Developing novel adjuvants for HIV vaccination

说明（申请人提供）：疫苗接种或自然感染引起的免疫反应不能有效预防和控制HIV感染。因此，探索替代免疫方法和新型佐剂以产生优于HIV感染天然免疫的保护性免疫反应是很重要的。抗原呈递细胞（APCs）如树突状细胞（DCs）在免疫应答的激活和维持中起着关键作用，它们受刺激和抑制信号的调节。最近，我们发现促炎信号的负调节因子，如锌指泛素修饰酶A20和细胞因子信号1抑制因子（SOCS1），在限制apc的免疫刺激效力和对自身抗原的自身反应性反应中起关键作用。我们证明A20或SOCS1的沉默极大地增强了TLR激动剂和DC疫苗诱导T细胞和抗体反应的刺激效力。在这项研究中，我们的目标是通过抑制促炎信号的关键负调节因子来开发新的强效佐剂，以促进HIV预防和治疗性疫苗的开发。中央本研究的假说是,组成的一种新型疫苗佐剂TLR配体和抑制剂的负面调节器的促炎信号触发和维持TLR信号级联可以创建在apc来增强细胞和体液反应的大小和持续时间对艾滋病毒水平较高,目前无法实现的描述或自然感染和疫苗接种方法也许能够克服艾滋病毒的免疫逃避和抑制。本研究的具体目的是：1)。测试可溶性鞭毛蛋白(FliC)-鱼精蛋白(P)融合蛋白/siA20复合物是否能作为新型佐剂刺激APCs诱导小鼠更强更广泛的hiv特异性CTL和Th反应；2)。在小鼠和家兔中，增强暴露于自然屏蔽的保护性表位的flicp /siA20复合物和突变HIV Env是否能更有效地诱导抗HIV的中和抗体；和3)。测试是否fli - p /sihA20复合物在激活人类dc刺激hiv特异性T细胞和B细胞方面优于TLR激动剂，用于潜在的临床应用。总之，迫切需要开发由TLR配体和A20抑制剂组成的新型佐剂，这些佐剂具有独特的刺激能力，可以将针对HIV的先天和适应性细胞和体液反应增强到更高水平，这是目前描述的佐剂无法实现的。在这项研究中，我们的目标是开发新的佐剂用于HIV疫苗接种，以诱导保护性细胞和体液免疫反应。具体来说，我们将生成并测试由TLR配体和A20抑制剂组成的新型佐剂，这些佐剂具有独特的刺激能力，可以将针对HIV的先天和适应性细胞和体液反应增强到更高水平，这是目前描述的佐剂无法实现的。

DESCRIPTION (provided by applicant): The immune responses induced by vaccination or natural infection fail to effectively prevent and control HIV infection. Thus, it is important to explore alternative immunization approaches and novel adjuvants to generate protective immune response that is superior to the natural immunity against HIV infection. Antigen-presenting cells (APCs) such as dendritic cells (DCs) play a critical role in the activation and maintenance of immune responses, and they are regulated by stimulatory as well as inhibitory signaling. Recently, we found that the negative regulators of proinflammatory signaling, such as the zinc-finger ubiquitin-modifying enzyme A20 and suppressor of cytokine signaling 1 (SOCS1), play critical roles in limiting the immunostimulatory potency of APCs and the autoreactive response against self-antigens. We demonstrated that silencing of A20 or SOCS1 drastically enhanced the stimulatory potency of TLR agonists and DC vaccines to induce both T cell and antibody responses. In this study we aim to develop novel and potent adjuvants by inhibiting key negative regulators of proinflammatory signaling to facilitate the development of prophylactic and therapeutic vaccines against HIV. The central hypothesis of this study is that a new type of vaccine adjuvants comprised of a TLR ligand and an inhibitor of the negative regulator of proinflammatory signaling for triggering and sustaining TLR signaling cascades in APCs can be created to enhance the magnitude and duration of cellular and humoral responses against HIV to higher levels that cannot be achieved by currently described vaccination approaches or natural infection and may be capable of overcoming HIV's immune evasion and suppression. The specific aims of this study are: 1). To test whether soluble flagellin (FliC)-protamine (P) fusion protein/siA20 complexes can potently stimulate APCs as novel adjuvants to induce stronger and broader HIV-specific CTL and Th responses in mice; 2). To investigate whether immunization of FliC-P/siA20 complexes and mutated HIV Env with enhanced exposure of naturally shielded, protective epitopes more efficiently induces neutralizing antibodies against HIV in mice and rabbits; and 3). To test whether FliC-P/sihA20 complexes are superior to TLR agonists in activating human DCs to stimulate HIV-specific T and B cells for potential clinic use. In summary, there is a compelling need for the development of the novel adjuvant comprised of TLR ligands and A20 inhibitors that have a unique stimulatory ability to enhance innate and adaptive cellular and humoral responses against HIV to higher levels that cannot be achieved by currently described adjuvants. In this proposed study, we aim to develop novel adjuvants for HIV vaccination to induce protective cellular and humoral immune responses. Specifically, we will generate and test the novel adjuvant comprised of TLR ligands and A20 inhibitors that have a unique stimulatory ability to enhance innate and adaptive cellular and humoral responses against HIV to higher levels that cannot be achieved by currently described adjuvants.