NOVEL DELIVERY SYSTEM FOR A HERPESVIRUS DNA VACCINE

疱疹病毒 DNA 疫苗的新型递送系统

• 批准号: 2634152
• 负责人: NANCY J BIGLEY
• 金额: $ 3.6万
• 依托单位: WRIGHT STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2634152
• 关键词: Herpesviridae vaccine; T cell receptor; active immunization; cytotoxic T lymphocyte; drug delivery systems; flow cytometry; gene expression; helper T lymphocyte; herpes simplex virus 1; histocompatibility antigens; immunocytochemistry; injection /infusion; laboratory mouse; lysine; monoclonal antibody; plasmids; polymerase chain reaction; proteoglycan; tissue /cell culture; vector vaccine; virus DNA; virus cytopathogenic effect; virus infection mechanism

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.

大多数成年人携带I型单纯疱疹病毒(HSV-1)，但只有少数 周期性地出现发烧起水泡的疼痛症状。T细胞限制病毒 通过在神经节细胞中诱导一种不允许的状态来致病，其中 这种病毒潜伏着。细胞毒性T淋巴细胞(CTL)，尤其是CD4+ CTL在抵抗复发疱疹皮损方面至关重要。病毒式传播 在HSV感染早期表达的糖蛋白(GP)是CTL靶标和 已经被用作治疗性疫苗，并取得了一些成功。虽然 通常需要活疫苗来刺激有效的CTL反应， 编码HSV gP的裸露DNA疫苗刺激CTL和抗体应答 试验性的。这项研究的假设是一种DNA疫苗 编码HSV-1糖蛋白D(GD)，不受核酸酶活性保护 靶向携带去唾液酸类粘蛋白受体(ASOR)的细胞，可以 激发小鼠对单纯疱疹病毒的特异性细胞免疫。 这一策略的新奇之处在于，配体，如多L赖氨酸-ASOR (ASOR-L)，保护DNA不受核酸酶的影响，同时将其引导到细胞 承载配体受体，缺乏裸露的HSV gp DNA 疫苗。为了验证这一假设，GDDNA要么与ASOR-L络合，要么 裸鼠将通过4种不同途径注射到BALB/c和C3H小鼠体内 静脉(Iv)、腹膜(Ip)、肌肉(Im)和口服 (PO)]。在注射10杯剂量的DNA后2周，脾T细胞 将针对组织相容靶标检查细胞的CTL活性 表达GD的细胞。由于肝细胞、巨噬细胞和肠细胞 熊的ASOR受体，更高水平的GD特异性CTL活性将 更有可能通过iv、ip和po途径接受GD-ASOR的小鼠发生。 激发最佳HSV CTL反应的条件将定义为 这项研究。这种新方法的实际应用是一个 减轻人类疱疹疼痛皮损复发的治疗。