Patent Publication Number: US-2010111935-A1

Title: Modulation of Toll-Like Receptor 2 Expression By Antisense Oligonucleotides

Description:
This application claims the benefit of priority from U.S. Provisional Patent Application No. 61/111,143, filed on Nov. 4, 2008, the disclosure of which is explicitly incorporated by reference herein. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to Toll-Like Receptor 2 (TLR2). In particular, the invention relates to antisense oligonucleotides that specifically hybridize with nucleic acids encoding TLR2, thus modulating TLR2 expression and activity, and their use in treating or preventing diseases associated with TLR2 or wherein modulation of TLR2 expression would be beneficial. 
     2. Summary of the Related Art 
     Toll-like receptors (TLRs) are present on many cells of the immune system and have been shown to be involved in the innate immune response (Hornung, V. et al., (2002) J. Immunol. 168:4531-4537). TLRs are a key means by which mammals recognize and mount an immune response to foreign molecules and also provide a means by which the innate and adaptive immune responses are linked (Akira, S. et al. (2001) Nature Immunol. 2:675-680; Medzhitov, R. (2001) Nature Rev. Immunol. 1:135-145). In vertebrates, this family consists of at least 11 proteins called TLR1 to TLR11, which are known to recognize pathogen associated molecular patterns (PAMP) from bacteria, fungi, parasites and viruses and induce an immune response mediated by a number of transcription factors. 
     Some TLRs are located on the cell surface to detect and initiate a response to extracellular pathogens and other TLRs are located inside the cell to detect and initiate a response to intracellular pathogens. Table 1 provides a representation of TLRs, the known agonists therefore and the cell types known to contain the TLR (Diebold, S. S. et al. (2004) Science 303:1529-1531; Liew, F. et al. (2005) Nature 5:446-458; Hemmi H et al. (2002) Nat Immunol 3:196-200; Jurk M et al., (2002) Nat Immunol 3:499; Lee J et al. (2003) Proc. Natl. Acad. Sci. USA 100:6646-6651); (Alexopoulou, L. (2001) Nature 413:732-738). 
     
       
         
           
               
               
               
             
               
                 TABLE 1 
               
               
                   
               
               
                 TLR 
                   
                 Cell Types 
               
               
                 Molecule 
                 Agonist 
                 Containing Receptor 
               
               
                   
               
             
            
               
                 Cell Surface 
                   
                   
               
               
                 TLRs: 
               
               
                 TLR2 
                 bacterial lipopeptides 
                 Monocytes/macrophages 
               
               
                   
                   
                 Myeloid dendritic cells 
               
               
                   
                   
                 Mast cells 
               
               
                 TLR4 
                 gram negative bacteria 
                 Monocytes/macrophages 
               
               
                   
                   
                 Myeloid dendritic cells 
               
               
                   
                   
                 Mast cells 
               
               
                   
                   
                 Intestinal epithelium 
               
               
                 TLR5 
                 motile bacteria 
                 Monocytes/macrophages 
               
               
                   
                   
                 Dendritic cells 
               
               
                   
                   
                 Intestinal epithelium 
               
               
                 TLR6 
                 gram positive bacteria 
                 Monocytes/macrophages 
               
               
                   
                   
                 Mast cells 
               
               
                   
                   
                 B lymphocytes 
               
               
                 Endosomal 
               
               
                 TLRs: 
               
               
                 TLR3 
                 double stranded RNA viruses 
                 Dendritic cells 
               
               
                   
                   
                 B lymphocytes 
               
               
                 TLR7 
                 single stranded RNA viruses; 
                 Monocytes/macrophages 
               
               
                   
                 RNA-immunoglobulin 
                 Plasmacytoid dendritic cells 
               
               
                   
                 complexes 
                 B lymphocytes 
               
               
                 TLR8 
                 single stranded RNA viruses; 
                 Monocytes/macrophages 
               
               
                   
                 RNA-immunoglobulin 
                 Dendritic cells 
               
               
                   
                 complexes 
                 Mast cells 
               
               
                 TLR9 
                 DNA containing unmethylated 
                 Monocytes/macrophages 
               
               
                   
                 “CpG” motifs; DNA- 
                 Plasmacytoid dendritic cells 
               
               
                   
                 immunoglobulin complexes 
                 B lymphocytes 
               
               
                   
               
            
           
         
       
     
     The signal transduction pathway mediated by the interaction between a ligand and a TLR is shared among most members of the TLR family and involves a toll/IL-1 receptor (TIR domain), the myeloid differentiation marker 88 (MyD88), IL-1R-associated kinase (IRAK), interferon regulating factor (IRF), TNF-receptor-associated factor (TRAF), TGFβ-activated kinasel, IκB kinases, IκB, and NF-κB (see for example: Akira, S. (2003) J. Biol. Chem. 278:38105 and Geller at al. (2008) Curr. Drug Dev. Tech. 5:29-38). More specifically, for TLRs 1, 2, 4, 5, 6, 7, 8, 9 and 11, this signaling cascade begins with a PAMP ligand interacting with and activating the membrane-bound TLR, which exists as a homo-dimer in the endosomal membrane or the cell surface. Following activation, the receptor undergoes a conformational change to allow recruitment of the TIR domain containing protein MyD88, which is an adapter protein that is common to all TLR signaling pathways except TLR3. MyD88 recruits IRAK4, which phosphorylates and activates IRAK1. The activated IRAK1 binds with TRAF6, which catalyzes the addition of polyubiquitin onto TRAF6. The addition of ubiquitin activates the TAK/TAB complex, which in turn phosphorylates IRFs, resulting in NF-κB release and transport to the nucleus. NF-κB in the nucleus induces the expression of proinflammatory genes (see for example, Trinchieri and Sher (2007) Nat. Rev. Immunol. 7:179-190). 
     The selective localization of TLRs and the signaling generated therefrom, provides some insight into their role in the immune response. The immune response involves both an innate and an adaptive response based upon the subset of cells involved in the response. For example, the T helper (Th) cells involved in classical cell-mediated functions such as delayed-type hypersensitivity and activation of cytotoxic T lymphocytes (CTLs) are Th1 cells. This response is the body&#39;s innate response to antigen (e.g. viral infections, intracellular pathogens, and tumor cells), and results in a secretion of IFN-gamma and a concomitant activation of CTLs. TLR2 is known to localize on the cell membrane and is activated by lipids, proteins and sugars present in the cell wall of pathogens, including but not limited to lipopolysaccharides and peptidoglycans. TLR2 has been shown to discriminate among pathogen types inside phagosomes (Underhill et al. (1999) Nature 401:811-815). This ability to distinguish among pathogens is believed to allow TLR2 to generate different cytokine and chemokine responses in different immune cells, which can in turn determine the nature of the innate and adaptive immune responses. 
     As a result of their involvement in regulating an inflammatory response, TLRs have been shown to play a role in the pathogenesis of many diseases, including autoimmunity, infectious disease and inflammation (Papadimitraki et al. (2007) J. Autoimmun. 29: 310-318; Sun et al. (2007) Inflam. Allergy Drug Targets 6:223-235; Diebold (2008) Adv. Drug Deliv. Rev. 60:813-823; Cook, D. N. et al. (2004) Nature Immunol. 5:975-979; Tse and Horner (2008) Semin. Immunopathol. 30:53-62; Tobias &amp; Curtiss (2008) Semin. Immunopathol. 30:23-27; Ropert et al. (2008) Semin. Immunopathol. 30:41-51; Lee et al. (2008) Semin. Immunopathol. 30:3-9; Gao et al. (2008) Semin. Immunopathol. 30:29-40; Vijay-Kumar et al. (2008) Semin. Immunopathol. 30:11-21). While activation of TLRs is involved in mounting an immune response, an uncontrolled or undesired stimulation of the immune system through TLRs may exacerbate certain diseases in immune compromised subjects or may cause unwanted immune stimulation. Thus, down-regulating TLR expression and/or activity may provide a useful means for disease intervention. 
     To date, investigative strategies aimed selectively at inhibiting TLR activity have involved small molecules (WO/2005/007672), antibodies (see for example: Duffy, K. et al. (2007) Cell Immunol. 248:103-114), catalytic RNAi technologies (e.g. small inhibitory RNAs), certain antisense molecules (Caricilli et al. (2008) J. Endocrinology 199:399), and competitive inhibition with modified or methylated oligonucleotides (see for example: Kandimalla et al. US2008/0089883; Banat and Coffman (2008) Immunol. Rev. 223:271-283). For example, chloroquine and hydroxychloroquine have been shown to block endosomal-TLR signaling by down-regulating the maturation of endosomes (Krieg, A. M. (2002) Annu Rev. Immunol. 20:709). Also, Huang et al. have shown the use of TLR4 siRNA to reverse the tumor-mediated suppression of T cell proliferation and natural killer cell activity (Huang et al. (2005) Cancer Res. 65:5009-5014), and the use of TLR9 siRNA to prevent bacterial-induced inflammation of the eye (Huang et al. (2005) Invest. Opthal. Vis. Sci. 46:4209-4216). 
     Additionally, several groups have used synthetic oligodeoxynucleotides having two triplet sequences, a proximal “CCT” triplet and a distal “GGG” triplet, a poly “G” (e.g. “GGGG” or “GGG”) or “GC” sequences that interact with certain intracellular proteins, resulting in the inhibition of TLR signaling and the concomitant production and release of pro-inflammatory cytokines (see for example: Lenert, P. et al. (2003) DNA Cell Biol. 22(10):621-631; Patole, P. et al. (2005) J. Am. Soc. Nephrol. 16:3273-3280), Gursel, I., et al. (J. Immunol., 171: 1393-1400 (2003), Shirota, H., et al., J. Immunol., 173: 5002-5007 (2004), Chen, Y., et al., Gene Ther. 8: 1024-1032 (2001); Stunz, L. L., Eur. J. Immunol. (2002) 32: 1212-1222; Kandimalla et al. WO2007/7047396). However, oligonucleotides containing guanosine strings have been shown to form tetraplex structures, act as aptamers and inhibit thrombin activity (Bock L C et al., Nature, 355:564-6, 1992; Padmanabhan, K et al., J Biol. Chem., 268(24):17651-4, 1993). Thus, the utility of these inhibitory oligodeoxynucleotide molecules may not be achievable in patients. 
     A potential approach to inhibiting, suppressing or down regulating expression of TLRs is antisense technology. The history of developing antisense technology indicates that while designing and testing antisense oligonucleotides that hybridize to target RNA is a relatively straight forward exercise, only a few antisense oligonucleotides work as intended and optimization of antisense oligonucleotides that have true potential as clinical candidates is not predictable. One skilled in the art would recognize that when optimizing antisense oligonucleotides, conceiving the correct oligonucleotide sequence and length, and utilizing the appropriate nucleic acid and oligonucleotide chemistries are not readily apparent. However, formulating these components is crucial to the utility of any antisense oligonucleotide (Stein and Cheng, 1993, Science 261: 1004-1012). One skilled in the art would further recognize that without conceiving the correct sequence, the correct length, and utilizing the appropriate nucleic acid and oligonucleotide chemistries, the antisense oligonucleotide can have off-target effects and can cause, among other things, the molecule to be unstable, inactive, non-specific, and toxic. As a result of the unpredictable nature of antisense oligonucleotides, to date only one antisense oligonucleotide has received approval for use in humans, and no antisense oligonucleotides are currently being marketed for human use. 
     Accordingly, there exists a need in the field for optimized antisense oligonucleotides that most efficiently down-regulate or inhibit gene expression. In particular, there exists a need in the field for antisense oligonucleotides that down-regulate TLR2 expression and that are stable, active, target specific, non-toxic, and do not activate an innate immune response. A molecule with such characteristics would overcome the problems that have previously prevented antisense oligonucleotides from being developed. 
     BRIEF SUMMARY OF THE INVENTION 
     The present invention is directed to, among other things, optimized synthetic antisense oligonucleotides that are targeted to a nucleic acid encoding TLR2 and that efficiently inhibit the expression of TLR2 through inhibition of mRNA translation and/or through an RNase H mediated mechanism. 
     In a first aspect, optimized antisense oligonucleotides according to the invention include those having SEQ ID NOs: 11, 23, 69, 94, 98, 111, 127 or 158. 
     In another aspect, the invention provides a composition comprising at least one optimized antisense oligonucleotide according to the invention and a physiologically acceptable carrier, diluent or excipient. 
     In another aspect, the invention provides a method of inhibiting TLR2 expression. In this method, an oligonucleotide or multiple oligonucleotides of the invention are specifically contacted or hybridized with TLR2 mRNA either in vitro or in a cell. 
     In another aspect, the invention provides methods for inhibiting the expression of TLR2 in a mammal, particularly a human, such methods comprising administering to the mammal a compound or composition according to the invention. 
     In another aspect, the invention provides a method for inhibiting a TLR2-mediated immune response in a mammal, the method comprising administering to the mammal a TLR2 antisense oligonucleotide according to the invention in a pharmaceutically effective amount. 
     In another aspect, the invention provides a method for therapeutically treating a mammal having a disease mediated by TLR2, such method comprising administering to the mammal, particularly a human, a TLR2 antisense oligonucleotide of the invention, or a composition thereof, in a pharmaceutically effective amount. 
     In another aspect, the invention provides methods for preventing a disease or disorder in a mammal, particularly a human, at risk of contracting or developing a disease or disorder mediated by TLR2. Such methods comprise administering to the mammal an antisense oligonucleotide according to the invention, or a composition thereof, in a prophylactically effective amount. 
     In another aspect, the invention provides a method for inhibiting TLR2 expression and activity in a mammal, comprising administering to the mammal an antisense oligonucleotide complementary to TLR2 mRNA and an antagonist of TLR2 protein, a kinase inhibitor or an inhibitor of signal transduction and transcription (STAT) protein. 
     The subject oligonucleotides and methods disclosed herein are also useful for examining the function of the TLR2 gene in a cell or in a control mammal or in a mammal afflicted with a disease or disorder associated with TLR2 or immune stimulation through TLR2. The cell or mammal is administered the oligonucleotide, and the expression of TLR2 mRNA or protein is examined. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a synthetic scheme for the linear synthesis of antisense oligonucleotides of the invention. DMTr=4,4′-dimethoxytrityl; CE=cyanoethyl. 
         FIG. 2  demonstrates that exemplary human TLR2 antisense oligonucleotides according to the invention are not immunostimulatory (Antisense Alone).  FIG. 2  also demonstrates the ability of exemplary oligonucleotides according to the invention to inhibit TLR2 expression and activation in HEK293 cells that were cultured and treated according to Example 2 (Agonist Plus Antisense). 
         FIG. 3  shows the nucleotide sequence of human TLR2 mRNA [SEQ ID NO: 171] (Genbank Accession No. NM 003264). 
     
    
    
     DETAILED DESCRIPTION 
     The invention relates to optimized TLR2 antisense oligonucleotides, compositions comprising such oligonucleotides and methods of their use for inhibiting or suppressing a TLR2-mediated immune response. More specifically, the antisense oligonucleotides according to the invention are stable, active, target specific, non-toxic, and do not activate an innate immune response. Pharmaceutical and other compositions comprising the compounds according to the invention are also provided. Further provided are methods of down-regulating the expression of TLR2 in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention alone or in combination with other prophylactic or therapeutic compositions. 
     Specifically, the invention provides antisense oligonucleotides designed to be complementary to a genomic region or an RNA molecule transcribed therefrom. These TLR2 antisense oligonucleotides are stable, target specific, and have unique sequences that result in the molecule being maximally effective at inhibiting or suppressing TLR2-mediated signaling in response to endogenous and/or exogenous TLR2 ligands or TLR2 agonists. 
     The TLR2 antisense oligonucleotides according to the invention inhibit immune responses induced by natural or artificial TLR2 agonists in various cell types and in various in vitro and in vivo experimental models. As such, the antisense compositions according to the invention are useful as tools to study the immune system, as well as to compare the immune systems of various mammals, such as humans and mice. 
     Further provided are methods of treating a mammal, particularly a human, having, suspected of having, or being prone to develop a disease or condition associated with TLR2 activation by administering a therapeutically or prophylactically effective amount of one or more of the antisense compounds or compositions of the invention. Since TLR2 has been identified as an important initiator of inflammatory responses (see for example: Leemans et al., (2005) J. Clin. Invest. 115:2894-2903), the optimized antisense oligonucleotides and compositions according to the invention can be used for immunotherapy applications such as, but not limited to, treatment of cancer, autoimmune disorders, asthma, respiratory allergies, food allergies, skin allergies, systemic lupus erythematosus (SLE), arthritis, pleurisy, chronic infections, inflammatory diseases, inflammatory bowel syndrome, sepsis, malaria, and bacteria, parasitic, and viral infections in adult and pediatric human and veterinary applications. In addition, TLR2 antisense oligonucleotides of the invention are useful in the prevention and/or treatment of various diseases, either alone, in combination with or co-administered with other drugs or prophylactic or therapeutic compositions, for example, DNA vaccines, antigens, antibodies, and allergens; and in combination with chemotherapeutic agents (both traditional chemotherapy and modern targeted therapies) and/or TLR2 antagonists for prevention and treatment of diseases. TLR2 antisense oligonucleotides of the invention are useful in combination with compounds or drugs that have unwanted TLR2-mediated immune stimulatory properties. 
     The objects of the present invention, the various features thereof, as well as the invention itself may be more fully understood from the following description, when read together with the accompanying drawings in which the following terms have the ascribed meaning. 
     The term “2′-O-substituted” means substitution of the 2′ position of the pentose moiety with an —O— lower alkyl group containing 1-6 saturated or unsaturated carbon atoms (for example, but not limited to, 2′-O-methyl), or with an —O-aryl or allyl group having 2-6 carbon atoms, wherein such alkyl, aryl or allyl group may be unsubstituted or may be substituted, (for example, with 2′-O-ethoxy-methyl, halo, hydroxy, trifluoromethyl, cyano, nitro, acyl, acyloxy, alkoxy, carboxyl, carbalkoxyl, or amino groups); or with a hydroxy, an amino or a halo group, but not with a 2′-H group. In some embodiments the oligonucleotides of the invention include four or five 2′-O-alkyl ribonucleotides at their 5′ terminus, and/or four or five 2′-O-alkyl ribonucleotides at their 3′ terminus. In exemplary embodiments, the nucleotides of the synthetic oligonucleotides are linked by at least one phosphorothioate internucleotide linkage. The phosphorothioate linkages may be mixed Rp and Sp enantiomers, or they may be stereoregular or substantially stereoregular in either Rp or Sp form (see Iyer et al. (1995) Tetrahedron Asymmetry 6:1051-1054). 
     The term “3′”, when used directionally, generally refers to a region or position in a polynucleotide or oligonucleotide 3′ (toward the 3′ end of the nucleotide) from another region or position in the same polynucleotide or oligonucleotide. The term “5′”, when used directionally, generally refers to a region or position in a polynucleotide or oligonucleotide 5′ (toward the 5′ end of the nucleotide) from another region or position in the same polynucleotide or oligonucleotide. 
     The term “about” generally means that the exact number is not critical. Thus, oligonucleotides having one or two fewer nucleoside residues, or from one to several additional nucleoside residues are contemplated as equivalents of each of the embodiments described above. 
     The term “agonist” generally refers to a substance that binds to a receptor of a cell and induces a response. An agonist often mimics the action of a naturally occurring substance such as a ligand. 
     The term “antagonist” generally refers to a substance that attenuates the effects of an agonist. 
     The term “airway inflammation” generally includes, without limitation, inflammation in the respiratory tract caused by allergens, including asthma. 
     The term “allergen” generally refers to an antigen or antigenic portion of a molecule, usually a protein, which elicits an allergic response upon exposure to a subject. Typically the subject is allergic to the allergen as indicated, for instance, by the wheal and flare test or any method known in the art. A molecule is said to be an allergen even if only a small subset of subjects exhibit an allergic (e.g., IgE) immune response upon exposure to the molecule. 
     The term “allergy” generally includes, without limitation, food allergies, respiratory allergies and skin allergies. 
     The term “antigen” generally refers to a substance that is recognized and selectively bound by an antibody or by a T cell antigen receptor. Antigens may include but are not limited to peptides, proteins, nucleosides, nucleotides and combinations thereof. Antigens may be natural or synthetic and generally induce an immune response that is specific for that antigen. 
     The term “autoimmune disorder” generally refers to disorders in which “self” antigen undergo attack by the immune system. Such term includes, without limitation, lupus erythematosus, multiple sclerosis, type I diabetes mellitus, irritable bowel syndrome, Chron&#39;s disease, rheumatoid arthritis, septic shock, alopecia universalis, acute disseminated encephalomyelitis, Addison&#39;s disease, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, Bullous pemphigoid, chagas disease, chronic obstructive pulmonary disease, coeliac disease, dermatomyositis, endometriosis, Goodpasture&#39;s syndrome, Graves&#39; disease, Guillain-Barré syndrome, Hashimoto&#39;s disease, hidradenitis suppurativa, idiopathic thrombocytopenic purpura, interstitial cystitis, morphea, myasthenia gravis, narcolepsy, neuromyotonia, pemphigus, pernicious anaemia, polymyositis, primary biliary cirrhosis, schizophrenia, Sjögren&#39;s syndrome, temporal arteritis (“giant cell arteritis”), vasculitis, vitiligo, vulvodynia and Wegener&#39;s granulomatosis autoimmune asthma, septic shock and psoriasis. 
     The term “cancer” generally refers to, without limitation, any malignant growth or tumor caused by abnormal or uncontrolled cell proliferation and/or division. Cancers may occur in humans and/or mammals and may arise in any and all tissues. Treating a patient having cancer may include administration of a compound, pharmaceutical formulation or vaccine according to the invention such that the abnormal or uncontrolled cell proliferation and/or division, or metastasis is affected. 
     The term “carrier” generally encompasses any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid containing vesicle, microspheres, liposomal encapsulation, or other material well known in the art for use in pharmaceutical formulations. It will be understood that the characteristics of the carrier, excipient, or diluent will depend on the route of administration for a particular application. The preparation of pharmaceutically acceptable formulations containing these materials is described in, for example,  Remington&#39;s Pharmaceutical Sciences,  18th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990. 
     The terms “co-administration” or “co-administered” generally refer to the administration of at least two different substances sufficiently close in time to modulate an immune response. Co-administration refers to simultaneous administration, as well as temporally spaced order of up to several days apart, of at least two different substances in any order, either in a single dose or separate doses. 
     The term “in combination with” generally means administering a compound according to the invention and another agent useful for treating the disease or condition that does not abolish TLR2 antisense activity of the compound in the course of treating a patient. Such administration may be done in any order, including simultaneous administration, as well as temporally spaced order from a few seconds up to several days apart. Such combination treatment may also include more than a single administration of the compound according to the invention and/or independently the other agent. The administration of the compound according to the invention and the other agent may be by the same or different routes. 
     The terms “individual” or “subject” or “patient” or “vertebrate” generally refer to a mammal, such as a human. 
     The terms “inhibit” or “suppress” or “down-regulate,” when used in reference to expression, generally refer to a decrease in a response or qualitative difference in a response, which could otherwise arise from eliciting and/or stimulation of a response. 
     The term “kinase inhibitor” generally refers to molecules that antagonize or inhibit phosphorylation-dependent cell signaling and/or growth pathways in a cell. Kinase inhibitors may be naturally occurring or synthetic and include small molecules that have the potential to be administered as oral therapeutics. Kinase inhibitors have the ability to rapidly and specifically inhibit the activation of the target kinase molecules. Protein kinases are attractive drug targets, in part because they regulate a wide variety of signaling and growth pathways and include many different proteins. As such, they have great potential in the treatment of diseases involving kinase signaling, including cancer, cardiovascular disease, inflammatory disorders, diabetes, macular degeneration and neurological disorders. Examples of kinase inhibitors include, but are not limited to, sorafenib (Nexavar®), Sutent®, dasatinib, Dasatinib™, Zactima™, Tykerb™ and STI571. 
     The term “linear synthesis” generally refers to a synthesis that starts at one end of an oligonucleotide and progresses linearly to the other end. Linear synthesis permits incorporation of either identical or non-identical (in terms of length, base composition and/or chemical modifications incorporated) monomeric units into an oligonucleotide. 
     The term “mammal” is expressly intended to include warm blooded, vertebrate animals, including, without limitation, humans, non-human primates, rats, mice, cats, dogs, horses, cattle, cows, pigs, sheep and rabbits. 
     The term “nucleoside” generally refers to compounds consisting of a sugar, usually ribose or deoxyribose, and a purine or pyrimidine base. 
     The term “nucleotide” generally refers to a nucleoside comprising a phosphorous-containing group attached to the sugar. 
     The term “modified nucleoside” generally is a nucleoside that includes a modified heterocyclic base, a modified sugar moiety, or any combination thereof. In some embodiments, the modified nucleoside is a non-natural pyrimidine or purine nucleoside, as herein described. For purposes of the invention, a modified nucleoside, a pyrimidine or purine analog or non-naturally occurring pyrimidine or purine can be used interchangeably and refers to a nucleoside that includes a non-naturally occurring base and/or non-naturally occurring sugar moiety. For purposes of the invention, a base is considered to be non-natural if it is not guanine, cytosine, adenine, thymine or uracil and a sugar is considered to be non-natural if it is not 13-ribo-furanoside or 2′-deoxyribo-furanoside. 
     The term “modified oligonucleotide” as used herein describes an oligonucleotide in which at least two of its nucleotides are covalently linked via a synthetic linkage, i.e., a linkage other than a phosphodiester linkage between the 5′ end of one nucleotide and the 3′ end of another nucleotide in which the 5′ nucleotide phosphate has been replaced with any number of chemical groups. The term “modified oligonucleotide” also encompasses oligonucleotides having at least one nucleotide with a modified base and/or sugar, such as a 2′-O-substituted, a 5-methylcytosine and a 3′-O-substituted ribonucleotide. 
     The term “nucleic acid” encompasses a genomic region or an RNA molecule transcribed therefrom. In some embodiments, the nucleic acid is mRNA. 
     The term “nucleotidic linkage” generally refers to a chemical linkage to join two nucleosides through their sugars (e.g. 3′-3′, 2′-3′,2′-5′, 3′-5′, 5′-5′) consisting of a phosphorous atom and a charged, or neutral group (e.g., phosphodiester, phosphorothioate, phosphorodithioate or methylphosphonate) between adjacent nucleosides. 
     The term “oligonucleotide” refers to a polynucleoside formed from a plurality of linked nucleoside units. The nucleoside units may be part of viruses, bacteria, cell debris or oligonucleotide-based compositions (for example, siRNA and microRNA). Such oligonucleotides can also be obtained from existing nucleic acid sources, including genomic or cDNA, but are preferably produced by synthetic methods. In certain embodiments each nucleoside unit includes a heterocyclic base and a pentofuranosyl, trehalose, arabinose, 2′-deoxy-2′-substituted nucleoside, 2′-deoxy-2′-substituted arabinose, 2′-O-substitutedarabinose or hexose sugar group. The nucleoside residues can be coupled to each other by any of the numerous known internucleoside linkages. Such internucleoside linkages include, without limitation, phosphodiester, phosphorothioate, phosphorodithioate, methylphosphonate, alkylphosphonate, alkylphosphonothioate, phosphotriester, phosphoramidate, siloxane, carbonate, carboalkoxy, acetamidate, carbamate, morpholino, borano, thioether, bridged phosphoramidate, bridged methylene phosphonate, bridged phosphorothioate, and sulfone internucleoside linkages. The term “oligonucleotide-based compound” also encompasses polynucleosides having one or more stereospecific internucleoside linkage (e.g., (R P )- or (S P )-phosphorothioate, alkylphosphonate, or phosphotriester linkages). As used herein, the terms “oligonucleotide” and “dinucleotide” are expressly intended to include polynucleosides and dinucleosides having any such internucleoside linkage, whether or not the linkage comprises a phosphate group. In certain exemplary embodiments, these internucleoside linkages may be phosphodiester, phosphorothioate or phosphorodithioate linkages, or combinations thereof. 
     The term “complementary to a genomic region or an RNA molecule transcribed therefrom” is intended to mean an oligonucleotide that binds to the nucleic acid sequence under physiological conditions, for example, by Watson-Crick base pairing (interaction between oligonucleotide and single-stranded nucleic acid) or by Hoogsteen base pairing (interaction between oligonucleotide and double-stranded nucleic acid) or by any other means, including in the case of an oligonucleotide, binding to RNA and causing pseudoknot formation. Binding by Watson-Crick or Hoogsteen base pairing under physiological conditions is measured as a practical matter by observing interference with the function of the nucleic acid sequence. 
     The term “peptide” generally refers to polypeptides that are of sufficient length and composition to affect a biological response, for example, antibody production or cytokine activity whether or not the peptide is a hapten. The term “peptide” may include modified amino acids (whether or not naturally or non-naturally occurring), where such modifications include, but are not limited to, phosphorylation, glycosylation, pegylation, lipidization and methylation. 
     The term “pharmaceutically acceptable” means a non-toxic material that does not interfere with the effectiveness of a compound according to the invention or the biological activity of a compound according to the invention. 
     The term “physiologically acceptable” refers to a non-toxic material that is compatible with a biological system such as a cell, cell culture, tissue, or organism. Preferably, the biological system is a living organism, such as a mammal, particularly a human. 
     The term “prophylactically effective amount” generally refers to an amount sufficient to prevent or reduce the development of an undesired biological effect. 
     The terms “therapeutically effective amount” or “pharmaceutically effective amount” generally refer to an amount sufficient to affect a desired biological effect, such as a beneficial result, including, without limitation, prevention, diminution, amelioration or elimination of signs or symptoms of a disease or disorder. Thus, the total amount of each active component of the pharmaceutical composition or method is sufficient to show a meaningful patient benefit, for example, but not limited to, healing of chronic conditions characterized by immune stimulation. Thus, a “pharmaceutically effective amount” will depend upon the context in which it is being administered. A pharmaceutically effective amount may be administered in one or more prophylactic or therapeutic administrations. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. 
     The term “treatment” generally refers to an approach intended to obtain a beneficial or desired result, which may include alleviation of symptoms, or delaying or ameliorating a disease progression. 
     The invention provides antisense oligonucleotides that are complementary to a nucleic acid that is specific for human TLR2 (SEQ ID NO: 171). The antisense oligonucleotides according to the invention are optimized with respect to (i) the targeted region of the TLR2 mRNA coding sequence, the 5′ untranslated region or the 3′ untranslated region, (ii) their chemical modification(s), or (iii) both. In some embodiments, the compounds are complementary to a region within nucleotides 220 through 2574 of the coding region, or nucleotides 1-219 of the 5′ untranslated region, or 2575-3417 of the 3′ untranslated region of TLR2 mRNA (SEQ ID NO: 171). 
     Antisense oligonucleotides according to the invention are useful in treating and/or preventing diseases wherein inhibiting a TLR2-mediated immune response would be beneficial. TLR2-targeted antisense oligonucleotides according to the invention that are useful include, but are not limited to, antisense oligonucleotides comprising naturally occurring nucleotides, modified nucleotides, modified oligonucleotides and/or backbone modified oligonucleotides. However, antisense oligonucleotides that inhibit the translation of mRNA encoded proteins may produce undesired biological effects, including but not limited to insufficiently active antisense oligonucleotides, inadequate bioavailability, suboptimal pharmacokinetics or pharmacodynamics, and immune stimulation. Thus, the optimal design of an antisense oligonucleotide according to the invention requires many considerations beyond simple design of a complementary sequence. Thus, preparation of TLR2-targeted antisense oligonucleotides according to the invention is intended to incorporate changes necessary to limit secondary structure interference with antisense activity, enhance the oligonucleotide&#39;s target specificity, minimize interaction with binding or competing factors (for example, proteins), optimize cellular uptake, stability, bioavailability, pharmacokinetics and pharmacodynamics, and/or inhibit, prevent or suppress immune cell activation. 
     It has been determined that the human TLR2 coding region is comprised of approximately 2.8 kB, that is most abundant in peripheral blood leukocytes, and corresponds to a 784 amino acid protein in humans (Chaudhary et al. (1998) Blood 91: 4020-4027). The oligonucleotides of the invention were designed to specifically hybridize with optimally available portions of the TLR2 nucleic acid sequence that most effectively act as a target for inhibiting TLR2 expression. These targeted regions of the TLR2 gene include portions of the known exons or 5′ untranslated region. In addition, intron-exon boundaries, 3′ untranslated regions and introns are potentially useful targets for antisense inhibition of TLR2 expression. The nucleotide sequences of some representative, non-limiting oligonucleotides specific for human TLR2 have SEQ ID NOS: 1-170. The nucleotide sequences of optimized oligonucleotides according to the invention include those having SEQ ID NOS: 11, 23, 69, 94, 98, 111, 127 or 158. 
     The oligonucleotides of the invention are at least 14 nucleotides in length, but are preferably 15 to 60 nucleotides long, preferably 20 to 50 nucleotides in length. In some embodiments, these oligonucleotides contain from about 14 to 28 nucleotides or from about 16 to 25 nucleotides or from about 18 to 22 nucleotides or 20 nucleotides. These oligonucleotides can be prepared by the art recognized methods such as phosphoramidate or H-phosphonate chemistry which can be carried out manually or by an automated synthesizer. The synthetic TLR2 antisense oligonucleotides of the invention may also be modified in a number of ways without compromising their ability to hybridize to TLR2 mRNA. Such modifications may include at least one internucleotide linkage of the oligonucleotide being an alkylphosphonate, phosphorothioate, phosphorodithioate, methylphosphonate, phosphate ester, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate or carboxymethyl ester or a combination of these and other internucleotide linkages between the 5′ end of one nucleotide and the 3′ end of another nucleotide in which the 5′ nucleotide phosphodiester linkage has been replaced with any number of chemical groups. 
     For example, U.S. Pat. No. 5,149,797 describes traditional chimeric oligonucleotides having a phosphorothioate core region interposed between methylphosphonate or phosphoramidate flanking regions. U.S. Pat. No. 5,652,356 discloses “inverted” chimeric oligonucleotides comprising one or more nonionic oligonucleotide region (e.g. alkylphosphonate and/or phosphoramidate and/or phosphotriester internucleoside linkage) flanked by one or more region of oligonucleotide phosphorothioate. Various oligonucleotides with modified internucleotide linkages can be prepared according to standard methods. Phosphorothioate linkages may be mixed Rp and Sp enantiomers, or they may be made stereoregular or substantially stereoregular in either Rp or Sp form according to standard procedures. 
     Oligonucleotides which are self-stabilized are also considered to be modified oligonucleotides useful in the methods of the invention (Tang et al. (1993) Nucleic Acids Res. 20:2729-2735). These oligonucleotides comprise two regions: a target hybridizing region; and a self-complementary region having an oligonucleotide sequence complementary to a nucleic acid sequence that is within the self-stabilized oligonucleotide. 
     Other modifications include those which are internal or at the end(s) of the oligonucleotide molecule and include additions to the molecule of the internucleoside phosphate linkages, such as cholesterol, cholesteryl, or diamine compounds with varying numbers of carbon residues between the amino groups and terminal ribose, deoxyribose and phosphate modifications which cleave, or crosslink to the opposite chains or to associated enzymes or other proteins which bind to the genome. Examples of such modified oligonucleotides include oligonucleotides with a modified base and/or sugar such as arabinose instead of ribose, or a 3′, 5′-substituted oligonucleotide having a sugar which, at both its 3′ and 5′ positions, is attached to a chemical group other than a hydroxyl group (at its 3′ position) and other than a phosphate group (at its 5′ position). 
     Other examples of modifications to sugars include modifications to the 2′ position of the ribose moiety which include but are not limited to 2′-O-substituted with an —O-alkyl group containing 1-6 saturated or unsaturated carbon atoms, or with an —O-aryl, or O-allyl group having 2-6 carbon atoms wherein such —O-alkyl, —O-aryl or O-allyl group may be unsubstituted or may be substituted, for example with halo, hydroxy, trifluoromethyl cyano, nitro acyl acyloxy, alkoxy, carboxy, carbalkoxyl or amino groups. None of these substitutions are intended to exclude the native 2′-hydroxyl group in the case of ribose or 2′1-H— in the case of deoxyribose. 
     The oligonucleotides according to the invention can comprise one or more ribonucleotides. For example, U.S. Pat. No. 5,652,355 discloses traditional hybrid oligonucleotides having regions of 2′-O-substituted ribonucleotides flanking a DNA core region. U.S. Pat. No. 5,652,356 discloses an “inverted” hybrid oligonucleotide which includes an oligonucleotide comprising a 2′-O-substituted (or 2′ OH, unsubstituted) RNA region which is in between two oligodeoxyribonucleotide regions, a structure that “inverted relative to the “traditional” hybrid oligonucleotides. Non-limiting examples of particularly useful oligonucleotides of the invention have 2′-O-alkylated ribonucleotides at their 3′, 5′, or 3′ and 5′ termini, with at least four or five contiguous nucleotides being so modified. Non-limiting examples of 2′-O-alkylated groups include 2′-O-methyl, 2′-O-ethyl, 2′-O-propyl, 2′-O-butyls and 2′-O-methoxy-ethyl. 
     Other modified oligonucleotides are capped with a nuclease resistance-conferring bulky substituent at their 3′ and/or 5′ end(s), or have a substitution in one non-bridging oxygen per nucleotide. Such modifications can be at some or all of the internucleoside linkages, as well as at either or both ends of the oligonucleotide and/or in the interior of the molecule. 
     The oligonucleotides of the invention can be administered in combination with one or more antisense oligonucleotides or other nucleic acid containing compounds that are not targeted to the same region as the antisense molecule of the invention. Such other nucleic acid containing compounds include, but are not limited to, ribozymes, RNAi molecules, siRNA, miRNA, and aptamers. In addition, the oligonucleotides of the invention can be administered in combination with one or more compounds or compositions that would activate a TLR2-mediated immune response but for the presence of the TLR2 antisense oligonucleotide according to the invention. In addition, the oligonucleotides of the invention can be administered in combination with one or more vaccines, antigens, antibodies, cytotoxic agents, allergens, antibiotics, TLR antagonists, siRNA, miRNA, antisense oligonucleotides, aptamers, peptides, proteins, gene therapy vectors, DNA vaccines, adjuvants, kinase inhibitors, inhibitors of STAT protein or co-stimulatory molecules or combinations thereof. 
     A non-limiting list of TLR2 antisense oligonucleotides are shown in SEQ ID NO. 1 through SEQ ID NO. 170 and Table 2 below. Optimized antisense oligonucleotides according to the invention include those having SEQ ID NOS: 11, 23, 69, 94, 98, 111, 127 or 158. In Table 2, the oligonucleotide-based TLR2 antisense compounds have all phosphorothioate (PS) linkages. Those skilled in the art will recognize, however, that phosphodiester (PO) linkages, or a mixture of PS and PO linkages can be used. 
     
       
         
           
               
               
               
             
               
                 TABLE 2 
               
               
                   
               
               
                 SEQ ID NO./ 
                 Position 
                 Antisense Sequence 
               
               
                 AS NO. 
                 of Binding 
                 Orientation is 5′-3′ 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 1 
                 1 
                 GCGCTTTCTCGCTGCCTCCG 
               
               
                   
               
               
                 2 
                 21 
                 CGCCGAGCAGCCGCCTGGCT 
               
               
                   
               
               
                 3 
                 41 
                 CGAGCAGTCACCTGAGAGAA 
               
               
                   
               
               
                 4 
                 61 
                 ACCAAACACTGGGAGAACTC 
               
               
                   
               
               
                 5 
                 81 
                 CTTTGGATCCTGCTTGCAAC 
               
               
                   
               
               
                 6 
                 101 
                 TGGGAGTCACTATAGGTCTC 
               
               
                   
               
               
                 7 
                 121 
                 ACTTGGTCACTAAGAGCTCC 
               
               
                   
               
               
                 8 
                 141 
                 TGAGCCCCACAGGTACCTTC 
               
               
                   
               
               
                 9 
                 161 
                 TGAAAGAGCAATGGGCACAA 
               
               
                   
               
               
                 10 
                 181 
                 CAACTACCAGTTGAAAGCAG 
               
               
                   
               
               
                 11 
                 207 
                   GUGG CATTGTCCAGTG CUUC   
               
               
                   
               
               
                 12 
                 221 
                 CATCCACAAAGTATGTGGCA 
               
               
                   
               
               
                 13 
                 241 
                 ATGACCCCCAAGACCCACAC 
               
               
                   
               
               
                 14 
                 261 
                 CTTCCTTGGAGAGGCTGATG 
               
               
                   
               
               
                 15 
                 281 
                 AGAAGCCTGATTGGAGGATT 
               
               
                   
               
               
                 16 
                 301 
                 CCATTGCGGTCACAAGACAG 
               
               
                   
               
               
                 17 
                 321 
                 CTGAGCTGCCCTTGCAGATA 
               
               
                   
               
               
                 18 
                 341 
                 GGGAATGGAGTTTAAAGATC 
               
               
                   
               
               
                 19 
                 361 
                 ACAGCTTCTGTGAGCCCTGA 
               
               
                   
               
               
                 20 
                 381 
                 TGGACAGGTCAAGGCTTTTT 
               
               
                   
               
               
                 21 
                 401 
                 AATGTAGGTGATCCTGTTGT 
               
               
                   
               
               
                 22 
                 421 
                 CTCTGTAGGTCACTGTTGCT 
               
               
                   
               
               
                 23 
                 440 
                   AGCC TGGAGGTTCACA CACC   
               
               
                   
               
               
                 24 
                 461 
                 TCCATTGGATGTCAGCACCA 
               
               
                   
               
               
                 25 
                 481 
                 TCTTCCTCTATTGTGTTAAT 
               
               
                   
               
               
                 26 
                 501 
                 TGCCCAGGGAAGAAAAAGAA 
               
               
                   
               
               
                 27 
                 521 
                 TAAGTCTAAATGTTCAAGAC 
               
               
                   
               
               
                 28 
                 541 
                 TTAGATAAGTAATTATAGGA 
               
               
                   
               
               
                 29 
                 561 
                 TGAACCAGGAAGACGATAAA 
               
               
                   
               
               
                 30 
                 581 
                 TGTTAAAGAAGAAAGGGGCT 
               
               
                   
               
               
                 31 
                 601 
                 TTTCCCAGTAAGTTTAAGAA 
               
               
                   
               
               
                 32 
                 621 
                 CCCCTAGGGTTTTGTAAGGA 
               
               
                   
               
               
                 33 
                 641 
                 ATGAGAAAAAAGAGATGTTT 
               
               
                   
               
               
                 34 
                 661 
                 AGGATTTGCAATTTTGTGAG 
               
               
                   
               
               
                 35 
                 681 
                 TGTCCATATTTCCCACTCTC 
               
               
                   
               
               
                 36 
                 701 
                 TCTTTGAATCTTAGTGAAGG 
               
               
                   
               
               
                 37 
                 721 
                 GTAAGTCCAGCAAAATCTTT 
               
               
                   
               
               
                 38 
                 741 
                 TCTCAAGTTCCTCAAGGAAG 
               
               
                   
               
               
                 39 
                 761 
                 CTGTAGATCTGAAGCATCAA 
               
               
                   
               
               
                 40 
                 781 
                 AAACTTTTTGGCTCATAGCT 
               
               
                   
               
               
                 41 
                 801 
                 TTACATTCTGAATTGACTTC 
               
               
                   
               
               
                 42 
                 821 
                 CATATGAAGGATCAGATGAC 
               
               
                   
               
               
                 43 
                 841 
                 AGCAGTAAAATATGCTGCTT 
               
               
                   
               
               
                 44 
                 861 
                 TAACATCTACAAAAATCTCC 
               
               
                   
               
               
                 45 
                 881 
                 CAAACATTCCACGGAACTTG 
               
               
                   
               
               
                 46 
                 901 
                 AAATCAGTATCTCGCAGTTC 
               
               
                   
               
               
                 47 
                 921 
                 CTGAAAAATGGAAAGTGTCC 
               
               
                   
               
               
                 48 
                 941 
                 TGTTTCACCAGTGGATAGTT 
               
               
                   
               
               
                 49 
                 961 
                 AACTTTTTAATCAATGAATT 
               
               
                   
               
               
                 50 
                 981 
                 TTTTCACATTTCTAAATGTA 
               
               
                   
               
               
                 51 
                 1001 
                 AAACAAACTTTCATCGGTGA 
               
               
                   
               
               
                 52 
                 1021 
                 TTCAAAAGTTTCATAACCTG 
               
               
                   
               
               
                 53 
                 1041 
                 CTAACAATCCAGAAATCTGA 
               
               
                   
               
               
                 54 
                 1061 
                 ACAGTCATCAAACTCTAATT 
               
               
                   
               
               
                 55 
                 1081 
                 TTACCAACTCCATTAAGGGT 
               
               
                   
               
               
                 56 
                 1101 
                 CATTATCAGATGCTCTAAAA 
               
               
                   
               
               
                 57 
                 1121 
                 ACCTGGATCTATAACTCTGT 
               
               
                   
               
               
                 58 
                 1141 
                 ATTGTTAACGTTTCCACTTT 
               
               
                   
               
               
                 59 
                 1161 
                 TTGGAATATGCAGCCTCCGG 
               
               
                   
               
               
                 60 
                 1181 
                 ATCATAAAATAAGTAAAACC 
               
               
                   
               
               
                 61 
                 1201 
                 AGTGAATATAAAGTGCTCAG 
               
               
                   
               
               
                 62 
                 1221 
                 TTCTTTTAACTCTTTCTGTA 
               
               
                   
               
               
                 63 
                 1241 
                 TTTACTGTTTTCTACTGTGA 
               
               
                   
               
               
                 64 
                 1261 
                 AAACAAGGAACCAGAAAAAC 
               
               
                   
               
               
                 65 
                 1281 
                 ATTTTAAATGTTGTGAAAGT 
               
               
                   
               
               
                 66 
                 1301 
                 GAGATCCAAGTATTCTAATG 
               
               
                   
               
               
                 67 
                 1321 
                 TCAACCATCAAATTTTCACT 
               
               
                   
               
               
                 68 
                 1341 
                 CTGAATTTTTCAAGTATTCT 
               
               
                   
               
               
                 69 
                 1357 
                   CAGG CATCCTCACAGG CUGA   
               
               
                   
               
               
                 70 
                 1381 
                 AAAATTAAAGTTTGTAGAGA 
               
               
                   
               
               
                 71 
                 1401 
                 ATGCCAAATGATTTTGCCTT 
               
               
                   
               
               
                 72 
                 1421 
                 CTCTCCGGTTTTTTCCAATG 
               
               
                   
               
               
                 73 
                 1441 
                 TTTTTCAGAGTGAGCAAAGT 
               
               
                   
               
               
                 74 
                 1461 
                 TGATATCAATGTTAGTCAAG 
               
               
                   
               
               
                 75 
                 1481 
                 AGAATGAAAACTATTCTTAC 
               
               
                   
               
               
                 76 
                 1501 
                 CACTGACAAGTTTCAGGCAT 
               
               
                   
               
               
                 77 
                 1521 
                 AATATTTCATCTTTTCTGGC 
               
               
                   
               
               
                 78 
                 1541 
                 TCGTGTGCTGGATAAGTTCA 
               
               
                   
               
               
                 79 
                 1561 
                 CAGCCTGTTACACTGTGTAT 
               
               
                   
               
               
                 80 
                 1581 
                 TTTCCAGTGTCTTGGGAATG 
               
               
                   
               
               
                 81 
                 1601 
                 GTTGTTGCTAACATCTAAAA 
               
               
                   
               
               
                 82 
                 1621 
                 AAAGAAAATAAATTGAGATT 
               
               
                   
               
               
                 83 
                 1641 
                 CTTTGAGTTGCGGCAAATTC 
               
               
                   
               
               
                 84 
                 1661 
                 ATTTCTGGAAATATAAAGTT 
               
               
                   
               
               
                 85 
                 1681 
                 TCTGGTAGAGTCATCAACTT 
               
               
                   
               
               
                 86 
                 1701 
                 ACATGGGTAAGAGGGAGGCA 
               
               
                   
               
               
                 87 
                 1721 
                 ACTGATTTTCAATACTAGTA 
               
               
                   
               
               
                 88 
                 1741 
                 AACGTAGTTATTGCATTCCT 
               
               
                   
               
               
                 89 
                 1761 
                 AGTCAAGTTGCTCCTTAGAA 
               
               
                   
               
               
                 90 
                 1781 
                 AGTCTTCAGTGTGTGAAATG 
               
               
                   
               
               
                 91 
                 1801 
                 TTATTGCCACCAGCTTCCAA 
               
               
                   
               
               
                 92 
                 1821 
                 ATTCACAGGAGCAAATGAAG 
               
               
                   
               
               
                 93 
                 1841 
                 CTCCTGAGTGAAGGAGAGGA 
               
               
                   
               
               
                 94 
                 1854 
                   CCAG TGCTTGCTGCTC CUGA   
               
               
                   
               
               
                 95 
                 1881 
                 TTGCTGGCCAATCAATCAAG 
               
               
                   
               
               
                 96 
                 1901 
                 TGGAGAGTCACACAGGTAAT 
               
               
                   
               
               
                 97 
                 1921 
                 TGCTGGCCACGCACATGGGA 
               
               
                   
               
               
                 98 
                 1934 
                   GACA TCCTGAACCTGC UGGC   
               
               
                   
               
               
                 99 
                 1941 
                 AGAGGCGGACATCCTGAACC 
               
               
                   
               
               
                 100 
                 1961 
                 CCTGTGACATTCCGACACCG 
               
               
                   
               
               
                 101 
                 1981 
                 ATGCCAGACACCAGTGCTGT 
               
               
                   
               
               
                 102 
                 2001 
                 GCAGGAACAGAGCACAGCAC 
               
               
                   
               
               
                 103 
                 2021 
                 GACCCCCGTGAGCAGGATCA 
               
               
                   
               
               
                 104 
                 2041 
                 CCATGGAAACGGTGGCACAG 
               
               
                   
               
               
                 105 
                 2061 
                 TCATTTTCATATACCACAGG 
               
               
                   
               
               
                 106 
                 2081 
                 GGCCTGGAGCCAGGCCCACA 
               
               
                   
               
               
                 107 
                 2101 
                 GCTTTCCTGGGCTTCCTTTT 
               
               
                   
               
               
                 108 
                 2121 
                 AGCAGATGTTCCTGCTGGGA 
               
               
                   
               
               
                 109 
                 2141 
                 GTAAGAAACAAATGCATCAT 
               
               
                   
               
               
                 110 
                 2161 
                 CAGTAGGCATCCCGCTCACT 
               
               
                   
               
               
                 111 
                 2181 
                   GGAC CATAAGGTTCTC CACC   
               
               
                   
               
               
                 112 
                 2201 
                 ATTGAAGTTCTCCAGCTCCT 
               
               
                   
               
               
                 113 
                 2221 
                 AGACACAACTTGAAGGGGGG 
               
               
                   
               
               
                 114 
                 2241 
                 GAATGAAGTCCCGCTTATGA 
               
               
                   
               
               
                 115 
                 2261 
                 GTCAATGATCCACTTGCCAG 
               
               
                   
               
               
                 116 
                 2281 
                 TCAATGGAGTCAATGATATT 
               
               
                   
               
               
                 117 
                 2301 
                 AGACAGTTTTGTGGCTCTTT 
               
               
                   
               
               
                 118 
                 2321 
                 AAAGTTTTCAGAAAGCACAA 
               
               
                   
               
               
                 119 
                 2341 
                 TTGCACCACTCACTCTTCAC 
               
               
                   
               
               
                 120 
                 2361 
                 GGGAGAAGTCCAGTTCATAC 
               
               
                   
               
               
                 121 
                 2381 
                 CTCATCAAAAAGACGGAAAT 
               
               
                   
               
               
                 122 
                 2401 
                 AGAATGGCAGCATCATTGTT 
               
               
                   
               
               
                 123 
                 2421 
                 CAATGGGCTCCAGAAGAATG 
               
               
                   
               
               
                 124 
                 2441 
                 CTGGGGAATGGCTTTTTTCT 
               
               
                   
               
               
                 125 
                 2461 
                 TTCCGCAGCTTGCAGAAGCG 
               
               
                   
               
               
                 126 
                 2481 
                 AGGTCTTGGTGTTCATTATC 
               
               
                   
               
               
                 127 
                 2489 
                   CUCC AGGTAGGTCTTG GUGU   
               
               
                   
               
               
                 128 
                 2501 
                 GTCCATGGGCCACTCCAGGT 
               
               
                   
               
               
                 129 
                 2521 
                 AATCCTTCCCGCTGAGCCTC 
               
               
                   
               
               
                 130 
                 2541 
                 CAGCTCTCAGATTTACCCAA 
               
               
                   
               
               
                 131 
                 2561 
                 GGGAACCTAGGACTTTATCG 
               
               
                   
               
               
                 132 
                 2581 
                 CAAAGACTGGTCTTAAATAT 
               
               
                   
               
               
                 133 
                 2601 
                 ACATAAAGATCCCAACTAGA 
               
               
                   
               
               
                 134 
                 2621 
                 GAACTTAACTATAACTAGTG 
               
               
                   
               
               
                 135 
                 2641 
                 TTATATAATTATGTCTGAAT 
               
               
                   
               
               
                 136 
                 2661 
                 ACGGTACATCCACGTAGTTT 
               
               
                   
               
               
                 137 
                 2681 
                 AGTAAGCAAGTCCTCAAATG 
               
               
                   
               
               
                 138 
                 2701 
                 ATTTGAAGTTTTGTAGTTTT 
               
               
                   
               
               
                 139 
                 2721 
                 AAAACAGCACCCCAGACAAA 
               
               
                   
               
               
                 140 
                 2741 
                 TAAATCTGGCATATGTTTAT 
               
               
                   
               
               
                 141 
                 2761 
                 AAAAACCAAAAACCAATTTT 
               
               
                   
               
               
                 142 
                 2781 
                 GGTTATCTCATAGAAAAAAG 
               
               
                   
               
               
                 143 
                 2801 
                 AGTAATAGACTTATGATCAT 
               
               
                   
               
               
                 144 
                 2821 
                 AGGGACTATATTCAGATATC 
               
               
                   
               
               
                 145 
                 2841 
                 ACCAATTCCCTTGGATACCA 
               
               
                   
               
               
                 146 
                 2861 
                 ATATCCACGAGGATCCTGCA 
               
               
                   
               
               
                 147 
                 2881 
                 TTGATCATCTATGAATTTTG 
               
               
                   
               
               
                 148 
                 2901 
                 ATGCCACTCTTATAAGGGAC 
               
               
                   
               
               
                 149 
                 2921 
                 ACAGGTTATATGCAAATACT 
               
               
                   
               
               
                 150 
                 2941 
                 AAGTATACAGGAGAATGTAC 
               
               
                   
               
               
                 151 
                 2961 
                 AGTAATCTAGAGATGATTTA 
               
               
                   
               
               
                 152 
                 2981 
                 ACATTGTATTGGGTATCATA 
               
               
                   
               
               
                 153 
                 3001 
                 CAACTATTTACATAGTATTT 
               
               
                   
               
               
                 154 
                 3021 
                 TATAAATAAAAAGACAGTA 
               
               
                   
               
               
                 155 
                 3041 
                 AATAAAAAATAACAATAATA 
               
               
                   
               
               
                 156 
                 3061 
                 TATGTTTTAAAAATTTTGAA 
               
               
                   
               
               
                 157 
                 3081 
                 AACCAACTGTGGATCAAAAG 
               
               
                   
               
               
                 158 
                 3097 
                   CUGC ATCCATGAAGTC AACC   
               
               
                   
               
               
                 159 
                 3121 
                 GTTGGCCCTCTATATCCATG 
               
               
                   
               
               
                 160 
                 3141 
                 GCCAGTTGCTACAGATTACA 
               
               
                   
               
               
                 161 
                 3161 
                 GCTGTTTCCTAATGAACTAA 
               
               
                   
               
               
                 162 
                 3181 
                 AGAATCTTAAGTTCATTTGT 
               
               
                   
               
               
                 163 
                 3201 
                 AAAGAATGACACAGTCATTG 
               
               
                   
               
               
                 164 
                 3221 
                 AGGAGTCTCTTAGCAGGAAG 
               
               
                   
               
               
                 165 
                 3241 
                 AATGCCTTTTGTGGCCACAG 
               
               
                   
               
               
                 166 
                 3261 
                 GACAGCTAGGTAGGACAGAG 
               
               
                   
               
               
                 167 
                 3281 
                 GATCAGCTGCACAGAGAAGT 
               
               
                   
               
               
                 168 
                 3301 
                 CTTTGCCTTGTTGCTCTTGA 
               
               
                   
               
               
                 169 
                 3321 
                 TTTGGGGAGTGCCCCAAATA 
               
               
                   
               
               
                 170 
                 3341 
                 TTCTAGGAATAGCAACAAGT 
               
               
                   
               
            
           
         
       
     
     AS is an abbreviation for antisense. Underlined nucleotides are 2′-O-methylribonucleotides; all others are 2′-deoxyribonucleotides. In the exemplary antisense oligonucleotides according to the invention, when a “CG” dinucleotide is contained in the sequence, such oligonucleotide is modified to remove or prevent the immune stimulatory properties of the oligonucleotide. 
     In another aspect, the invention provides a composition comprising at least one optimized antisense oligonucleotide according to the invention and a physiologically acceptable carrier, diluent or excipient. The characteristics of the carrier will depend on the route of administration. Such a composition may contain, in addition to the synthetic oligonucleotide and carrier, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The pharmaceutical composition of the invention may also contain other active factors and/or agents which enhance inhibition of TLR2 expression. For example, combinations of synthetic oligonucleotides, each of which is directed to different regions of the TLR2 mRNA, may be used in the pharmaceutical compositions of the invention. The pharmaceutical composition of the invention may further contain nucleotide analogs such as azidothymidine, dideoxycytidine, dideoxyinosine, and the like. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic, additive or enhanced effect with the synthetic oligonucleotide of the invention, or to minimize side-effects caused by the synthetic oligonucleotide of the invention. The pharmaceutical composition of the invention may be in the form of a liposome in which the synthetic oligonucleotides of the invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers which are in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. One particularly useful lipid carrier is lipofectin. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Pat. Nos. 4,235,871; 4,501,728; 4,837,028; and 4,737,323. The pharmaceutical composition of the invention may further include compounds such as cyclodextrins and the like that enhance delivery of oligonucleotides into cells or slow release polymers. 
     In another aspect, the invention provides a method of inhibiting TLR2 expression. In this method, an oligonucleotide or multiple oligonucleotides of the invention are specifically contacted or hybridized with TLR2 mRNA either in vitro or in a cell. 
     In another aspect, the invention provides methods for inhibiting the expression of TLR2 in a mammal, particularly a human, such methods comprising administering to the mammal a compound or composition according to the invention. One skilled in the art would recognize that the antisense compounds and compositions according to the invention can be administered through a variety of means. One such means for administration is according to Example 3. The antisense activity of a compound or composition according to the invention can be determined by measuring TLR2 mRNA and TLR2 protein concentration. The data is anticipated to demonstrate that administration of an exemplary TLR2 antisense oligonucleotide according to the invention can cause down-regulation of TLR2 expression in vivo. 
     In another aspect, the invention provides a method for inhibiting a TLR-mediated immune response in a mammal, the method comprising administering to the mammal a TLR2 antisense oligonucleotide according to the invention in a pharmaceutically effective amount, wherein routes of administration include, but are not limited to, parenteral, intramuscular, subcutaneous, intraperitoneal, intraveneous, mucosal delivery, oral, sublingual, transdermal, topical, inhalation, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, by gene gun, dermal patch or in eye drop or mouthwash form. One skilled in the art would recognize that one such administration can be accomplished according to Example 3, or by known methods. The antisense activity of compound or composition according to the invention can be determined by measuring biomarkers related to TLR2 signaling, for example, but not limited to, measuring IL-12. The data is anticipated to demonstrate that administration of an exemplary TLR2 antisense oligonucleotide according to the invention can cause down-regulation of TLR2 expression in vivo and prevent the induction of IL-12 by a TLR2 agonist. More generally, the data is anticipated to demonstrate the ability of a TLR2 antisense oligonucleotide according to the invention to inhibit the induction of pro-inflammatory cytokines by a TLR2 agonist. 
     In another aspect, the invention provides a method for therapeutically treating a mammal having a disease mediated by TLR2, such method comprising administering to the mammal, particularly a human, a TLR2 antisense oligonucleotide of the invention in a pharmaceutically effective amount. 
     In certain embodiments, the disease is cancer, an autoimmune disorder, airway inflammation, inflammatory disorders, infectious disease, malaria, Lyme disease, ocular infections, conjunctivitis, skin disorders, psoriasis, scleroderma, cardiovascular disease, atherosclerosis, chronic fatigue syndrome, sarcoidosis, transplant rejection, allergy, asthma or a disease caused by a pathogen. Preferred autoimmune disorders include without limitation lupus erythematosus, multiple sclerosis, type I diabetes mellitus, irritable bowel syndrome, Chron&#39;s disease, rheumatoid arthritis, septic shock, alopecia universalis, acute disseminated encephalomyelitis, Addison&#39;s disease, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, Bullous pemphigoid, chagas disease, chronic obstructive pulmonary disease, coeliac disease, dermatomyositis, endometriosis, Goodpasture&#39;s syndrome, Graves&#39; disease, Guillain-Barré syndrome, Hashimoto&#39;s disease, hidradenitis suppurativa, idiopathic thrombocytopenic purpura, interstitial cystitis, morphea, myasthenia gravis, narcolepsy, neuromyotonia, pemphigus, pernicious anaemia, polymyositis, primary biliary cirrhosis, schizophrenia, Sjögren&#39;s syndrome, temporal arteritis (“giant cell arteritis”), vasculitis, vitiligo, vulvodynia and Wegener&#39;s granulomatosis. In certain embodiments, inflammatory disorders include without limitation airway inflammation, asthma, autoimmune diseases, chronic inflammation, chronic prostatitis, glomerulonephritis, Behçet&#39;s disease, hypersensitivities, inflammatory bowel disease, reperfusion injury, rheumatoid arthritis, transplant rejection, ulcerative colitis, uveitis, conjunctivitis and vasculitis. 
     In another aspect, the invention provides methods for preventing a disease or disorder in a mammal, particularly a human, at risk of contracting or developing a disease or disorder mediated by TLR2. Such method comprises administering to the mammal a prophylactically effective amount of an antisense oligonucleotide or composition according to the invention. Such diseases and disorders include, without limitation, cancer, an autoimmune disorder, airway inflammation, inflammatory disorders, infectious disease, malaria, Lyme disease, ocular infections, conjunctivitis, skin disorders, psoriasis, scleroderma, cardiovascular disease, atherosclerosis, chronic fatigue syndrome, sarcoidosis, transplant rejection, allergy, asthma or a disease caused by a pathogen in a vertebrate. Autoimmune disorders include, without limitation, lupus erythematosus, multiple sclerosis, type I diabetes mellitus, irritable bowel syndrome, Chron&#39;s disease, rheumatoid arthritis, septic shock, alopecia universalis, acute disseminated encephalomyelitis, Addison&#39;s disease, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, Bullous pemphigoid, chagas disease, chronic obstructive pulmonary disease, coeliac disease, dermatomyositis, endometriosis, Goodpasture&#39;s syndrome, Graves&#39; disease, Guillain-Barré syndrome, Hashimoto&#39;s disease, hidradenitis suppurativa, idiopathic thrombocytopenic purpura, interstitial cystitis, morphea, myasthenia gravis, narcolepsy, neuromyotonia, pemphigus, pernicious anaemia, polymyositis, primary biliary cirrhosis, schizophrenia, Sjögren&#39;s syndrome, temporal arteritis (“giant cell arteritis”), vasculitis, vitiligo, vulvodynia and Wegener&#39;s granulomatosis. Inflammatory disorders include, without limitation, airway inflammation, asthma, autoimmune diseases, chronic inflammation, chronic prostatitis, glomerulonephritis, Behçet&#39;s disease, hypersensitivities, inflammatory bowel disease, reperfusion injury, rheumatoid arthritis, transplant rejection, ulcerative colitis, uveitis, conjunctivitis and vasculitis. 
     In another aspect, the invention provides a method for inhibiting TLR2 expression and activity in a mammal, comprising administering to the mammal an antisense oligonucleotide complementary to TLR2 mRNA and an antagonist of TLR2 protein, a kinase inhibitor or an inhibitor of STAT protein. Accordingly, TLR2 expression is inhibited by the antisense oligonucleotide, while any TLR2 protein residually expressed is inhibited by the antagonist. Preferred antagonists include anti-TLR2 antibodies or binding fragments or peptidomimetics thereof, RNA-based compounds, oligonucleotide-based compounds, and small molecule inhibitors of TLR2 activity or of a signaling protein&#39;s activity. 
     In the various methods according to the invention, a therapeutically or prophylactically effective amount of a synthetic oligonucleotide of the invention and effective in inhibiting the expression of TLR2 is administered to a cell. This cell may be part of a cell culture, a neovascularized tissue culture, or may be part or the whole body of a mammal such as a human or other mammal. Administration of the therapeutic compositions of TLR2 antisense oligonucleotide can be carried out using known procedures at dosages and for periods of time effective to reduce symptoms or surrogate markers of the disease, depending on the condition and response, as determined by those with skill in the art. It may be desirable to administer simultaneously, or sequentially a therapeutically effective amount of one or more of the therapeutic TLR2 antisense oligonucleotides of the invention to an individual as a single treatment episode. In some exemplary embodiments of the methods of the invention described above, the oligonucleotide is administered locally and/or systemically. The term “administered locally” refers to delivery to a defined area or region of the body, while the term “systemic administration” is meant to encompass delivery to the whole organism. 
     In any of the methods according to the invention, one or more of the TLR2 antisense oligonucleotide can be administered alone or in combination with any other agent useful for treating the disease or condition that does not diminish the immune modulatory effect of the TLR2 antisense oligonucleotide. In any of the methods according to the invention, the agent useful for treating the disease or condition includes, but is not limited to, one or more vaccines, antigens, antibodies, cytotoxic agents, allergens, antibiotics, antisense oligonucleotides, TLR agonists, TLR antagonists, siRNA, miRNA, aptamers, peptides, proteins, gene therapy vectors, DNA vaccines, adjuvants or kinase inhibitors to enhance the specificity or magnitude of the immune response, or co-stimulatory molecules such as cytokines, chemokines, protein ligands, trans-activating factors, peptides and peptides comprising modified amino acids. For example, in the treatment of autoimmune disease, it is contemplated that the TLR2 antisense oligonucleotide may be administered in combination with one or more targeted therapeutic agents and/or monoclonal antibodies. Alternatively, the agent can include DNA vectors encoding for antigen or allergen. In these embodiments, the TLR2 antisense oligonucleotide of the invention can produce direct immune modulatory or suppressive effects. When co-administered with one or more other therapies, the synthetic oligonucleotide of the invention may be administered either simultaneously with the other treatment(s), or sequentially. 
     In the various methods according to the invention the route of administration may be, without limitation, parenteral, mucosal delivery, oral, sublingual, transdermal, topical, inhalation, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, by gene gun, dermal patch or in eye drop or mouthwash form. 
     When a therapeutically effective amount of synthetic oligonucleotide of the invention is administered orally, the synthetic oligonucleotide will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet form, the pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95% synthetic oligonucleotide and preferably from about 25 to 90% synthetic oligonucleotide. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains from about 0.5 to 90% by weight of the synthetic oligonucleotide or from about 1 to 50% synthetic oligonucleotide. 
     When a therapeutically effective amount of synthetic oligonucleotide of the invention is administered by parenteral, mucosal delivery, oral, sublingual, transdermal, topical, inhalation, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, by gene gun, dermal patch or in eye drop or mouthwash form, the synthetic antisense oligonucleotide will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A pharmaceutical composition for parenteral, mucosal delivery, oral, sublingual, transdermal, topical, inhalation, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, by gene gun, dermal patch or in eye drop or mouthwash form should contain, in addition to the synthetic oligonucleotide, an isotonic vehicle such as Sodium Chloride Injection, Ringer&#39;s Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer&#39;s Injection or other vehicle as known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants or other additives known to those of skill in the art. 
     When administered parenteral, mucosal delivery, oral, sublingual, transdermal, topical, inhalation, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, by gene gun, dermal patch or in eye drop or mouthwash form, doses ranging from 0.01% to 10% (weight/volume) may be used. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, sesame oil or synthetic oils may be added. Topical administration may be by liposome or transdermal time-release patch. 
     The amount of synthetic oligonucleotide in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 10 micrograms to about 20 mg of synthetic oligonucleotide per kg body or organ weight. 
     The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient. 
     Some diseases lend themselves to acute treatment while others require longer term therapy. Both acute and long term intervention in diseases are worthy goals. Injections of antisense oligonucleotides against TLR2 can be an effective means of inhibiting certain diseases in an acute situation. However for long term therapy over a period of weeks, months or years, systemic delivery (intraperitoneal, intramuscular, subcutaneous, intravenous) either with carriers such as saline, slow release polymers or liposomes are likely to be considered. 
     In some chronic diseases, systemic administration of oligonucleotides may be preferable. The frequency of injections is from continuous infusion to once a month, several times per month or less frequently will be determined based on the disease process and the biological half life of the oligonucleotides. 
     The oligonucleotides and methods of the invention are also useful for examining the function of the TLR2 gene in a cell or in a control mammal or in a mammal afflicted with a disease associated with TLR2 or immune stimulation through TLR2. In such use, the cell or mammal is administered the oligonucleotide, and the expression of TLR2 mRNA or protein is examined. 
     Without intending to be limited to any theory or mechanism, it is generally believed that the activity of oligonucleotides according to the invention depends on the hybridization of the oligonucleotide to the target nucleic acid (e.g. to at least a portion of a genomic region, gene or mRNA transcript thereof), thus disrupting the function of the target. Such hybridization under physiological conditions is measured as a practical matter by observing interference with the function of the nucleic acid sequence. Thus, an exemplary oligonucleotide used in accordance with the invention is capable of forming a stable duplex (or triplex in the Hoogsteen or other hydrogen bond pairing mechanism) with the target nucleic acid; activating RNase H or other in vivo enzymes thereby causing effective destruction of the target RNA molecule; and is capable of resisting nucleolytic degradation (e.g. endonuclease and exonuclease activity) in vivo. A number of the modifications to oligonucleotides described above and others which are known in the art specifically and successfully address each of these exemplary characteristics. 
     The patents and publications cited herein reflect the level of knowledge in the art and are hereby incorporated by reference in their entirety. Any conflict between the teachings of these patents and publications and this specification shall be resolved in favor of the latter. Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific substances and procedures described herein. For example, antisense oligonucleotides that overlap with the oligonucleotides may be used. Such equivalents are considered to be within the scope of this invention. 
     The following examples illustrate the exemplary modes of making and practicing the present invention, but are not meant to limit the scope of the invention since alternative methods may be utilized to obtain similar results. 
     EXAMPLES 
     Example 1 
     Preparation of TLR2-Specific Antisense Oligonucleotides 
     Chemical entities according to the invention were synthesized on a 1 μmol to 0.1 mM scale using an automated DNA synthesizer (OligoPilot II, AKTA, (Amersham) and/or Expedite 8909 (Applied Biosystem)), following the linear synthesis procedure outlined in  FIG. 1 . 
     5′-DMT dA, dG, dC and T phosphoramidites were purchased from Proligo (Boulder, Colo.). 5′-DMT 7-deaza-dG and araG phosphoramidites were obtained from Chemgenes (Wilmington, Mass.). DiDMT-glycerol linker solid support was obtained from Chemgenes. 1-(2′-deoxy-β-D-ribofuranosyl)-2-oxo-7-deaza-8-methyl-purine amidite was obtained from Glen Research (Sterling, Va.), 2′-O-methylribonuncleoside amidites were obtained from Promega (Obispo, Calif.). All compounds according to the invention were phosphorothioate backbone modified. 
     All nucleoside phosphoramidites were characterized by  31 P and  1 H NMR spectra. Modified nucleosides were incorporated at specific sites using normal coupling cycles recommended by the supplier. After synthesis, compounds were deprotected using concentrated ammonium hydroxide and purified by reverse phase HPLC, detritylation, followed by dialysis. Purified compounds as sodium salt form were lyophilized prior to use. Purity was tested by CGE and MALDI-TOF MS. Endotoxin levels were determined by LAL test and were below 1.0 EU/mg. 
     Example 2 
     Cell Culture Conditions and Reagents 
     HEK293 Cell Culture Assays for TLR2 Antisense Activity 
     HEK293 cells stably expressing human TLR2/TLR6 (Invivogen, San Diego, Calif.) were plated in 48-well plates in 250 μL/well DMEM supplemented with 10% heat-inactivated FBS in a 5% CO2 incubator. At 80% confluence, cultures were transiently transfected with 400 ng/mL of the secreted form of human embryonic alkaline phosphatase (SEAP) reporter plasmid (pNifty2-Seap) (Invivogen) in the presence of 4 μL/mL of lipofectamine (Invitrogen, Carlsbad, Calif.) in culture medium. The SEAP reporter plasmid is inducible by NF-κB. Plasmid DNA and lipofectamine were diluted separately in serum-free medium and incubated at room temperature for 5 min. After incubation, the diluted DNA and lipofectamine were mixed and the mixtures were incubated further at room temperature for 20 min. Aliquots of 25 μL of the DNA/lipofectamine mixture containing 100 ng of plasmid DNA and 1 μL of lipofectamine were added to each well of the cell culture plate, and the cells were transfected for 6 h. After transfection, medium was replaced with fresh culture medium (no antibiotics), antisense compounds were added to the wells, and incubation continued for 18-20 h. Cells were then stimulated with the TLR2/TLR6 agonist, FSL-1, at 10 ng/ml for 6 h. 
     At the end of the treatment, 20 μL of culture supernatant was taken from each well and assayed for SEAP by the Quanti Blue method according to the manufacturer&#39;s protocol (Invivogen). The data are depicted in  FIG. 2 . The data in  FIG. 2  depict NF-κB activity compared to control and demonstrate (i) that exemplary human TLR2 antisense oligonucleotides according to the invention are not immunostimulatory (Antisense Alone); and (ii) that exemplary human TLR2 antisense oligonucleotides according to the invention inhibit TLR2 expression and activation (Agonist Plus Antisense). 
     Example 3 
     In Vivo Activity of TLR2 Antisense Oligonucleotide 
     Female C57BL/6 mice of 5-6 weeks age (N=3/group) are injected with exemplary murine TLR2 antisense oligonucleotides according to the invention at 5 mg/kg, or PBS, subcutaneously once a day for three days. Subsequent to administration of the TLR2 antisense oligonucleotide, mice are injected with 0.25 mg/kg of a TLR2 agonist subcutaneously. Two hours after administration of the TLR2 agonist, blood is collected and TLR2 mRNA, TLR2 protein, and IL-12 concentrations are determined by ELISA.