Isolation and characterization of human cells resistant to retrovirus infection
<p>Abstract</p> <p>Background</p> <p>Identification of host cell proteins required for HIV-1 infection will add to our knowledge of the life cycle of HIV-1 and in the development of therapeutics to combat viral infection. We and other investigators have mutagenized rode...
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| Format: | Article |
| Language: | English |
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BMC
2007-07-01
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| Series: | Retrovirology |
| Online Access: | http://www.retrovirology.com/content/4/1/45 |
| _version_ | 1819123465375973376 |
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| author | Somia Nikunj V Lech Patrycja |
| author_facet | Somia Nikunj V Lech Patrycja |
| author_sort | Somia Nikunj V |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Identification of host cell proteins required for HIV-1 infection will add to our knowledge of the life cycle of HIV-1 and in the development of therapeutics to combat viral infection. We and other investigators have mutagenized rodent cells and isolated mutant cell lines resistant to retrovirus infection. Since there are differences in the efficiency of single round infection with VSVG pseudotyped HIV-1 on cells of different species, we conducted a genetic screen to isolate human cells resistant to HIV-1 infection. We chemically mutagenized human HeLa cells and validated our ability to isolate mutants at test diploid loci. We then executed a screen to isolate HeLa cell mutants resistant to infection by an HIV-1 vector coding for a toxic gene product.</p> <p>Results</p> <p>We isolated two mutant cell lines that exhibit up to 10-fold resistance to infection by HIV-1 vectors. We have verified that the cells are resistant to infection and not defective in gene expression. We have confirmed that the resistance phenotype is not due to an entry defect. Fusion experiments between mutant and wild-type cells have established that the mutations conferring resistance in the two clones are recessive. We have also determined the nature of the block in the two mutants. One clone exhibits a block at or before reverse transcription of viral RNA and the second clone has a retarded kinetic of viral DNA synthesis and a block at nuclear import of the preintegration complex.</p> <p>Conclusion</p> <p>Human cell mutants can be isolated that are resistant to infection by HIV-1. The mutants are genetically recessive and identify two points where host cell factors can be targeted to block HIV-1 infection.</p> |
| first_indexed | 2024-12-22T07:08:47Z |
| format | Article |
| id | doaj.art-4acf1bf87fc547c9a7404199ae102a58 |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-12-22T07:08:47Z |
| publishDate | 2007-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-4acf1bf87fc547c9a7404199ae102a582022-12-21T18:34:36ZengBMCRetrovirology1742-46902007-07-01414510.1186/1742-4690-4-45Isolation and characterization of human cells resistant to retrovirus infectionSomia Nikunj VLech Patrycja<p>Abstract</p> <p>Background</p> <p>Identification of host cell proteins required for HIV-1 infection will add to our knowledge of the life cycle of HIV-1 and in the development of therapeutics to combat viral infection. We and other investigators have mutagenized rodent cells and isolated mutant cell lines resistant to retrovirus infection. Since there are differences in the efficiency of single round infection with VSVG pseudotyped HIV-1 on cells of different species, we conducted a genetic screen to isolate human cells resistant to HIV-1 infection. We chemically mutagenized human HeLa cells and validated our ability to isolate mutants at test diploid loci. We then executed a screen to isolate HeLa cell mutants resistant to infection by an HIV-1 vector coding for a toxic gene product.</p> <p>Results</p> <p>We isolated two mutant cell lines that exhibit up to 10-fold resistance to infection by HIV-1 vectors. We have verified that the cells are resistant to infection and not defective in gene expression. We have confirmed that the resistance phenotype is not due to an entry defect. Fusion experiments between mutant and wild-type cells have established that the mutations conferring resistance in the two clones are recessive. We have also determined the nature of the block in the two mutants. One clone exhibits a block at or before reverse transcription of viral RNA and the second clone has a retarded kinetic of viral DNA synthesis and a block at nuclear import of the preintegration complex.</p> <p>Conclusion</p> <p>Human cell mutants can be isolated that are resistant to infection by HIV-1. The mutants are genetically recessive and identify two points where host cell factors can be targeted to block HIV-1 infection.</p>http://www.retrovirology.com/content/4/1/45 |
| spellingShingle | Somia Nikunj V Lech Patrycja Isolation and characterization of human cells resistant to retrovirus infection Retrovirology |
| title | Isolation and characterization of human cells resistant to retrovirus infection |
| title_full | Isolation and characterization of human cells resistant to retrovirus infection |
| title_fullStr | Isolation and characterization of human cells resistant to retrovirus infection |
| title_full_unstemmed | Isolation and characterization of human cells resistant to retrovirus infection |
| title_short | Isolation and characterization of human cells resistant to retrovirus infection |
| title_sort | isolation and characterization of human cells resistant to retrovirus infection |
| url | http://www.retrovirology.com/content/4/1/45 |
| work_keys_str_mv | AT somianikunjv isolationandcharacterizationofhumancellsresistanttoretrovirusinfection AT lechpatrycja isolationandcharacterizationofhumancellsresistanttoretrovirusinfection |