Medical Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Clinical Services Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
Vaccine. 2019 Jan 7;37(2):333-342. doi: 10.1016/j.vaccine.2018.08.043. Epub 2018 Dec 3.
A human cytomegalovirus (HCMV) vaccine to prevent infection and/or reduce disease associated with congenital infection or visceral disease in transplant recipients is a high priority, but has remained elusive. We created a disabled infectious single cycle rhesus CMV (RhCMV) deleted for glycoprotein L (gL) and the MHC class I immune evasion genes Rh178 and Rh182-189, and restored its epithelial cell tropism by inserting the Rh128-131A genes. The resulting virus, RhCMVRΔgL/178/182-189, was used to vaccinate rhesus monkeys intramuscularly and was compared with vaccination of animals with soluble RhCMV glycoprotein B (gB) in alum/monophosphoryl lipid A or with PBS as a control. At 4 weeks after the second vaccination, an increased frequency of RhCMV-specific CD8 T cells was detected in animals vaccinated with the RhCMVRΔgL/178/182-189 vaccine compared to animals vaccinated with soluble gB. In contrast, monkeys vaccinated with soluble gB had 20-fold higher gB antibody titers than animals vaccinated with RhCMVRΔgL/178/182-189. Titers of neutralizing antibody to RhCMV infection of fibroblasts were higher in animals vaccinated with gB compared with RhCMVRΔgL/178/182-189. Following vaccination, monkeys were challenged subcutaneously with RhCMV UCD59, a low passage virus propagated in monkey kidney epithelial cells. All animals became infected after challenge; however, the frequency of RhCMV detection in the blood was reduced in monkeys vaccinated with soluble gB compared with those vaccinated with RhCMVRΔgL/178/182-189. The frequency of challenge virus shedding in the urine and saliva and the RhCMV copy number shed at these sites was not different in animals vaccinated with RhCMVRΔgL/178/182-189 or soluble gB compared with those that received PBS before challenge. Although the RhCMVRΔgL/178/182-189 vaccine was superior in inducing cellular immunity to RhCMV, it induced lower titers of neutralizing antibody and antibody to gB than the soluble gB vaccine; after challenge, animals vaccinated with soluble gB had a lower frequency of virus detection in the blood than those vaccinated with RhCMVRΔgL/178/182-189.
一种用于预防感染和/或降低移植受者中与先天性感染或内脏疾病相关的疾病的人巨细胞病毒(HCMV)疫苗是当务之急,但一直难以实现。我们构建了一种失活的传染性单周期恒河猴 CMV(RhCMV),其缺失糖蛋白 L(gL)和 MHC Ⅰ类免疫逃逸基因 Rh178 和 Rh182-189,并通过插入 Rh128-131A 基因恢复其上皮细胞嗜性。由此产生的病毒,RhCMVRΔgL/178/182-189,被用于肌肉内接种恒河猴,并与用铝佐剂/单磷酰脂质 A 中的可溶性 RhCMV 糖蛋白 B(gB)或 PBS 接种的动物进行比较。在第二次接种后 4 周,与用可溶性 gB 接种的动物相比,用 RhCMVRΔgL/178/182-189 疫苗接种的动物中检测到 RhCMV 特异性 CD8 T 细胞的频率增加。相比之下,用可溶性 gB 接种的猴子的 gB 抗体滴度比用 RhCMVRΔgL/178/182-189 接种的猴子高 20 倍。接种 gB 的动物对成纤维细胞感染 RhCMV 的中和抗体滴度高于 RhCMVRΔgL/178/182-189。接种后,猴子被皮下接种 RhCMV UCD59,这是一种在猴肾上皮细胞中传代的低传代病毒。所有动物在挑战后均被感染;然而,与用 RhCMVRΔgL/178/182-189 接种的动物相比,用可溶性 gB 接种的动物血液中 RhCMV 的检测频率降低。与接种前接受 PBS 的动物相比,用 RhCMVRΔgL/178/182-189 或可溶性 gB 接种的动物在尿液和唾液中挑战病毒脱落的频率以及这些部位脱落的 RhCMV 拷贝数没有差异。尽管 RhCMVRΔgL/178/182-189 疫苗在诱导针对 RhCMV 的细胞免疫方面更优越,但它诱导的中和抗体和 gB 抗体滴度低于可溶性 gB 疫苗;在挑战后,用可溶性 gB 接种的动物血液中病毒检测频率低于用 RhCMVRΔgL/178/182-189 接种的动物。