Early antigen-specific B cell responses as markers of oxycodone vaccine efficacy

描述（由申请人提供）：药物成瘾疫苗在临床前研究中显示出令人鼓舞的疗效，但由于免疫受试者血清抗体浓度的可变性，转化为临床的速度减慢。建立疫苗免疫原性的早期标记将有助于预测有效性，加快疫苗设计的筛选，并在临床环境中个性化疫苗设计选择。本研究的总体假设是，对药物成瘾疫苗（药物半抗原蛋白偶联物）的临床显著反应可以从未接种（未接种）和接种的受试者中半抗原特异性B细胞的数量来预测。特异性羟考酮（OXY）特异性B细胞将在不同的OXY疫苗之间进行比较，并将与OXY疫苗免疫前后的OXY特异性血清抗体和OXY分布相关。最近开发的一种策略检测罕见的抗原特异性B细胞在幼稚和免疫受试者。抗原特异性B细胞通过荧光抗原富集方法从未免疫或免疫宿主的B细胞库中选择，然后用流式细胞术进行表征。通过将药物半抗原偶联到用于检测半抗原特异性B细胞的荧光载体蛋白上，该方法可适用于药物偶联疫苗。我们的实验室已经开发了两种羟氧基疫苗，显示出一系列减少羟氧基大脑分布和行为影响的效果，羟氧基是最常被滥用的处方阿片类药物之一。这些疫苗将用于测试氧特异性B细胞是否可以用来解释结构不同的疫苗之间的差异以及免疫后受试者之间的个体差异。我们将验证以下假设：1)免疫前和免疫后的氧特异性B细胞数量将会更高，小鼠和大鼠的氧疫苗效率更高；2)免疫前和免疫后的氧特异性B细胞数量将与氧特异性血清抗体滴度相关，并与免疫对氧在大鼠大脑分布的影响相关。这将通过：目的1A)表征氧特异性B细胞和氧特异性抗体对小鼠两种氧疫苗反应的时间过程来确定B细胞反应；目的1B)在第二个物种（大鼠）中评估相同的方法；目的2A)免疫前激活的氧特异性B细胞与氧特异性抗体滴度的关系及其对大鼠脑内氧分布的影响；目的2B)免疫后早期激活的氧特异性B细胞与氧特异性血清抗体滴度的关系及其对大鼠脑内氧分布的影响。处方药滥用是美国增长最快的毒品问题，羟考酮和羟吗啡酮是滥用最多的药物。药物成瘾疫苗为目前治疗药物依赖的药物疗法提供了一个有希望的优势。免疫原性的早期筛查将通过使疫苗与患者相匹配和快速调整治疗来实现个体化治疗。这种方法可以扩展到其他药物结合疫苗。

DESCRIPTION (provided by applicant): Drug addiction vaccines show encouraging efficacy in pre-clinical studies, yet translation to the clinic has been slowed by the variability of serum antibody concentrations reported in immunized subjects. Establishing early markers of vaccine immunogenicity would help predict efficacy, accelerate screening of vaccine designs, and individualize vaccine design selection in clinical settings. The overall hypothesis of this study i that clinically significant responses to drug addiction vaccines (drug hapten-protein conjugate) can be predicted from the number of hapten-specific B cells in naive (unimmunized) and immunized subjects. Specifically oxycodone (OXY)-specific B cells will be compared between different OXY vaccines and will correlate with OXY-specific serum antibodies and OXY distribution before and after immunization with OXY vaccines in rodents. A recently developed strategy detects rare antigen-specific B cells in naive and immunized subjects. Antigen-specific B cells are selected from the total B cell repertoire present in an unimmunized or immunized host by a fluorescent antigen-based enrichment method and then characterized by flow cytometry. This method can be adapted to drug conjugate vaccines by conjugating drug haptens to a fluorescent carrier protein used to detect hapten-specific B cells. Our lab has developed two OXY vaccines showing a range of effects in reducing brain distribution and behavioral effects of OXY, one of the most commonly abused prescription opioids. These vaccines will be used to test if OXY-specific B cells can be used to explain variability between structurally different vaccines and individual variability between subjects after immunization. We will test the hypotheses that: 1) the numbers of pre- and post-immunization OXY-specific B cells will be higher for the more efficient OXY vaccine in both mice and rats 2) the numbers of pre- and post- immunization OXY-specific B cells will correlate with OXY-specific serum antibody titers and with the effect of immunization on OXY distribution to the brain in rats. This will be tested by: aim 1A) characterizing the time course of OXY-specific B cell and OXY-specific antibody responses to two OXY vaccines in mice to determine B cell responses; aim 1B) assessing the same method in a second species (rats); aim 2A) correlating pre- immunization na¿ve OXY-specific B cells to OXY-specific antibody titers and their effects on OXY distribution to the brain in rats, and aim 2B) correlating early post- immunization activated OXY-specific B cells to OXY- specific serum antibody titers and their effects on OXY distribution to brain in rats PUBLIC HEALTH SIGNIFICANCE. Prescription drug abuse is the fastest-growing drug problem in the US, with oxycodone and oxymorphone among the most abused. Drug addiction vaccines offer a promising advantage to current pharmacotherapies to treat dependence. Early screening of immunogenicity will enable individualized treatment by matching the vaccine to patient and rapid treatment adjustment. This approach could be expanded to other drug-conjugate vaccines. PUBLIC HEALTH RELEVANCE: Prescription drug abuse is the fastest-growing drug problem in the US, with oxycodone and oxymorphone among the most abused. Drug addiction vaccines offer a promising advantage to current pharmacotherapies to treat dependence. Early screening of immunogenicity will enable individualized treatment by matching the vaccine to patient and rapid treatment adjustment. This approach could be expanded to other drug-conjugate vaccines.