Impact of Human STING Variants on Pneumococcal Vaccine Effectiveness

摘要 这项提案的目标是确定肺炎球菌疫苗的有效性，该疫苗受 人类常见的刺痛(干扰素基因刺激物)变异体R71H-G230A-R293Q(HAQ)。肺炎双球菌 疾病导致的死亡人数比所有其他疫苗可预防疾病的总和还要多。Pappovax®23和Prevnar 13®是提供血清型特异性保护的多糖疫苗。自1983年批准以来， 仅有约57%的总体疾病减少率为Patiovax 23®。最近，布鲁斯·比特勒博士的团队展示了 多糖银在体内产生一种名为2‘5’-3‘5’-cGMP-AMP的环二核苷酸(CDN)。 B细胞激活刺激型I型干扰素途径。STING是CDN的受体。被叮咬的小鼠 已经减少了对Pappovax®23的抗体产生。我们自己的数据显示，被叮咬的老鼠也有 抗体产量降至Prevnar®13。我们之前发现了一种人类刺变异体HAQ，它是 在非非洲人中很常见。约3%的高加索人和约16%的东亚人是HAQ/HAQ。事实上，R232(重量)/Haq， 而不是R232(Wt)/R232(Wt)，是东亚人最常见的刺痛基因。检测来源的B细胞 从人类HAQ/HAQ个体中，我们发现HAQ B细胞显著减少了刺痛 表情。我们假设肺炎病毒®23和Prevnar 13®疫苗的效力受以下因素影响 哈克·斯汀变种。为了验证这一假设并揭示体内机制，我们生成了HAQ，AQ 和Q293叮咬敲入鼠。在目标1中，我们将确定(A)HAQ刺痛对有效性的影响 (B)HAQ在体内的作用机制 STING会影响Pappovax®23和Prevnar13®的有效性(目标1.2)。Pappovax®23和Prevnar 13®不能引起较强的粘膜免疫。一种由一种常见蛋白Ag组成的粘膜肺炎球菌疫苗 一种有效的粘膜佐剂可以提供不依赖于血清型的粘膜保护。最近，一种合成的 CDN，RpRp-c-di-AMPss，被报道能激活HEK293T细胞中所有的人刺突变体。我们的预赛 数据表明，RpRp-c-di-AMPss具有黏膜疫苗佐剂活性，可诱导产生强大的抗体 平衡Th1/Th2/Th17反应，这依赖于刺痛。在目标2中，我们将确定(A) HAQ刺痛RpRp-c-di-AMPss在体内的粘膜佐剂活性(Aim 2.1)；(B)体内机制 HAQ刺痛通过其影响RpRp-c-di-AMPss粘膜佐剂活性(目标2.2)。我们将评估 RpRp-c-di-AMPss佐剂肺炎球菌表面蛋白A疫苗诱导的体液、细胞免疫 我们的敲入小鼠的免疫反应和对肺炎链球菌感染的保护性免疫。 实现这些目标将为人类刺靶精准医学的发展铺平道路。

Abstract The goal of this proposal is to determine the effectiveness of pneumococcal vaccines influenced by the common human STING (stimulator of interferon genes) variant R71H-G230A-R293Q (HAQ). Pneumococcal diseases kill more people than all other vaccine-preventable diseases combined. Pneumovax® 23 and Prevnar 13® are polysaccharide vaccines providing serotype-specific protection. Since its approval in 1983, Pneumovax 23® results in only ~57% of overall disease reduction. Recently, Dr. Bruce Beutler’s group showed that polysaccharide Ag generates a cyclic dinucleotide (CDN) called 2’5’-3’5’-cyclic GMP-AMP (2’3’-cGAMP) in B cells leading to the activation of STING-Type I IFN pathway. STING is a receptor for CDN. STING-/- mice have decreased Ab production to Pneumovax® 23. Our own data showed that STING-/- mice also have decreased Ab production to Prevnar® 13. We previously identified a human STING variant HAQ that is common in non-Africans. ~3% of Caucasians and ~16% of East Asians are HAQ/HAQ. In fact, R232(wt)/HAQ, not the R232(wt)/R232(wt), is the most common STING genotype in East Asians. Examining B cells derived from human HAQ/HAQ individuals, we found that HAQ B cells have dramatically decreased STING expression. We hypothesize that Pneumovax ® 23 and Prevnar 13® vaccine effectiveness is influenced by HAQ STING variant. To test this hypothesis and uncover the in vivo mechanism, we generated the HAQ, AQ and Q293 STING knock-in mice. In Aim 1, we will determine (a) the impact of HAQ STING on the effectiveness of Pneumovax® 23 and Prevnar13® vaccines in vivo (Aim 1.1); (b) the in vivo mechanism by which HAQ STING influences Pneumovax® 23 and Prevnar13® effectiveness (Aim 1.2). Pneumovax® 23 and Prevnar 13® do not elicit strong mucosal immunity. A mucosal pneumococcal vaccine consists of a common protein Ag and a potent mucosal adjuvant can provide serotype-independent mucosal protection. Recently, a synthetic CDN, RpRp-c-di-AMPss, was reported to activate all human STING variants in HEK293T cells. Our preliminary data demonstrated that RpRp-c-di-AMPss has mucosal vaccine adjuvant activity eliciting strong Ab production and balanced Th1/Th2/Th17 response that depends on STING. In Aim 2, we will determine (a) the impact of HAQ STING on the mucosal adjuvant activity of RpRp-c-di-AMPss in vivo (Aim 2.1); (b) the in vivo mechanism by which HAQ STING influences RpRp-c-di-AMPss mucosal adjuvant activity (Aim 2.2). We will evaluate RpRp-c-di-AMPss adjuvanted pneumococcal surface protein A (PspA) vaccine induced humoral, cellular immune responses, and the protective immunity against S. pneumoniae infection in our knock-in mice. Achieving these Aims will pave the way for the development of STING-targeting precision medicine in humans.