Mosca Joseph D, Chang Yung-Nien, Williams Gregory
JDM Technologies, Inc., Ellicott City, MD 21042, USA.
Retrovirology. 2007 May 15;4:32. doi: 10.1186/1742-4690-4-32.
Developments in cell-based and gene-based therapies are emerging as highly promising areas to complement pharmaceuticals, but present day approaches are too cumbersome and thereby limit their clinical usefulness. These shortcomings result in procedures that are too complex and too costly for large-scale applications. To overcome these shortcomings, we described a protein delivery system that incorporates over-expressed proteins into viral particles that are non-infectious and stable at room temperature. The system relies on the biological process of viral egress to incorporate cellular surface proteins while exiting their host cells during lytic and non-lytic infections.
We report here the use of non-infectious surface-engineered virion particles to modulate immunity against three infectious disease agents--human immunodeficiency virus type 1 (HIV-1), herpes simplex virus (HSV), and Influenza. Surface-engineering of particles are accomplished by genetic modification of the host cell surface that produces the egress budding viral particle. Human peripheral blood lymphocytes from healthy donors exposed to CD80/B7.1, CD86/B7.2, and/or antiCD3 single-chain antibody surface-engineered non-infectious HIV-1 and HSV-2 particles stimulate T cell proliferation, whereas particles released from non-modified host cells have no T cell stimulatory activity. In addition to T cell proliferation, HIV-based particles specifically suppress HIV-1 replication (both monocytotropic and lymphocytotropic strains) 55 to 96% and HSV-based particles specifically induce cross-reactive HSV-1/HSV-2 anti-herpes virus antibody production. Similar surface engineering of influenza-based particles did not modify the intrinsic ability of influenza particles to stimulate T cell proliferation, but did bestow on the engineered particles the ability to induce cross-strain anti-influenza antibody production.
We propose that non-infectious viral particles can be surface-engineered to produce antigen-presenting particles that mimic antigen-presenting cells to induce immune responses in human peripheral blood lymphocytes. The viral particles behave as "biological carriers" for recombinant proteins, thereby establishing a new therapeutic paradigm for molecular medicine.
基于细胞和基于基因的疗法正成为补充药物治疗的极有前景的领域，但目前的方法过于繁琐，从而限制了它们的临床实用性。这些缺点导致程序对于大规模应用来说过于复杂且成本过高。为了克服这些缺点，我们描述了一种蛋白质递送系统，该系统将过度表达的蛋白质整合到在室温下无感染性且稳定的病毒颗粒中。该系统依赖于病毒释放的生物学过程，在裂解性和非裂解性感染期间，在离开宿主细胞时将细胞表面蛋白整合进去。
我们在此报告使用无感染性的表面工程化病毒粒子来调节针对三种传染病病原体——1型人类免疫缺陷病毒（HIV-1）、单纯疱疹病毒（HSV）和流感病毒的免疫。粒子的表面工程是通过对产生释放出芽病毒粒子的宿主细胞表面进行基因改造来实现的。来自健康供体的人外周血淋巴细胞暴露于CD80/B7.1、CD86/B7.2和/或抗CD3单链抗体表面工程化的无感染性HIV-1和HSV-2粒子会刺激T细胞增殖，而从未修饰的宿主细胞释放的粒子则没有T细胞刺激活性。除了T细胞增殖外，基于HIV的粒子特异性抑制HIV-1复制（嗜单核细胞和嗜淋巴细胞毒株）55%至96%，基于HSV的粒子特异性诱导交叉反应性HSV-1/HSV-2抗疱疹病毒抗体产生。基于流感病毒粒子的类似表面工程并没有改变流感病毒粒子刺激T细胞增殖 的内在能力，但确实赋予了工程化粒子诱导跨毒株抗流感抗体产生的能力。
我们提出，可以对无感染性病毒粒子进行表面工程改造，以产生模拟抗原呈递细胞的抗原呈递粒子，从而在人外周血淋巴细胞中诱导免疫反应。病毒粒子充当重组蛋白的“生物载体”，从而为分子医学建立一种新的治疗模式。
