Understanding internalization of rotavirus VP6 nanotubes by cells: towards a recombinant vaccine
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Rotavirus VP6 nanotubes are an attractive option for a recombinant vaccine against rotavirus disease. Protection against rotavirus infection and an adjuvant effect have been observed upon immunization with VP6 nanotubes. However, little information exists on how VP6 nanotubes interact with cells and trigger an immune response. In this work, the interaction between VP6 nanotubes and different cell lines was characterized. VP6 nanotubes were not cytotoxic to any of the animal or human cell lines tested. Uptake of nanotubes into cells was cell-line-dependent, as only THP1 and J774 macrophage cells internalized them. Moreover, the size and spatial arrangement of VP6 assembled into nanotubes allowed their uptake by macrophages, as double-layered rotavirus-like particles also displaying VP6 in their surface were not taken up. The internalization of VP6 nanotubes was inhibited by methyl-β-cyclodextrin, but not by genistein, indicating that nanotube entry is specific, depends on the presence of cholesterol in the plasma membrane, and does not require the activity of tyrosine kinases. The information generated here expands our understanding of the interaction of protein nanotubes with cells, which is useful for the application of VP6 nanotubes as a vaccine.
KeywordsGenistein THP1 Cell Recombinant Vaccine J774 Cell J774 Macrophage
Financial support was provided by SEP-Conacyt 101847 and PAPIIT-UNAM IN223210 and IT200113. MR received financial support from Conacyt during her graduate studies. Technical support was provided by A. R. Pastor and V. Hernández and by G. Zavala and A. N. Lecona at the Electron Microscopy Units of Instituto de Biotecnología UNAM and Instituto Nacional de Salud Pública, Mexico, and J. A. Pimentel at the Laboratorio Nacional de Microscopía Avanzada UNAM, México. Materials were provided by Dr. Y. Rosenstein, Dr. A. Valdez, and Dr. C. Treviño.
Conflict of interest
The authors declare that they do not have any direct financial relationship with the commercial entities mentioned in this manuscript.
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