title: Efficacy, but not antibody titer or affinity, of a heroin hapten conjugate vaccine correlates with increasing hapten densities on tetanus toxoid, but not on CRM197 carriers.
abstract: Vaccines against drugs of abuse have induced antibodies in animals that blocked the biological effects of the drug by sequestering the drug in the blood and preventing it from crossing the blood-brain barrier. Drugs of abuse are too small to induce antibodies and, therefore, require conjugation of drug hapten analogs to a carrier protein. The efficacy of these conjugate vaccines depends on several factors including hapten design, coupling strategy, hapten density, carrier protein selection, and vaccine adjuvant. Previously, we have shown that 1 (MorHap), a heroin/morphine hapten, conjugated to tetanus toxoid (TT) and mixed with liposomes containing monophosphoryl lipid A [L(MPLA)] as adjuvant, partially blocked the antinociceptive effects of heroin in mice. Herein, we extended those findings, demonstrating greatly improved vaccine induced antinociceptive effects up to 3% mean maximal potential effect (%MPE). This was obtained by evaluating the effects of vaccine efficacy of hapten 1 vaccine conjugates with varying hapten densities using two different commonly used carrier proteins, TT and cross-reactive material 197 (CRM197). Immunization of mice with these conjugates mixed with L(MPLA) induced very high anti-1 IgG peak levels of 400-1500 μg/mL that bound to both heroin and its metabolites, 6-acetylmorphine and morphine. Except for the lowest hapten density for each carrier, the antibody titers and affinity were independent of hapten density. The TT carrier based vaccines induced long-lived inhibition of heroin-induced antinociception that correlated with increasing hapten density. The best formulation contained TT with the highest hapten density of ≥30 haptens/TT molecule and induced %MPE of approximately 3% after heroin challenge. In contrast, the best formulation using CRM197 was with intermediate 1 densities (10-15 haptens/CRM197 molecule), but the %MPE was approximately 13%. In addition, the chemical synthesis of 1, the optimization of the conjugation method, and the methods for the accurate quantification of hapten density are described.
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.|‡U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Jalah
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.|‡U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
authors.forename: Oscar B
authors.initials: OB
authors.lastname: Torres
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.|‡U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Mayorov
authors.affiliation: §Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, 9800 Medical Drive, Bethesda, Maryland 20892, United States.|¶National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, 9800 Medical Drive, Bethesda, Maryland 20892, United States.
authors.forename: Fuying
authors.initials: F
authors.lastname: Li
authors.affiliation: §Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, 9800 Medical Drive, Bethesda, Maryland 20892, United States.|¶National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, 9800 Medical Drive, Bethesda, Maryland 20892, United States.
authors.forename: Joshua F G
authors.initials: JF
authors.lastname: Antoline
authors.affiliation: §Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, 9800 Medical Drive, Bethesda, Maryland 20892, United States.|¶National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, 9800 Medical Drive, Bethesda, Maryland 20892, United States.
authors.forename: Arthur E
authors.initials: AE
authors.lastname: Jacobson
authors.affiliation: §Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, 9800 Medical Drive, Bethesda, Maryland 20892, United States.|¶National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, 9800 Medical Drive, Bethesda, Maryland 20892, United States.
authors.forename: Kenner C
authors.initials: KC
authors.lastname: Rice
authors.affiliation: ||Center for Biomolecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, United States.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Deschamps
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.|‡U.S. Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Beck
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.
authors.forename: Carl R
authors.initials: CR
authors.lastname: Alving
authors.affiliation: †Laboratory of Adjuvant and Antigen Research, US Military HIV Research Program, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.
authors.forename: Gary R
authors.initials: GR
authors.lastname: Matyas
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology; D000701:Analgesics, Opioid / Q000276:immunology* / Q000494:pharmacology; D000818:Animals; D000915:Antibody Affinity; D001426:Bacterial Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004337:Drug Carriers / Q000737:chemistry*; D005260:Female; D006241:Haptens / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D003932:Heroin / Q000276:immunology* / Q000494:pharmacology; D006556:Heroin Dependence / Q000276:immunology / Q000517:prevention & control; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D008050:Lipid A / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000276:immunology; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D013745:Tetanus Toxoid / Q000737:chemistry*; D018074:Vaccines, Conjugate / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology
chemical_list: D000276:Adjuvants, Immunologic; D000701:Analgesics, Opioid; D001426:Bacterial Proteins; D004337:Drug Carriers; D006241:Haptens; D007074:Immunoglobulin G; D008050:Lipid A; D013745:Tetanus Toxoid; D018074:Vaccines, Conjugate; C028581:CRM197 (non-toxic variant of diphtheria toxin); D003932:Heroin; C048436:monophosphoryl lipid A