunostimulatory nucleic acid comprises a semi-soft backbone. 
     
     
         15 . A method of treating a subject having an HCV infection and likely to be non-responsive to a non-CpG therapy comprising
 administering to a subject in need of such treatment a CpG immunostimulatory nucleic acid in an amount effective to treat the infection.   
     
     
         16 . The method of  claim 15 , further comprising identifying a subject likely to be non-responsive to a non-CpG therapy. 
     
     
         17 . The method of  claim 16 , wherein the subject is identified as likely to be non-responsive based on an assay of interferon-alpha produced per dendritic cell. 
     
     
         18 . The method of  claim 16 , wherein the subject is identified as likely to be non-responsive based on HCV genotype. 
     
     
         19 . The method of  claim 15 , wherein the non-CpG therapy includes interferon-alpha. 
     
     
         20 . The method of  claim 15 , wherein the non-CpG therapy includes interferon-alpha and Ribavirin. 
     
     
         21 . The method of  claim 20 , further comprising administering to the subject an anti-viral agent. 
     
     
         22 . The method of  claim 21 , wherein the anti-viral agent is interferon-alpha. 
     
     
         23 . The method of  claim 22 , wherein the interferon-alpha is interferon-alpha-2b, interferon-alpha-2a or consensus interferon alpha. 
     
     
         24 . The method of  claim 21 , wherein interferon-alpha administered in a sub-therapeutic amount. 
     
     
         25 . The method of  claim 15 , wherein the CpG immunostimulatory nucleic acid is a C class CpG immunostimulatory nucleic acid. 
     
     
         26 . The method of  claim 15 , wherein the CpG immunostimulatory nucleic acid comprises a semi-soft backbone. 
     
     
         27 . A method for screening CpG immunostimulatory nucleic acids useful in the treatment of chronic hepatitis C viral infection comprising
 contacting peripheral blood mononuclear cells from a subject having a chronic hepatitis C viral infection, with a CpG immunostimulatory nucleic acid, and   measuring a test response of the blood mononuclear cells after exposure, wherein the subject was not successfully treated using a previous therapy.   
     
     
         28 . The method of  claim 27 , wherein the test response is selected from the group consisting of B cell stimulation, secretion of IL-6, secretion of IL-10, secretion of IL-12, secretion of interferon-gamma, secretion of type 1 interferons (alpha +beta), secretion of IP-10, NK activity, expression of CD80, expression of CD 86, expression of CD83, and upregulation of class II MHC expression. 
     
     
         29 . The method of  claim 27 , wherein the peripheral blood mononuclear cells comprise dendritic cells. 
     
     
         30 . The method of  claim 29 , wherein the dendritic cells comprise plasmacytoid dendritic cells. 
     
     
         31 . The method of  claim 29 , wherein the test response is selected from the group consisting of secretion of IL-12, secretion of type 1 interferons, expression of CD80, expression of CD 86, expression of CD83, and upregulation of class II MHC expression. 
     
     
         32 . The method of  claim 29 , wherein the contacting occurs in vitro. 
     
     
         33 . The method of  claim 32 , wherein the peripheral blood mononuclear cells are cultured. 
     
     
         34 . The method of  claim 33 , wherein the CpG immunostimulatory nucleic acid is added to the cultured peripheral blood mononuclear cells. 
     
     
         35 . The method of  claim 29 , wherein the previous therapy is a non-CpG therapy. 
     
     
         36 . The method of  claim 29 , wherein the previous therapy is therapy with a CpG nucleic acid of a different sequence or class. 
     
     
         37 . The method of  claim 29 , further comprising screening the CpG immunostimulatory nucleic acid for the ability to stimulate a control response from peripheral blood mononuclear cells from a normal subject. 
     
     
         38 . The method of  claim 29 , further comprising contacting peripheral blood mononuclear cells to interferon-alpha substantially simultaneously with the CpG immunostimulatory nucleic acid. 
     
     
         39 . The method of  claim 29 , wherein the CpG immunostimulatory nucleic acid is a C class CpG immunostimulatory nucleic acid. 
     
     
         40 . A method for identifying a subject having an HCV infection and likely to be non-responsive to a non-CpG therapy comprising
 exposing peripheral blood mononuclear cells harvested from a subject having a hepatitis C viral infection to a CpG immunostimulatory nucleic acid,   measuring interferon-alpha produced from the cells, and   determining an amount of interferon-alpha produced per dendritic cell, wherein an amount that is below 1.0 pg/ml is indicative of a subject that is likely to be non-responsive to a non-CpG therapy.   
     
     
         41 . The method of  claim 40 , wherein an amount that is below 0.5 pg/ml is indicative of a subject that is likely to be non-responsive to a non-CpG therapy. 
     
     
         42 . The method of  claim 40 , wherein the non-CpG therapy comprises interferon-alpha. 
     
     
         43 . The method of  claim 42 , wherein the non-CpG therapy comprises Ribavirin. 
     
     
         44 . The method of  claim 42 , wherein the IFN-alpha is pegylated interferon-alpha. 
     
     
         45 . The method of  claim 40 , wherein the CpG immunostimulatory nucleic acid is an A class or a C class CpG immunostimulatory nucleic acid. 
     
     
         46 . The method of  claim 40 , wherein the peripheral blood mononuclear cells are further exposed to an anti-viral agent together with a CpG immunostimulatory nucleic acid. 
     
     
         47 . The method of  claim 46 , wherein the anti-viral agent is interferon-alpha. 
     
     
         48 . The method of  claim 47 , wherein interferon-alpha is interferon-alpha-2b, interferon-alpha-2a or consensus interferon alpha. 
     
     
         49 . The method of  claim 40 , wherein the peripheral blood mononuclear cells comprise dendritic cells. 
     
     
         50 . The method of  claim 49 , wherein the dendritic cells comprise plasmacytoid dendritic cells. 
     
     
         51 . The method of  claim 40 , wherein the hepatitis C viral infection is an acute hepatitis C viral infection. 
     
     
         52 . The method of  claim 40 , further comprising determining a genotype of the HCV. 
     
     
         53 . A method of treating a subject having a hepatitis C viral infection comprising
 administering to a subject identified according to the method of  claim 40  a CpG immunostimulatory nucleic acid molecule in an amount effective to treat the infection.   
     
     
         54 . The method of  claim 53 , further comprising administering to the subject interferon-alpha. 
     
     
         55 . The method of  claim 54 , wherein the interferon-alpha is interferon-alpha-2b, interferon-alpha-2a or consensus interferon-alpha. 
     
     
         56 . The method of  claim 53 , wherein the CpG immunostimulatory nucleic acid is an A class CpG immunostimulatory nucleic acid. 
     
     
         57 . The method of  claim 53 , wherein the CpG immunostimulatory nucleic acid is a B class CpG immunostimulatory nucleic acid. 
     
     
         58 . The method of  claim 53 , wherein the CpG immunostimulatory nucleic acid is a C class CpG immunostimulatory nucleic acid. 
     
     
         59 . The method of  claim 53 , wherein the CpG immunostimulatory nucleic acid comprises a backbone modification. 
     
     
         60 . The method of  claim 59  wherein the backbone modification is a phosphorothioate backbone modification. 
     
     
         61 . The method of  claim 53 , wherein the CpG immunostimulatory nucleic acid comprises a semi-soft backbone. 
     
     
         62 . The method of  claim 53 , wherein the hepatitis C viral infection is a chronic hepatitis C viral infection. 
     
     
         63 . The method of  claim 53 , wherein the hepatitis C viral infection is an acute hepatitis C viral infection. 
     
     
         64 . A method of treating a subject having an HCV infection that was not successfully treated using a previous non-CpG therapy comprising
 administering to a subject in need of such treatment a C class CpG immunostimulatory nucleic acid having a semi-soft backbone in an amount effective to treat the infection.   
     
     
         65 . A method of treating a subject having an HCV infection and likely to be non-responsive to a non-CpG therapy comprising
 administering to a subject in need of such treatment a C class CpG immunostimulatory nucleic acid having a semi-soft backbone in an amount effective to treat the infection.   
     
     
         66 . A method for identifying a subject having an HCV infection and likely to be non-responsive to a non-CpG therapy comprising
 exposing peripheral blood mononuclear cells harvested from a subject having a hepatitis C viral infection to a A class or a C class CpG immunostimulatory nucleic acid,   measuring interferon-alpha produced from the cells, and   determining an amount of interferon-alpha produced per dendritic cell, wherein an amount that is below 1.0 pg/ml is indicative of a subject that is likely to be non-responsive to a non-CpG therapy.   
     
     
         67 . A method of treating a subject having an HCV infection that was not successfully treated using a previous non-CpG therapy comprising
 contacting peripheral blood mononuclear cells from a subject in need of such treatment, with a CpG immunostimulatory nucleic acid in an amount effective to stimulate an immune response, and   re-infusing the cells into the subject.   
     
     
         68 . The method of  claim 67 , wherein the peripheral blood mononuclear cells comprise dendritic cells. 
     
     
         69 . The method of  claim 68 , wherein the dendritic cells comprise plasmacytoid dendritic cells. 
     
     
         70 . The method of  claim 67 , wherein the CpG immunostimulatory nucleic acid is a C class immunostimulatory nucleic acid. 
     
     
         71 . The method of claim70, wherein the C class immunostimulatory nucleic acid has a semi-soft backbone.