NOVEL DELIVERY SYSTEM FOR A HERPESVIRUS DNA VACCINE

大多数成年人携带单纯疱疹病毒I型（HSV-1），但只有一些 患有周期性发作的疼痛性发热水泡。 T细胞限制病毒 通过诱导神经节细胞中的非允许状态来发病，其中 病毒潜伏着。 细胞毒性T淋巴细胞（CTL），尤其是CD 4 + CTL在抵抗复发性疱疹损伤中至关重要。 病毒 在HSV感染早期表达的糖蛋白（gP）是CTL靶标， 已经被用作治疗性疫苗并取得了一定的成功。 虽然 通常需要活疫苗来刺激有效的CTL应答， 编码HSV gP的裸DNA疫苗刺激CTL和抗体应答 实验性的 这项研究的假设是， 编码HSV-1糖蛋白D（gD），保护其不受核酸酶活性影响， 靶向携带无唾液酸类粘蛋白受体（ASOR）的细胞， 刺激小鼠对单纯疱疹病毒特异性细胞免疫。 这种策略的新奇在于，配体，如聚-L-赖氨酸-ASOR， （ASOR-L），保护DNA免受核酸酶的侵害，同时将其引导至细胞 带有配体的受体，具有裸HSV gP DNA所缺乏的特征 疫苗。 为了检验这一假设，gD DNA与ASOR-L复合， 通过4种不同的途径将裸的注射到BALB/c和C3 H小鼠中 [静脉内（iv）、腹腔内（ip）、肌内（im）和口服 （po）]。 在注射10 μ g剂量的DNA后2周， 将检查细胞针对组织相容性靶标的CTL活性 表达gD的细胞。 因为肝细胞，巨噬细胞和肠细胞 携带ASOR受体，更高水平的gD特异性CTL活性将 更可能在通过iv、ip和po途径接受gD-ASOR的小鼠中发展。 刺激最佳HSV CTL应答的条件将由以下定义： 本研究 这种新方法的实际应用是一种 用于减轻人类疼痛性疱疹损伤复发的疗法。

Most adults harbor herpes simplex virus type I(HSV-1) but only some suffer periodic episodes of painful fever blisters. T cells limit viral pathogenesis by inducing a nonpermissive state in ganglion cells, where the virus lies latent. Cytotoxic T lymphocytes (CTL), especially CD4+ CTL, are critical in resistance against recurrent herpes lesions. Viral glycoproteins (gP), expressed early in HSV infection are CTL targets and have been used as therapeutic vaccines with some success. Although living vaccines are usually needed to stimulate effective CTL responses, naked DNA vaccines encoding HSV gP stimulate CTL and antibody responses experimentally. The hypothesis of this study is that a DNA vaccine encoding HSV-1 glycoprotin D (gD), protected from nuclease activity and targeted to cells bearing receptors for asialoorosomucoid (ASOR), can stimulate specific cellular immunity to herpes simplex virus in mice. The novelty of this strategy is that ligands, such as poly-L-lysine-ASOR (ASOR-L), protect the DNA from nucleases while directing it to cells bearing receptors for the ligand, features lacking in naked HSV gP DNA vaccines. To test the hypothesis, gD DNA either complexed to ASOR-L or naked will be injected into BALB/c and C3H mice by 4 different routes [intravenous (iv), intraperitoneal (ip), intramuscular (im), and oral (po)]. At 2 weeks after injection of 10 mug doses of DNA, splenic T cells will be examined for CTL activity against histocompatible target cells expressing gD. Since hepatocytes, macrophages and enterocytes bear receptors for ASOR, higher levels of gD-specific CTL activity will more likely develop in mice receiving gD-ASOR by iv, ip and po routes. Conditions for stimulating optimal HSV CTL responses will be defined by this study. The practical application of this novel approach is a therapy for alleviating recurrences of painful herpes lesions in humans.