Fig. 7: Extension of lysoTRAP for clearance of H1N1 in vitro, in vivo and on human lung organoids.
From: Lysosomal “TRAP”: a neotype modality for clearance of viruses and variants
a Schematic for the construction of sialic acid (SA) engineering lysosome (lysoTRAP (SA)). Upon 2,6-siayltransferase (SIAT1) and Cytidine 5’-monophosphate-N-acetylneuraminic acid (CMP-SA), the surface of the lysosome was modified with SA, which served as a H1N1 entry-point. b CLSM images of lysoTRAP (SA). c TEM lysoTRAP (SA) showing the typical lysosome morphology. d QCM analysis of the interaction between H1N1-PR8 virions and lysoTRAP (SA). e TEM imaging of lysoTRAP (SA) after a 4-hour co-incubation with H1N1-PR8 virions. The internalized H1N1-PR8 virions were indicated by red arrows in lysoTRAP (SA). f ELISA measurement of viral N protein level and q-PCR analysis of H1N1-PR8 mRNA expression after a 24-hour incubation with lysoTRAP (SA). g q-PCR analysis of H1N1-PR8 mRNA expression (left) and representative CLSM images of N protein expression (right) in MDCK cells upon treatment with PBS or lysoTRAP (SA). h Schematic for the detection of in vivo clearance of H1N1 virions by lysoTRAP (SA) in C57BL/6 mice. i q-PCR analysis of H1N1-PR8 mRNA levels in the lung tissues. j Representative H&E images of the lung tissue after H1N1-PR8 infection. k–l Similar data as that presented in i,j following infection with H1N1-CA07. m Schematic for the anti-infection experiment on human lung organoid model. 1 d after inoculation with 100 TCID50 H1N1-CA07 or H1N1-PR8 and treatment with PBS or h-lysoTRAP (SA), the human lung organoids were collected for q-PCR analysis and immunofluorescence analysis. n q-PCR analysis of H1N1-PR8 RNA copies and representative immunofluorescence images of N protein of H1N1-PR8 in human lung organoids. o Similar data as that presented in n following infection with H1N1-CA07. The data in f, g, n, and o represent the mean ± S.D. (n = 5 biologically independent experiments). The data in i and k represent the mean ± S.D. (n = 10 biologically independent mice). Statistical significance was calculated using two-tailed unpaired t-test in f, g i, k, n, and o. Source data were provided in the Source data file.
