US008470342B2 (12) United States Patent (10) Patent N0.: US 8,470,342 B2 Klinman et a]. (45) Date of Patent: Jun. 25, 2013 (54) METHODS OF ALTERING AN IMMUNE OTHER PUBLICATIONS RESPONSE INDUCED BY CPG H d t 1 “Th D. t . l.k M t 11 t . ADAMIO OLIGODEOXYNUCLEOTIDES I un ausene a ., ‘e I1s1n egnn- 1 e e a opro einase I is Involved in Constitutive Cleavage of CX3CLl (fractalkine) and (75) Inventors: Dennis M_ Klinman’ Potomac, MD Regulates CXBCLl-mediated Cell-CellAdhesion,”Blood 10211186 (US); Mayda Gursel, Ankara (TR); 1195 (2003) Ihsan Gursel’ Ankara (TR) Wuttge et al., “CXCLl6/SR-PSOX Is an Interferon-y-Regulated Chemokine and Scavenger Receptor Expressed in Atherosclerotic (73) Assigneez The United states OfAmeriCa, as Lesions,” A(rterio)sclerosis, Thrombosis, and Vascular Biology Represented by the Secretary of the 24:750'755 2004 ~ _ _ _ Department of Health and Human Abel et al., The Transmembrane CXC-Chemokine Ligand 16 1s services Washington DC (Us) Induced by IFN-yand TNF-otand Shed by the Activity of the ’ ’ Disintergrin-Like Metalloproteinase ADAMlO,” Journal of Immu * N t- . S b- u d~ 1 - th t fthi nology 172(10):6362-6372 (May 15, 2004). ( ) 0 Ice :teJITtC isoeilgn 2x11251115; de ilnllioer 3 2 Gijbeles et al., “Reversal of Experimental Autoimmune % S C 154(1)) b 214 da JS Encephalomyelitis With a Hydroxamate Inhibitor of Matrix Metal ' ' ' y y ' loproteases,”Journal of Clinical Investigation 94:2177-2182 (1994). _ Gough et al., “A Disintegrin and Metalloproteinase lO-Mediated (21) Appl' NO" 13/005’488 Cleavage and Shedding Regulates the Cell Surface Expression of (22) Flled.. _ Jan. 12, 2011 CXC Chemokine Ligand 16,” Journal ofImmunology 17213678 3685 (2004). Gursel et al., “CXCLl6 In?uences the Nature and Speci?cally of (65) Pnor Pubhcatlon Data CpG-Induced Immune Activation,” Journal of Immunology US 2011/0110883 A1 May 12, 2011 l77(3):l575-l580 (Aug. 3, 2006). International Search report from PCT Application No. PCT/US2006/ Related US. Application Data 033774, dated Feb. 5, 2007. _ _ _ _ _ Tobata et al., “Distribution and Kinetics of SR-PSOX/CXCL16 and (62) Dlvlsloll of apphcanon NO‘ 12/065’085’ ?led as CXCR6 Expression on Human Dendritic cell Subsets and CD4+ T apphcanon NO‘ PCT/US2006/033774 on Aug‘ 28’ Cells,” Journal ofLeukocyte Biology 77:777-786 (2005). 2006’ HOW Pat' NO' 7>892>569~ Verthelyi et al., “Human Peripheral Blood Cells Differentially Rec (60) Provisional application NO‘ 60/713 547 ?led on Aug‘ ogniZe and Respond to Two Distinct CPG Motifs,”Journal ofImmu 31 2005 ’ ’ nology 166(4):2372-2377 (Feb. 15, 2001). (51) Int_ CL Primary Examiner * Patricia A Duffy A61]; 45/00 (200601) (74) Attorney, Agent, or Firm * Klarquist Sparkman, LLP A61K 39/12 (2006.01) A61K 39/02 (2006.01) (57) ABSTRACT (52) U-S- Cl- It is disclosed herein that agents that affect the activity and/or . .. expression of can be used to alter the uptake of 424/2341; 424/2741 D-type CpG oligodeoxynucleotides (D ODNs). Methods of (58) Field of Classi?cation Search inducing an immune response are disclosed that include None _ _ _ administering agents that increase the activity and/or expres See aPPhCaUOn ?le for Complete Search hlstory- sion of CXCL16 and a D ODN. Methods of decreasing an _ immune response to a CpG ODN are also disclosed. These (56) References Clted methods include administering an agent that decreases the FOREIGN PATENT DOCUMENTS activity and/or expression of CXCL1 6. Compositions includ JP 2 138868 5/1990 ing one or more D-type ODNs and an agent that modulates W0 W0 9556755 11/1999 that activity and/ or expression of CXCL1 6 are provided. W0 W0 00/61 151 10/2000 W0 WO 01/22990 4/2001 17 Claims, 7 Drawing Sheets US. Patent Jun. 25, 2013 Sheet 1 of7 US 8,470,342 B2 NCIO G NCIO >1 NGO Cl 0 O .M ow wo/ ,..... w. o L8 N r 2.u W03 mON 00.?omdowdovd :23cozgcmucoo200 6 U M m. w m0?@E8 4;.0E 00 cu LO O "I m. ‘ omd- v0.0- o o 0 (W 9017) GO US. Patent Jun. 25, 2013 Sheet 2 of7 US 8,470,342 B2 G NUumO 0O Q 8 n,}a)G n 9OGMW GM w,Wd.3mNi ...................N OMO GWm.m mm,é >3nuH amé, numiaoM ,. wm. QNGE Mx w d 2»$6,,mg»,, 869.3 am m .. .. w;~.N21.2, mw , ._, , m . ZOOZQO“cacaomxmm.GE mw zmomo2on750xQ8 n, <mt®¢ . a @8011! US. Patent Jun. 25, 2013 Sheet 4 of7 US 8,470,342 B2 QFIOXO ‘m @0502._. .GEmw US. Patent Jun. 25,2013 Sheet 5 of7 US 8,470,342 B2 ZOOQ v333 £2, HGM mm.P m»... 5“ “NAMES 33 US. Patent Jun. 25, 2013 Sheet 6 of7 US 8,470,342 B2 (2) NCIO (1 (L) NGOCI NCIO )1 @.UE IOJlUOO-G mum->1 Lumpew 0 O O om? om? CD (.0 00 (SHOHIHH) ma US. Patent Jun. 25,2013 Sheet 7 of7 US 8,470,342 B2 5282682023S9a. m5839a68% .v\w?EH55o\‘ Qowd.M EHmw§mmawér&mU»,‘musam‘. W,w?E a o., mvNF %m962w2 gEitIt.. ‘U‘U‘Uwa.U o l @mw .oZSE5 .2.‘mm..wn\ UK{K . o0 750! a wN .W..r wzoomO2‘ mKGE QSQ M8 owB w.3 .3..................................... US 8,470,342 B2 1 2 METHODS OF ALTERING AN IMMUNE Methods to alter the uptake and subsequent immune activa RESPONSE INDUCED BY CPG tion triggered by CpG ODN are disclosed herein. OLIGODEOXYNUCLEOTIDES SUMMARY PRIORITY CLAIM, It is disclosed herein that agents that affect the activity CROSS REFERENCE TO RELATED and/or expression of CXCL16 can be used to alter the uptake APPLICATIONS of CpG oligodeoxynucleotides (ODN), speci?cally D-type CpG oligodeoxynucleotides (D ODN). Thus, agents that This application is a divisional of US. patent application affect the activity and/ or expression of CXCL1 6 can be used Ser. No. 12/065,085, ?led Feb. 27, 2008 now US. Pat. No. to alter an immune response induced by D ODN. In one 7,892,569, Which is the US. National Stage of International example, the agent increases the activity and/or expression of CXCL16, thereby increasing the uptake of D ODN. Agents Application No. PCT/US2006/033774, ?led Aug. 28, 2006, that increase the activity and/or expression of CXCL1 6 can be Which Was published in English under PCT Article 21(2), used to increase an immune response induced by a D ODN. Which in turn claims the bene?t of US. Provisional Applica Agents that decrease the activity and/or expression of tion No. 60/713,547, ?led Aug. 31, 2005. The prior applica CXCL16 can be used to decrease an immune response tions are incorporated herein by reference in their entirety. induced by D ODN. Speci?c compositions including one or more D-type FIELD 20 ODNs and an agent that modulates that activity and/or expression of CXCL1 6 are provided herein. These composi This application relates to the ?eld of immunology, spe tions are of use to induce an immune response, such as to a ci?cally to agents that can be used to alter the uptake of speci?c antigen. immunostimulatory oligodeoxynucleotides (ODNs). The foregoing and other features and advantages Will 25 become more apparent from the folloWing detailed descrip BACKGROUND tion of several embodiments, Which proceeds With reference to the accompanying ?gures. DNA is a complex macromolecule Whose activities are in?uenced by its base composition and base modi?cation, as BRIEF DESCRIPTION OF THE FIGURES Well as helical orientation. Bacterial DNA, as Well as certain 30 synthetic oligodeoxynucleotides (ODNs) containing unm FIGS. 1A-1C are graphs and digital images illustrating that ethylated CpG sequences, can induce proliferation and CXCL16 selectively recognizes D ODN. FIG. 1A is a line immunoglobulin production by murine B cells. Unmethy graph shoWing CpG ODN binding to recombinant CXCL16. lated CpG dinucleotides are more frequent in the genomes of 96-Well ?at bottom ELISA plates Were coated With 0.4 ug/ml bacteria and viruses than vertebrates. Studies have suggested 35 anti-CXCL16 antibody and then incubated Without (dotted that immune recognition of these motifs may contribute to the lines) or With 200 ng/ml of recombinant CXCL1 6 (full lines). host’s innate immune response. (Klinman et al., Proc. Natl. FolloWing Washing, 1, 0.2 or 0.04 uM biotin-conjugated K or D oligodeoxynucleotide (ODN) Were added. After Washing, Acad. Sci. USA 9312879, 1996;Y1 et al., .I. Immun. 15715394, ODN binding Was detected colorimetrically using phos 1996; Liang et al., J. Clin. Invest. II 9:89, 1996; Krieg et al., 40 phatase-conjugated avidin folloWed by a phosphatase spe Nature 3741546, 1995). ci?c colorimetric substrate. Results are presented as A CpG oligodeoxynucleotide (ODN) is an oligodeoxy averagezstandard deviation (SD) of three independent read nucleotide including a CpG motif, Wherein the pyrimidine ings. FIG. 1B is a set of digital images and plots shoWing that ring of the cytosine is unmethylated. Three types of CpG D but not K ODN colocaliZe With CXCL16 and its uptake is ODNs have been identi?ed1 C-type, K-type and D-type 45 enhanced in transfected HEK293 cells. CXCL16 transfected ODNs. Generally, CpG ODNs range from about 8 to 30 bases HEK293 cells Were incubated With 3 uM of FITC conjugated in siZe. D- and K-type nucleic acid sequences have been CpG ODN at 37° C. for 20 minutes. Cells Were stained for described in the published PCT Publication No. W0 CXCL16 expression (left) and colocaliZation (bright cells, 98/ 18810A1 (K-type) and published PCT Publication No.