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 . (canceled)  
     
     
         37 . A method for stimulating an immune response in a subject comprising 
 administering to a subject by intravenous or intraperitoneal route of administration a composition comprising a oligonucleotide delivery complex having an immunostimulatory CpG-containing oligonucleotide associated with a sterol or a lipid, in an amount effective to stimulate an immune response,    wherein the oligonucleotide is 8-100 bases in length.    
     
     
         38 . The method of  claim 37 , wherein the oligonucleotide is synthesized de novo.  
     
     
         39 . The method of  claim 37 , wherein the oligonucleotide has a phosphorothioate modified phosphate backbone.  
     
     
         40 . The method of  claim 37 , wherein the composition is administered by intravenous route of administration.  
     
     
         41 . The method of  claim 37 , wherein the subject has an immune system deficiency.  
     
     
         42 . The method of  claim 37 , wherein the method is a method for treating an immune system deficiency.  
     
     
         43 . The method of  claim 41  or  42 , wherein the immune system deficiency is a viral infection.  
     
     
         44 . The method of  claim 41  or  42 , wherein the immune system deficiency is cancer or a tumor.  
     
     
         45 . The method of  claim 44 , wherein the tumor or cancer is eliminated.  
     
     
         46 . The method of  claim 37 , wherein the immune response comprises increased expression of IFN-gamma.  
     
     
         47 . The method of  claim 37 , wherein the immune response comprises induction of NK activity.  
     
     
         48 . The method of  claim 37 , wherein the subject is a human, a dog, cat, horse, cow, pig, sheep, goat, chicken, monkey, rat or mouse.  
     
     
         49 . The method of  claim 37 , wherein the subject is a human.  
     
     
         50 . The method of  claim 37 , wherein the oligonucleotide comprises the formula  
         5′ X 1 X 2 CGX 3 X 4  3′ wherein C and G are unmethylated, X 1 , X 2 , X 3  and X 4  are nucleotides and a GCG trinucleotide sequence is not present at the 5′ or 3′ termini.    
     
     
         51 . A method for stimulating an immune response in a subject comprising 
 administering to a subject a composition comprising an oligonucleotide delivery complex having an immunostimulatory CpG-containing oligonucleotide associated with a sterol or a lipid, in an amount effective to stimulate an immune response,    wherein the oligonucleotide is 8-100 bases in length.    
     
     
         52 . The method of  claim 51 , wherein the oligonucleotide has a phosphodiester backbone.  
     
     
         53 . A method for stimulating an immune response in a subject having an immune system deficiency that is cancer or a tumor comprising 
 administering to a subject a composition comprising a oligonucleotide delivery complex having an immunostimulatory CpG-containing oligonucleotide associated with a sterol or a lipid, in an amount effective to stimulate an immune response to treat, prevent or ameliorate the immune system deficiency,    wherein the oligonucleotide is 8-100 bases in length.    
     
     
         54 . The method of  claim 53 , wherein the composition is administered by intravenous or intraperitoneal route of administration.  
     
     
         55 . A method for stimulating an immune response in a subject having an immune system deficiency that is a viral infection comprising 
 administering to a subject a composition comprising a oligonucleotide delivery complex having an immunostimulatory CpG-containing oligonucleotide associated with a sterol or a lipid, in an amount effective to stimulate an immune response to treat, prevent or ameliorate the immune system deficiency,    wherein the oligonucleotide is 8-100 bases in length.    
     
     
         56 . The method of  claim 37 ,  51 ,  52 ,  53  or  55 , wherein the oligonucleotide is 8-40 bases in length.  
     
     
         57 . The method of  claim 37 ,  51 ,  52 ,  53  or  55 , wherein the lipid is a cationic lipid, virosome or liposome.  
     
     
         58 . The method of  claim 57 , wherein the oligonucleotide is ionically or covalently bound to or encapsulated within the cationic lipid, virosome or liposome.  
     
     
         59 . The method of  claim 37 ,  51 ,  52 ,  53  or  55 , wherein the sterol is a cholesterol.  
     
     
         60 . The method of  claim 37 ,  51 ,  52 ,  53  or  55 , wherein the oligonucleotide comprises a CpG flanked by two 5′ purines and two 3′ pyrimidines.  
     
     
         61 . The method of  claim 37 ,  51 ,  52 ,  53  or  55 , wherein the oligonucleotide does not contain a palindrome.  
     
     
         62 . A method to elicit a systemic, non-antigen-specific immune response in a mammal, comprising administering to said mammal a therapeutic composition by a route of administration selected from the group consisting of intravenous and intraperitoneal, said therapeutic composition comprising: 
 a. a liposome delivery vehicle; and    b. an isolated nucleic acid molecule selected from the group consisting of: 
 i. an isolated nucleic acid molecule consisting of a nucleic acid molecule that does not express a peptide or protein; and  
 ii. an isolated nucleic acid vector without a gene insert, or a fragment thereof;  
 wherein said therapeutic composition elicits a systemic, non-antigen-specific immune response in said mammal.  
   
     
     
         63 . The method of  claim 62 , wherein said route of administration is intravenous.  
     
     
         64 . The method of  claim 62 , wherein said liposome delivery vehicle comprises cationic liposomes.  
     
     
         65 . The method of  claim 62 , wherein administration of said therapeutic composition elicits a systemic, anti-viral immune response in said mammal.  
     
     
         66 . The method of  claim 62 , wherein administration of said therapeutic composition elicits a systemic, anti-tumor immune response in said mammal.  
     
     
         67 . The method of  claim 62 , wherein administration of said therapeutic composition results in a reduction in a tumor in said mammal.  
     
     
         68 . The method of  claim 62 , wherein administration of said therapeutic composition increases production of IFN-gamma in said mammal.  
     
     
         69 . The method of  claim 62 , wherein administration of said therapeutic composition increases natural killer (NK) cell activity in said mammal.  
     
     
         70 . The method of  claim 62 , wherein said mammal is selected from the group consisting of humans, dogs, cats, mice, sheep, cattle, horses and pigs.  
     
     
         71 . The method of  claim 62 , wherein said mammal is a human.  
     
     
         72 . A method to elicit a systemic, non-antigen-specific immune response in a mammal, comprising administering to said mammal a therapeutic composition comprising: 
 a. a liposome delivery vehicle; and    b. an isolated non-coding nucleic acid sequence, wherein said therapeutic composition elicits a systemic, non-antigen-specific immune response in said mammal.    
     
     
         73 . The method of  claim 62 , wherein said isolated nucleic acid molecule of (i) is selected from the group consisting of: 
 1) an isolated nucleic acid molecule consisting of a nucleic acid sequence from the coding strand of a DNA molecule, wherein said molecule does not express a peptide or protein;    2) an isolated nucleic acid molecule consisting of a nucleic acid sequence from an RNA molecule, wherein said molecule does not express a peptide or protein; and,    3) a chemically synthesized nucleic acid molecule consisting of a nucleic acid sequence that is not a sequence from a naturally occurring nucleic acid molecule.    
     
     
         74 . The method of  claim 62 , wherein said isolated nucleic acid molecule is an oligonucleotide.  
     
     
         75 . The method of  claim 62 , wherein said isolated nucleic acid molecule contains CpG moieties.  
     
     
         76 . A method to elicit a systemic, non-antigen specific, immune response in a mammal that has cancer, wherein said immune response inhibits or reduces cancer growth in said mammal, said method comprising administering to said mammal a therapeutic composition comprising: 
 a. a liposome delivery vehicle; and    b. an isolated nucleic acid molecule selected from the group consisting of: 
 i. an isolated nucleic acid molecule consisting of a nucleic acid molecule that does not express a peptide or protein; and  
 ii. an isolated nucleic acid vector without a gene insert, or a fragment thereof.  
   
     
     
         77 . The method of  claim 76 , wherein said composition is administered by a route selected from the group consisting of intravenous administration, intraperitoneal administration, and direct administration to the site of said cancer.  
     
     
         78 . A method to elicit a systemic, non-antigen-specific, anti-viral immune response in a mammal, comprising administering to said mammal a therapeutic composition comprising: 
 a. a liposome delivery vehicle; and    b. an isolated nucleic acid molecule selected from the group consisting of: 
 i. an isolated nucleic acid molecule consisting of a nucleic acid molecule that does not express a peptide or protein; and  
 ii. an isolated nucleic acid vector without a gene insert, or a fragment thereof.  
   
     
     
         79 . A method to elicit an immune response in a mammal, comprising administering to said mammal a therapeutic composition, said composition comprising: 
 a. a cationic liposome delivery vehicle; and    b. at least two nucleotides joined together by a phosphodiester linkage,    wherein said nucleotides elicit said immune response by a non-antigen specific pathway.