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Identification of cellular factors associated with HIV-1 virions isolated by filtration and ultracentrifugation. (Viral particles)

ABSTRACT: The aim of this project was to identify cellular factors enriched with HIV-1 virions after filtration and ultracentrifugation of culture surpernatants of HIV-1 producer cells compared to a control. To this end, HeLa cells were transfected with HIV-1 NL4-3 provirus and, 48 hours after transfection, culture surpernatants of HIV-1 producer cells were collected, filtered and ultracentrifugated on a 20% sucrose cushion to pellet viruses released in the supernatants. As a control, supernatant from mock-transfected HeLa cells was collected and prepared as the supernatant of HIV-1 producer cells. The same volume of filtered and ultracentrifugated supernatants from mock- and HIV-1-producing cells were then subjected to LC-MS analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Marjorie Leduc

LAB HEAD: Clarisse Berlioz-Torrent

PROVIDER: PXD057129 | Pride | 2025-10-08

REPOSITORIES: Pride

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			Action	DRS
	ClBE230314_PVVIH_NI_P_125A_A49923_BM1_1_9889.d.7z	Other
	ClBE230314_PVVIH_NI_P_125B_A49929_BM7_1_9913.d.7z	Other
	ClBE230314_PVVIH_NI_P_125C_A49935_BM13_1_9977.d.7z	Other
	ClBE230314_PVVIH_NI_P_167_A49941_BM19_1_10001.d.7z	Other
	ClBE230314_PVVIH_U_168_A49948_BN2_1_10029.d.7z	Other

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Project description:Replicating viruses have broad applications in biomedicine, notably in cancer virotherapy and in the design of attenuated vaccines, however uncontrolled virus replication in vulnerable tissues can give pathology and often restricts the use of potent strains. Increased knowledge of tissue-selective microRNA expression now affords the possibility of engineering replicating viruses that are attenuated at the RNA level in sites of potential pathology, but retain wild type replication activity at sites not expressing the relevant microRNA. To assess the usefulness of this approach for the DNA virus, adenovirus, we have engineered a hepatocyte-safe wild type adenovirus 5 (Ad5), which normally mediates significant toxicity and is potentially lethal in mice. To do this we have included binding sites for hepatocyte-selective microRNA122a within the 3' UTR of the E1A transcription cassette. Imaging versions of these viruses, produced by fusing E1A with luciferase, showed that inclusion of microRNA122a binding sites caused up to 80 fold decreased hepatic expression of E1A following intravenous delivery to mice. Animals administered a ten-times lethal dose of wild type Ad5 (5 x 1010 viral particles/mouse) showed substantial hepatic genome replication and extensive liver pathology, while inclusion of 4 microRNA binding sites decreased replication 50-fold and virtually abrogated liver toxicity. This modified wild type virus retained full activity within cancer cells and provides a potent, liver-safe oncolytic virus. In addition to providing many potent new viruses for cancer virotherapy, microRNA-control of virus replication should provide a new strategy for designing safe attenuated vaccines applied across a broad range of viral diseases. four groups which we had were: WT: RNA extracted from livers of mice treated with wild type Ad5, 3 Replicates miR: RNA extracted from livers of mice treated with wild type Ad5 bearing mir122-binding sites in the 3'UTR of E1A (3 Replicates) Luc: RNA extracted from livers of mice treated with non-replicating Ad5 (3 Replicates) PBS: RNA extracted from livers of mice treated with PBS only (3 Replicates)

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Identification of cellular factors associated with HIV-1 virions isolated by filtration and ultracentrifugation. (Viral particles)

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