tetano
Editor, Senior Moderator
Front Cell Infect Microbiol
. 2022 Mar 3;12:802147.
doi: 10.3389/fcimb.2022.802147. eCollection 2022.
Development of an Adeno-Associated Virus-Vectored SARS-CoV-2 Vaccine and Its Immunogenicity in Mice
Xi Qin[SUP] 1 [/SUP], Shanhu Li[SUP] 2 [/SUP], Xiang Li[SUP] 1 [/SUP], Dening Pei[SUP] 1 [/SUP], Yu Liu[SUP] 3 [/SUP], Youxue Ding[SUP] 1 [/SUP], Lan Liu[SUP] 1 [/SUP], Hua Bi[SUP] 1 [/SUP], Xinchang Shi[SUP] 1 [/SUP], Ying Guo[SUP] 1 [/SUP], Enyue Fang[SUP] 4 [/SUP], Fang Huang[SUP] 2 [/SUP], Lei Yu[SUP] 1 [/SUP], Liuqiang Zhu[SUP] 1 [/SUP], Yifang An[SUP] 1 [/SUP], C Alexander Valencia[SUP] 3 [/SUP], Yuhua Li[SUP] 4 [/SUP], Biao Dong[SUP] 3 [/SUP], Yong Zhou[SUP] 1 [/SUP]
Affiliations
Abstract
Owing to the outbreak of the novel coronavirus (SARS-CoV-2) worldwide at the end of 2019, the development of a SARS-CoV-2 vaccine became an urgent need. In this study, we developed a type 9 adeno-associated virus vectored vaccine candidate expressing a dimeric receptor binding domain (RBD) of the SARS-CoV-2 spike protein (S protein) and evaluated its immunogenicity in a murine model. The vaccine candidate, named AAV9-RBD virus, was constructed by inserting a signal peptide to the N-terminus of two copies of RBD, spaced by a linker, into the genome of a type 9 adeno-associated virus. In vitro assays showed that HeLa cells infected by the recombinant AAV virus expressed high levels of the recombinant RBD protein, mostly found in the cell culture supernatant. The recombinant AAV9-RBD virus was cultured and purified. The genome titer of the purified recombinant AAV9-RBD virus was determined to be 2.4 × 10[SUP]13[/SUP] genome copies/mL (GC/mL) by Q-PCR. Balb/c mice were immunized with the virus by intramuscular injection or nasal drip administration. Eight weeks after immunization, neutralizing antibodies against the new coronavirus pseudovirus were detected in the sera of all mice; the mean neutralizing antibody EC[SUB]50[/SUB] values were 517.7 ± 292.1 (n=10) and 682.8 ± 454.0 (n=10) in the intramuscular injection group and nasal drip group, respectively. The results of this study showed that the recombinant AAV9-RBD virus may be used for the development of a SARS-CoV-2 vaccine.
Keywords: AAV9; SARS-CoV-2; immune; long term protection; vaccine.
. 2022 Mar 3;12:802147.
doi: 10.3389/fcimb.2022.802147. eCollection 2022.
Development of an Adeno-Associated Virus-Vectored SARS-CoV-2 Vaccine and Its Immunogenicity in Mice
Xi Qin[SUP] 1 [/SUP], Shanhu Li[SUP] 2 [/SUP], Xiang Li[SUP] 1 [/SUP], Dening Pei[SUP] 1 [/SUP], Yu Liu[SUP] 3 [/SUP], Youxue Ding[SUP] 1 [/SUP], Lan Liu[SUP] 1 [/SUP], Hua Bi[SUP] 1 [/SUP], Xinchang Shi[SUP] 1 [/SUP], Ying Guo[SUP] 1 [/SUP], Enyue Fang[SUP] 4 [/SUP], Fang Huang[SUP] 2 [/SUP], Lei Yu[SUP] 1 [/SUP], Liuqiang Zhu[SUP] 1 [/SUP], Yifang An[SUP] 1 [/SUP], C Alexander Valencia[SUP] 3 [/SUP], Yuhua Li[SUP] 4 [/SUP], Biao Dong[SUP] 3 [/SUP], Yong Zhou[SUP] 1 [/SUP]
Affiliations
- PMID: 35310850
- PMCID: PMC8927296
- DOI: 10.3389/fcimb.2022.802147
Abstract
Owing to the outbreak of the novel coronavirus (SARS-CoV-2) worldwide at the end of 2019, the development of a SARS-CoV-2 vaccine became an urgent need. In this study, we developed a type 9 adeno-associated virus vectored vaccine candidate expressing a dimeric receptor binding domain (RBD) of the SARS-CoV-2 spike protein (S protein) and evaluated its immunogenicity in a murine model. The vaccine candidate, named AAV9-RBD virus, was constructed by inserting a signal peptide to the N-terminus of two copies of RBD, spaced by a linker, into the genome of a type 9 adeno-associated virus. In vitro assays showed that HeLa cells infected by the recombinant AAV virus expressed high levels of the recombinant RBD protein, mostly found in the cell culture supernatant. The recombinant AAV9-RBD virus was cultured and purified. The genome titer of the purified recombinant AAV9-RBD virus was determined to be 2.4 × 10[SUP]13[/SUP] genome copies/mL (GC/mL) by Q-PCR. Balb/c mice were immunized with the virus by intramuscular injection or nasal drip administration. Eight weeks after immunization, neutralizing antibodies against the new coronavirus pseudovirus were detected in the sera of all mice; the mean neutralizing antibody EC[SUB]50[/SUB] values were 517.7 ± 292.1 (n=10) and 682.8 ± 454.0 (n=10) in the intramuscular injection group and nasal drip group, respectively. The results of this study showed that the recombinant AAV9-RBD virus may be used for the development of a SARS-CoV-2 vaccine.
Keywords: AAV9; SARS-CoV-2; immune; long term protection; vaccine.