Immunomodulatory oligonucleotides

Abstract

Oligonucleotides containing unthylated CpG dinucleotides and therapeutic utilities based on their ability to stimulate an immune response in a subject are disclosed. Also disclosed are therapies for treating diseases associated with immune system activation that are initiated by unthylated CpG dinucleotides in a subject comprising administering to the subject oligonucleotides that do not contain unmethylated CpG sequences (i.e. methylated CpG sequences or no CpG sequence) to outcompete unmethylated CpG nucleic acids for binding. Further disclosed are methylated CpG containing dinucleotides for use antisense therapies or as in vivo hybridization probes, and immunoinhibitory oligonucleotides for use as antiviral therapeutics.

1 - 36 . (Cancelled).  
     
     
         37 . A method for boosting an immune response of a subject comprising administering to the subject an isolated immunostimulatory oligonucleotide comprising X1X2CGX3×4, 
 wherein C and G are unmethylated and X1, X2, X3 and X4 are nucleotides and wherein the immunostimulatory oligonucleotide is between 6 and 100 bases in length, and  
 wherein an increase in activation of the subject's lymphocytes or NK cells indicates that the subject's immune response has been boosted.  
 
     
     
         38 . The method of  claim 37 , wherein the activation of the subject's lymphocytes or NK cells is lymphocyte proliferation.  
     
     
         39 . The method of  claim 37 , wherein the activation of the subject's lymphocytes or NK cells is IgM secretion.  
     
     
         40 . The method of  claim 37 , wherein the activation of the subject's lymphocytes or NK cells is increased expression of IL-12 and IFN-gamma.  
     
     
         41 . The method of  claim 37 , wherein the subject has an immune system deficiency.  
     
     
         42 . The method of  claim 41 , wherein the immune system deficiency is an infection.  
     
     
         43 . The method of  claim 42 , wherein the infection is a bacterial, viral, fungal or parasitic infection.  
     
     
         44 . The method of  claim 41 , wherein the immune system deficiency is a bacterial infection by bacteria having bacterial antigens and wherein the increase in lymphocyte or NK activation is activated B cell with antigen receptors specific for the bacterial antigens.  
     
     
         45 . The method of  claim 41 , wherein the immune system deficiency is cancer.  
     
     
         46 . The method of  claim 37 , wherein the immunostimulatory oligonucleotide is 8-100 bases in length.  
     
     
         47 . The method of  claim 37 , wherein the immunostimulatory oligonucleotide is 8-40 bases in length.  
     
     
         48 . The method of claim  37 - 46  or  47 , wherein the immunostimulatory oligonucleotide is administered in conjunction with a vaccine.  
     
     
         49 . The method of claim  37 - 46  or  47 , wherein the immunostimulatory oligonucleotide is not administered in conjunction with a vaccine.  
     
     
         50 . A method for stimulating an immune response in a subject, wherein increases in IFN-gamma and IL-12 expression are indicators of the immune response, comprising 
 administering to the subject an isolated immunostimulatory oligonucleotide comprising X1X2CGX3×4, wherein C and G are unmethylated and X1, X2, X3 and X4 are nucleotides and wherein the immunostimulatory oligonucleotide is between 6 and 100 bases in length, in an amount whereby IFN-gamma and IL-12 expression is increased.    
     
     
         51 . The method of  claim 50 , wherein the subject has an immune system deficiency.  
     
     
         52 . The method of  claim 51 , wherein the immune system deficiency is an infection.  
     
     
         53 . The method of  claim 52 , wherein the infection is a bacterial, viral, fungal or parasitic infection.  
     
     
         54 . The method of  claim 51 , wherein the immune system deficiency is cancer.  
     
     
         55 . The method of  claim 50 , wherein the immunostimulatory oligonucleotide is 8-100 bases in length.  
     
     
         56 . The method of  claim 50 , wherein the immunostimulatory oligonucleotide is 8-40 bases in length.  
     
     
         57 . The method of  claim 50 , wherein the immunostimulatory oligonucleotide is administered in conjunction with a vaccine.  
     
     
         58 . The method of  claim 50 , wherein the immunostimulatory oligonucleotide is not administered in conjunction with a vaccine.  
     
     
         59 . The method of  claim 37  or  50 , wherein the immunostimulatory oligonucleotide includes a nucleotide sequence consisting of 5′-purine-purine-CG-pyrimidine-pyrimidine-3′.  
     
     
         60 . The method of  claim 59 , wherein the nucleotide sequence consists of AACGTT.  
     
     
         61 . The method of  claim 37  or  50 , wherein the immunostimulatory oligonucleotide comprises GACGTC and GACGTT.  
     
     
         62 . The method of  claim 43  or  53 , wherein the infection is a viral infection.  
     
     
         63 . A method for boosting the immune responsiveness of a subject to a sensitizing antigen without immunization of the subject by the sensitizing antigen comprising 
 administering an immunostimulatory oligonucleotide to the subject,    wherein an increase in the magnitude of the subject's immune response to the sensitizing antigen indicates that the desired boost to the subject's immune responsiveness has been achieved.    
     
     
         64 . The method of  claim 63 , wherein the antigen is presented by a pathogen and the subject's immune responsiveness to an intracellular infection by the pathogen is boosted.  
     
     
         65 . A method for shifting the immune response of a subject to a sensitizing antigen toward a Th1 phenotype comprising 
 administering an immunostimulatory oligonucleotide to the subject,    wherein detection of a Th1 type immune response by the subject indicates that the desired shift to the Th1 phenotype has been achieved.    
     
     
         66 . The method of  claim 63  or  65 , wherein the immunostimulatory oligonucleotide includes a hexameric nucleotide sequence consisting of 5′-purine-purine-CG-pyrimidine-pyrimidine-3′.  
     
     
         67 . The method of  claim 66 , wherein the hexameric nucleotide sequence consists of AACGTT.  
     
     
         68 . The method of  claim 63  or  65 , wherein the hexameric nucleotide sequence is selected from the group of sequences consisting of GACGTC and GACGTT.  
     
     
         69 . The method of  claim 65 , wherein the subject is suffering from an intracellular infection by a pathogen and the shift to the Th1 phenotype strengthens the subject's immune response to the pathogen.  
     
     
         70 . The method of  claim 64  or  69 , wherein the pathogen is a virus.  
     
     
         71 . The method of  claim 63  or  65 , wherein the desired result is measured by detecting in a sample containing lymphocytes obtained from the immunostimulatory oligonucleotide treated subject higher levels of IL-12 and/or IFN-gamma in the immunostimulatory oligonucleotide treated subject as compared to an antigen-challenged control.