tetano
Editor, Senior Moderator
Antiviral Res
. 2023 Nov 7:105744.
doi: 10.1016/j.antiviral.2023.105744. Online ahead of print. Functional assessments of SARS-CoV-2 single-round infectious particles with variant-specific spike proteins on infectivity, drug sensitivity, and antibody neutralization
Wen-Chi Su[SUP] 1 [/SUP], Zan-Yu Chen[SUP] 2 [/SUP], Young-Sheng Chang[SUP] 3 [/SUP], King-Song Jeng[SUP] 4 [/SUP], Uyen Nguyen Phuong Le[SUP] 5 [/SUP], Yu-Chi Chou[SUP] 6 [/SUP], Li-Lan Kuo[SUP] 7 [/SUP], Ivonne Melano[SUP] 8 [/SUP], Jesse[SUP] 8 [/SUP], Wei-Jan Wang[SUP] 9 [/SUP], Ying-Chyi Song[SUP] 10 [/SUP], Sin-Rong Li[SUP] 11 [/SUP], Mien-Chie Hung[SUP] 12 [/SUP], Michael M C Lai[SUP] 13 [/SUP], Cheng-Wen Lin[SUP] 14 [/SUP]
Affiliations
Working with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is restricted to biosafety level III (BSL-3) laboratory. The study used a trans-complementation system consisting of virus-like particles (VLPs) and DNA-launched replicons to generate SARS-CoV-2 single-round infectious particles (SRIPs) with variant-specific spike (S) proteins. S gene of Wuhan-Hu-1 strain (S[SUB]WH1[/SUB]) or Omicron BA.1 variant (S[SUB]BA.1[/SUB]), along with the envelope (E) and membrane (M) genes, were cloned into a tricistronic vector, co-expressed in the cells to produce variant-specific S-VLPs. Additionally, the replicon of the WH1-like strain without S, E, M and accessory genes, was engineered under the control by a CMV promoter to produce self-replicating RNAs within VLP-producing cells, led to create S[SUB]WH1[/SUB]- and S[SUB]BA.1[/SUB]-based SARS-CoV-2 SRIPs. The S[SUB]BA.1[/SUB]-based SRIP showed lower virus yield, replication, N protein expression, fusogenicity, and infectivity compared to S[SUB]WH1[/SUB]-based SRIPs. S[SUB]BA.1[/SUB]-based SRIP also exhibited intermediate resistance to neutralizing antibodies produced by S[SUB]WH1[/SUB]-based vaccines, but were effective at infecting cells with low ACE2 expression. Importantly, both S-based SRIPs responded similarly to remdesivir and GC376, with EC50 values ranging from 0.17 to 1.46 μM, respectively. The study demonstrated that this trans-complementation system is a reliable and efficient tool for generating SARS-CoV-2 SRIPs with variant-specific S proteins. SARS-CoV-2 SRIPs, mimicking authentic live viruses, facilitate comprehensive analysis of variant-specific virological characteristics, including antibody neutralization, and drug sensitivity in non-BSL-3 laboratories.
Keywords: ACE2; Antibody neutralization; Drug sensitivity; Replicon; SARS-CoV-2; Single-round infectious particle; Spike; Variant; Virus yield; Virus-like particle.
. 2023 Nov 7:105744.
doi: 10.1016/j.antiviral.2023.105744. Online ahead of print. Functional assessments of SARS-CoV-2 single-round infectious particles with variant-specific spike proteins on infectivity, drug sensitivity, and antibody neutralization
Wen-Chi Su[SUP] 1 [/SUP], Zan-Yu Chen[SUP] 2 [/SUP], Young-Sheng Chang[SUP] 3 [/SUP], King-Song Jeng[SUP] 4 [/SUP], Uyen Nguyen Phuong Le[SUP] 5 [/SUP], Yu-Chi Chou[SUP] 6 [/SUP], Li-Lan Kuo[SUP] 7 [/SUP], Ivonne Melano[SUP] 8 [/SUP], Jesse[SUP] 8 [/SUP], Wei-Jan Wang[SUP] 9 [/SUP], Ying-Chyi Song[SUP] 10 [/SUP], Sin-Rong Li[SUP] 11 [/SUP], Mien-Chie Hung[SUP] 12 [/SUP], Michael M C Lai[SUP] 13 [/SUP], Cheng-Wen Lin[SUP] 14 [/SUP]
Affiliations
- PMID: 37944823
- DOI: 10.1016/j.antiviral.2023.105744
Working with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is restricted to biosafety level III (BSL-3) laboratory. The study used a trans-complementation system consisting of virus-like particles (VLPs) and DNA-launched replicons to generate SARS-CoV-2 single-round infectious particles (SRIPs) with variant-specific spike (S) proteins. S gene of Wuhan-Hu-1 strain (S[SUB]WH1[/SUB]) or Omicron BA.1 variant (S[SUB]BA.1[/SUB]), along with the envelope (E) and membrane (M) genes, were cloned into a tricistronic vector, co-expressed in the cells to produce variant-specific S-VLPs. Additionally, the replicon of the WH1-like strain without S, E, M and accessory genes, was engineered under the control by a CMV promoter to produce self-replicating RNAs within VLP-producing cells, led to create S[SUB]WH1[/SUB]- and S[SUB]BA.1[/SUB]-based SARS-CoV-2 SRIPs. The S[SUB]BA.1[/SUB]-based SRIP showed lower virus yield, replication, N protein expression, fusogenicity, and infectivity compared to S[SUB]WH1[/SUB]-based SRIPs. S[SUB]BA.1[/SUB]-based SRIP also exhibited intermediate resistance to neutralizing antibodies produced by S[SUB]WH1[/SUB]-based vaccines, but were effective at infecting cells with low ACE2 expression. Importantly, both S-based SRIPs responded similarly to remdesivir and GC376, with EC50 values ranging from 0.17 to 1.46 μM, respectively. The study demonstrated that this trans-complementation system is a reliable and efficient tool for generating SARS-CoV-2 SRIPs with variant-specific S proteins. SARS-CoV-2 SRIPs, mimicking authentic live viruses, facilitate comprehensive analysis of variant-specific virological characteristics, including antibody neutralization, and drug sensitivity in non-BSL-3 laboratories.
Keywords: ACE2; Antibody neutralization; Drug sensitivity; Replicon; SARS-CoV-2; Single-round infectious particle; Spike; Variant; Virus yield; Virus-like particle.