HERPES SIMPLEX VIRUS IMMUNE EVASION IN HIV SUBJECTS

描述：(申请人提供)在HIV感染者中，HSV 随着CD4T细胞计数的减少，感染的频率和严重程度都会增加。这 应用提出了一种通过改进控制HSV-1感染的新策略 抗体和补体(C)在宿主防御中的有效性。单纯疱疹病毒1型的用途 躲避抗体和C攻击的隐形策略。单纯疱疹病毒1型糖蛋白GC 在C级联的几个步骤中干扰C的激活，呈现C 对抗病毒效果不佳。然而，如果结合C3的关键GC结构域 从病毒中删除，C将HSV-1的毒力降低50%至100倍。单纯疱疹病毒1型 糖蛋白GE通过结合Ig G Fc结构域和封闭来逃避抗体 Fc介导的活性，包括C激活和抗体依赖的细胞 细胞毒性。一株不能结合Ig G Fc结构域的HSV-1 gE变异病毒是50 由于抗体增强，毒力比野生型病毒低100倍 有效性。申请者构建了一种突变病毒，在GC和 GE，并证明这些糖蛋白协同作用以逃避抗体 和C攻击，因为GC-GE双突变病毒的数量减少了1,000到1,000 比野生型病毒毒力强。作为宾夕法尼亚大学CFAR倡议的一部分，一种艾滋病毒 病人登记簿和标本资料库于1999年开发，用于登记 受试者在宾夕法尼亚大学的诊所接受治疗。申请者将分析血清样本 从储存库获取以确定HIV对象是否保持 足够水平的HSV抗体和C来中和突变的HSV-1毒株 这在GC和GE免疫逃避方面是有缺陷的。他们假定大多数人 HIV/HSV-1混合感染者将保持足够的抗体和C 滴度，以中和至少100倍的GC-GE突变比野生型病毒。 根据初步结果，他们推测在 自然感染HSV-1不能防止GC和GE免疫逃逸，因为 所涉及的域对主机是“隐藏”的。因此，他们建议修改 GC和GE免疫逃逸结构域，提高免疫原性。他们将对这些进行测试 在小鼠模型中改变蛋白质作为候选疫苗以确定 产生的抗体阻断了GC和GE的免疫逃避，减少了疾病 严肃性。CD4基因敲除小鼠将被用来研究抗体和C 如果病毒免疫缺陷，可以弥补CD4T细胞免疫功能的下降 逃避。防止病毒隐形可能会极大地提高抗体和C活性 可能为发展HSV和其他病毒提供一种新的策略 病毒疫苗。

DESCRIPTION: (Provided by the Applicant) In HIV infected individuals, HSV infections increase in frequency and severity as CD4 T cells counts wane. This application proposes a novel strategy to control HSV-1 infection by improving the effectiveness of antibody and complement (C) in host defense. HSV-1 uses stealth strategies to evade antibody and C attack. HSV-1 glycoprotein gC interferes with C activation at several steps in the C cascade, rendering C ineffective against the virus. However, if a critical gC domain that binds C3 is deleted from the virus, C reduces HSV-1 virulence 50- to 100-fold. HSV-1 glycoprotein gE evades antibody by binding the IgG Fc domain and blocking Fc-mediated activities, including C activation and antibody-dependent cellular cytotoxicity. An HSV-1 gE mutant virus unable to bind the IgG Fc domain is 50 to 100-fold less virulent than wild-type virus because of enhanced antibody effectiveness. The applicants constructed a mutant virus altered in both gC and gE and demonstrate that these glycoproteins act in synergy to evade antibody and C attack, since the gC-gE double mutant virus is 1,000- to 10,000-fold less virulent than wild-type virus. As part of the CFAR initiative at Penn, an HIV patient registry and specimen repository was developed in 1999 to enroll subjects cared for at Penn clinics. The applicants will analyze serum samples obtained from the repository to determine whether HIV subjects maintain adequate levels of HSV antibodies and C to neutralize a mutant HSV-1 strain that is defective in gC and gE immune evasion. They postulate that the majority of HIV/HSV-1 co-infected subjects will maintain sufficient antibody and C titers to neutralize at least 100-fold more gC-gE mutant than wild-type virus. Based on preliminary results, they postulate that antibodies produced during natural HSV-1 infection cannot prevent gC and gE immune evasion because the domains involved are "hidden" from the host. Therefore, they propose to modify gC and gE immune evasion domains to improve immunogencity. They will test these altered proteins as vaccine candidates in murine models to determine whether the antibodies produced block gC and gE immune evasion and reduce disease severity. CD4-knockout mice will be used to investigate whether antibody and C can compensate for waning CD4 T cell immunity if virus is defective in immune evasion. Preventing viral stealth may greatly improve antibody and C activities in HIV subjects, and may offer a novel strategy for developing HSV and other viral vaccines.