Liu Ruikang, Americo Jeffrey L, Cotter Catherine A, Earl Patricia L, Erez Noam, Peng Chen, Moss Bernard
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda MD 20892 USA.
bioRxiv. 2021 Jan 1:2020.12.30.424878. doi: 10.1101/2020.12.30.424878.
Replication-restricted modified vaccinia virus Ankara (MVA) is a licensed smallpox vaccine and numerous clinical studies investigating recombinant MVAs (rMVAs) as vectors for prevention of other infectious diseases have been completed or are in progress. Two rMVA COVID-19 vaccine trials are at an initial stage, though no animal protection studies have been reported. Here, we characterize rMVAs expressing the S protein of CoV-2. Modifications of full length S individually or in combination included two proline substitutions, mutations of the furin recognition site and deletion of the endoplasmic retrieval signal. Another rMVA in which the receptor binding domain (RBD) flanked by the signal peptide and transmembrane domains of S was also constructed. Each modified S protein was displayed on the surface of rMVA-infected human cells and was recognized by anti-RBD antibody and by soluble hACE2 receptor. Intramuscular injection of mice with the rMVAs induced S-binding and pseudovirus-neutralizing antibodies. Boosting occurred following a second homologous rMVA but was higher with adjuvanted purified RBD protein. Weight loss and lethality following intranasal infection of transgenic hACE2 mice with CoV-2 was prevented by one or two immunizations with rMVAs or by passive transfer of serum from vaccinated mice. One or two rMVA vaccinations also prevented recovery of infectious CoV-2 from the lungs. A low amount of virus was detected in the nasal turbinates of only one of eight rMVA-vaccinated mice on day 2 and none later. Detection of subgenomic mRNA in turbinates on day 2 only indicated that replication was abortive in immunized animals.
复制受限的安卡拉痘苗病毒(MVA)是一种已获许可的天花疫苗,许多将重组MVA(rMVA)作为预防其他传染病载体的临床研究已经完成或正在进行。两项rMVA COVID-19疫苗试验正处于初始阶段,不过尚未有动物保护研究的报告。在此,我们对表达新冠病毒-2(CoV-2)刺突蛋白(S蛋白)的rMVA进行了表征。对全长S蛋白单独或组合进行的修饰包括两个脯氨酸取代、弗林蛋白酶识别位点的突变以及内质网回收信号的缺失。还构建了另一种rMVA,其中S蛋白的受体结合域(RBD)两侧分别为信号肽和跨膜域。每种修饰后的S蛋白都展示在rMVA感染的人细胞表面,并能被抗RBD抗体和可溶性人血管紧张素转换酶2(hACE2)受体识别。给小鼠肌肉注射rMVA可诱导产生与S蛋白结合及中和假病毒的抗体。第二次同源rMVA接种后出现了抗体增强现象,但使用佐剂化纯化RBD蛋白后抗体增强效果更高。用rMVA进行一次或两次免疫接种,或通过被动转移接种疫苗小鼠的血清,可预防转基因hACE2小鼠经鼻感染CoV-2后的体重减轻和致死情况。一次或两次rMVA疫苗接种还可防止在肺部检测到传染性CoV-2。仅在第2天,在8只接种rMVA的小鼠中有1只的鼻甲中检测到少量病毒,之后未再检测到。仅在第2天在鼻甲中检测到亚基因组mRNA表明,在免疫动物中病毒复制是流产性的。