Establishment of a MAGI-derived indicator cell line that detects the Nef enhancement of HIV-1 infectivity with high sensitivity
| Autor: | Michiyuki Matsuda, Minoru Tobiume, Masashi Tatsumi, Mikako Takahoko |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Gene Expression Regulation
Viral viruses Biology Virus Replication medicine.disease_cause Sensitivity and Specificity Gene Products nef Virus Virology medicine Humans nef Gene Products Human Immunodeficiency Virus Serial Passage Infectivity Wild type virus diseases Simian immunodeficiency virus biology.organism_classification In vitro Clone Cells Viral replication Cell culture CD4 Antigens Mutation Lentivirus HIV-1 HeLa Cells |
| Zdroj: | Journal of Virological Methods. 97:151-158 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/s0166-0934(01)00349-4 |
| Popis: | The Nef protein of the simian and human immunodeficiency viruses (SIV and HIV) is regarded as one of the critical determinants of the pathogenicity of HIV-1 in vivo. The positive effect of Nef on viral replication is examined most easily in vitro by the use of indicator cells such as HeLa-CD4-LTR-beta-gal cells (MAGI) or MAGIC5 cells, which are MAGI-derived, CCR5-expressing cells. However, Nef increases the infectivity of many HIV-1 strains no more than 10-fold in these indicator cells. It was noted that MAGI cells expressing a lower level of CD4 enabled us to discriminate more clearly between wild-type and Nef-defective virions. A MAGIC5-derived cell line, MAGNEF, which stably expressed a low level of CD4, was established. The infectivity of the Nef-defective HIV-1 NL4-3 strain was consistently less than one-twentieth of that of the wild type in MAGNEF cells. By using MAGNEF cells, it was shown that Nef enhanced the infectivity of a subtype C HIV-1, Indie-C1 strain, although the effect of Nef on Indie-C1 was significantly less than that on the subtype B strains NL4-3 and SF2. These results validate the versatility of MAGNEF cells for use in the simple and sensitive assay for the level of Nef dependence of various HIV-1 isolates. |
| Databáze: | OpenAIRE |
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