{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T18:30:44Z","timestamp":1790620244510,"version":"4.1.0"},"reference-count":33,"publisher":"American Chemical Society (ACS)","issue":"3","license":[{"start":{"date-parts":[[2015,2,11]],"date-time":"2015-02-11T00:00:00Z","timestamp":1423612800000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000026","name":"National Institute on Drug Abuse","doi-asserted-by":"publisher","award":["DA026625"],"award-info":[{"award-number":["DA026625"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,3,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Active immunization is an effective means of blocking the pharmacodynamic effects of drugs and holds promise as a treatment for heroin addiction. Previously, we demonstrated the efficacy of our first-generation vaccine in blocking heroin self-administration in rats, however, many vaccine components can be modified to further improve performance. Herein we examine the effects of varying heroin vaccine injection route and adjuvant formulation. Mice immunized via subcutaneous (sc) injection exhibited inferior anti-heroin titers compared to intraperitoneal (ip) and sc\/ip coadministration injection routes. Addition of TLR9 agonist cytosine-guanine oligodeoxynucleotide 1826 (CpG ODN 1826) to the original alum adjuvant elicited superior antibody titers and opioid affinities compared to alum alone. To thoroughly assess vaccine efficacy, full dose\u2013response curves were generated for heroin-induced analgesia in both hot plate and tail immersion tests. Mice treated with CpG ODN 1826 exhibited greatly shifted dose\u2013response curves (10\u201313-fold vs unvaccinated controls) while non-CpG ODN vaccine groups did not exhibit the same robust effect (2\u20137-fold shift for ip and combo, 2\u20133-fold shift for sc). Our results suggest that CpG ODN 1826 is a highly potent adjuvant, and injection routes should be considered for development of small molecule\u2013protein conjugate vaccines. Lastly, this study has established a new standard for assessing drugs of abuse vaccines, wherein a full dose\u2013response curve should be performed in an appropriate behavioral task.<\/jats:p>","DOI":"10.1021\/mp400631w","type":"journal-article","created":{"date-parts":[[2014,2,11]],"date-time":"2014-02-11T18:46:17Z","timestamp":1392144377000},"page":"1075-1080","source":"Crossref","is-referenced-by-count":88,"title":["Injection Route and TLR9 Agonist Addition Significantly\nImpact Heroin Vaccine Efficacy"],"prefix":"10.1021","volume":"11","author":[{"given":"Paul T.","family":"Bremer","sequence":"first","affiliation":[{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joel E.","family":"Schlosburg","sequence":"additional","affiliation":[{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenny M.","family":"Lively","sequence":"additional","affiliation":[{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim D.","family":"Janda","sequence":"additional","affiliation":[{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]},{"name":"The Scripps Research Institute , , 10550 North Torrey Pines Road , , ,","place":["La Jolla, California, United States, 92037"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,2,11]]},"reference":[{"issue":"2","key":"2026090121194183200_cit1","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S0376-8716(00)00162-9","article-title":"The economic costs\nof heroin addiction in the United States","volume":"61","author":"Mark","year":"2001","journal-title":"Drug\nAlcohol Depend."},{"issue":"3","key":"2026090121194183200_cit2","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.psychres.2013.01.014","article-title":"The economic\ncost of heroin dependency and quality of life among heroin users in\nTaiwan","volume":"209","author":"Lin","year":"2013","journal-title":"Psychiatry Res."},{"key":"2026090121194183200_cit3","volume-title":"Results from the 2010 National Survey on Drug Use and Health: Summary\nof National Findings","author":"Substance Abuse and Mental Health Services Administration.","year":"2011"},{"key":"2026090121194183200_cit4","volume-title":"WONDER (database)","author":"Centers for Disease Control and Prevention.","year":"2012"},{"issue":"5485","key":"2026090121194183200_cit5","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1038\/252708a0","article-title":"Changes\nin heroin self-administration by a rhesus monkey after morphine immunisation","volume":"252","author":"Bonese","year":"1974","journal-title":"Nature"},{"issue":"16","key":"2026090121194183200_cit6","doi-asserted-by":"crossref","first-page":"3232","DOI":"10.1016\/j.vaccine.2006.01.047","article-title":"A novel bivalent morphine\/heroin vaccine that prevents relapse to\nheroin addiction in rodents","volume":"24","author":"Anton","year":"2006","journal-title":"Vaccine"},{"issue":"31","key":"2026090121194183200_cit7","doi-asserted-by":"crossref","first-page":"4617","DOI":"10.1016\/j.vaccine.2012.04.101","article-title":"Co-administration of morphine and\noxycodone vaccines\nreduces the distribution of 6-monoacetylmorphine and oxycodone to\nbrain in rats","volume":"30","author":"Pravetoni","year":"2012","journal-title":"Vaccine"},{"issue":"14","key":"2026090121194183200_cit8","doi-asserted-by":"crossref","first-page":"5195","DOI":"10.1021\/jm200461m","article-title":"A vaccine\nstrategy that induces protective\nimmunity against heroin","volume":"54","author":"Stowe","year":"2011","journal-title":"J. Med. Chem."},{"issue":"22","key":"2026090121194183200_cit9","doi-asserted-by":"crossref","first-page":"9036","DOI":"10.1073\/pnas.1219159110","article-title":"Dynamic\nvaccine blocks relapse to compulsive intake\nof heroin","volume":"110","author":"Schlosburg","year":"2013","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"issue":"6813","key":"2026090121194183200_cit10","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1038\/35047123","article-title":"A Toll-like\nreceptor recognizes bacterial\nDNA","volume":"408","author":"Hemmi","year":"2000","journal-title":"Nature"},{"issue":"2","key":"2026090121194183200_cit11","doi-asserted-by":"crossref","first-page":"944","DOI":"10.4049\/jimmunol.164.2.944","article-title":"Mechanism and function\nof a newly identified CpG DNA\nmotif in human primary B cells","volume":"164","author":"Hartmann","year":"2000","journal-title":"J. Immunol."},{"issue":"17","key":"2026090121194183200_cit12","doi-asserted-by":"crossref","first-page":"1755","DOI":"10.1016\/S0264-410X(99)00526-5","article-title":"CpG DNA induces\nstronger immune responses with less toxicity than other adjuvants","volume":"18","author":"Weeratna","year":"2000","journal-title":"Vaccine"},{"issue":"26","key":"2026090121194183200_cit13","doi-asserted-by":"crossref","first-page":"15553","DOI":"10.1073\/pnas.95.26.15553","article-title":"CpG DNA can induce strong Th1 humoral and cell-mediated\nimmune responses against hepatitis B surface antigen in young mice","volume":"95","author":"Brazolot Millan","year":"1998","journal-title":"Proc. Natl. Acad. Sci. 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Cell Biol."},{"issue":"1","key":"2026090121194183200_cit15","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.vaccine.2006.06.084","article-title":"Immunogenicity of a receptor-binding\ndomain of SARS coronavirus spike protein in mice: implications for\na subunit vaccine","volume":"25","author":"Zakhartchouk","year":"2007","journal-title":"Vaccine"},{"issue":"6","key":"2026090121194183200_cit16","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1007\/s10875-004-6244-3","article-title":"CPG 7909,\nan immunostimulatory TLR9 agonist oligodeoxynucleotide, as adjuvant\nto Engerix-B HBV vaccine in healthy adults: a double-blind phase I\/II\nstudy","volume":"24","author":"Cooper","year":"2004","journal-title":"J. Clin. Immunol."},{"issue":"8","key":"2026090121194183200_cit17","doi-asserted-by":"crossref","first-page":"e2940","DOI":"10.1371\/journal.pone.0002940","article-title":"Phase 1 trial\nof AMA1-C1\/Alhydrogel plus CPG 7909:\nan asexual blood-stage vaccine for Plasmodium falciparum malaria","volume":"3","author":"Mullen","year":"2008","journal-title":"PloS One"},{"issue":"1","key":"2026090121194183200_cit18","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1086\/653112","article-title":"Improving\nthe immunogenicity of pneumococcal conjugate vaccine in HIV-infected\nadults with a toll-like receptor 9 agonist adjuvant: a randomized,\ncontrolled trial","volume":"51","author":"Sogaard","year":"2010","journal-title":"Clin. Infect. Dis."},{"issue":"21","key":"2026090121194183200_cit19","doi-asserted-by":"crossref","first-page":"8947","DOI":"10.1073\/pnas.0703395104","article-title":"Vaccination with NY-ESO-1 protein\nand CpG in Montanide induces integrated antibody\/Th1 responses and\nCD8 T cells through cross-priming","volume":"104","author":"Valmori","year":"2007","journal-title":"Proc. Natl.\nAcad. Sci. U.S.A."},{"issue":"28","key":"2026090121194183200_cit20","doi-asserted-by":"crossref","first-page":"4324","DOI":"10.1200\/JCO.2010.28.9793","article-title":"In situ vaccination with a TLR9 agonist induces systemic lymphoma\nregression: a phase I\/II study","volume":"28","author":"Brody","year":"2010","journal-title":"J. Clin. Oncol."},{"issue":"23","key":"2026090121194183200_cit21","doi-asserted-by":"crossref","first-page":"10776","DOI":"10.1021\/jm301262z","article-title":"Investigating\nthe Effects of a Hydrolytically Stable\nHapten and a Th1 Adjuvant on Heroin Vaccine Performance","volume":"55","author":"Bremer","year":"2012","journal-title":"J. Med. Chem."},{"issue":"7","key":"2026090121194183200_cit22","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1097\/FBP.0b013e3283584794","article-title":"Evaluation\nof the endogenous cannabinoid system in mediating the behavioral effects\nof dipyrone (metamizol) in mice","volume":"23","author":"Schlosburg","year":"2012","journal-title":"Behav. Pharmacol."},{"issue":"1","key":"2026090121194183200_cit23","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.vaccine.2012.10.069","article-title":"High immunogenicity\nof nicotine vaccines obtained by intradermal delivery with safe adjuvants","volume":"31","author":"Chen","year":"2012","journal-title":"Vaccine"},{"issue":"12","key":"2026090121194183200_cit24","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1016\/j.medmal.2008.09.012","article-title":"[Are there differences in immunogenicity\nand safety of vaccines according to the injection method?]","volume":"38","author":"Ajana","year":"2008","journal-title":"Med. Mal. Infect."},{"issue":"7198","key":"2026090121194183200_cit25","doi-asserted-by":"crossref","first-page":"1122","DOI":"10.1038\/nature06939","article-title":"Crucial role for the Nalp3 inflammasome in the immunostimulatory\nproperties of aluminium adjuvants","volume":"453","author":"Eisenbarth","year":"2008","journal-title":"Nature"},{"issue":"Part 3","key":"2026090121194183200_cit26","first-page":"369","article-title":"Patterns\nof lymphatic drainage in the adult laboratory rat","volume":"109","author":"Tilney","year":"1971","journal-title":"J. Anat."},{"issue":"20","key":"2026090121194183200_cit27","doi-asserted-by":"crossref","first-page":"3588","DOI":"10.1016\/j.vaccine.2010.02.085","article-title":"Mechanism of immunopotentiation\nby aluminum-containing adjuvants elucidated by the relationship between\nantigen retention at the inoculation site and the immune response","volume":"28","author":"Noe","year":"2010","journal-title":"Vaccine"},{"issue":"11","key":"2026090121194183200_cit28","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1038\/nri2868","article-title":"Vaccine delivery: a matter of size, geometry, kinetics\nand molecular patterns","volume":"10","author":"Bachmann","year":"2010","journal-title":"Nat. Rev. Immunol."},{"issue":"1","key":"2026090121194183200_cit29","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.intimp.2013.03.021","article-title":"Enhancing immunogenicity of a 3\u2032aminomethylnicotine-DT-conjugate\nanti-nicotine vaccine with CpG adjuvant in mice and non-human primates","volume":"16","author":"McCluskie","year":"2013","journal-title":"Int. Immunopharmacol."},{"issue":"2","key":"2026090121194183200_cit30","doi-asserted-by":"crossref","first-page":"870","DOI":"10.4049\/jimmunol.160.2.870","article-title":"CpG DNA is a potent enhancer of specific immunity in\nmice immunized with recombinant hepatitis B surface antigen","volume":"160","author":"Davis","year":"1998","journal-title":"J. Immunol."},{"issue":"4","key":"2026090121194183200_cit31","doi-asserted-by":"crossref","first-page":"1544","DOI":"10.1073\/pnas.94.4.1544","article-title":"Opiate receptor\nknockout mice define mu receptor roles in endogenous nociceptive responses\nand morphine-induced analgesia","volume":"94","author":"Sora","year":"1997","journal-title":"Proc. Natl.\nAcad. Sci. U.S.A."},{"issue":"2","key":"2026090121194183200_cit32","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1021\/mp900213u","article-title":"A critical evaluation of a nicotine\nvaccine within a self-administration behavioral model","volume":"7","author":"Moreno","year":"2010","journal-title":"Mol. Pharmaceutics"},{"issue":"22","key":"2026090121194183200_cit33","doi-asserted-by":"crossref","first-page":"2981","DOI":"10.1016\/j.vaccine.2009.02.105","article-title":"Immune responses to methamphetamine\nby active immunization with peptide-based, molecular adjuvant-containing\nvaccines","volume":"27","author":"Duryee","year":"2009","journal-title":"Vaccine"}],"container-title":["Molecular\nPharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/mp400631w","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/mpohbp\/article-pdf\/11\/3\/1075\/14367069\/mp400631w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/mpohbp\/article-pdf\/11\/3\/1075\/14367069\/mp400631w.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T01:19:52Z","timestamp":1788311992000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/mpohbp\/article\/11\/3\/1075\/1379184\/Injection-Route-and-TLR9-Agonist-Addition"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,2,11]]},"references-count":33,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2014,2,11]]},"published-print":{"date-parts":[[2014,3,3]]}},"URL":"https:\/\/doi.org\/10.1021\/mp400631w","relation":{},"ISSN":["1543-8384","1543-8392"],"issn-type":[{"value":"1543-8384","type":"print"},{"value":"1543-8392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,2,11]]}}}