A highly quantitative detection system for cell entry of human norovirus-like particles based on the complementation of NanoLuc luciferase

• Published in: Virology (New York, N.Y.) Vol. 573; pp. 23 - 28
• Main Authors: Kimura, Miyuki, Sekiguchi, Kazuki, Okitsu, Shoko, Ushijima, Hiroshi, Tani, Hideki
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2022
• Subjects: Cell entry; gastroenteritis; heat treatment; HiBiT tag; Human norovirus; humans; HuNoV; Infectious Disease; luciferase; NanoLuc; Norovirus; virology; Virus-like particle; VLP; Virus-like particle; Cell entry; HuNoV; HiBiT tag; Human norovirus; VLP; NanoLuc
• ISSN: 0042-6822; 1096-0341; 1096-0341
• DOI: 10.1016/j.virol.2022.06.001

Human norovirus (HuNoV) is the common cause of acute gastroenteritis worldwide. Norovirus-like particles (VLPs) have been applied to analyze virus–host cell interactions. This study established a highly quantitative detection system for HuNoV VLP entry into live cells based on the NanoLuc luciferase complementation. First, we generated a HiBiT-tagged VLP and examined VLP-HiBiT entry into Vero cells expressing LgBiT. The entry was saturable and competed by a non-labeled VLP, indicating the VLP-specific entry into the host cells. Upon comparison of the VLP-HiBiT entry among some cell lines, Caco-2 cells were found to be the most susceptible. The VLP-HiBiT entry was unaffected by treatment at an acidic pH value above 3.0 and was significantly reduced by heat treatment at 75°C for more than 2 min. These results reflected those of previous studies, indicating the stability of VLPs and binding to histo-blood group antigens at various pH ranges and temperatures and cellular tropism. •This study established quantitative detection system for the entry of norovirus.•HiBiT-tagged VLP (VLP-HiBiT) entered into LgBiT-expressing live cells.•VLP-HiBiT entry was unaffected by treatment at pH value above 3.0.•VLP-HiBiT entry was significantly reduced by heat treatment at 75°C for 2 min.•Caco-2 cells were the most susceptible among some cell lines examined.