Immunostimulatory nucleic acid molecules

Abstract

Nucleic acids containing unmethylated CpG dinucleotides and therapeutic utilities based on their ability to stimulate an immune response and to redirect a Th2 response to a Th1 response in a subject are disclosed. Methods for treating atopic diseases, including atopic dermatitis, are disclosed.

1 .- 18 . (canceled)  
     
     
         19 . A method for stimulating an immune response in a subject comprising 
 administering to a subject a composition comprising a nucleic acid delivery complex having a CpG containing immunostimulatory nucleic acid associated with a sterol or a lipid, in an amount effective to prevent or treat an allergy,    wherein the nucleic acid is 8-100 bases in length.    
     
     
         20 . The method of  claim 19 , wherein the composition is administered by intravenous or intraperitoneal route of administration.  
     
     
         21 . The method of  claim 19  or  20 , wherein the nucleic acid is 8-40 bases in length.  
     
     
         22 . The method of  claim 19  or  20 , wherein the lipid is a cationic lipid, virosome or liposome.  
     
     
         23 . The method of  claim 22 , wherein the nucleic acid is ionically or covalently bound to or encapsulated within the cationic lipid, virosome or liposome.  
     
     
         24 . The method of  claim 19  or  20 , wherein the sterol is a cholesterol.  
     
     
         25 . The method of  claim 19  or  20 , wherein the subject is a human.  
     
     
         26 . The method of  claim 19  or  20 , wherein the nucleic acid is synthetic.  
     
     
         27 . The method of  claim 19  or  20 , wherein the nucleic acid contains a CpG motif of  
         5′ X1X2CGX3X4 3′ wherein X1, X2, X3 and X4 are nucleotides.    
     
     
         28 . The method of  claim 27 , wherein X1X2 is selected from the group consisting of GpT, GpG, GpA and ApA and/or the X3X4 is selected from the group consisting of TpT, CpT and GpT.  
     
     
         29 . The method of  claim 19  or  20 , wherein the nucleic acid does not include a GCG trinucleotide sequence at a 5′ and/or a 3′ terminal.  
     
     
         30 . The method of  claim 27 , wherein the nucleic acid does not include a GCG trinucleotide sequence at a 5′ and/or a 3′ terminal.  
     
     
         31 . The method of  claim 19  or  20 , wherein the nucleic acid does not contain a 5′ X1X2CGX3X4 3′ palindrome.  
     
     
         32 . The method of  claim 27 , wherein the nucleic acid does not contain a 5′ X1X2CGX3X4 3′ palindrome.  
     
     
         33 . The method of  claim 19  or  20 , wherein the nucleic acid comprises a CpG flanked by two 5′ purines and two 3′ pyrimidines.  
     
     
         34 . The method of  claim 27 , wherein the nucleic acid comprises a CpG flanked by two 5′ purines and two 3′ pyrimidines.  
     
     
         35 . The method of  claim 19  or  20 , wherein the nucleic acid is a stabilized nucleic acid.  
     
     
         36 . The method of  claim 19  or  20 , wherein the nucleic acid is a phosphorothioate modified nucleic acid.  
     
     
         37 . The method of  claim 27 , wherein the nucleic acid is a phosphorothioate modified nucleic acid.  
     
     
         38 . The method of  claim 19  or  20 , wherein the allergy is allergic rhinitis.  
     
     
         39 . The method of  claim 19  or  20 , wherein the allergy is food allergy.  
     
     
         40 . A method to elicit a systemic, non-antigen-specific, immune response in a mammal, wherein said immune response reduces allergic inflammation in said 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.  
   
     
     
         41 . A method comprising administering a composition comprising a nucleic acid delivery complex to a subject, wherein the nucleic acid delivery complex comprises a CpG containing immunostimulatory nucleic acid associated with a targeting means, wherein the nucleic acid is 8-100 nucleotides in length.  
     
     
         42 . The method of  claim 41 , further comprising modulating an immune response in the subject.  
     
     
         43 . The method of  claim 42 , wherein modulating the immune response comprises stimulating an immune response.  
     
     
         44 . The method of  claim 42 , wherein modulating the immune response comprises suppression of an immune response.  
     
     
         45 . The method of  claim 41 , further comprising activating lymphocytes in said subject.  
     
     
         46 . The method of  claim 41 , further comprising boosting an immune response in the subject.  
     
     
         47 . The method of  claim 41 , wherein the nucleic acid is 8-40 nucleotides in length.  
     
     
         48 . The method of  claim 41 , wherein the nucleic acid is represented by the following formula:  
         5′X1X2CGX3X4 3′ 
       wherein C and G are unmethylated, X1, X2, X3, and X4 are nucleotides.  
     
     
         49 . The method of  claim 48 , wherein the nucleic acid does not include a GCG trinucleotide sequence at or near the 5′ and 3′ termini.  
     
     
         50 . The method of  claim 43 , wherein the subject has an immune system deficiency.  
     
     
         51 . The method of  claim 50 , wherein the immune system deficiency is cancer.  
     
     
         52 . The method of  claim 51 , wherein the cancer is a tumor of the brain, lung, ovary, breast, prostate or colon, or carcinoma or sarcoma.  
     
     
         53 . The method of  claim 50 , wherein the immune system deficiency is an infection.  
     
     
         54 . The method of  claim 53 , wherein the infection is a viral, fungal, bacterial or parasitic infection.  
     
     
         55 . The method of  claim 42 , wherein the subject has an allergy.  
     
     
         56 . The method of  claim 55 , wherein the allergy is caused by an allergen.  
     
     
         57 . The method of  claim 56 , wherein the allergen is a pollen, an insect venom, animal dander, dust or a drug.  
     
     
         58 . The method of  claim 42 , wherein the subject has an autoimmune disease.  
     
     
         59 . The method of  claim 58 , wherein the autoimmune disease involves an immune response to self antigens.  
     
     
         60 . The method of  claim 41 , wherein the composition is administered by oral, transdermal, subcutaneous, intravenous, parenteral, intraperitoneal or intrathecal route of administration.  
     
     
         61 . The method of  claim 41 , wherein the targeting means is a sterol, a lipid, or a target cell specific binding agent.  
     
     
         62 . The method of  claim 61 , wherein the sterol is cholesterol.  
     
     
         63 . The method of  claim 61 , wherein the lipid is a cationic lipid, virosome or liposome.  
     
     
         64 . The method of  claim 41 , wherein the nucleic acid is ionically or covalently bound to or encapsulated within the targeting means.  
     
     
         65 . The method of claim  1 , wherein the nucleic acid is a stabilized nucleic acid.  
     
     
         66 . The method of claim  1 , wherein the nucleic acid is a phosphorothioate modified nucleic acid.  
     
     
         67 . The method of claim  1 , wherein the nucleic acid does not contain a 5′ X1X2CGX3X4 3′ palindrome.  
     
     
         68 . A composition comprising a nucleic acid delivery complex, wherein the nucleic acid delivery complex comprises a CpG containing immunostimulatory nucleic acid associated with a targeting means, wherein the nucleic acid is 8-100 nucleotides in length, and wherein said composition is capable of boosting an immune response in a subject.  
     
     
         69 . The composition of  claim 68 , wherein the nucleic acid is 8-40 nucleotides in length.  
     
     
         70 . The composition of  claim 68 , wherein the nucleic acid is represented by the following formula:  
         5′ X1X2CGX3X4 3′ 
       wherein C and G are unmethylated, X1, X2, X3, and X4 are nucleotides.  
     
     
         71 . The composition of  claim 70 , wherein the nucleic acid does not include a GCG trinucleotide sequence at or near the 5′ and 3′ termini.  
     
     
         72 . The composition of  claim 68 , wherein the nucleic acid is synthetic.  
     
     
         73 . The composition of  claim 68 , wherein the nucleic acid is a phosphorothioate modified nucleic acid.  
     
     
         74 . The composition of  claim 68 , wherein the immune response comprises induction of NK activity, activation of macrophages and/or activation of dendritic cells.  
     
     
         75 . The composition of  claim 68 , wherein the nucleic acid is ionically or covalently bound to or encapsulated within the targeting means.  
     
     
         76 . The composition of  claim 68  or  75 , wherein the targeting means is a sterol, a lipid or a target cell specific binding agent.  
     
     
         77 . The composition of  claim 76 , wherein the lipid is a cationic lipid, a virosome or a liposome.  
     
     
         78 . The composition of  claim 68 , further comprising a vaccine that comprises an antigen.  
     
     
         79 . The composition of  claim 78 , further comprising an adjuvant.  
     
     
         80 . The composition of  claim 78 , wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, allergens, and antigens derived from an infectious organism.  
     
     
         81 . The composition of  claim 80 , wherein the nucleic acid and the antigen are encapsulated.  
     
     
         82 . The composition of  claim 78 , wherein the targeting means is a lipid that is a cationic lipid, virosome or liposome.  
     
     
         83 . The composition of  claim 78 , wherein the nucleic acid is synthetic.  
     
     
         84 . The composition of  claim 68  or  78 , wherein the nucleic acid is bacterial DNA.  
     
     
         85 . The composition of  claim 68  or  78 , wherein the nucleic acid is a ribonucleotide or an oligodeoxyribonucleotide, that is single stranded or double stranded.  
     
     
         86 . A method for stimulating an immune response in a subject comprising administering to a subject a composition comprising a vaccine and a nucleic acid delivery complex, wherein the nucleic acid delivery complex comprises a CpG containing immunostimulatory nucleic acid associated with a targeting means, wherein the nucleic acid is 8-100 nucleotides in length, and wherein the vaccine comprises an antigen.  
     
     
         87 . The method of  claim 86 , further comprising an adjuvant.  
     
     
         88 . The method of  claim 86 , wherein the nucleic acid is 8-40 nucleotides in length.  
     
     
         89 . The method of  claim 86 , wherein the nucleic acid is ionically or covalently bound to, or encapsulated within the targeting means.  
     
     
         90 . The method of  claim 86 , wherein the composition is administered by oral, transdermal, subcutaneous, intravenous, parenteral, intraperitoneal or intrathecal route of administration.  
     
     
         91 . The method of  claim 86 , wherein the subject has an immune system deficiency.  
     
     
         92 . The method of  claim 91 , wherein the immune system deficiency is cancer or infection.  
     
     
         93 . The method of  claim 92 , wherein the cancer is a tumor of the brain, lung cancer, breast, ovary, prostate, or colon, or a carcinoma or a sarcoma.  
     
     
         94 . The method of  claim 92 , wherein the infection is a viral, bacterial, fungal or parasitic infection.  
     
     
         95 . A method for treating a subject having cancer comprising administering to the subject a cancer treatment and a nucleic acid delivery complex comprising a CpG containing immunostimulatory nucleic acid associated with a targeting means, wherein an immune response in the subject is boosted.  
     
     
         96 . The method of  claim 95 , wherein the cancer treatment is chemotherapy or immunotherapy.  
     
     
         97 . The method of  claim 96 , wherein the nucleic acid delivery complex is administered before the chemotherapy or immunotherapy.  
     
     
         98 . The method of  claim 95 , wherein the targeting means is a sterol, a lipid, or a target cell specific binding agent.  
     
     
         99 . The method of  claim 98 , wherein the lipid is a cationic lipid, virosome or liposome.  
     
     
         100 . The method of  claim 98 , wherein the nucleic acid is an oligodeoxyribonucleotide or bacterial DNA.  
     
     
         101 . The method of  claim 98 , wherein the nucleic acid does not contain a 5′ X1X2CGX3X4 3′ palindrome.  
     
     
         102 . A method comprising 
 administering to a subject a nucleic acid delivery complex comprising a CpG containing immunostimulatory nucleic acid associated with a targeting means, wherein the nucleic acid is 8-100 nucleotides in length, in an amount effective to speed recovery of bone marrow after chemotherapy or immunotherapy.    
     
     
         103 . The method of  claim 68 , wherein the nucleic acid does not contain a 5′ X1X2CGX3X4 3′ palindrome.  
     
     
         104 . The method of  claim 86 , wherein the nucleic acid and the antigen are encapsulated.  
     
     
         105 . A method comprising: administering a composition comprising at least one ligand for a pattern recognition receptor molecule and a delivery vehicle to a subject.  
     
     
         106 . The method of  claim 105 , wherein a ligand for a pattern recognition receptor comprises a ligand for a signaling pattern recognition receptor.  
     
     
         107 . The method of  claim 106 , wherein said signaling pattern recognition receptor comprises at least one receptor selected from the group consisting of Toll-like receptors TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9, TLR-10, TLR-11 and TLR-12 and mannan-binding lectins, and macrophage mannose receptor and scavenger receptors.  
     
     
         108 . The method of  claim 107 , wherein said ligand comprises a ligand for TLR-2, TLR-3 and/or TLR-9.  
     
     
         109 . The method of  claim 106 , further comprising modulating an immune response in said subject.  
     
     
         110 . The method of  claim 109 , wherein modulating an immune response comprises augmenting an immune response.  
     
     
         111 . The method of  claim 109 , wherein modulating an immune response comprises down regulating an immune response.  
     
     
         112 . The method of  claim 109 , wherein modulating an immune response comprises augmenting an immune response in a subject disposed of cancer.  
     
     
         113 . The method of  claim 112 , wherein cancer comprises one or more selected from the group consisting of lung cancer, skin cancer, liver cancer, bone marrow cancer, leukemia, ovarian cancer, breast cancer, prostate cancer, colon cancer, lymphoma, brain cancer, renal cell cancer, and cancers of mesenchymal tissues.  
     
     
         114 . The method of  claim 109 , wherein modulating an immune response comprises augmenting an immune response in a subject disposed of an infectious disease.  
     
     
         115 . The method of  claim 114 , wherein said infectious disease is caused by one or more organisms selected from the group consisting of a viral pathogen, a fungal pathogen, a bacterial pathogen, a rickettsial pathogen, a parasitic pathogen and a prion pathogen.  
     
     
         116 . The method of  claim 109 , wherein modulating an immune response comprises augmenting or suppressing an immune response in a subject disposed of an allergic disease.  
     
     
         117 . The method of  claim 116 , wherein said allergic disease is caused by an abnormal immune response against an endogenous non-self antigen.  
     
     
         118 . The method of  claim 117 , wherein said non-self antigens comprise at least one of the group consisting of inhaled allergens, cutaneous allergens and oral allergens.  
     
     
         119 . The method of  claim 109 , wherein modulating an immune response comprises modulating an immune response in a subject disposed of an autoimmune disease.  
     
     
         120 . The method of  claim 119 , wherein said autoimmune disease is caused by an abnormal immune response against self antigens.  
     
     
         121 . The method of  claim 105 , wherein said administration comprises administration by at least one route selected from the group consisting of intravenously, intraperitoneally, by inhalation, subcutaneously, intradermally, intranodally, intramuscularly, intranasally, orally, rectally, intravaginally, intravesicularly, intraocularly, and topically.  
     
     
         122 . A method comprising: administering an agent capable of inducing an immune response against a specific cell type wherein said agent is capable of inducing an immune response against that cell type and inhibiting the normal or abnormal function of that cell type.  
     
     
         123 . A composition comprising: a ligand for the pattern recognition molecule family of receptors; and a delivery vehicle wherein said composition is capable of inducing an immune response in a subject.  
     
     
         124 . The composition of  claim 123 , wherein inducing an immune response comprises inducing an innate immune response.  
     
     
         125 . The composition of  claim 124 , wherein the innate immune response comprises an innate immune response by macrophages, neutrophils, NK cells, and/or dendritic cells.  
     
     
         126 . The composition of  claim 123 , wherein the delivery vehicle comprises a liposome.  
     
     
         127 . The composition of  claim 126 , wherein said liposome comprises at least one liposome selected from the group consisting of a positively charged liposome; a negatively charged liposome; and a neutral liposome.  
     
     
         128 . The composition of  claim 127 , wherein said positively charged liposome is complexed to a ligand for the pattern recognition molecule family of receptors.  
     
     
         129 . The composition of  claim 123 , wherein the delivery vehicle is non-liposomal.  
     
     
         130 . The composition of  claim 129 , wherein the non-liposomal delivery vehicle comprises at least one vehicle selected from the group consisting of polypeptides, polyamines, chitosan, PEI, polyglutamic acid, protamine sulfate, and microspheres.  
     
     
         131 . The composition of  claim 126 , wherein said ligand comprises a TLR ligand.  
     
     
         132 . The composition of  claim 131 , wherein the TLR ligand comprises any portion of a bacterium.  
     
     
         133 . The composition of  claim 132 , wherein any portion of a bacterium further comprises any portion of a bacterium that associates with a TLR.  
     
     
         134 . The composition of  claim 133 , wherein said TLR ligand comprises any portion of a a bacterium that associates with at least one of the following consisting of TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9, TLR-10, TLR-11 and TLR-12.  
     
     
         135 . The composition of  claim 131 , wherein the TLR ligand comprises any portion of a unicellular organism.  
     
     
         136 . The composition of  claim 123 , wherein said ligand comprises at least one of the following consisting of a glycoprotein, lipoprotein, glycolipid, carbohydrate, lipid, nucleic acid and/or protein or peptide sequence derived from any portion of a bacterial pathogen.  
     
     
         137 . The composition of  claim 136 , further comprising any portion of a bacterial pathogen that binds at least one receptor selected from the group consisting of TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9, TLR-10, TLR-11 and TLR-12.  
     
     
         138 . The composition of  claim 123 , wherein said ligand comprises a nucleic acid encoding a TLR ligand.  
     
     
         139 . The composition of  claim 138 , wherein said nucleic acid comprises at least one molecule selected from the group consisting of bacterial DNA, eukaryotic DNA, dsDNA, ssDNA a synthetic oligonucleotide, RNA, and synthetic RNA.  
     
     
         140 . The composition of  claim 123 , wherein said ligand consists of any molecule that associates with and/or stimulates a pattern recognition receptor.  
     
     
         141 . The composition of  claim 123 , wherein said ligand comprises a synthetically generated ligand that binds to and stimulates a pattern recognition receptor.  
     
     
         142 . The composition of  claim 123 , further comprising any molecule with a steroid backbone.  
     
     
         143 . A composition comprising: at least one antigen; and an adjuvant composition comprising a delivery vehicle; and at least one ligand for a pattern recognition receptor molecule.  
     
     
         144 . The composition of  claim 143 , further comprising said antigen and said ligand for a pattern recognition molecule receptor are complexed to said delivery vehicle.  
     
     
         145 . The composition of  claim 143 , wherein the ligand for the pattern recognition molecule receptor comprises a TLR receptor ligand.  
     
     
         146 . The composition of  claim 143 , wherein said antigen comprises an intact microorganism.  
     
     
         147 . The composition of  claim 146 , wherein said microorganism comprises at least one organism selected from the group consisting of viral organism, bacterial organism, fungal organism, protozoan organism, parasitic pathogenic organisms, rickettsial organisms, and arthropod organisms.  
     
     
         148 . The composition of  claim 143 , wherein said antigen comprises at least one molecule selected from the group consisting of a protein, a peptide, a carbohydrate, a lipoprotein; a glycopeptide, a glycoprotein, a glycolipid and a lipid.  
     
     
         149 . The composition of  claim 143 , wherein said antigen is a cell.  
     
     
         150 . The composition of  claim 143 , wherein said delivery vehicle comprises a liposome.  
     
     
         151 . The composition of  claim 150 , wherein said liposome comprises at least one of the group consisting of a positively charged liposome, a modified multilamellar liposome, a cationic liposome, a neutral liposome and a negatively charged liposome.  
     
     
         152 . The composition of  claim 143 , wherein said delivery vehicle comprises a non-liposomal delivery vehicle.  
     
     
         153 . The composition of  claim 152 , wherein the delivery vehicle comprises at least one vehicle selected from the group consisting of polypeptides, polyamines, chitosan, PEI, polyglutamic acid, protamine sulfate and triclosan.  
     
     
         154 . A method for vaccinating comprising: administering to a subject a composition of an antigen; and an adjuvant composition including a delivery vehicle; and a TLR ligand to a subject.  
     
     
         155 . The method of  claim 154 , further comprising said antigen and said TLR ligand complexed to said delivery vehicle.  
     
     
         156 . The method of  claim 154 , further comprising administering said composition by a route selected from the group consisting of: intravenously, intraperitoneally, by inhalation, subcutaneously, intradermally, intranodally, intramuscularly, intranasally, orally, rectally, intravaginally, intravesicularly, intraocularly, and topically.  
     
     
         157 . The method of  claim 154 , further comprising augmenting an immune response in a subject disposed of cancer.  
     
     
         158 . The method of  claim 157 , wherein the cancer comprises at least one cancer selected from of the group consisting of lung cancer, skin cancer, liver cancer, bone marrow cancer, ovarian cancer, breast cancer, prostate cancer, colon cancer, lymphoma, brain cancer, renal cell cancer, and cancers of mesenchymal tissues.  
     
     
         159 . The method of  claim 157 , further comprising augmenting an immune response in a subject disposed of infectious disease.  
     
     
         160 . The method of  claim 159 , wherein said infectious disease comprises at least one disease selected from the group consisting of a disease due to a viral pathogen; a fungal pathogen, a bacterial pathogen, a ricketssial pathogen, a parasitic pathogen, an arthrorpod pathogen, and a prion pathogen.  
     
     
         161 . A composition comprising: an adjuvant composition comprising at least one antigen, a delivery vehicle; and at least one ligand for a pattern recognition receptor molecule.  
     
     
         162 . The composition of  claim 161 , further comprising said antigen incorporated into said delivery vehicle and then mixed with a ligand for a pattern recognition molecule receptor.  
     
     
         163 . The composition of  claim 162 , wherein said ligand for a pattern recognition molecule receptor comprises a TLR ligand.  
     
     
         164 . The composition of  claim 161 , wherein the delivery vehicle comprises a liposome.  
     
     
         165 . The composition of  claim 164 , wherein said liposome consists of at least one molecule selected from the group consisting of a positively charged liposome, a negatively charged liposome; a cationic liposome; and a modified multilamellar liposome.  
     
     
         166 . The composition of  claim 165 , wherein said cationic liposome further comprises said cationic liposomes complexed to bacterial DNA.  
     
     
         167 . The composition of  claim 161 , wherein said TLR ligand comprises any portion of a bacterium that associates with a TLR.  
     
     
         168 . The composition of  claim 161 , wherein said TLR ligand comprises a bacterial cell wall component.  
     
     
         169 . The composition of  claim 161 , wherein said TLR ligand binds at least one receptor selected from the group consisting of TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9, TLR-10, TLR-11 and TLR-12.  
     
     
         170 . The composition of  claim 161 , wherein said TLR ligand binds TLR 2, TLR 5 or TLR 9.  
     
     
         171 . The composition of  claim 161 , wherein said TLR ligand comprises a nucleic acid.  
     
     
         172 . The composition of  claim 171 , wherein said nucleic acid comprises at least one molecule selected from the group consisting of bacterial DNA, eukaryotic DNA, synthetic oligonucleotide, and RNA.  
     
     
         173 . The composition of  claim 172 , wherein said RNA comprises at least one molecule selected from the group of double-stranded RNA, single-stranded RNA and synthetic RNA.  
     
     
         174 . The composition of  claim 161 , wherein said TLR ligand consists of any ligand that associates with and/or stimulates a TLR.  
     
     
         175 . A method of treating a subject with cancer comprising: administering at least one ligand for a pattern recognition receptor and a delivery vehicle; in conjunction with at least one cancer therapy wherein said method elicits a response in a subject disposed of cancer.  
     
     
         176 . The method of  claim 175 , wherein said cancer therapy comprises at least one therapy consisting of hyperthermia therapy, radiation therapy, chemotherapy, photodynamic therapy (PDT), surgery, ultrasound, and focused ultrasound.  
     
     
         177 . The method of  claim 175 , wherein the order of administering the therapy generates different responses.  
     
     
         178 . The method of  claim 175 , wherein the pattern recognition receptor ligand comprises a nucleic acid molecule.  
     
     
         179 . The method of  claim 175 , wherein the pattern recognition receptor ligand comprises bacterial DNA.  
     
     
         180 . The method of  claim 175 , wherein the delivery vehicle comprises a liposome.  
     
     
         181 . The method of  claim 175 , wherein the delivery vehicle comprises a non-liposomal delivery vehicle.