Impact of Human STING Variants on Pneumococcal Vaccine Effectiveness

摘要 该提案的目的是确定肺炎球菌疫苗的有效性， 常见的人STING（干扰素基因刺激因子）变体R71 H-G230 A-R293 Q（HAQ）。肺炎球菌 这些疾病造成的死亡人数超过了所有其他疫苗可预防疾病的总和。Pneumovax® 23和Prevnar 13®是多糖疫苗，可提供针对特定类型的保护。自1983年批准以来， Pneumovax 23®仅能减少约57%的总体疾病。最近，布鲁斯·伯特勒博士的研究小组发现， 该多糖Ag产生称为2 '5'-3 '5 '-环状GMP-AMP（2' 3 '-cGAMP）的环状二核苷酸（CDN）， B细胞导致STING-I型IFN途径的激活。STING是CDN的受体。STING-/-小鼠 减少了Pneumovax® 23的抗体产生。我们自己的数据显示，STING-/-小鼠也有 将Ab产量降低至Prevnar® 13。我们先前鉴定了一种人类STING变体HAQ， 在非洲以外的地方很常见。约3%的白人和约16%的东亚人是HAQ/HAQ。事实上，R232（wt）/HAQ， 而不是R232（wt）/R232（wt），是东亚人中最常见的STING基因型。检查B细胞来源 从人类HAQ/HAQ个体中，我们发现HAQ B细胞具有显著降低的STING， 表情我们假设Pneumovax ® 23和Prevnar 13®疫苗的有效性受到以下因素的影响： HAQ STING变体。为了验证这一假设并揭示体内机制，我们生成了HAQ，AQ 和Q293 STING敲入小鼠。在目标1中，我们将确定（a）HAQ STING对有效性的影响 Pneumovax® 23和Prevnar 13 ®疫苗的体内作用（目的1.1）;（B）HAQ STING影响Pneumovax® 23和Prevnar 13 ®的有效性（目标1.2）。Pneumovax® 23和Prevnar 13®不引起强粘膜免疫。粘膜肺炎球菌疫苗由一种常见的蛋白质Ag 并且有效的粘膜佐剂可以提供非依赖于粘膜类型的粘膜保护。最近，一种合成的 据报道，CDN RpRp-c-di-AMPss激活HEK 293 T细胞中的所有人STING变体。我们的初步 数据表明RpRp-c-di-AMPss具有粘膜疫苗佐剂活性，引起强Ab产生 以及依赖于STING的平衡的Th 1/Th 2/Th 17应答。在目标2中，我们将确定（a） HAQ STING对RpRp-c-di-AMPss的体内粘膜佐剂活性的影响（目的2.1）;（B）体内机制 HAQSTING通过其影响RpRp-c-di-AMPss粘膜佐剂活性（目的2.2）。我们将评估 RpRp-c-di-AMPss佐剂肺炎球菌表面蛋白A（PspA）疫苗诱导的体液、细胞免疫应答 免疫应答，以及对S. pneumoniae肺炎infection感染in our knockin敲入mice小鼠. 实现这些目标将为人类靶向STING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.