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效果，但由于免疫受试者血清抗体浓度的变化，临床应用进展缓慢。建立疫苗免疫原性的早期标记将有助于预测疗效，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。这项研究I的总体假设是，从未免疫的和免疫的受试者中半抗原特异性B细胞的数量可以预测临床上对药物成瘾疫苗(药物半抗原-蛋白结合物)的显著反应。具体地说，将比较不同Oxy疫苗之间的羟考酮(Oxy)特异性B细胞，并将与Oxy特异性血清抗体和Oxy在啮齿动物中的分布相关。最近开发的一种策略在幼稚和免疫的受试者中检测罕见的抗原特异性B细胞。通过基于荧光抗原的富集法从未免疫或免疫的宿主体内的全部B细胞库中选择抗原特异的B细胞，然后用流式细胞术对其进行表征。这种方法可以通过将药物半抗原连接到用于检测半抗原特异性B细胞的荧光载体蛋白来适应药物结合疫苗。我们的实验室已经开发出两种Oxy疫苗，显示了在减少Oxy的大脑分布和行为影响方面的一系列效果，Oxy是最常被滥用的处方阿片之一。这些疫苗将被用来测试是否可以用Oxy特异的B细胞来解释免疫后结构不同的疫苗之间的差异和受试者之间的个体差异。我们将检验以下假设：1)更有效的Oxy疫苗对小鼠和大鼠免疫前后Oxy特异性B细胞的数量都会更高；2)免疫前后Oxy特异性B细胞的数量与Oxy特异性血清抗体效价以及免疫对Oxy在大鼠脑内分布的影响有关。这将通过以下方面进行测试：目的1A)表征小鼠体内Oxy特异性B细胞和Oxy特异性抗体对两种Oxy疫苗的反应时间进程，以确定B细胞反应；目的1B)在第二个物种(大鼠)中评估相同方法；目的2A)将免疫前的Oxy特异性B细胞与Oxy特异性抗体效价相关联，以及它们对Oxy在大鼠脑内分布的影响，以及目的2B)将免疫后早期激活的Oxy特异性B细胞与Oxy特异性血清抗体效价相关联，及其对Oxy在大鼠脑内分布的影响，以及对公共卫生意义。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严重的药物之一。与目前治疗依赖的药物疗法相比，药物成瘾疫苗提供了一个有希望的优势。及早筛查免疫原性将使疫苗与患者相匹配并快速调整治疗，从而实现个体化治疗。这种方法可以扩展到其他药物结合疫苗。 与公共健康相关：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严重的药物之一。与目前治疗依赖的药物疗法相比，药物成瘾疫苗提供了一个有希望的优势。及早筛查免疫原性将使疫苗与患者相匹配并快速调整治疗，从而实现个体化治疗。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.